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Wolf Prize in
Agriculture
Edited by
Ilan Chet
The Hebrew University of Jerusalem, Israel
World Scientific
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for their assistance and permission to reproduce the selected reprints found in this volume:
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American Society of Plant Biologists
Atlanta Magazine
Avian Pathology
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Blackwell Ltd
Elsevier Ltd
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Journal of Animal Science
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ISBN-13 978-981-283-584-0
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Disclaimer
Most of the material from this volume was extracted from old archives of papers containing almost illegible
text. Hence, they have been reproduced to the best of the Editor’s and publisher’s ability.
Printed in Singapore.
CONTENTS
Preface xiii
John C. Walker 1
About Dr. John Charles Walker 1
List of Publications 7
George F. Sprague 33
Curriculum Vitae 34
List of Publications 37
Jay L. Lush 67
Curriculum Vita 68
List of Publications 71
Karl Maramorosch 81
Biographical Information 81
Recent Publications (since 1996) 85
Viruses, Vectors, and Vegetation: An Autobiography 90
v
vi Wolf Prize in Agriculture
The Wolf Foundation began its activities in 1976, with an initial endowment fund
donated by the Wolf family. The Foundation’s founders and major donors were
Dr. Ricardo Subirana y Lobo Wolf and his wife Francisca. The annual income from
investments is used to award prizes and scholarships, as well as fund the operating
expenses of the Foundation.
One of the aims of the Wolf Foundation, as stated by the law is “to award
prizes to outstanding scientists and artists — irrespective of nationality, race, color,
religion, sex or political views — for achievements in the interest of mankind
and friendly relations among peoples” in the fields of agriculture, chemistry,
mathematics, medicine and the arts. The official prize award ceremony takes place
at the Knesset building (Israel’s Parliament) in Jerusalem and the President of the
State of Israel hands the awards to the winners. Through the years, the Wolf Prize
has become one of the most prestigious prizes. In agriculture, it is probably the
most highly esteemed prize in the world.
The list of laureates in agricultural sciences from 1978 to 2008 contains 41
scientists in fields such as genetics, bio-control, ecology, plant sciences, animal
breeding, soil chemistry and physics, and plant biochemistry. Because of the
depletion in natural resources, agricultural development and human well-being
became one of the major concerns in the world. The Wolf Prize winners in the
field of agriculture are awarded for their remarkable innovative and pioneering
discoveries, development of new technologies and/or extraordinary contribution
to agricultural research, ecological conservation and food produce.
Now, at the end of 2008, there is an extremely high interest in agriculture, as
the concern of the world’s population is not only food supply but also alternative
sources of energy and neutraceuticals.
In this volume, we documented the Wolf Prize winners’ bibliography,
curriculum vitae (CV), autobiographical accounts and/or reports on their work
and achievements by others, important papers, lectures and other relevant
information. We did our best to include the data of prize winners who are no
longer with us.
The diverse fields of interest of the laureates provide a unique overview of the
work done by exceptional scientists in different institutes around the world. I wish
to share with the readers some of the deliberations and excitement involved in
discovering novel ideas and approaches, and reassessing the old ones. In rare
disclosures, updated findings alongside their views and speculations on various
xiii
xiv Wolf Prize in Agriculture
subjects in agriculture are offered in this volume. I sincerely hope that the readers,
be they researchers, students, biotechnologists, or extension specialists, find the
following pages stimulating and exciting as I did.
I would like to thank my talented secretary Mrs. Nili Ben-Yehezkel for putting
this volume together with all of her enthusiasm and devotion, and to the prize
winners and colleagues who helped in collecting the material for the volume.
I would also like to thank my publisher, World Scientific Publishing Co.,
especially Ms. Joy Quek, for her cooperation and assistance.
Ilan Chet
Rehovot, Israel
John C. Walker
University of Wisconsin
Madison, Wisconsin, USA
k
1893–1994
This is a story of a local industry and a Racine man. The industry is one of the
prime industries of Racine and Kenosha counties; the man has contributed more
basic knowledge to that industry here and throughout the world than any other
living person. This is the story of a man who attended Beebe grade school, Racine
High School, and graduated from the University of Wisconsin. Today he is world-
renowned in his field, a member of the National Academy of Science (membership
in this society is the highest recognition possible in the United States for scientific
research) and an honored member in many international scientific societies.
John Charles Walker, because of his inherent modesty, is hardly known in his
home town. He, who has contributed so very much toward the economic and
physical welfare of ourselves and the world, has never been formally recognized
by his fellow townspeople.
In the early 1800’s a few rugged individualists from Northern Europe
settled near Racine. These individuals founded the type of specialized agriculture
1
2 Wolf Prize in Agriculture
for which Racine and the surrounding area has long been noted, namely the
production of truck crops — cabbage — onions — potatoes — etc.
About the time of the Civil War, Racine had become the major source of
supply for cabbage in the Midwest. In the middle 1800’s, Ben Bones operated
a farm just east of Lathrop Road, and south of Chicory Road. He found, in this
particular year, that he had a surplus of cabbage, more than could be marketed
through the local trade channels. Learning that the beer industry of St. Louis had
started using a refrigerated freight car to ship beer, Mr. Bones conceived the idea
of loading one of these cars with cabbage and shipping this cabbage under
refrigeration to the German settlement of St. Louis. It is believed that this was the
first time that perishable produce was shipped under refrigeration. Two carloads
were shipped this first year which proved so profitable that Mr. Bones increased
his acreage the following year. Thus Wisconsin’s Cabbage Industry was born.
A disease (cabbage “yellows”) was noted in the area during the late 1800’s. It
was serious enough that the growers called a special meeting at the Berryville
School in 1890. Professor H. L. Russell, later Dean of the College of Agriculture,
was present at this meeting. The disease problem was discussed and reviewed but
nothing constructive was accomplished. By 1910, the disease was epidemic, resulting
in a complete crop failure. The Durand farm (now Case South Works) operated by
the Gunthers was replanted twice, and each time the crop died as a result of this
new disease. Those who were fortunate enough to have a partial crop found yields
reduced to one or two tons per acre. Today, a yield of 30 tons per acre is not
exceptional.
Dr. L. R. Jones, a native of Wisconsin and a graduate of Ripon College, had
been teaching botany at Vermont. In 1910 the University of Wisconsin enticed
L. R. Jones to join its staff, and at the second cabbage meeting held at the Berryville
school, 1910, Dr. Jones was present. This time definite plans were made to learn
more about the disease, to get it under control if possible. All growers offered full
cooperation with Dr. Jones and the University. This disease had to be controlled, or
farmers would have to change to different crops, and the kraut packers would
have to move into an area in which the disease was not a factor.
The few cabbage plants that could survive the disease were interesting to these
men. Healthy plants from the most severely infected field were to be selected for
seed production — stored over winter and replanted the following spring. Three
plants were selected from a field of over 25,000, they were carefully stored and
replanted in the same infected field of Matt Broesch. One night Mr. Broesch’s cow
got loose and partially destroyed two of the plants! The cabbage industry was
saved, however, by that single plant that was spared.
John Charles Walker was born on July 6, 1893 in a house on Lathrop Road
that still stands just north of 21st Street. He entered Beebe grade school, attended
Racine High School where he graduated with the highest scholastic record of any
boy in his class. In 1910 he entered the University of Wisconsin. In the spring of
1912 he went to Dr. L. R. Jones and stated that he thought he would like to major
in plant diseases, the new science recently named Plant Pathology. Dr. Jones’ answer
to this was, “That is fine, Charley. You have quite a problem in your own backyard,
a disease in cabbage that we know very little about. You can start working on that
this summer during your vacation” — and for more than 45 years J. C. Walker has
been working with cabbage.
ACCOMPLISHMENTS
Cabbage
Seed of the first yellows-resistant variety of cabbage, Wisconsin Hollander
No. 8, was being increased locally as rapidly as possible. Slightly less than
100 pounds were produced in 1916, hardly enough to meet the needs of the local
growers. Seed production in 1917 exceeded 800 pounds. (In 1957, forty years
later, total production of yellows resistant cabbage seed in this country exceeded
135,000 pounds!) At this critical time another disease, Blackleg hit Wisconsin.
This was a disease that was transmitted from one crop to another through seed,
unlike cabbage yellows which was soilborne. The success of the new yellows-
resistant program was doomed unless blackleg could be controlled. J. C. Walker
was handed this assignment and in short order he had the disease under control.
He showed that if cabbage seed was produced in the Pacific Northwest instead of
in Wisconsin it was disease-free. This not only saved the day for the cabbage
4 Wolf Prize in Agriculture
industry in Wisconsin but was a prime factor in the shift of the cabbage seed
industry from Northern Europe to the United States.
Walker next turned his attention to the yellows disease which had been brought
under control by the development of the Wisconsin Hollander No. 8 variety. This
variety was only partially resistant and was not suitable for all uses. Walker
discovered a more desirable type of resistance and has given to the growers some
12 or 15 different varieties of resistant cabbage.
The Clubroot disease of cabbage is as ancient as the crop and control of it has
defied man through the centuries. Walker turned his plant breeding skill to this
problem and has developed resistance to it. He took resistance from curly kale and
transferred it to cabbage. Control of this most serious disease is now certain.
Onion
Walker did some of the early work showing the effect of artificial heat on the
drying of onions in storage and the relationship of their preservation to disease
control. This work was done about 40 years ago. With the recent change in onion
production, mechanical harvesting and storage in bulk, with large amounts of air
being forced through these bulk piles of onions, every grower today is referring to
Dr. Walker’s paper and is using heat to cure and control the diseases of onion in
storage. He made the original technical descriptions of two of the neckrot diseases.
Smut is Wisconsin’s worst disease of onions. Our basic knowledge on the effect
of soil temperature on the occurrence of this disease was made by Walker. He took
the idea of dripping formaldehyde solution on the onion seed as they are planted
and worked out a practical application that saved the onion industry in Wisconsin.
John C. Walker 5
Onion smudge is a disease of white but not of yellow or red onions. Walker
showed that the pigments that form the yellow color in onions acted as a chemical
substance forming resistance to smudge in onions. White onions, without this
chemical substance, are susceptible to smudge. This is the most classical piece of
research on the nature of resistance due to chemicals that has ever been worked out.
Walker was instrumental in starting a national program for the development
of onions resistant to pink root, a disease of increasing importance in Wisconsin.
Peas
The story of peas differs little from that of cabbage. Pea wilt was ravaging the
fields of southern Wisconsin. Walker tested some 250 varieties of peas and found
some to be naturally resistant. With Professor E. J. Deiwiche, he developed resistant
varieties suitable to Wisconsin’s needs. Today, Wisconsin’s 150,000 acres of peas
are planted to these resistant varieties.
Beets
As Wisconsin’s beet canning industry developed, it was faced with a serious disease
known as black spot or heart rot. This disease was shown to be due not to a
parasite but to a lack of boron in the soil. Walker showed that the disease could be
controlled by adding boron in minute amounts to the soil or by spraying it on the
leaves.
A few years ago Wisconsin’s great pickle industry was threatened with extinction
by epidemic occurrences of the scab disease and by severe losses from the mosaic
disease. Walker crossed a scab-resistant slicing variety with a mosaic-resistant
pickle variety and developed varieties of the pickle-type resistant to both diseases.
These varieties are saving Wisconsin’s agriculture over one million dollars each year.
Bean
The growers and canners of beans have also been on the receiving end of J. C.
Walker’s service. In the 1920’s common bean mosaic was causing great losses to
the canning industry. In 1930 Walker and one of his students developed 2 varieties
of bean resistant to mosaic as well as making available to bean breeders everywhere,
information on the inheritance of the resistance.
ACADEMIC RECOGNITION
Few scientists have received greater acclaim for their contributions than J. C.
Walker. He is a member of The Wisconsin Academy of Science, The American
Association for the Advancement of Science, The American Institute of Biological
Sciences, The American Phytopathological Society (president in 1943), the Genetics
Society, The Botanical Society of America, and The American Society of Naturalists.
He has earned the coveted honor of election to the National Academy of
Sciences consisting of one hundred of the leading scientists of this country.
Dr. Walker was also elected to the College of Electors Hall of Fame of New York
University and was named “Man of the Year” by the National Vegetable Growers
Association in 1953.
Walker has been invited to present papers at several International Congresses.
He was a guest lecturer in 1953 at the Agricultural Institute of Sao Paulo, Brazil.
He has published well over 300 technical papers and has guided some 60 students
through their doctorate in Plant Pathology.
National recognition services for J. C. Walker have been held by the American
Seed Trade Association, The National Kraut Packers Association, The Vegetable
Growers of America, and the Manufacturers of Processing Equipment.
LIST OF PUBLICATIONS
1. Walker, J. C. Control of potato diseases in Wisconsin. Wis. Country Magazine
7: 34. 1913.
2. Walker, J. C. Studies upon the anthracnose of onion. Phytopathology
7: 59. 1917.
3. Walker, J. C. Control of neck rot and anthracnose of onion sets. Phytopathology
8: 70. 1918.
4. Walker, J. C. Notes on the resistance of onions to anthracnose. Phytopathology
8: 70-71. 1918.
5. Walker, J. C. Onion diseases and their control. U.S.D.A. Farm. Bul. 1060, 23
pp. 1919.
6. Vaughan, R. E., and J. C. Walker. Onion smut. Wis. Agr. Expt. Sta. Circ. 114,
4 pp. 1919.
7. Walker, J. C. Occurrence and control of black leg of cabbage. Phytopathology
10: 64. 1920.
8. Walker, J. C. Experiments upon formaldehyde-drip control of onion smut.
Phytopathology 10: 323-327. 1920.
9. Walker, J. C., and W. B. Tisdale. Observations on seed transmission of the
cabbage black rot organism. Phytopathology 10: 175-177. 1920.
10. Jones, L. R., J. C. Walker, and W. B. Tisdale. Fifth progress report on Fusarium-
resistant cabbage. Phytopathology 10: 64. 1920.
11. Jones, L. R., J. C. Walker, and W. B. Tisdale. Fusarium resistant cabbage. Wis.
Agr. Expt. Sta. Res. Bul. 148, 314 pp. 1920.
12. Vaughan, R. E., and J. C. Walker. Onion smut. Wis. Horticulture 10:
133-145. 1920.
13. Walker, J. C. A Macrosporium rot of onion. Phytopathology 11: 53. 1921.
14. Walker, J. C. The occurrence of dodder on onions. Phytopathology 11: 53.
1921.
15. Walker, J. C. Onion smudge. Jour. Agr. Res. 20: 685-722. 1921.
16. Walker, J. C. Rust of onion followed by a secondary parasite. Phytopathology
11: 87-90. 1921.
8 Wolf Prize in Agriculture
17. Walker, J. C., and L. R. Jones. The relation of soil temperature and other
factors to onion smut and infection. Phytopathology 11: 52-53. 1921.
18. Walker, J. C., and L. R. Jones. Relation of soil temperature and other factors
to onion smut infection. Jour. Agr. Res. 22: 235-262. 1921.
19. Walker, J. C. Onion diseases and their control. U.S.D.A. Farm. Bul. 1060 (1st
rev.), 28 pp. 1922.
20. Walker, J. C. Seed treatment and rainfall in relation to the control of cabbage
black leg. U.S.D.A. Bul. 1029, 27 pp. 1922.
21. Walker, J. C., and W. B. Tisdale. Further notes on the occurrence of cabbage
black leg. Eiiytppathology 12: 143. 1922.
22. Walker, J. C. The hot water treatment of cabbage seed, Phytopathology 13:
251-253. 1923.
23. Walker, J. C. Disease resistance to onion smudge. Jour. Agr. Res. 214: 1019-
1040. 1923.
24. Jones, L. R., and J. C. Walker. Yellows-resistant cabbage varieties. Seed World
(Feb. 2, 1923): 20-21. 1923.
25. Jones, L. R., J. C. Walker, and E. C. Tims. Work upon Fusarium-resistant
cabbage in 1922. Phytopathology 13: 57. 1923.
26. Harter, L. L., and L. R. Jones. Cabbage diseases. (Revised by J. C. Walker.)
U.S.D.A. Farm. Bul. 1351, 29 pp. 1923.
27. Walker, J. C. Occurrence of white rot of Alliuin (Sclerotium opivorum Berk.)
in Europe and America. Phytopathology 114: 26. 19214.
28. Tims, E. C., and J. C. Walker. A Fusarium bulb rot of onion. Phytopathology
114: 26-27. 1924.
29. Walker, J. C., and F. L. Wellman. Temperature relations of Urocystis cepulae
(Frost). Phytopathology 114: 26. 1924.
30. Walker, J. C. White rot of Allium in Europe and America. Phytopathology
114: 315-322. 1924.
31. Walker, J. C. Cabbage—seed treatment. U.S.D.A. Circ. 311, 4 pp. 1924.
32. Walker, J. C. Resistant varieties and disease-free seed save cabbage industry.
Wis. Horticulture 114: 117-118; 122. 1924.
33. Walker, J. C. Observations on the cultivation and diseases of cabbage arid
onions in Europe, 1922. Plant Disease Reporter, Suppl. 32, 314 pp. 1924.
34. Walker, J. C. On the nature of disease resistance in plants. Trans. Wis. Acad.
Science, Arts, and Letters 21: 225-2147, 19214.
35. Walker, J. C., and C. C. Lindegren. Further studies on the relation of onion scale
pigmentation to disease resistance. Jour. Agr. Research 29: 507-514. 1924.
36. Walker, J. C., and E. C. Tims. A Fusariun bulb rot of onion and the relation of
environment to its development. Jour. Agr. Research 28: 683-694. 1924.
37. Walker, J. C. An investigation into the nature of disease resistance in plants.
Science 59: 448. 1924.
John C. Walker 9
38. Walker, J. C. Onion diseases and their control. U.S.D.A. Farm. Bul. 1066 (2nd
rev.), 18 pp. 1925.
39. Walker, J. C. Two undescribed species of Botrytis associated with the neck
rot diseases of onion bulbs. Phytopathology 15: 708-713. 1925.
40. Walker, J. C. Control of mycelial neck rot of onion by artificial curing. Jour.
Agr. Research 30: 365-373. l925.
41. Walker, J. C. Studies on disease resistance in the onion. Proc. Nat. Acad. Sci.
11: 183-189. 1925.
42. Walker, J. C., C. C. Lindegren, and Freda N. Bachmann. Further studies on
the toxicity of juice extracted from succulent onion scales. Jour. Agr. Research
30: 175-187. 1925.
43. Jones, L. R., J. C. Walker, and J. Monteith, Jr. Fusarium resistant cabbage:
progress with second early varieties. Jour. Agr. Research 30: 1027-1034.
1925.
44. Walker, J. C. Studies upon the inheritance of Fusarium-resistance in cabbage.
Phytopathology 16: 87. 1926.
45. Walker, J. C. The influence of soil temperature and soil moisture upon white
rot of Allium. Phytopathology 16: 697-710. 1926.
46. Walker, J. C. Botrytis neck rots of onions. Jour. Agr. Research 33:
893-928. 1926.
47. Walker, J. C. Three new yellows-resistant mid-season cabbage varieties. Seed
World (Dec. 31, 1926.). 126.
48. Walker, J. C., and F. L. Wellman. Relation of temperature to spore germination
and growth of Urocystis cepulae. Jour. Agr. Research 32: 133-146. 1926.
49. Walker, J. C., John Monteith, Jr., and F. L. Tellman. A new Fusarium resistant
cabbage. Phytopathology 16: 72-73. 1926.
50. Walker, J. C. Newer aspects of breeding yellows-resistant cabbage. Nkt. Grow.
Jour. 38: 412-414. Also in The Canner 62 (10-2): 158. 1926.
51. Walker, J. C. Diseases of cabbage and related plants. U.S.D.A. Farm. Bul.
1439, 30 pp. 1927.
52. Walker, J. C. Onion curing to prevent decay while in storage. U.S.D.A. Yearbook
1926: 558-559. 1927.
53. Walker, J. C., John Monteith, Jr., and F. L. Wellman. Development of three
midseason varieties of cabbage resistant to yellows (Fusarium conglutinans
(Woil.). Jour. Agr. Research 35: 785-809. 1927.
54. Walker, J. C. Yellows-resistant midseason cabbage varieties now available.
Wis. Horticulture 17: 113-114. 1927.
55. Walker, J. C. An onion disease under successful control. Wis. Horticulture
17: 155-156. 1927.
56. Walker, J. C. Spraying and dusting calendar for North Central States
(Minnesota, Wisconsin, and Michigan). Amer. Produce Grower, April, 1927.
10 Wolf Prize in Agriculture
95. Walker, J. C., J. F. Adams, and W. H. Pierce. Results of planting tests with
Wisconsin Refugee and Idaho Refugee beans. The Canner 80 (12): 7-8: 22.
1935.
96. Walker, J. C. Diseases of vegetable crops. 65 pp. Ann Arbor. 1935.
97. Walker, J. C., and K. P. Link. Toxicity of phenolic compounds to certain
onion bulb parasites. Bot. Gaz. 96: 468-484. 1935.
98. Anderson, N. E., and J. C. Walker. Histological studies of Wisconsin Hollander
and Wisconsin Ballhead cabbage in relation to resistance to yellows. Jour.
Agr. Research 50: 823-826. 1935.
99. Walker, J. C., and R. H. Larson. Calcium cyanamide in relation to control of
clubroot of cabbage. Jour. Agr. Research 51: 183-189. 1935.
100. Walker, J. C. Types of disease resistance. Proc. Sixth int. Bot. Cong.
(Amsterdam) 2: 206-208. 1935.
101. Walker, J. C. A study of resistance to Fusarium wilt in Alaska peas. Amer.
Jour. Botany 22: 849-857. 1935.
102. Walker, J. C. Resistance to clubroot in Brassica. Phytopathology 26: 112.
1936.
103. Walker, J. C. Discussion of disease-free and disease-resistant beans. The
Canning Age 17: 171-172; 178. 1936.
104. Walker, J. C., and Otis C. Whipple. Tomato-seed certification. In Reports and
observations of some 1936 tomato growers etc. Crop Service Dept., Campbell
Soup Co., Chicago. Jan., 1937.
105. Walker, J. C. The fungicidal value of mustard oils. Phytopathology 27: 142.
1937.
106. Walker, J. C., and R. H. Larson. The increasing importance of cabbage mosaic.
Phytopathology 27: 142. 1937.
107. Walker, J. C. Varieties of beans for canning. Canning Trade 59 (30): 20-21.
1937.
108. Walker, J. C., Sam Morell, and H. H. Foster. Toxicity of mustard oils
and related sulfur compounds to certain fungi. Amer. Jour. Botany 214:
536-541. 1937.
109. Walker, J. C. Injury to -cabbage by lightning. Phytopathology 27: 858-861.
1937.
110. Walker, J. C. Onion diseases and their control. U.S.D.A. Farm. Bul. 1060 (5th
rev.), 24 pp. 1937.
111. Larson, R. H., and J. C. Walker. Properties and host range of a cabbage
mosaic virus. Phytopathology 28: 13. 1938.
112. Walker, J. C., and R. H. Larson. Soil temperature in relation to potato yellow
dwarf. Phytopathology 28: 21. 1938.
113. Whipple, O. C., and J. C. Walker. Two strains of cucumber virus on pea and
bean. Phytopathology 28: 22. 1938.
John C. Walker 13
114. Walker, J. C. Diseases of cabbage and related plants. U.S.D.A. Farm. Bul.
1439 (2nd rev.), 35 pp. 1938.
115. Walker, J. C., R. H. Larson, and A. Ft. Albert. Studies of resistance to potato
scab in Wisconsin. Amer. Potato Jour. 15: 246-252. 1938.
116. Walker, J. C. Cabbage disease controlled in South. Southern Seedsman. Nov.
1938, pp. 8, 20. 1938.
117. Larson, R. H., A. R. Albert, and J. C. Walker. Soil reaction in relation to potato
scab. Amer. Potato Jour. 15: 325-330. 1938.
118. Walker, J. C., J. P. Jolivette, and J. G. McLean. Internal black spot of canning
beets and its control. Canning Age 19: 489-491; 508. 1938.
119. Albert, A. R., R. H. Larson, and J. C. Walker. The comparative productiveness
of seed potatoes grown oh sandy and on peat soils in central Wisconsin.
Amer. Potato Jour. 16: 16-23. 1939.
120. Walker, J. C. Diseases of vegetable crops. 2nd rev., 67 pp. Ann Arbor.
1939.
121. Walker, J. C. Disease resistant pea varieties. Canning Trade 61 (27): 10.
1939.
122. Walker, J. C. Use of boron to control internal black spot of beets. Canning
Trade 61 (27): 16. 1939.
123. Walker, J. C. Internal black spot of garden beet. Phytopathology 29:
120-128. 1939.
124. Walker, J. C. Freezing injury to canning peas in Wisconsin. Phytopathology
29: 188-194. 1939.
125. Delwiche, E. J., F. L. Musbach, W. B. Sarles, Emil Truog, J. C. Walker, and H. F.
Wilson. Canning peas in Wisconsin. Wis. Agr. Expt. Sta. Bul. 444, 24 pp.
1939.
126. Pryor, Dean E., and J. C. Walker. A method for testing the toxicity of volatile
compounds. Phytopathology 29: 641-643. 1939.
127. Walker, J. C., and R. H. Larson. Yellow dwarf of potato in Wisconsin. Jour.
Agr, Research 59: 259-280. 1939.
128. Larson, R. H., and J. C. Walker. A mosaic disease of cabbage. Jour. Agr.
Research 59: 367-392. 1939.
129. Walker, J. C., and F. L. Musbach. Effect of moisture, fertility and fertilizer
placement on root rot of canning peas in Wisconsin. Jour. Agr. Research 59:
579-590. 1939.
130. Virgin, W. J., and J. C. Walker. Relation of temperature and moisture to near-
wilt of pea. Jour, Agr. Research 59: 591-600. 1939.
131. Walker, J. C. Resistance to clubroot in varieties of turnip and rutabaga. Jour.
Agr. Research 59: 815-828. 1939.
132. Larson, R. H., and J. C. Walker. A necrotic virosis of cabbage. Phytopathology 30:
15. 1940.
14 Wolf Prize in Agriculture
191. Walker, J. C., and J. P. Jolivette. Relation of boron to sugar beet production in
Wisconsin. Amer. Soc. Sugar Beet Tech. East U.S. and Canada Proc. 2nd
Region Meet. 1941: 30-31. 1941.
192. Walker, J. C. Soil management and plant nutrition in relation to disease
development. Soil Science 61: 47-54. 1946.
193. Smith, F. O., and J. C. Walker. Relation of environmental and hereditary
factors to ascorbic acid in cabbage. Amer. Jour. Bot. 33: 120-129. 1946.
194. Felton, H. W., and J. C. Walker. Environal factors affecting downy mildew of
cabbage. Jour. Agr. Res. 72: 69-81. 1946.
195. Jones, H. A., J. C. Walker, T. H. Little, and R. H. Larson. Relation of
color-inhibiting factor to smudge resistance in onion. Jour. Agr. Res. 72:
259-264. 1946.
196. Walker, J. C., and R. E. Foster. Plant nutrition in relation to disease development.
III. Fusarium wilt of tomato. Amer. Jour. Bot. 33: 259-264. 1946.
197. Smith, F. G., J. C. Walker, and W. J. Hooker. Effect of hydrogen-ion
concentration on the toxicity to Colletotrichum circinans (Berk.) Vogl. of
some carboxylic acids, phenols, and crucifer extracts. Amer. Jour. Bot. 33:
351-356. 1946.
198. Kuntz, J. E., and J. C. Walker. Virus inhibitors in spinach extract. Phytopath.
36: 404. 1946.
199. Foster, R. E., and J. C. Walker. Improvement of ascorbic acid content in
yellows-resistant cabbage. Phytopath. 36: 398. 1946.
200. Walker, J. C. James Peter Jolivette. Phytopath. 36: 415-417. 1946.
201. Walker, J. C., W. C. Edmundson, and H. A. Jones. Onion-set production. U. S.
Dept. Agr. Farmers’ Bul. 1955. 21 p. Rev. 1946.
202. Walker, J. C., and R. E. Foster. The inheritance of ascorbic acid content in
cabbage. Amer. Jour. Bot. 33: 758-76l. 1946.
203. Gorenz, A. H., and J. C. Walker. Influence of method sticker on the effectiveness
of Arasan for onion-smut control. Phytopath. 37: 7-8. 1947.
204. Walker, J. C., and Glenn S. Pound. Improvement of cabbage for disease
resistance. Phytopath. 37: 23. 1947.
205. Foster, R. E., and J. C. Walker. Predisposition of tomato to Fusarium wilt. Jour.
Agr. Res. 74: 165-185. 1947.
206. Smith, F. G., K. P. Link, and J. C. Walker. Acidic and phenolic fractions of
crucifer roots in relation to clubroot. Jour. Agr. Res. 74: 193-204. 1947.
207. Kuntz, J. E., and J. C. Walker. Virus inhibition by extracts of spinach. Phytopath.
37: 561-579. 1947.
208. Walker, J. C. Onion diseases and their control. U.S. Dept. Agr. 1060. 26 p.
7th Rev. 1947.
209. Walker, J. C. Seed-borne pea diseases and their control. Can. Seed Growers’
Assoc. Ann. Rpt. 1946-1947: 12-13. 1947.
18 Wolf Prize in Agriculture
210. Pound, G. S., and J. C. Walker. Strains of cucumber mosaic virus pathogenic
on crucifers. Phytopath. 38: 21-22. 1948.
211. Walker, J. C., and R. H. Larson. Development of clubroot resistance in cabbage.
Ibid. 38: 28. 1948.
212. Kendrick, J. B., Jr., and J. C. Walker. Anthracnose of tomato. Phytopath. 38:
247-260. 1948.
213. Walker, J. C., and J. B. Kendrick, Jr. Plant nutrition in relation to
disease development. IV. Bacterial canker of tomato. Amer. Jour. Bot. 35:
186-192. 1948.
214. Walker, J. C., and J. P. Jolivette. Yellows-resistant varieties of cabbage in the
early and midseason roundhead groups. U. S. Dept. Agr. Circ. 775. 20 p.
1948.
215. Walker, J. C., R. H. Larson, R. E. Foster, and J. E. Kuntz. Yellows- resistant
cabbage varieties in the Danish Ballhead-Hollander group. U.S. Dept. Agr.
Circ. 776. 12 p. 1948.
216. Grogan, R. G., and J. C. Walker. Interrelation of bean virus 1 and bean virus
2 as shown by cross-protection tests, Phytopath. 38: 1489-1493. 1948.
217. Pound, O. S., and J. C. Walker. Strains of cucumber mosaic virus pathogenic
on crucifers. Jour. Agr. Res. 77: 1-12. 1948.
218. Hatfield, W. C., J. C. Walker, and J. H. Owen. Antibiotic substances in onion
in relation to disease resistance. Jour. Agr. Res. 77: 115-135. 1948.
219. Kendrick, J. B., Jr., and J. C. Walker. Predisposition of tomato to bacterial
canker. Jour. Agr. Res. 77: 169-186. 1948.
220. Gorenz, A. N., J. C. Walker, and R. H. Larson. Morphology and taxonomy of
the onion pink-root fungus. Phytopath. 38: 831-840. 1948.
221. Walker, J. C. Diseases of cabbage and related plants. U. S. Dept. Agr. Farmers’
Bul. 1439. 38 p. 4th Rev. 1948.
222. Walker, J. C., G. S. Pound, and J. E. Kuntz. Development of Wisconsin 55
tomato. Wis. Agr. Expt. Sta. Bul. 478, 20 p. 1948.
223. Walker, J. C. Vegetable seed treatment. Bot. Rev. 14: 588-601. 1948.
224. Grogan, R. G., and J. C. Walker. A pod-distorting strain of the yellow mosaic
virus of bean. Jour. Agr. Res. 77: 301-314. 1948.
225. Grogan, R. G., and J. C. Walker. The relation of common mosaic to black root
of bean. Jour. Agr. Res. 77: 315-331. 1948.
226. Gorenz, A. N., R. H. Larson, and J. C. Walker. Factors affecting pathogenicity
of pink root fungus of onions. Jour. Agr. Res. 78: 1-18. 1949.
227. Hagedorn, D. J., and J. C. Walker. Cross protection tests between Wisconsin
pea streak virus and several strains of bean virus 2 from pea. Phytopath. 39:
11. 1949.
228. Walker, J. C. Resistance in cucumber to scab. Phytopath. 39: 27. 1949.
229. Chiu, W. F., and J. C. Walker. Morphology and variability of the cucurbit
black rot fungus. Jour. Agr. Res. 78: 81-102. 1949.
John C. Walker 19
230. Hare, W. W., J. C. Walker, and E. J. Delwiche. Inheritance of a gene for near-
wilt resistance in the garden pea. Jour. Agr. Res. 78: 239-250. 1949.
231. Walker, J. C., G. S. Pound, and J. E. Kuntz. Wisconsin 55 tomato. Seed World
64(10): 8-10; 15. 1949.
232. Calrert, O. H., G. S. Pound, J. C. Wa1ker, M. A. Stahmann, and J. F. Stauffer.
Induced variability in Phoma lingam. Jour. Agr. Res. 78: 571-588. 1949.
233. Chiu, N. F., and J. C. Walker. Physiology and pathogenicity of the cucurbit
black-rot fungus. Jour. Agr. Res. 51: 589-615. 1949.
234. Hagedorn, D. J., and J. C. Walker. Wisconsin pea stunt, a newly described
disease. Jour. Agr. Res. 78: 617-626. 1949.
235. Wells, D. G., N. W. Hare, and J. C. Walker. Evaluation of resistance and
susceptibility in garden pea to near-wilt in the greenhouse. Phytopath. 39:
771-779. 1949.
236. Hagedorn, D. J., and J. C. Walker. Wisconsin pea streak. Phytopath. 39: 837-
847. 1949.
237. Gallegly, M. E., Jr., and J. C. Walker. Plant nutrition in relation to disease
development. V. Bacterial wilt of tomato. Amer. Jour. Bot. 36: 613-623.
1949.
238. Wells, D. G., J. C. Walker, and W. W. Hare. A study of linkage between factors
for resistance t5 wilt and near-wilt in garden peas. Phytopath. 39: 907-912.
1949.
239. Gallegly, M. E., Jr., and J. C. Walker. Relation of environmental factors to
bacterial wilt of tomato. Phytopath. 39: 936-946. 1949.
240. Ladeburg, R. C., R. H. Larson, and J. C. Walker. The ringspot type of potato
virus X. Amer. Potato Jour. 26: 432-435. 1949.
241. Ladeburg, R. C., R. H. Larson, and J. C. Walker. Origin interrelation and
properties of ringspot strains of virus X in American potato varieties. Wis.
Agr. Expt. Sta. Res. Bul. 165. 1950.
242. Walker, J. C. The mode of seed infection by the cabbage blackrot organism.
Phytopath. 40: 30. 1950.
243. Walker, J. C., J. H. Owen, and M. A. Stahmann. Relative importance of
phenols and volatile sulfides in disease resistance in the onion, Phytopath.
40: 30. 1950.
244. Walker, J. C. The breeding and development of pea varieties for processing.
Canner 110(7): 50, 52. 1950.
245. Owen, J. H., J. C. Walker, and M. A. Stahmann. Pungency, color, and moisture
supply in relation to disease resistance in the onion. Phytopath. 40: 292-
297. 1950.
246. Hagedorn, D. J., and J. C. Walker. The relation of bean virus 2 to pea mosaic
in Wisconsin. Phytopath. 40: 684-698. 1950.
247. Owen, J. H., J. C. Walker, and M. A. Stahmann. Variability in onion neck- rot
fungi. Phytopath. 40: 749-768. 1950.
20 Wolf Prize in Agriculture
248. Walker, J. C. Can a staff member do research and teach at the same time?
Yes. P1. Dis. Rptr. 34: 281-282. 1950.
249. Larson, R. H., R. E. F. Mathews, and J. C. Walker. Relationships between
certain viruses affecting the genus Brassiea. Phytopath. 40: 955-962. 1950.
250. Stahmann, M. A., and J. C. Walker. Synthesized chemical can stop virus
action. Wis. Agr. Expt. Sta. Bul. 491: 60-62. 1950.
251. Walker, J. C. History of plant disease control. Ann. Rpt. Veg. Growers. Assoc.
Amer. 1950: 209-212. 1950.
252. Walker, J. C. Environment and host resistance in relation to cucumber scab.
Phytopath. 40: 1094-1102. 1950.
253. Walker, J. C., and R. H. Larson. Progress in the development of clubroot-
resistant cabbage. Phytopath. 41: 37. 1951. (Abst,) Jan.
254. Walker, J. C., and A. B. Wiles. Development of a scab-resistant pickling
cucumber for Wisconsin. Phytopath. 41: 37. 1951. Jan.
255. Walker, J. C. Plant pathology. 699 p. McGraw-Hill, New York, 1950.
256. Walker, J. C. Genetics and plant pathology. In Genetics in the Twentieth
Century, pp. 527-554. McMillan Co., NewYork, 1951. Mar.
257. Wellman, F. L., J. C. Walker, Allyn Cook, and M. E. Gallegly, Jr. Effects of
temperature on vegetative growth of five coffee disease fungi. In: Primera
Asamblea Latinoamericana de Fitoparasitologia. Misc. Folleto Mex. Dept. of
Agr, No. 4, pp. 251-257. 1951.
258. Webb, R. E., R. H. Larson, and J. C. Walker. Naturally occurring strains of the
potato leaf roll virus. Amer. Potato Jour. 28: 667-671. 1951. July.
259. Darby, J. F., R. H. Larson, and J. C. Walker. Variation in virulence and properties
of potato virus I strains. Wis. Agr. Expt. Sta. Res. Bul. 177. 1951. Aug.
260. Bridgmon, G. H., and J. C. Walker. The relation of southern bean mosaic to
black root. Phytopath. 41: 865-871. 1951. Oct.
261. Walker, J. C., and M. E. Gallegly, Jr. Plant nutrition in relation to disease
development. VI. Black rot of’ cabbage and ring rot of tomato. Amer. Jour.
Bot. 38: 663-665. 1951. Oct.
262. Wiles, A. B., and J. C. Walker. The relation of Pseudomonas lachrymans to
cucumber fruits and seeds. Phytopathology 41: 1059-1064. 1951.
263. Walker, J. C. Potato diseases: past, present, and future. In Ann. Rpt. Veg.
Growers Assn. America. 1951: 36-40. 1951.
264. Pound, G. S., and J. C. Walker. Mosaic resistance in cabbage. Phytopath. 41:
1083-1090. 1951.
265. Stahmann, M. A., L. H. Graf, E. L. Patterson, J. C. Walker, and D. W. Watson.
The inhibition of tobacco mosaic virus by synthetic lysine polypeptides. Jour,
Biol. Chem. 189: 45-52. 1951.
266. Sill, W. H., Jr., W. C. Burger, M. A. Stahmann, and J. C. Walker. Electron
microscopy of cucumber virus 1. Phytopathology 42: 19-20. 1952.
John C. Walker 21
286. Scheffer, H. P., and J. C. Walker. The physiology of Fusarium wilt of tomato.
Phytopathology 43: 116-125. 1953.
287. Walker, J. C., C. F. Pierson, and A. B. Wiles. Two new scab-resistant cucumber
varieties. Phytopathology 43: 215-217. 1953.
288. Scheffer, R. P., S. S. Gothoskar, M. A. Stahmann, and J. C. Walker. Tomato wilt
caused by fungus enzymes. Wis. Agr. Expt. Sta. Bul. 500: 11. 1953.
289. MacLachlan, D. S., R. H. Larson, and J. C. Walker. Strain interrelationships in
potato virus A. Wis. Agr. Expt. Sta. Res. Bul. 180. 1953.
290. Larson, R. H., and J. C. Walker. Arasan controls onion smut. Wis. Agr. Expt.
Sta. Bul. 509: 34. 1953.
291. Walker, J. C. Wisconsin SR6 cucumber resists scab disease. Wis. Agr. Expt.
Sta. Bul. 509: 36. 1953.
292. Pound, G. S., and. J. C. Walker. Autogenous necrosis of cabbage. Phytopathology
43: 415-418. 1953.
293. Winstead, N. N., and J. C. Walker. Metabolites of vascular Fusaria
of cabbage and cotton in relation to resistance. Phytopathology 43:
489-490. 1953.
294. Winstead, N. N., and J. C. Walker. Production of vascular browning by
metabolites from several pathogens. Phytopathology 43: 490. 1953.
295. Gothoskar, S. S., R. P. Scheffer, J. C. Walker, and M. A. Stahmann. The
role of pectic enzymes in Fusarium wilt of tomato. Phytopathology 43:
535-536. 1953.
296. Walker, J. C. Cauliflower, cabbage, and others. U. S. Dept. Agr. Yearbook
1953: 425-430. 1953.
297. Walker, J. C. Hazards to onions in many areas. U. S. Dept. Agr. Yearbook
1953: 43l-435. 1953.
298. Larson, R. H., and J. C. Walker. Thiram for smut control in onion set plantings.
Phytopathology 43: 596-597. 1953.
299. Walker, J. C. Disease resistance in the vegetable crops. II. Bot. Rev. 19: 606-
614. 1953.
300. Walker, J. C., R. H. Larson, and G. S. Pound. Badger Market, a new disease
resistant cabbage. Phytopathology 43: 649-650. 1953.
301. Walker, J. C., and E. K. Wade. The potato rot nematode. Special Leaflet Dept.
Plant Pathology, University of Wisconsin. 1954.
302. Walker, J. C. A few recollections and observations from thirty years
of battling vegetable diseases. Ann. Rept. Veg. Growers Assn. of America
1953: 75-78. 1954.
303. Scheffer, R. P., and J. C. Walker. 1954. Distribution and nature of Fusarium
resistance in the tomato plant. Phytopathology 44: 94-101.
304. Winstead, N. N., and J. C. Walker. 1954. Production of vascular browning by
metabolites from several pathogens. Phytopathology 44: 153-159.
John C. Walker 23
305. Winstead, N. N., and J. C. Walker. 1954. Toxic metabolites of the athogen in
relation to Fusarium resistance. Phytopathology 44: 159-166.
306. Hagedorn, D. J., and J. C. Walker. Virus diseases of canning peas in isconsin.
Wis. Agr. Expt. Sta. Res. Bul. 185. 1954.
307. MacLachlan, D. S., R. H. Larson, and J. C. Walker. Potato virus A. Amer.
Potato Jour. 31: 67-72. 195)-i.
308. MacLachlan, D. S., R. H. Larson, and J. C. Walker. Interveinal mosaic of
potato. Amer. Potato Jour. 31: 101-105. 1954.
309. Walker, J. C., and C. F. Pierson. SMR12 cucumber combines scab, mosaic
resistance. In What’s New in Farm Science. Wis. Agr. Expt. Sta. Bul. 511. 1954.
310. Pierson, C. F., and J. C. Walker. Relation of Cladosporium cucumerinum to
susceptible and resistant cucumber tissue. Phytopathology 44: 459-465. 1954.
311. Walker, J. C. Methods of controlling diseases of cauliflower and broccoli. In
Cauliflower and broccoli, varieties and culture. U.S.D.A. Farm. Bul. 1957.
312. Rice, R. V., M. A. Stahmann, G. D. Lindberg, and J. C. Walker. Some physical
biochemical characteristics of squash mosaic virus. Phytopathology 144: 503.
1954.
313. Sinclair, J. B., and J. C. Walker. Inheritance of resistance to common cucumber
mosaic virus in cowpea. Phytopathology 144: 506. 19514.
314. Walker, J. C., M. E. Gallegly, Jr., J. R. Bloom, and R. P. Scheffer. Plant nutrition
in relation to disease development. VIII. Verticillium wilt of tomato. Amer.
Jour. Bot. 141: 760-762. 1954.
315. Walker, J. C. 19514. Better cabbage through breeding. Vegetable Growers’
Messenger. 6 (3): 30.
316. Walker, J. C. 1955. Competition, cooperation, and evolution. Second Annual
Forty Miner Service Award Presentation.
317. Rice, R. V., G. D. Lindberg, P. Kaesberg, J. C. Walker, and M. A. Stahmann.
The three components of squash mosaic virus. Phytopathology 45: 145-148.
1955.
318. Gothoskar, S. S., R. P. Scheffer, M. A. Stahmann, and J. C. Walker. Further
studies on the nature of Fusarium resistance in tomato. Phytopathology 45:
303-307. 1955.
319. Walker, J. C., and M. A. Stahmann. Chemical nature of disease resistance in
plants. Ann. Rev. Plant Physiology 6: 351-366. 1955.
320. Gothoskar, S. S., R. P. Scheffer, J. C. Walker, and M. A. Stahmann. The role of
enzymes in the development of Fusarium wilt of tomato. Phytopathology
145: 381-387. 1955.
321. Bloom, J. R., and J. C. Walker. Effect of nutrient sprays on Fusarium wilt of
tomato. Phytopathology 45: 443-444. 1955.
322. Walker, J. C., and C. F. Pierson, Two new cucumber varieties resistant to scab
and mosaic. Phytopathology 145: 1451-1453. 1955.
24 Wolf Prize in Agriculture
361. Walker, J. C. Two new pickling varieties resistant to scab and mosaic. Plant
Dis. Reptr. 42: 1337-1338. 1958.
362. Walker, J. C. Benjamin Minge Duggar. Nat. Acad. Sci. U. S. Biog, Memoirs 32:
113-131. 1958.
363. Walker, J. C., R. H. Larson, and A. L. Taylor. Diseases of cabbage and related
plants. U. S. Dept. Agr. Handbook 144. 1958.
364. Tomlinson, J. A., R. J. Shepherd, and J. C. Wa1ker. Purification and serology
of cucumber mosaic virus. Nature 182: 1616. 1958.
365. Walker, J. C. Investigation of virus diseases of Brassica crops, by L. Broadbent,
Cambridge Univ. Press, New York, 94 p. 1957. A review. Quarterly Rev. of
Biology 33: 280. 1958.
366. Walker, J. C. Plant Pathology 2nd Edit. Review by Hassebrauk in Angewandte
Botanik No. 31. 1957.
367. Walker, J. C. Plant Pathology 2nd Edit. Review by S. D. Garrett in The New
Phytologist Vol 57, No. 1, March, 1958.
368. Srivastava, D. N., E. Echandi, and J. C. Walker. Pectolytic and cellulytic enzymes
produced by Rhizopus stolonifer. Phytopathology 49: 145-148. 1959.
369. Walker, J. C. Enfermedades de las hortizalizas. Translation by Antonia Arnal
Verderol. Salvat Editores, Barcelona. 624 pp. 1959.
370. Tomlinson, J., R. J. Shepherd, and J. C. Walker. Purification, properties, and
serology of cucumber mosaic virus. Phytopathology 49: 293-299. 1959.
371. Srivastava, D. N., and J. C. Walker. Mechanisms of infection of sweet potato
roots by Rhizopus stolonifer. Phytopathology 49: 400-406. 1959.
372. Walker, J. C. Disease resistance in crucifers. Proc. IXth Int. Botanical Congress
(Montreal) Vol. II, p. 420. 1959.
373. Walker, J. C. Progress and problems in controlling plant diseases by host
resistance. Plant Pathology. Problems and Progress 1908-1958: 32-41. Univ.
of Wis. Press, 1959. 1959.
374. Walker, J. C. Disease resistance. Sigma Xi lecture, Univ. of Nebraska, Nov. 18,
1959.
375. Walker, J. C., and R. H. Larson. 1959. Badger shipper cabbage resists clubroot
and yellows. Wis. Agr. Expt. Sta. Bul. 538: 100.
376. Heitefuss, R., D. J. Buchanan-Davidson, M. A. Stahmann, and J. C. Walker.
Electrophoretic and immunochemical studies of proteins in cabbage infected
with Fusarium oxysporuza f. conglutinans. Phytopathology 50: 198-205. 1960.
377. Nayudu, M. V., and J. C. Walker, 1960. Bacterial spot of tomato as influenced
by temperature and by age and nutrition of the host. Phytopathology 50:
360-364.
378. Heitefuss, R., M. A. Stahmann, and J. C. Walker. Production of pectolytic
enzymes and fusaric acid by Fusarium oxysporum f. conglutinans in relation
to cabbage yellows. Phytopathology 50: 367-370. 1960.
John C. Walker 27
414. Walker, J. C. Control of plant diseases through breeding for host resistance,
lecture delivered at Yale Forestry School, May 14, 1963.
415. Deverall, B. J., and J. C. Walker. A physiological difference between bean
leaves (Phaseolus vulgaris), resistant and susceptible to halo blight, caused
by Pseudomonas phaseolicola. Ann. Appl. Biology 52: 105-115. 1963.
416. Williams, P. H., and J. C. Walker. Races of clubroot in North America. Plant
Dis. Reptr. 47: 608-611. 1963.
417. Menke, G. H. Physiology of Rhizopus infection on carrot. Phytopathology 53:
882. 1963.
418. Patel, P. N., and J. C. Walker. Free amino acid and amide content
of tobacco and oats infected by wildfire and halo blight bacteria.
Phytopathology 53: 885. 1963.
419. Menke, G. H., and J. C. Walker. Metabolism of resistant and susceptible
cucumber varieties infected with cucumber mosaic virus. Phytopathology
53: 1349-1355. 1963.
420. Walker, J. C. Bacterial plant pathogen. By C. Stapp: translated by
A. Schoenfeld. Oxford Univ. Press. 1961. A Review. Quar. Rev. Biol. 38: 265-
266. 1963.
421. Walker, J. C. Diseases of sorghum, sudan grass and broom corn. By S. A. J.
Tazrr. Commonwealth Myco. Inst. 1962. A review. Quart. Rev. Biology 38:
266. 1963.
422. Seaman, W. L., J. C. Walker, and R. H. Larson. A new race of Plasmodiophora
brassicae affecting Badger Shipper cabbage. Phytopathology 53: 1426-1429.
1963.
423. Walker, J. C. The future of plant pathology. Ann. Rev. of Phytopathology 1:
1-4. 1963.
424. Walker, J. C. The physiology of disease resistance. W. Va. Agr. Exp. Sta. Bull.
488T: l-25. 1963.
425. Walker, J. C., and P. N. Patel. Changed bean disease may cause trouble. Wis.
Agr. Expt. Sta. Bull. 566: 44. 1963.
426. Walker, J. C., G. S. Pound, and P. H. Williams. TBR Globe arid Globelle
cabbages which resist tipburn. Wis. Agr. Expt. Sta. Bull. 566: 46. 1963.
427. Chand, J. N., and J. C. Walker. Relation of age of leaf and varietal resistance
to bacterial multiplication in cucumber inoculated with Pseudomonas
lachrymans. Phytopathology 54: 49-50. 1964.
428. Chand, J. N., and J. C. Walker. Inheritance of resistance to angular leafspot
of cucumber. Phytopathology 54: 51-53. 1964.
429. Walker, J. C., and P. N. Patel. Splash dispersal and wind as factors in
epidemiology of halo blight of bean. Phytopathology 54: 140-141. 1964.
430. Menke, G. H., P. N. Patel, and J. C. Walker. Physiology of Rhizopus stolonifer
infection on carrot. Z. f. Pflanzenkrankheiten und Pflanzenschutz 71:
128-140. 1964.
30 Wolf Prize in Agriculture
466. Walker, J. C. Plant Pathology. 3rd. edition. McGraw-Hill Co., New York.
1969. Not bound herein.
467. Chand, J. N., and J. C. Walker. Relation of free amino acids in cucumber
leaves to the development of angular leaf spot. Phyt. Ztschr. 64: 94-97.
1969.
468. Walker, J. C. Plant Pathology, its background and importance. BSA McGraw-
Hill Tape. 1969.
469. Walker, J. C. The story of disease resistance in cabbage. Tape I. 1969.
470. Walker, J. C. The story of disease resistance in cabbage. Tape II. 1969.
471. Walker, J. C. Six decades with cabbage clubroot. Tape 1969.
472. Halloin, J. M., J. C. Walker, G. A. de Zoeten, and G. Gaard. Effects of tentoxin
on plastids of cucumber and cabbage. Phytopathology 59: 1028-1029. 1969.
473. Strandberg, J. O., J. F. Darby, J. C. Walker, and P. H. Williams. Black speck, a
nonparasitic disease of cabbage. Phytopathology 59: 1879-1883. 1969.
474. Halloin, J. M., G. A. de Zoeten, G. Gaard, and J. C. Walker. The effects of
tentoxin on chlorophyll synthesis and plastid structure in cuctumber and
cabbage. Plant Physiol. 45: 310-3l4. 1970.
475. Yoshii, K., and J. C. Walker. Production of indolacetic acid in culture by
Pythium debaryanum. Phytopathology 60: 1321. 1970.
476. Main, Charles E., and J. C. Walker. Physiological responses of susceptible and
resistant cucumber to Erwinia tracheiphila. Phytopathology 61: 518-522.
1971.
477. Walker, J. C. Fusarium wilt of tomato. Amer. Phytopathol. Soc. Monograph
No. 6. 1971.
George F. Sprague
University of Illinois
Urbana, Illinois, USA
k
1902–1998
33
34 Wolf Prize in Agriculture
gene pool of maize germ plasm. Professor Sprague’s genetic research laid the
ground work, for improvement in nutritional quality in maize. A fact, which holds
great promise to maize-eating nations. He conducted investigations, which
demonstrated that protein quality of maize was genetically modifiable.
In summary, few people in the history of agriculture have had such
a profound impact on the improvement of a major crop as has Professor
Sprague.
CURRICULUM VITAE
PERSONAL DATA:
DEGREES
B.S. University of Nebraska 1924
M.S. University of Nebraska 1926
Ph.D. Cornell University (Genetics) 1930
SIGNIFICANCE OF CONTRIBUTIONS
Dr. Sprague’s accomplishments have been primarily in three categories: development
of new breeding methodology and the consequent production of exceedingly
important inbreds widely used in commercial hybrid production, the training of
graduate students, and the elaboration of genetic models dealing with such diverse
traits in maize as endosperm color, leaf and scutellum conformation and chemical
characteristics as oil content and protein quality.
We particularly cite Dr. Sprague’s untiring effort to join theoretical quantitative
genetic theory and practical plant breeding. Throughout his career he has had, by
his own wish, responsibility both for basic research and applied plant breeding
Unquestionably he is preeminent in his success in simultaneously conducting basic
studies on the nature of heterosis and in developing hybrids for the farmer. Basic
studies on the mathematics of selection led to the development of a gene pooi,
“Super Stiff Stalk Synthetic”, from which he isolated two lines, perhaps more
widely grown than any others, B14 and B37. The nominators believe that 70% of
the corn acres in the central Corn Belt in 1973 were planted to hybrids carrying
either B14 or B37, or both.
Sprague’s interest in nutritional quality led him to pioneer in the development
of waxy (straight chain starch) hybrids which were used during World War II to
produce a substitute for tapioca. In the 1940’s, he and his students began a series
of investigations which demonstrated that protein quality in corn was genetically
modifiable. Unfortunately, the group failed to analyze the opaque-2 mutant and it
remained for Purdue scientists to discover high lysine corn 20 years later. Oil
content of the kernel also came under his study and he developed breeding schemes
which rapidly increased it. Most of the research conducted by Sprague has been of
long-range interest and fundamental to the successes of corn agriculture.
Most important and consequential to the development of modern plant breeding
have been the following principles for which he was primarily responsible:
Even though Sprague was deeply involved with research, he devoted a great deal
of time and interest to graduate teaching. He taught a widely acclaimed course in
corn breeding during his 18-year stay at Iowa State. He attracted students from
around the world and served as major advisor for more than 50 M.S. and Ph.D.
candidates. His excellence as a teacher was recognized by the Gamma Sigma Delta
Superior Teaching Award, an unusual award for teachers of graduate courses.
Sprague’s influence has not been confined to the U. S. He has long been a
consultant with the Rockefeller Foundation and has traveled widely in that
role. He also was involved with the Marshall Plan (ECA) after the war and
was instrumental in the rapid spread of hybrid corn in Europe. Since 1963
he has played a central role in the U. S. Department of Agriculture activity in
improvement of cereal production in Africa. He has represented agriculture ably
in the National Academy of Science as chairman of the Section of Applied Biology.
Sprague’s monumental contributions to theoretical plant breeding and to the
improvement of maize are now made most evident by the wide use of his lines and
of the basic breeding pools that arose under his direction. Breeding methodologies
that he pioneered are widely used by commercial corn breeders in the U. S. and
around the world. The success of corn improvement programs in East Africa, Latin
America and Europe trace largely to the procedures he worked out and the carefully
planned selection experiments which generated data to support his basic theories.
Few people in the history of agriculture have had such a profound impact on the
improvement of a major crop.
We particularly wish to emphasize the impact Sprague’s research has had on
practical agriculture, particularly that of the US Corn Belt. For example, inbred
B37 was developed by Sprague from Iowa Stiff Stalk Synthetic, a variety developed
by recurrent selection particulary to serve as a source of superior inbred. Because
of the superiority of B37, hybrids carrying it in their pedigree have dominated the
commercial market because of their superiority in yield, standability and early
maturity. In 1970, it was the most extremely important, but other lines developed
by Sprague himself and by others using his synthetics have been almost as
consequential and continue to dominate corn agriculture.
George F. Sprague 37
LIST OF PUBLICATIONS
Sprague, G.F. 1927. Heritable characters of maize XXVII colored scutellum.
J. Hered. XVIII: 41-44.
Sprague, G.F. 1929. Heterofertilization in maize. Science LXIX: 526-527.
Richey, F.D., and G.F. Sprague. 1931. Experiments on hybrid vigor and convergent
improvement in corn. USDA Tech. Bull. 267. 22 p.
Sprague, G.F. 1932. The inheritance of colored scutellum in maize. USDA Tech.
Bull. 292. 43 p.
Sprague, G.F., and M.N. Pope. 1932. A case of two simultaneous mutations for
virescent seedlings in maize. Amer. Nat. LXVI: 284-85.
Sprague, G.F. 1932. The nature and extent of hetero-fertilization in maize. Genetics
17: 58-68.
Richey, F.D., and G.F. Sprague. 1932. Some factors affecting the reversal of sex
expression in the tassels of maize. Amer. Nat. LXVI: 433-443.
Sprague, G.F. 1933. Pollen tube establishment and the deficiency of waxy seeds in
certain maize crosses. Proc. Nat. Acad. Sci. 19: 838-841.
Sprague, G.F. 1934. Experiments on iarovising corn. J. Agr. Res. 48:1113-1120.
Richey, F.D., G.H. Stringfield, and G.F. Sprague. 1934. The loss in yield that may be
expected from planting second generation double-crossed seed corn.
J. Amer. Soc. Agron. 26: 196-199.
Sprague, G.F. 1935. Random sampling and the distribution of phenotypes on ears
of backcrossed maize. J. Agr. Res. 51: 751-758.
Sprague, G.F. 1936. The relation of moisture content and time of harvest to
germination of immature corn. J. Amer. Soc. Agron. 28: 472-478.
Stadler, L.J., and G.F. Sprague. 1936. Genetic effects of ultra-violet radiation in
maize. I. Unfiltered radiation. Proc. Nat. Acad. Sci. 22: 572-578.
Stadler, L.J., and G.F. Sprague. 1936. Genetic effects of ultra-violet radiation in
maize. II. Filtered radiation. Proc. Nat. Acad. Sci. 22: 579-583.
Stadler, L.J., and G.F. Spraque. 1936. Genetic effects of ultra-violet monochromatic
2537, and comparison-of effects of X-ray and ultra-violet treatment. Proc.
Nat. Acad. Sci. 22: 584-591.
Sprague, G.F. 1936. Hybrid vigor and growth rates in a maize cross and its
reciprocal. J. Agr. Res. 53: 819-830.
Perry, H.S., and G.F. Sprague. 1936. A second chromosome gene, Y3, producing
yellow endosperm color in maize. J. Amer. Soc. Agron. 28: 990-996.
Stadler, L.J., and G.F. Sprague. 1937. Contrasts in the genetic effects of ultra-violet
radiation and X-rays. Science 85: 57-58.
Sprague, G.F. 1939. Corn hybrids for Missouri. Mo. Agr. Exp. Sta. Circ. 201. 27 p.
Sprague, G.F. 1939. An estimation of the number of top-crosses plants required
for adequate representation of a corn variety. J. Amer. Soc. Agron. 31:
11-16.
38 Wolf Prize in Agriculture
Sprague, G.F. 1939. Heritable characters in maize 50 vestigal glume. J. Hered. 30:
143-145.
Bryan, A.A., R.C. Eckhardt, and G.F. Sprague. 1940. Spacing experiments with
corn. J. Amer. Soc. Agron. 32:707-714.
Millang, Amy, and G.F. Sprague. 1940. The use of punched card equipment in
predicting the performance of corn double crosses. J. Amer. Soc. Agron. 32:
815-816.
Sprague, G.F. 1941. The location of dominant favorable genes in maize by means
of an inversion. Genetics 26: 170.
Sprague, G.F., and A.A. Bryan. 1941. The segregation of genes affecting yield
prepotency, lodging and disease resistance in F3 and- F4 lines-of corn.
J. Amer. Soc. Agron.- 33: 207-214.
Sprague, G.F. 1941. Transmission tests of maize mutants induced by ultra-violet
radiation. Ia. Agr. Exp. Sta. Res. Bull. 292. p. 389-407.
Sprague, G.F. Building new corn hybrids. Ia. Farm Science Reporter 2: 7-9.
Sprague, G.F. 1942. Production of hybrid corn. Ia. Agr. Exp. Sta. Bull. P48.
p. 556-582.
Sprague, G.F. 1942. Trends in corn breeding. 33rd Ann. Report; Nebr. Crop Imp.
Assc. p. 50-57.
Sprague, G F., and L.A. Tatum. General vs. combining ability in single crosses of
corn. J. Amer. Soc. Agron. 34: 923-932.
Hixon, R.M., and G.F. Sprague. 1942. Waxy starch of maize and other cereals. Ind.
Eng. Chem. 34: 959-962.
Sprague, G.F., and R.M. Hixon. 1942. Waxy corn, a new crop. Ia. Farm Science
Reporter 3: 10-11.
Sprague, G.F., and M.T. Jenkins. 1943. A comparison of synthetic varieties, multiple
crosses and double crosses in corn. J. Amer. Soc. Agron. 35: 137-147.
Sprague, G.F., B. Brimhall, and R.M. Hixon. 1943. Some effects of the waxy
gene in corn on properties of the endosperm starch. J. Amer. Soc. Agron. 35:
817-822.
Sprague, G.F. 1943. The problem of heterosis. Chronica Botanica 7: 418-419.
Hixon, R.M., G.S. Shepherd, and G.F. Sprague. 1943. Technological problems as
related to war demands on corn. 8th Ann. Ia. Corn Res. Inst. Rpt. p. 7-10.
Sprague, G.F. 1944. The value of hybrid corn to the Iowa farmer. 9th Ann. Ia. Corn
Res. Inst. Rpt. p. 7-11.
Kinman, M.L., and G.F. Sprague. 1945. Relation between number of parental lines
and theoretical performance of synthetic varieties of corn. J. Amer. Soc. Agron.
37: 341-351.
Brimhall, B., G.F. Sprague, and J.E. Sass. 1945. A new waxy allele in corn and its
effect on the properties of the endosperm starch. J. Amer. Soc. Agron. 37:
937-944.
George F. Sprague 39
Sprague, G.F. 1946. Early testing of inbred lines of corn. J. Amer. Soc. Agron. 38:
108-117.
Sprague, G.F. 1946. The experimental basis for hybrid maize. Biol. Review 21:
101-120.
Hansen, D.W., Brimhall, B., and G.F. Sprague. 1946. Relationship of zein to the
total protein of corn. Cereal Chem. 23: 329-335.
Sprague, G.F., and J.C. Cunningham. 1946. Growing the bumper corn crop. Ch. 3
in A century of farming in Iowa — 1846–l946. p. 32–44.
Sprague, G.F., and J.C. Cunningham. 1946. One hundred years of corn growing in
Iowa. Ia. Yearbook of Agr. (1945) 46: 258-274.
Federer, W.T., and G.F. Sprague. 1947. A comparison of variance components in
corn yield trials: 1. Error, tester x line and line components in top cross
experiments. J. Amer. Soc. Agron. 39: 453-463.
Sprague, G.F., and M.T. Jemkins. 1947. The development of waxy corn for industrial
use. Ia. State Colil. Jour. Sci. 22: 205-213.
Sprague, G.F. Breeding for improved nutritional and industrial use. p. 42-49. In
Growth and development of the corn plant. Published by Amer. Seed Trade
Assn.
Groszmann, A., and G.F. Sprague. 1948. Comparative growth rates in a reciprocal
maize cross. The kernel and its component parts. J. Amer. Soc. Agron. 40:
88-98.
Sprague, G.F. 1948. Les Bases Experirnentales de la production de Maiz Hybrid.
p. 77-127. In L’Ameliorat.ion du Mais. Rabat, Morocco.
Sprague, G.F. 1948. Better protein, more oil in your corn. Iowa Farm Science 2:
3-4.
Sprague, G.F., and M.T. Jenkins. 1949. O Desenvolvimento do Metho Ceraceo para
uso Industrial. Ceres 8: 112-123.
Frey, K.J., B. Brimhall, and G.F. Sprague. 1949. The effect of selection upon protein
quality in the corn kernel. Agron. J. 41: 399-403.
Sass, J.E., and G.F. Sprague. 1949. Histological development of “accessory
blade” and associated abnormalities in maize. Ia. State Coll. J. Sci. 23:
301-309.
Rossman, E.C., and G.F. Sprague. 1949. Effect of 2,4-D on yields of maize in the
succeeding. generation after treatment. Plant Physiol. 24: 770-773.
Sprague, G.F., and B. Brimhall. 1949. Quantitative inheritance of oil in the corn
kernel. Agron. J. 41: 30-33.
Sprague, G.F. 1949. Feeding quality of corn improved by breeding. Crops and Soils.
Sass, J.E., and G.F. Sprague. 1950. The embryology of “germiess” maize. Ia. State
Coll. J. Sci. 24: 209-218.
Sprague, G.F., and B. Brimhall. 1950. Relative effectiveness of two systems of
selection for oil content of the corn kernel. Agron. J. 42: 83-88.
40 Wolf Prize in Agriculture
Sprague, G.F., and P.A. Miller. 1950. A suggestion for evaluating current concepts
of the genetic mechanism of heterosis in corn. Agron. J. 42: 161-162.
Sprague, G.F., J.H. Lilly, and D.D. Rubis. 1950. Can planting dates beat borers. Ia.
Farm Sci. 4: 150-151.
Brimhall, B., and G.F. Sprague. 1951. Unsaturates of corn oil-inheritance and
maturity studies. Cereal Chem. 28: 225-231.
Sprague, G.F., and W.T. Federer. 1951. A comparison of variance components in
corn yield trials. II. Error, year x variety, location x variety and variety
components. Agron. J. 43: 535-541.
Sprague, G.F., and P.A. Miller. 1951. New corn hybrids for Iowa. Ia. Farm Sci.
5: 137-138.
Sprague, G.F., and A.L. Lang. 1951. Further progress in hybrid maize production in
European countries. O.E.E.C.
Sprague, G.F., P.A. Miller, and B. Brimhall. 1952. Additional studies of the relative
effectiveness of two systems of selection for oil content of the corn kernel.
Agron. J. 44: 329-331.
Sprague, G.F. 1952. Early testing and recurrent selection. Chap. 26, p. 400-417. In
Heterosis. Ia. State Coil. Press.
Sprague, G.F., and P.A. Miller. 1952. The influence of visual selection during
inbreeding on combining ability in corn. Agron. J. 44: 258-261.
Rojas, B.A., and G.F. Sprague. 1952. A comparison of variance components in corn
yield trials. III. General and specific combining ability and their interaction
with locations and years. Agron. J. 44: 462-466.
Sprague, G.F. 1953. Heterosis. Chapt. 7. In Growth and differentiation in plants.
p. 113-136.
Sprague, G.F. 1954. Breeding for resistance to stalk rot. Ninth Ann. Hybrid Corn
Industry Research Conf., Proc. p. 38-43.
Sprague, G.F. 1955. Corn breeding. Chapt. 5. In Corn., and corn improvement.
p. 221-292. Academic Press. New York.
Sprague, G.F. 1955. Industrial utilization. Chapt. 14. In Corn and corn improvement.
p. 613-636. Academic Press. New York.
Sprague, G.F. 1955. The world production of corn. Chap. 16. In Corn and corn
improvement. p. 679-688. Academic Press. New York.
Sprague, G.F. 1954. The use of variance components as a guide in designing an
adequate testing program. 2nd Pan Amer. Agron. Congress.
Sprague, G.F. 1954. Factors affecting choice of tester for the evaluation of inbred
lines of corn. 2nd Pan Amer. Agron. Congress.
Sprague, G.F. 1954. A comparison of theoretical genetic implications of continuous
self-fertilization and recurrent selection as methods of corn breeding. 2nd Pan
Amer. Agron Congress.
LeRoux, P M., J.G. Dickson, and G.F. Sprague. 1954. A genetic basis for rust resistance
in corn. Phytopathology 44: 496.
George F. Sprague 41
Hooker, A.L., G.F. Sprague, and W.A. Russell. 1954. Correlation of resistance to
several stalk rots, ear rots and seedling blights and root necrosis in corn.
Agron. Abstracts. p. 70.
Hooker, A.L., G.F. Sprague, and W.A. Russell. 1955. Resistance to rust Puccinia
sorghi in corn. Agron. J. 47: 388.
Schuler, J.F., and G.F. Sprague. 1955. Natural mutations in inbred lines of maize
and their heterotic effect. II. Comparisons of mother line vs. mutant when
outcrosses to unrelated inbrecls. Genetics 41: 281-291.
Clem, Mary A., C.C. Mosier, and G.F. Sprague. 1956. Simplified punched
card procedures for predicting double cross performance. Agron. J. 48:
319-320.
Sprague, G.F. 1955. Problems in the estimation and utilization of genetic variability.
Cold Spring Harbor Symposia on Quantitative Biology, XX. p. 87-92.
Sprague, G.F., and A. Tavcar. 1956. Nais (Zea mays). General considerations and
American breeding work. Handbuch der Pflanzenzuchtung II Band 2:
103-143.
Sprague, G.F. 1955. Replications vs. location. 10th Ann. Hybrid Corn Industiry-
Research Conf., Proc. p. 10-13.
Pesek, J., G.F. Sprague, John Hanway, and F.A. Loeffel. 1955. The uptake of nitrogen,
phosphorus and potassium by different corn single crosses as affected by
population and nitrogen levels. Agron. Abst. 47: 32.
Sprague, G.F., and W.A. Russell. 1956. Some evidence on type of gene action
involved in yield heterosis in maize. Int. Genenetics Symposia, Proc. (Tokyo).
p. 522-526.
Sprague, G.F., W.A. Russell, and L.H. Penny. 1959. Further studies on convergent
improvement in corn. Genetics 44: 341-346.
Sprague, G.F. 1969. Germplasm manipulations of the future. In Physiological aspects
of crop yield. p. 375-396.
Sprague, G.F. 1970. World research needs for increasing corn production. Agr.
Eng. 51: 71, 91 and 108.
Sprague, G.F. 1970. High lysine corn. Span. 13: 1-4.
Sprague, G.F. 1970. A method of breeding for resistance. 6th Indian Phytopathological
Cof. Vol 23, June.
Russell, W.A., L.H. Peny, G.F. Sprague, W.D. Guthrie, and F.F. Dicke. 1970. Registration
of maize (PL1-13) parental lines. Crop Sci. 11: 143.
Sprague, G.F., and H.H. McKinney. 1971. Further evidence of the behavior of AR in
maize. Genetics 67: 533-543.
Sprague, G.F., and R.G. Dahms. 1971. Development of crop resistance to insects.
J. Envir. Quality 1: 28-34.
Sprague, G.F. 1971. Genetic vulnerability in corn and sorghum. 26th Ann. Corn
and Sorghum Res. Conf., Proc. p. 96-104.
42 Wolf Prize in Agriculture
1919–1991
BIOGRAPHICAL SKETCH
Sir Kenneth was trained in agricultural science at the University of Reading and in
1939 he joined the staff of the National Institute for Research in Dairying to work
on aspects of the nutrition of cattle. After a period in the Armed Services he
returned to Reading and later moved to the Central Veterinary Laboratory at
Weybridge. Prom there, after a period spent working with the late Dr H.H. Mitchell
at the University of Illinois, he was appointed Head of the Nutrition Department
of the Hannah Research Institute, Ayr, where he remained for 17 years until, in
1995, he was appointed Director of the Rowett Research Institute, Aberdeen.
Sir Kenneth has made outstanding contributions to knowledge of nutrition,
particularly ruminant nutrition. In his earlier days he did important work on the
nutritive value of feeds for dairy cattle, on protein metabolism and on the protein
requirements of cattle. In the fields of endocrinology he tackled the then
controversial subject of iodinated proteins. Some of the work on protein
requirements formed part of an extensive series of experiments on the metabolism
and nutritional needs of the very young calf and its relation to maternal feeding
which Sir Kenneth began soon after going to the Hannah Research Institute in
1948. This work is so outstanding in its originality and elegance that it is now
regarded as classic; it provided a basis for the development of systems of artificial
rearing of calves with specially formulated diets which have revolutionized
commercial calf production.
43
44 Wolf Prize in Agriculture
CURRICULUM VITAE
ACADEMIC TRAINING:
1930-1935 - City of Norwich School
1936-1939 - University of Reading
ACADEMIC TTRAINING:
1939 - B.Sc. (Agric.) University of Reading
1939 - N.D.A. (Hons.)
1944 - Ph.D. University University of Reading
1952 - D.Sc. University of Reading
1974 - D.Sc. (honoris causa) Queen’s University, Belfast
1975 - D.Sc. Agr. (honoris causa) University, Norway
1977 - D.Sc. Agr. (honoris causa) University of Leeds
PROFESSIONAL QUALIFICATIONS:
1962 - Fellowship of Royal Society of Arts (F.R.S.A.)
1963 - Fellowship of Institute of Biology (F. Inst. Biol.)
1964 - Fellowship of Royal Agricultural Society of England (F.R.Agr.E.)
1965 - Fellowship of Royal Society of Edinburgh (F.R.S.E.)
1967 - Fellowship of Royal Society (F.R.S.)
1970 - Foreign Member of the Lenin Academy of Agricultural Sciences (USSR)
46 Wolf Prize in Agriculture
CAREER:
1939-1940 - Research Assistant at The National Institute for Research in Dairying
1941-1944 - -"-
1940-1941 - War Service (Royal Artillery)
1944-1946 - Research Officer, Ministry of Agriculture, Veterinary Laboratory,
Weybridge
1947-1948 - -"-
1946-1947 - Commonwealth Fund Fellow at the University of Illinois, Division of
Nutrition under Professor H. H. Mitchell
1948-1965 - PSO then SPSO, Head, Nutrition Department, Hannah Dairy Research
Institute, Kirkhill, Ayr, Scotland, DCSO, Special Merit
Since 1965 - CSO, Director, Rowett Research Institute, Bucksburn, Aberdeen, and
Consultant
1975 - Director, Commonwealth Bureau of Nutrition, Under Secretary,
Special Merit
HONORARY APPOINTMENTS:
1958-1961 - Olive Belirens Lecturer in Faculty of Agriculture, University of Leeds
1962 - Visiting Lecturer, Berlin Academy, D.D.R.
1964-1965 - Visitiiag Lecturer, The National Research Council of Canada, to
Universities of British Columbia
1965 - Manitoba, Saskatchewan. and Alberta, Corresponding member of
the French Academy of Sciences
PROFESSIONAL ACTIVITIES:
1970-1971 - President, British Society of Animal Production
1974-1977 - President, The Nutrition Society
1965 - Editor, Journal of Agricultural Science
NOMINATED LECTURES:
1962 - Fernhurst Lecture, Royal Society of Arts
1964 - Samuel Brody Memorial Lectures, University of Missouri, U.S.A.
1965 - Scott Robertson Memorial Lecture, Queen’s University, Belfast
1972 - Blackman Lecture, University of Oxford
1973 - Wooldridge Memorial Lecture (British Veterinary Association)
1974 - Jubilee Lecture of the Agricultural Research Council of Norway
1976 - Hammond Lecture, British Society of Animal Production
1976 - Jubilee Lec1ture of the University of Reading
1977 - Wi1liam Dick Memorial Lecture, Edinburgh
1978 - Storer Lectures, University of California, Davis, California, U.S.A.
LIST OF PUBLICATIONS
1. Straw pulp: Recent experiments. K. L. Blaxter, S. Bartlett. Journal of the
Ministry of Agriculture, 50, 224-226 (1943).
2. The normal variation in the heart rate of dairy cows. K. L. Blaxter. British
Veterinary Journal, 99, 2-4 (1943).
3. Stimulation of the milk production of dairy cows by feeding thyroid-active
iodinated proteins. K. L. Blaxter. Nature, London, 152, 751-752 (1943).
4. Experiments on the use of homegrown foods for milk production. 1. The
effect of war-time can change food supply on the nutrient intake and milk
production of dairy cows. K. L. Blaxter. Journal of Agricultural Science, 34,
22-26 (1944).
5. Experiments on the use of homegrown foods for mill: production. 2. The
effect of feeding concentrated and bulky foods prior to calving on subsequent
milk production. K. L. Blaxter. Journal of Agricultural Science, 34, 37-44
(1944).
6. Experiments on the use of homegrown foods for milk production. 3. The
effect of over and under feeding in mid lactation. K. L. Blaxter. Journal of
Agricultural Science, 34, 213-216 (1944).
48 Wolf Prize in Agriculture
23. The protein and energy nutrition of the young calf. K. L. BLaxter. Agricultural
Progress, 25, 1-9 (1949).
24. Iodinated, protein and lactation. K. L. Blaxter. Journal of the Ministry of
Agriculture, 57, 413-417 (1950).
25. Lead as a nutritional hazard to farm animals. 1. The determination of lead in
biological material. K. L. Blaxter, R. Allcroft. Journal of comparative Pathology,
60, 133-139 (1950).
26. Lead as a nutritional hazard to farm livestock. 2. The absorption and excretion
of lead by sheep and rabbits. K. L. Blaxter. Journal of Comparative Pathology
(1950).
27. Lead as a nutritional hazard to farm livestock. 3. Factors affecting the
distribution of lead in the tissues. K. L. Blaxter. Journal of Comparative
Pathology, 60, 177-189 (1950).
28. Lead as a nutritional hazard to farm livestock. 5. The toxicity of lead to cattle
and sheep and an evaluation of the lead hazard under farm conditions.
R. Allcroft, K. L. Blaxter. Journal of Comparative Pathology, 60, 209-218
(1950).
29. Energy feeding standards for dairy cattle. Nutrition Abstracts & Reviews, 20,
1-21 (1950).
30. The nutrition of the young Ayrshire calf. I. The endogenous nitrogen and
basal energy metabolism in the calf. K. L. Blaxter, W. A. Wood. British Journal
of Nutrition, 5, 11-25 (1951).
31. The nutrition of the young Ayrshire calf. II. A spirometer for the determination
of the respiratory exchange of the calf. K. L. Blaxtar, A. Howells. British
Journal of Nutrition, 5, 25-29 (1951).
32. The nutrition of the young Ayrshire calf. III. The metabolism of the calf
during starvation and. subsequent realimentation. K. L. Blaxter, W. A. Wood.
British Journal of Nutrition, 5, 29-55 (1951).
33. The nutrition of the young Ayrshire calf. IV. Some factors affecting the
biological value of protein determined by nitrogen-balance methods.
K. L. Blaxter, W. A. Wood. British Journal of Nutrition, 5, 55-67 (1951).
34. Separation of the tocopherols by paper chromatography. P. Brown,
K. L. Blaxter. Chemistry & Industry, pp. 633-634 (1951).
35. The importance of fat-soluble vitamins for calves. K. L. Blaxter. Journal of
the Royal Agricultural Society of England, 112, 25-35 (1951).
36. Conversion factors for vegetable and animal foods for human consumption.
K. L. Blaxter. British Journal of Nutrition, 5, 250-255 (1951).
37. The nutrition of the young Ayrshire calf. V. The nutritive value of whole
cows’ milk. K. L. Blaxter, W. A. Wood. British Journal of Nutrition, 6, 1-11
(1952).
50 Wolf Prize in Agriculture
38. The nutrition of the young Ayrshire calf. VI. The utilization of the energy of
whole milk. K. L. Blaxter. British Journal of Nutrition, 6, 12-18 (1952).
39. The nutrition of the young Ayrshire calf. VII. The biological value of gelatin
and casein when given as the sole source of protein. K. L. Blaxter, W. A.
Wood. British Journal of Nutrition, 6, 56-70 (1952).
40. The nutrition of the young Ayrshire calf. VIII. Muscular dystrophy in the
growing calf. K. L. Blaxter, P. S. Watts, W. A. Wood. British Journal of Nutrition,
6, 125-143 (1952).
41. The nutrition of the young Ayrshire calf. IX. The composition of the tissues
of normal and dystrophic calves. K. .L. Blaxter, W. A. Wood. British Journal
of Nutrition, 6, 144-163 (1952).
42. The nutrition of the young Ayrshire calf. X. The histopathology of muscular
dystrophy and its relation to muscle chemistry. A. M. MacDonald, K. L.
Blaxter, P. S. Watts, W. A. Wood. British Journal of Nutrition, 6, 164-169
(1952).
43. Some effects of thyroxine and iodinated casein on dairy cows and their
practical significance. K. L. Blaxter. “Vitamins & Hormones”, 10, 218-250
(1952).
44. Vitamin B in the nutrition of farm animals. K. L. Blaxter, P. Brown. Nutrition
Abstracts & Reviews, 22, 1-21 (1952).
45. The utilization of the minerals, vitamins and other constituents of grass.
K. L. Blaxter. British Journal of Nutrition, 6, 110-117 (1952).
46. The influence of diet on the development of the alimentary tract of the calf.
K. L. Blaxter, M. K. Hutcheson, J. N. Robertson, A. L. Wilson. British Journal
of Nutrition, 6, i-ii (1952).
47. The effect of Mg deficiency on the energy exchange of calves. K. L. Blaxter,
J. A. F. Rook. Journal of Physiology, 121, 48-49 (1953).
48. The nutrition of the young Ayrshire calf. XI. The toxicity of cod-liver oil.
K. L. Blaxter, W. A. Wood, A. N. MacDonald. British Journal of Nutrition, 7,
34-50 (1953).
49. The nutrition of the young Ayrshire calf. XII. Factors affecting the tocopherol
reserves, muscle composition and muscle histology of 4-day old calves. K. L.
Blaxter, P. Brown, A. M. MacDonald. British Journal of Nutrition 7, 105-123
(1953).
50. The nutrition of the young Ayrshire calf. XIII. The toxicity of the unsaturated
acids of codliver oil. K. L. Blaxter, P. Brown, A. M. MacDonald. British Journal
of Nutrition, 7, 287-298 (1953).
51. The nutrition of the young Ayrshire calf. XIV. Some effects of natural and
synthetic antioxidants on the incidence of muscular dystrophy induced by
codliver oil. K. L. Blaxter, F. Brown, W. A. Wood, A. M. MacDonald. British
Journal of Nutrition, 7, 337-349 (1953).
Sir Kenneth Blaxter 51
132. The heat increment in fasting sheep of acetic acid partially neutralized with
sodium hydroxide. D. G. Armstrong, K. L. Blaxter, N. McC. Graham. British
Journal of Nutrition, 15, 169-175 (1961).
133. The utilization of food by sheep end cattle. K. L. Blaxter, F. W. Wainman.
Journal of Agricultural Science, 57, 419-425 (1961).
134. The regulation of food intake by sheep. K. L. Blaxter, F. W. Wainman,
R. S. Wilson. Animal Production, 3, 51-61 (1961).
135. The utilization of food by sheep and cattle. K. L. Blaxter, F. W. Wainman.
Journal of Agricultural Science, 57, 419-425 (1961).
136. Protein as a source of energy for synthesis of fat in sheep. A. K. Martin,
K. L. Blaxter. Proceedings of the Nutrition Society, 20, pp. vii-viii (1961).
137. The utilization of the energy of the same food by cattle and sheep.
K. L. Blaxter, F. W. Wainman. Proceedings of the Nutrition Society, 20,
pp. xxxiii-xxxiv (1961).
138. The utilization of the energy of carbohydrate by ruminants. D. G. Armstrong,
K. L. Blaxter. Proceedings of the 2nd Symposium on Energy Metabolism of
Farm Animals, Wageningen, pp. 187-197 (1961).
139. The utilization of the energy of food by ruminants. K. L. Blaxter. Proceedings
of the 2nd Symposium on Energy Metabolism of Farm Animals, Wageningen,
pp. 211-224 (1961).
140. The utilization of the energy of protein by ruminants. A. K. Martin,
K. L. Blaxter. Proceeding of the 2nd Symposium on Energy Metabolism of
Farm Animals, Wageningen, pp. 200-209 (1961).
141. Environmental temperature and the energy metabolism and heat emission of
steers. K. L. Blaxter, F. W. Wainman. Journal of Agricultural Science, 56,
81-90 (1961).
142. Lactation and the growth of the young. Chapter in ‘Milk: The Mammary
Gland and its Secretion’. Edited S. K. Kon and A. T. Cowie, Vol. II,
pp. 305-361 (1961).
143. The nutritional physiology of livestock under natural conditions. K. L. Blaxter.
Journal of the Royal Society of Arts, 109, 764-781 (1961).
144. 144 Minerals in relation to disease. K. L. Blaxter. Veterinary Annual,
pp. 230-238 (1961).
145. The influence of the environment on animal production and health under
housing conditions. W. Bianca, K. L. Blaxter. Pestschrift zum VII Intern.
Tierzuchtkongress, Hamburg, pp. 113-147 (1961).
146. Efficiency of feed conversion by different classes of livestock in relation to
food production. K. L. Blaxter. Federation proceedings 20, Supplement No. 7,
268-274 (1961).
147. Economics and animal husbandry. K. L. Blaxter. Journal of Agricultural
Economics, 14, 308-313 (1961).
Sir Kenneth Blaxter 57
197. The metabolism of formic acid in sheep. J. E. Vercoe, K. L. Blaxter. Br. J. Nutr.
19, 523-530 (1965).
198. The effect of artificial drying on the energy value of grass. A. Ekern,
K. L. Blaxter, D. Sawers. Brit. J. Nutr., 19, 417-434 (1965).
199. Prediction of the amount of methane produced by ruminants. K. L. Blaxter,
J. L. Clapperton. Brit. J. Nutr., 19, 511-522 (1965).
200. Climatic requirements for optimum economic production: current research.
K. L. Blaxter. In Pull Rep. of Proc. Beef Housing Conf., Kenilworth, pp. 2-7.
London: Royal Agricultural Society of England (1965).
201. Animal production in adverse environments. K. L. Blaxter. J. Univ. Newcastle
upon Tyne Agric. Soc., 19, 10-15 (1965).
202. The heat of combustion of the urine of sheep and cattle in relation
to its chemical composition and to diet. K. L. Blaxter, J. L. Clapperton,
A. K. Martin. Br. J. Nutr., 20, 449-460 (1966).
203. The extent of differences between six British breeds of sheep in their
metabolism, feed intake and utilization, and resistance to climatic stress.
K. L. Blaxter, J. L. Clapperton, F. W. Wainman. Br. J. Nutr., 20, 283-294 (1966).
204. The fasting metabolism of cattle. K. L. Blaxter, F. W. Waimnan. Br. J. Nutr.,
20, 103-111 (1966).
205. The voluntary intake of food by sheep and cattle in relation to their energy
requirements for maintenance. K. L. Blaxter, F. W. Waimnan, J. L. Davidson.
H.D.R.I. Reprint No. 585. Anim. Production, 8, 75-83 (1966).
206. The metabolism of oleic, linoleic and linolenic acids by sheep with reference
to their effects on aiethane products. J. W. Czerkawski, K. L. Blaxter, F. W.
Wainman. Br. J. Nutr., 20, 349-362 (1966).
207. The effect of linseed oil and of linseed oil fatty acids incorporated in
the diet on the metabolism of sheep. J. W. Czerkawski, K. L. Blaxter,
F. W. Waimnan. Br. J. Nutr., 20, 485-494 (1966).
208. The effect of functional groups other than carboxyl on the metabolism of
C18 and C12 alkyl compounds by sheep. J. W. Czerkawski, K. L. Blaxter,
F. W. Wainman. Br. J. Nutr., 20, 495-508 (1966).
209. The effect of natural outdoor environments on the energy requirements of
sheep. J. P. Joyce, K. L. Blaxter, C. Park. Res. Vet. Sci., 7 (1966).
210. The voluntary intake of food by sheep and cattle in relation to their energy
requirements for maintenance. K. L. Blaxter, F. W. Wainman, J. L. Davidson.
Anim. Prod., 8, 75-83 (1966).
211. Modifications of the methane production of the sheep by supplementation
of its diet. K. L. Blaxter, J. Czerkawski. J. Sci. Fd. Agric., 17, 417-421 (1966).
212. Utilization of the energy and protein of the same diet by cattle of different
ages. K. L. Blaxter, J. L. Clapperton, F. W. Wainman. J. Agric. Sci., Camb., 67,
67-75 (1966).
Sir Kenneth Blaxter 61
246. A review of shelter research to date and some ideas for future development.
K. L. Blaxter. Rep. 3rd Symp. Shelter Res. Cambr., pp. 113-120 (1970).
247. The utilization of volatile acids in the energy metabolism of ruminants. K. L.
Blaxter. Proc. 8th mt. Congr. Nutr., Prague, pp. 299-303 (1970).
248. The comparative biology of’ lactation. K. L. Blaxter. In Lactation. (University
of Nottingham 17th Easter School in Agricultural Science 1970. Ed. Ian R.
Falconer: London: Butterworths, pp. 51-69 (1970).
249. Current research and future problems. K. L. Blaxter. J. Ass. Agric. 5-10
(1970).
250. The nutritive value of artificially dried herbage. Report of 20th Annual
Convention: Association of Green Crop Driers, pp. 44-57. K. L. Blaxter
(1970).
251. The effects of nitrogenous fertilizer on the nutritive value of artificially dried
grass. K. L. Blaxter, F. W. Wainman, P. J. S. Dewey, J. Davidson, H. Denerley,
J. B. Gunn. J. Agric. Sci., Camb., 76, 307-319 (1971).
252. Methods of measuring the energy metabolism of animals and interpretation
of results obtained. K. L. Blaxter. Fedn. Prod. Pedn. Am. Socs Exp. Biol., 30,
1436-1443 (1971).
253. Voluntary intake and energy metabolism of sheep fed chopped dried grass
and the same material milled and pelleted. F. W. Wainman, J. S. Smith, K. L.
Blaxter. Proc. Nutr. Soc., 30, 23-24A (1971).
254. Some effects of poor reproductive performance in cattle. K. L. Blaxter. In
‘Reproduction and Breeding Management of Cattle’; Proc. 7th Conf. North of
Scotland Division, Br. Vet. Ass., pp. 1-8. Ed. By Sharman, G.A.M. Aberdeen:
North of Scotland College. of Agriculture (1971).
255. Bioenergetics of ruminant animals. K. L. Blaxter. In ‘Bioenergetics: Proceedings
of the International Symposium on Environmental Physiology’, pp. 35-41.
Ed. by Smith, R. E. Bethesda: Federation of American Societies for Experimental
Biology (1972).
256. Deer Farming. K. L. Blaxter. Scot. Agric., 51, 225-230 (1972).
257. Fasting metabolism and the energy required by animals for maintenance. In
‘Pestskrift til professor Dr. agr. Dr. h.c. Knut Breirem til hans 70 ars 20 April
1972’, pp. 19-36. Gjǿvik: Mariendals Boktrykkeri A.s. (1972).
258. Plants for animals. K. L. Blaxter. 51st Rep. Scott. Pl. Breed. Stn., p. 37 (1972).
259. Relevance of animal feeding trials to human dietary requirements. K. L.
Blaxter. J. Sci. Fd Agric., 23, 941-948 (1972).
260. A new method for estimating the heat production of animals. K. L. Blaxter,
J. M. Brockway, A. W. Boyne. Q. J. exp. Physiol., 57, 60-72 (1972).
261. The effect of grinding and pelleting on the nutritive value of poor quality
roughages. F. W. Wainman, K. L. Blaxter. J. Agric. Sci., Camb., 79, 435-445
(1972).
64 Wolf Prize in Agriculture
262. The utilization of the energy of artificially dried grass prepared in different
ways. F. W. Wainman, K. L. Blaxter, J. S. Smith. J. Agric. Sci., Camb., 78,
441-447 (1972).
263. Animal. production and substitute foods. K. L. Blaxter. In Modern Animal
Production and Veterinary Problems: Papers given at the Peter Wilson Bequest
Course, pp. 147-152. Edinburgh: Faculty of Veterinary Medicine, Royal (Dick)
School of Veterinary Studies, University of Edinburgh (1972).
264. Approaches to the problem of augmentation of animal protein food sources
in Asia. K. L. Blaxter. In Proceedings of the First Asian Congress of Nutrition
pp. 83-92. Ed. by Tulpule, P. G. & Rao, K. S. J. Hyderabad: Nutrition Society
of India (1972).
265. John Boyd Orr: Animal Experimentation and the human situation. Rep. Rowett
Inst. 28, 108-115. K. L. Blaxter (1972).
266. Efficiency — the nutritional basis. K. L. Blaxter. In Breeding for Beef:
Proceedings of a National Conference held at Peebles on 1, 2 and 3 November
1971, pp. 3-11. Bletchley: Meat and Livestock Commission (1973).
267. Intensive animal production. K. L. Blaxter. Vet. Rec., 92, 383-386 (1973).
268. The purpose of protein production. K. L. Blaxter. In The Biological Efficiency
of Protein Production, pp. 3-11. Ed. by Jones, J. G. W. Cambridge: Cambridge
University Press (1973).
269. Review lecture. The nutrition of ruminant animals in relation to intensive
methods of agriculture. K. L. Blaxter. Proc. R. Soc. Lond., B. 183, 321-336
(1973).
270. Increasing output of animal production: measures for increasing productivity.
In Man, Food and Nutrition, pp. 127-146. Ed. by Rechcigl, M. Cleveland:
C.R.C. Press (1973).
271. The Individual and. the Information Problem. K. L. Blaxter, M. L. Blaxter.
Nature, Lond. 246, 335-339 (1973).
272. Adjustments of the metabolism of the sheep to confinement. K. L. Blaxter. In
Energy Metabolism of Farm Animals: Proceedings of the 6th Symposium of
the European Association for Animal Production, pp. 115-118. Ed. by Menke,
K. H., Lantzsch, H.-J. & Reichl, J. R. Stuttgart: Universität Hohenheim (1974).
273. The nutritive value of different combinations of oats, oat husk and
soy-bean meal when fed to sheep. F. W. Wainman, K. L. Blaxter, M. S.
McDDonald, P. J. S. Dewey, J. S. Smith. Proc. 6th Symp. Energy Metab. Fm
Anim., pp. 217-219 (1974).
274. Artificially dried herbage as a source of energy for ruminant animals. K. L.
Blaxter. Proc. 1st mt. Green Crop Dry. Congr., Oxford, pp. 64-72 (1974).
275. The limits to agricultural improvement. K. L. Blaxter. J. Univ. Newcastle-
upon-Tyne Agric. Soc., 25, 3-12 (1974).
276. Metabolisable energy and feeding systems: for ruminants. K. L. Blaxter. Proc.
7th Nutr. Conf. Feed Mfrs, Univ. Nottm., pp. 3-25 (1974).
Sir Kenneth Blaxter 65
277. Power and the agricultural revolution. K. L. Blaxter. New Scientist, 61,
400-403 (1974).
278. Deer Farming. K. L. Blaxter. Mammals Rev. 4, 119-122 (1974).
279. The technical possibilities for animal production from grassland. K. L. Blaxter.
Rep. Grassld. Res. Inst. Hurley, pp. 141-146 (1974).
280. Farming the red deer. The first report of the investigations by the Rowett
Research Institute and the Hill Farming Research Organjsation. K. L. Blaxter,
R. N. B. Kay, G. A. M. Sharman, J. M. M. Cunningham, W. J. Hamilton.
Edinburgh: HMSO (1974).
281. A new challenge to agriculture. K. L. Blaxter. Arsmeld, Norg. landbrvit.
Porskrad, pp. 91-100 (1974).
282. Energy-protein relationships in rwninants. K. L. Blaxter. Proc. 9th mt. Congr.
Nutr., Mexico, 3, 122-127 (1975).
283. Self-sufficient Britain — Food. K. L. Blaxter. New Scientist, 65, 697-702
(1975).
284. Conventional and unconventional farmed animals. K. L. Blaxter. Proc. Nutr.
Soc., 34, 51-56 (1975).
285. The energetics of British agriculture. K. L. Blaxter. Biologist, 22, 14-18 (1975).
286. Protein from non-domesticated herbivores. K. L. Blaxter. Food Protein Sources,
International Biological Programme, Vol. 4. Cambridge University Press.
pp. 147-156 (1975).
287. Concluding remarks. K. L. Blaxter. In ‘Bread: Social, Nutritional and
Agricultural Aspects of Wheaten Bread’, pp. 339-350. Ed. by Spicer, A.
London: Applied Science Publishers Ltd. (1975).
288. Nutrients required for animal production as related to the world food supply.
K. L. Blaxter. In ‘Agriculture in the Whirlpool of Change: Papers presented at
the Centennial Symposium, Ontario Agricultural College, University of
Guelph, pp. 201-213. Guelph: University of Guelph (1975).
289. The energetics of British agriculture. K. L. Blaxter. Journal of the Science of
Food and Agriculture, 26, 1055-1064 (1975).
290. The Rowett Institute: Its current work and the farming industry. K. L. Blaxter.
Journal of the Royal Agricultural Society of England, 136, 86-92 (1975).
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Jay L. Lush
Iowa State University
Ames, Iowa, USA
k
Jay L. Lush has one more than any agricultural scientist both to investigate the
genetic basis of traits important for animal production, to assess their value and to
improve them by initiating breeding programmes of proven practical effectiveness.
He has been responsible for inspiring a great band of graduate students and others
who have disseminated his ideas and practical suggestions throughout the world
and so led to the improvement of poultry, pigs, beef and dairy cattle. His
contributions are of several kinds. First, he pioneered the value to the scientific
breeder of adequate measurement of the performance of individuals, their progeny
and their pedigree in order to assess the genetic worth of an animal. Secondly, he
pioneered the assessment of the ‘heritability’ of a trait, i.e. the degree to which its
expression reflected a genetic basis rather than the effects of a favorable or
unfavorable environment. Thirdly, having demonstrated how the inherited
potentialities could be measured and distinguished from those due to non-heritable
causes, he synthesized these ideas into practical and workable animal breeding
plans to promote an inherited increase in the yield of meat, milk or eggs. He has
exploited his ideas as a University teacher, at a diversity of national and international
gatherings and enshrined much of it in his major and most influential text ‘Animal
Breeding Plans’. He may truly be described as the ‘father’ of scientific animal
breeding in the twentieth century.
67
68 Wolf Prize in Agriculture
CURRICULUM VITA
Date and place of Birth: January 3, 1896, Shambaugh, Iowa
QUALIFICATIONS OF NOMINEE:
a) Degrees received
Kansas State College B.S. 1916
Kansas State College M.S. 1918
University of Wisconsin Ph.D. 1921
which permit maximum rates of genetic improvement in farm animals. His clear
insight into the roles of heredity, gene action and interaction, and the effects of
environment permitted him to develop methods of measuring the relative
importance of these sources. These results have been applied to poultry, swine,
beef cattle and dairy cattle in ways most suitable to the particular species and to
the economically important traits of each species. For example, family selection
has been of considerable use in improving egg production, progeny testing, and
artificial insemination in milk production, individual selection in growth rate for
beef and swine.
Dr. Lush has worked closely with breed organizations and public institutions in
developing plans for improved animal production. Some of the foreign assignments
to which he has devoted time and effort are as follows:
1934 - Denmark. Study and Analysis of the Danish Pig Progeny Testing System.
1948 - Great Britain. Survey of research in genetics and animal husbandry for
U.S.D.A.
1948 - Australia. Survey of research in animal production for C.S.I.R.O.
1951 - Paraguay. Developed cattle breeding plans in Paraguay for A.I.D.
1961 - Argentina. Survey and plans for more efficient beef industry in Argentina.
LIST OF PUBLICATIONS
1. Inheritance in Swine. J. Hered. 12: 57-71 (1921).
2. An Hereditary Notch in the Ears of Jersey Cattle. J. Hered. 13: 8-13 (1922).
3. The Influence of Age and Individuality upon the Yield of Wool. Proc. Amer.
Soc. An. Prod. 1922, 105-109 (1922).
4. The Practicability of the Milking Machine. Texas Sta. Circular 30. Inheritance
in Swine. (with E. N. Wentworth). J. Agric. Res. 23: 557-582 (1923).
5. I. Fattening Steers on Cottonseed Meal and Hulls with and without Corn. II.
The Influence of Age on Fattening Steers. (with J. M. Jones and J. H. Jones).
Texas Sta. Bull. 309 (1923).
6. The Influence of Individuality, Age and Season Upon the Weight of Fleeces
Produced by Range Sheep. (with J. H. Jones). Texas Sta. Bull. 31 (1923).
7. Twinning In Brahma Cattle. J. Hered. 15: 93-96 (1924).
8. “Double Ears” in Brahma Cattle. J. Hered. 15: 93-96 (1924).
9. The Influence of Individuality, Age and Season Upon the Weights of Fleeces
Produced by Angora Goats Under Range Conditions. (with J. M. Jones). Texas
Sta. Bull. 320 (1924).
10. Nature’s Score Card for Feeder Steers. Proc. Amer. Soc. An. Prod. 1924,
98-101 (1924).
11. The Possibility of Sex Control by Artificial Insemination with Centrifuged
Spermatozoa. J. Agric. Res. 30: 893-913 (1925).
12. Methods of Selecting Wool Samples in Shrinkage Studies. (with J. M. Jones).
Proc. Amer. Soc. An. Prod. 1925, 115-117 (1925).
13. Practical Methods of Estimating the Proportions of Fat and Bone in Cattle
Slaughtered in Commercial Packing Plants. J. Agric. Res. 32: 727-755 (1926).
14. Inheritance of Horns, Wattles, and Color in Grade Toggenburg Goats.
J. Hered. 17: 72-91 (1926).
15. How Much Accuracy is Gained by Weighing Cattle Three Days Instead of
One at the Beginning and End of Feeding Experiments. (with W. H. Black).
Proc. Amer. Soc. An. Prod. 1926, 206-210 (1926).
16. Rice Bran as a Feed for Dairy Cows. (with Fred Hale). Texas Sta. Bull. 352
(1927).
17. “Percentage of Blood” and Mendelism. J. Hered. 18: 351-367 (1927).
18. The Production of Clean Milk. (with Fred Hale). Texas Sta. Circular 48
(1927).
19. Practices and Problems Involved in Crossbreeding Cattle in the Coastal Plain
of Texas. Proc. Amer. Soc. An. Prod. 1927, 58-61 (1927).
20. A Statistical Interpretation of Some Texas Lamb Feeding Data. (with J. M.
Jones). Proc. Amer. Soc. An. Prod. 1927, 167-170 (1927).
21. Changes in Body Measurements of Steers During Intensive Fattening. Texas
Sta. Bull. 385 (1928).
72 Wolf Prize in Agriculture
44. The Relation of Body Shape of Feeder Steers to Rate of Gain, to Dressing
Percent, and to Value of Dressed Carcass. Texas Sta. Bull. 471 (1932).
45. The Amount and Kind of Inbreeding which has Occurred in the Development
of Breeds of Livestock. Proc. Sixth International Congress Genetics. 2:
123-126 (1932).
46. Mutton, and How it Gets that Way. (A review of John Hammond’s
“Growth and the Development of Mutton Qualities in the Sheep”). J. Hered.
23: 312-314 (1932).
47. Inbreeding and the Genetics History of the Rambouillet Sheep in America.
(with W. F. Dickson). J. Hered. 24: 19-33 (1933).
48. The Use of Statistical Methods in Animal Husbandry. Proc. Amer. Soc. An.
Prod. 1932, 15-19 (1933).
49. Linebreeding. Iowa Agric. Exp. Sta. Bull. 301 (1933).
50. A Linebreeding Program for Horse Breeding. (with P. B. Pearson). J. Hered.
24: 185-191 (11. Paper No. B62) (1933).
51. The Bull Index Problem in the Light of Modern Genetics. J. Dairy Sci. 16:
501-522 (1933).
52. The Reliability of Some Measures of Productiveness in Brood Sows. (with
A. L. Anderson, C. C. Culbertson and W. E. Hammond). Proc. Amer. Soc. An.
Prod. 1933, 282-287 (1934).
53. Freshening Ages of Purebred Cows in Iowa Cow Testing Associations. (with
Mogens Plum). J. Dairy Sci. 17: 625-636 (1934).
54. Progress Report on Comparison of Lactation and Yearly Records. (with
C. N. Harris and E. N. Schultz). J. Dairy Sci. 17: 737-742 (1934).
55. Beef Production arid Quality as Influenced by Crossing Brahman with
Hereford and Shorthorn Cattle. (with W. H. Black and A. T. Semple) U.S.D.A.
Tech. Bull. 417 (1934).
56. A Herd of Cattle Bred for Twenty Years without New Blood. J. Hered. 25:
209-216. ( J. Paper No. J-l33) (1934).
57. Factors Affecting Birth Weights of Swine. (with H. O. Hetzer and C. C.
Culbertson). Genetics 19: 329-343 (1934).
58. Progeny Test and Individual Performance as Indicators of an Animal’s Breeding
Value. J. Dairy Sci. 18: 1-19 (1935).
59. The Inheritance of Productivity in Farm Livestock. V. Discussion of Preceding
Contributions. Empire J. Exp. Agric. 3: 25-30 (1935).
60. Genetics History of the Hoistein-Friesian Cattle in the United States. (with
J. C. Holbert and 0. S. Wiliharn). 3. Hered. 27: 61-72 (1936).
61. Genetics and Animal Breeding. (A review of C. Kronacher’s “Genetik und
Tierzuchtung”). J. Hered. 27: 201-203 (1936).
62. Genetic Aspects of the Danish System of Progeny-testing Swine. Iowa Agric.
Exp. Sta. Res. Bull. 204 (1936).
74 Wolf Prize in Agriculture
123. Proving Sires and Dams. (with Lon D. McGilliard). J. Dairy Sci. 38: 163-180
(1955).
124. Relations Between Parts of Lactations and Producing Ability of Holstein Cows.
(with D. E. Madden and L. D. McGilliard). J. Dairy Sci. 38(11): 1264-71
(1955).
125. Statistics in Investigations in Animal Production. J. of the Indian Society of
Agricultural Statistics. 7: 7-22 (1955).
126. Dairy Cattle Genetics. J. Dairy Sci. 39(6): 693-694 (1956).
127. Changes in Type Classifications of Dairy Cattle. J. Dairy Sci. 39(7):
1015-1026 (1956).
128. Theoretical Consequences of Breeding for the Heterozygote in Intra and
Interbreeding Populations. p. 3-26 in Fifth Poultry Breeders Roundtable
(1956).
129. Studies on Bovine Ocular Squamous Carcinoma (“Cancer Eye”). (with David
E. Anderson and Doyle Chambers) (1957).
130. II. Relationship Between Eyelid Pigmentation and Occurrence of Cancer Eye
Lesions. J. An. Sci. 16: 739-746 (1957).
131. III. Inheritance of Eyelid Pigmentation. J. An. Sci. 16: 1007-1016 (1957).
132. Twinning in Dairy Cattle and its Relation to Production. J. Dairy Sci.
40: 1430-1436. (with C. E. Meadows) (1957).
133. Effect of Inbreeding on Production in Holsteins. J. Dairy Sci. 41: 105-113.
(with C. N. von Krosigk) (1958).
134. Practical Application of Performance Testing. The Shorthorn World. 43(11):
44 and 318-321 (1958).
135. Genetics in Plant and Animal Breeding. (Translated Title). Tolvmandsbladet
30(10): 403-409. Copenhagen (1958).
136. Genetic Relations between Body Measurements at Three ages in Holsteins.
J. Dairy Sci. 41: 1045-1049. (with D. W. Blackmore and L. D. McGilliard)
(1958).
137. Relationships between Body Measurements, Meat Conformation, and
Milk Production. J. Dairy Sci. 41: 1050-1056. (with D. W. Blackmore and
L. D. McGilliard) (1958).
138. Genetic and Environmental Portions of the Variation among Herds in Butterfat
Production. J. Dairy Sci. 42: 115-122. (with F. Pirchner) (1959).
139. Making Use of New Knowledge about Basic Principles. pp. 141-155, Eighth
Poultry Breeders’ Roundtable (1959).
140. Controls for Selection Experiments. pp. 46-55 in Proc. NC-1 Technical
Committee Meeting at Ames, July 23-24. Mimeo (no reprints) (1959).
141. Improving Dairy Cattle by Breeding. I. Current Status and Outlook.
J. Dairy Sci. 43: 702-706 (1960).
142. Accuracy of Partial Trapnest Records. Poultry Science. (with John D. Wheat)
(1960).
78 Wolf Prize in Agriculture
161. Animal Breeding in the Age of A.I. “Animal Breeding and Application, present,
and future.” pp. 8-25. (North America). Mimeo. (Sponsored jointly with
Univ. Wisconsin) (1968).
162. Research in Animal Production, its Past Accomplishments and Present
Prospects. pp. 45-65. (In Bull. 160. 38th Jahrgang 38. The centenary of the
Abteilung fur Landwirtschaft an der Eidegenossischen Technischen
Hochschule, Zurich. June 25-26, 1971.
163. Early Statistics at Iowa State University. Chap. 13 (p. 221-226 in the book,
“Statistical Papers in Honor of George W. Snedecor.” ISU press (1972).
164. Teaching Animal Breeding and Teaching Graduate Students. An. Breeding
and Genetics Symposium. pp. 78-88. (By the American Society of Animal
Science and the American Dairy Science Assoc.) (1972).
165. Response to the Dedication of the Jay L. Lush Auditorium. Iowa State Journal
of Science. 48(4): 281-284 (1974).
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Karl Maramorosch
Rutgers University
New Brunswick, N.J., USA
k
BIOGRAPHICAL INFORMATION
Positions
Rockefeller University, New York City, from Assistant to Associate Professor,
1949-1961.
Boyce Thompson Institute for Plant Research, Yonkers, New York from
Senior Entomologist to program Director on Insect Physiology and Virology,
1961-1974.
Rutgers- The State University of New Jersey, Professor II (Distinguished Professor)-
1974 - until now; Named Professor (Robert L. Starkey Professor of Microbiology)
1983 - until now; Adjunct Professor of Entomology 1985 - until now.
81
82 Wolf Prize in Agriculture
Visiting Professorship
1) Wageningen Agric. University, Netherlands 1953.
2) Cornell University 1957.
3) Agric. University, Bucharest, Romania 1964.
4) Agric. University, Skierniewice and Warsaw (SGGW), Poland 1964.
5) Rutgers University 1968.
6) Fordham University, NY 1972.
7) Fulbright Senior Prof. Yugoslavia 1972 and 1977.
8) Agric. University, Bangalore, India 1979.
9) Dist Senior Prof. Hokkaido University, Sapporo, Japan 1980.
10) Fudan University, Shanghai, China 1982.
11) Justus Liebig University, Giessen, Germany 1983.
Professional Affiliations
1) American Phytopathological Society (Fellow).
2) Entomological Society of America (Fellow and Honorary Member).
3) Indian Virological Society (Honorary Fellow).
4) American Association for the Advancement of Science, AAAS (Fellow).
5) New York Academy of Sciences (Fellow).
6) New York Academy of Sciences (Recording Secretary, 1960-1962).
7) New York Academy of Sciences (Vice-President, 1961-1963).
8) National Academy of Sciences, India (Honorary Fellow).
9) Leopold ma Academy, Germany.
10) Microscopy Society of America.
11) International Organization for Mycoplasmology.
12) Society for Invertebrate Pathology.
13) Society of In Vitro Biology.
14) American Council on Science and Health (Board of Science and Policy
Advisors).
15) International Association for Research on Medicinal Forest Plants (President).
16) Phi Beta Kappa Award in Science Committee.
17) Tropical Medicine and Parasitology USPH-NIH Panel (1971-1976).
18) American Institute of Biological Sciences
19) American Society of Virology.
Visiting Professorships
During several visits to the International Center for Insect Physiology (ICIPE) in
Nairobi and Mbita Point, Kenya, Professor Maramorosch taught about novel insect
control measures using insect cell culture.
In the former Soviet Union, he lectured and consulted in the Republics of
Russia, Armenia and Uzbekistan.
As an invited Guest Professor at Fudan University in Shanghai, China, he lectured
on applications of invertebrate cell culture.
Maramorosch’s fluency in seven languages was very helpful in his international
activities, When the Justus Leibig University in Giessen, Germany invited him, he
lectured there in German. In Romania, as a guest of their Academy of Sciences, he
made use of knowledge of Romanian, acquired during his years as civilian internee
during World War II in that country. In St. Petersburg, Moscow, Yerevan and
Tashkent, he lectured in Russian, and in Poland in Polish.
Maramorosch’s creativity has been demonstrated throughout his career. His
lecturing commitments often took him outside the U.S. borders. He has tremendous
energy to meet people from around the world, to learn new cultures, and to pass
on his knowledge wherever he steps. His passion for science and for learning is his
inspiration. Numerous postgraduate associates from around the globe owe their
own enthusiasm to having been fortunate enough to spend time in his laboratory.
Professor Karl Maramorosch has served the discipline of entomology and plant
pathology with uninterrupted distinction across a span of seven decades. His
hundreds of scholarly publications document the remarkable scientific journey of
one of the giants and icons of entomology. He has gained true world renown in
insect pathology, plant-insect vectors, insect cell culture, and virology. To have
contributed to such a breadth of disciplines is notable but to have had substantial
impact in each is extraordinary.
21. Vectors of Disease Agents: Interactions with Plants, Animals and Man.
McKelvey, J.J., Eldridge, B.F. and Maramorosch, K., eds. Praeger. 243 pp. 1980.
22. Plant Diseases and Vectors: Ecology and Epidemiology. Maramorosch, K. and
Harris, K.F., eds. Academic Press. 368 pp. 1981.
23. Mycoplasma Diseases of Trees and Shrubs. Maramorosch, K. and Raychaudhuri,
S.P., eds. Academic Press. 362 pp. 1981.
24. Pathogens, Vectors, and Plant Diseases: Approaches to Control. Harris, K.F.
and Maramorosch, K., eds. Academic Press. 310 pp. 1982.
25. Advances in Cell Culture. 5 volumes, (I–V) Maramorosch, K., ed. Academic
Press. 1981-1987.
26. Advances in Cell Culture. Vols. VI and VII. Maramorosch, K. and Sato, G.H.,
eds. Academic Press. 1988-1989.
27. Mycoplasma and Allied Pathogens of Plants, Animals and Human Beings.
Govindu, H.C., Maramorosch, K., Raychaudhuri, S.P. and Munlyappa, V., eds.
Univ. Agric. Sciences, Bangalore, India. 118 pp. 1981.
28. Invertebrate Cell Culture Applications. Maramorosch, K. and Mitsuhashi, J.,
eds. Academic Press. 245 pp. 1982.
29. Advances in Virus Research. Two volumes (Nos. 28 and 29). Lauffer, A.M.
and Maramorosch, K., eds. Academic Press. 1983 and 1984.
30. Subviral Pathogens of Plants and Animals: Viroids and Prions. Maramorosch,
K. and McKelvey, J.J., eds. Academic Press. 550 pp. 1985.
31. Viral insecticides for biological control. Maramorosch, K. and Sherman, K.E.,
eds. Academic Press. 809 pp. 1985.
32. Biotechnology in Insect Pathology and Cell Culture. Maramorosch, K., ed.
Academic Press. 511 pp. 1987.
33. Mycoplasma Diseases of Crops: Basic and Applied Aspects. Maramorosch, K.
and Raychaudhuri, S.P., eds. Springer Verlag. 456 pp. 1987.
34. Invertebrate and Fish Tissue Culture. Kurstak, E. Kuroda, Y. and Maramorosch,
K., eds. Springer 1988.
35. Biotechnology for Biological Control of Pests and Vectors. Maramorosch, K.
ed. CRC Press. 278 pp. 1991.
36. Viroids and Satellites: Molecular Parasites at the Frontier of Life. Maramorosch,
K., ed. CRC Press. 174 pp. 1991.
37. Plant Diseases of Viral, Viroid, Mycoplasma and Uncertain Etiology.
Maramorosch, K., ed. Oxford and IBH. 190 pp. 1992.
38. Arthropod Cell Culture Systems. Maramorosch, K. and McIntosh, A.H., eds.
CRC Press. 236 pp. 1994.
39. Insect Cell Biotechnology. Maramorosch, K. and McIntosh, A.H., eds. CRC
Press. 190 pp. 1994.
40. Forest Trees and Palms: Disease and Control. Raychaudhuri, S.P. and
Maramorosch, K., eds. Science Publishers, New Hampshire. 334 pp. 1996.
Karl Maramorosch 89
41. Advances in Virus Research. Vol. 46 and 47. Maramorosch, K., Murphy, F.A.
and Shatkin, A.J., eds. 1996.
42. Advances in Virus Research. Vol. 48. Maramorosch, K., Murphy, F.A. and
Shatkin, A.J., eds. 1997.
43. Invertebrate Cell Culture: Novel Directions and Biotechnology Applications.
Maramorosch, K. and Mitsuhashi, J., eds. Science Publishers, New Hampshire.
296 pp. 1997.
44. Invertebrate Cell Culture: Looking Toward The Twenty First Century.
Proceedings of the IX International Conference on Invertebrate Cell Culture.
Maramorosch, K. and Loeb, M.J., eds. Society for In Vitro Biology, Columbia,
MD. 1997.
45. Advances in Virus Research. Vols. 49. Maramorosch, K., Murphy, F.A. and
Shatkin, A.J., eds. Academic Press, New York. 1998.
46. Advances in Virus Research. Vols. 50 and 51. Maramorosch, K., Murphy, F.A.
and Shatkin, A.J., eds. Academic Press, New York. 1998.
47. Advances in Virus Research. Vols. 52, 53, and 54. Maramorosch, K., Murphy,
F.A. and Shatkin, A.J., eds. Academic Press, New York. 1999.
48. Maintenance of Human, Animal, and Plant Pathogen Vectors. Maramorosch,
K. and Mahmood, F., eds. Science Publishers, Enfield, NH. 1999.
49. Biotechnology and Plant Protection in Forestry Science. Raychaudhuri, S.P.
and Maramorosch, K., eds. Science Publishers, Enfield, NH. 1999.
50. Advances in Virus Research. Forty volumes, Nos. 30-70. Maramorosch, K.,
Murphy, F.A. and Shatkin, A.J., eds. Academic Press/Elsevier. 1987-2007.
90 Wolf Prize in Agriculture
CHAPTER 1
Viruses, Vectors, and Vegetation:
An Autobiography
Karl Maramorosch
Department of Entomology, Rutgers-The State University of New Jersey, New Brunswick, New Jersey 08901
(maramors@rci.rutgers.edu)
This article was published in Advances in Virus Research Vol. 70, Maramorosch, K., viruses, vectors,
and vegetation: an autobiography, pp. 1-31, Copyright Elsevier Inc. (2007).
Karl Maramorosch 91
2 Karl Maramorosch
the Austro-Hungarian Monarchy. There I was born in 1915. The farm did
not move, but the borders moved many times. The family estate found
itself under no less than seven different regimes: Austria, Poland,
Petlura’s Ukraine, Romania, again Poland, USSR, Nazi Germany, USSR,
and currently Ukrainian Republic.
My father, a graduate of the Vienna Agricultural University, started
Ph.D. studies in Halle/Saal, Germany in 1898 but after 1 year returned
home to manage a 4000 acres estate, Kamionki Wielkie near Kolomyja,
owned by my grandfather. Around 1900, the estate was sold and the
smaller farm, Soroki, was purchased. My father considered himself a
Pole of Jewish creed. My mother, born in Zagreb, Croatia, was an accom-
plished pianist and a linguist, fluent in German, English, French, Italian,
and Serbo-Croatian. My siblings, Alfred, 6 years older, and Karla Bronia,
5 years older, spoke only Polish with my father and only German with my
mother. I grew up into this system, not realizing that it was not usual for
everyone to speak only Polish to one’s father and only German to one’s
mother. I grew up bilingual and only realized this clearly when I left
home and started writing letters to my parents—my thoughts were in
Polish when I addressed my father, and German toward my mother, and
I had to write not one, but two letters during my studies in Warsaw. I was
often asked how my parents spoke to each other. They spoke German
because, despite the great language skills of my mother, she could not
speak Polish without an accent, and it was, unfortunately, customary
in Poland to make fun of everybody who mispronounced Polish
words. My mother used Polish only when she went shopping or when
she spoke with people who helped at home, but never with friends or
visitors.
My third language was Ukrainian, which was spoken by all peasants
in the village where our farm was located. In high school I had 4 years of
Ukrainian and learned the Cyrillic script and some Ukrainian poetry by
Taras Shevchenko and Ivan Franko.
When I was 14 years old, my brother came home for his winter
vacation from Lwow (Lviv), where he was studying medicine. He told
me how his biology professor, Rudolf Weigl, invented a vaccine during
World War I that protected against exanthematous typhus. I was
completely fascinated, hearing how Professor Weigl was giving enemas
to individual body lice. Weigl infected the lice with Rickettsia prowazekii,
inserting glass micropipettes into their anal openings. Afterward he
maintained the inoculated lice on human volunteers for several days.
Subsequently, he removed the intestines from batches of 140 inoculated
lice, crushed the intestines in a tiny glass mortar with a few drops of
formalin, and obtained a single doze of his vaccine. Later I found out
that this was in Europe the only available vaccine against trench fever
until the end of World War II (Szybalski, 1999). The information about the
92 Wolf Prize in Agriculture
4 Karl Maramorosch
6 Karl Maramorosch
leafhoppers alive but are interested in destroying the pests. I was shocked
but my mentor consoled me and suggested to resubmit my paper to the
Journal of the New York Entomological Society. It was accepted and pub-
lished in 1951 (Maramorosch, 1951a). Twenty years later, I became Editor
in Chief of the Journal of the New York Entomological Society and remained
in that capacity for a dozen years.
After a few weeks, Dr. Black suggested that I should continue my
doctoral studies at Columbia University, and he gave me time off to take
courses and laboratory sessions. One day he suggested that I should
apply to the American Cancer Society for a predoctoral fellowship that
would pay $200 per month plus tuition at the university. When I read the
application form, I noticed immediately that it specified that the applicant
must be a US citizen. I was less than 1 year in the United States and thus
was at least 4 years from applying for US citizenship. Therefore, I put the
form aside and did nothing about it. A few days later Dr. Black asked me
whether I have filled out the form and when I replied that I could not do
this, he said, with a poker face: ‘‘Karl, I thought that you wanted to
become a scientist, but now I am disappointed.’’ I explained that I could
not fill out the form because it specified that the applicant must be a US
citizen. I was quite surprised when Dr. Black said: ‘‘If you want to be a
scientist, you have to be accurate and logical. Filling out the form is one
thing, while being a US citizen is another. I can help you in filling out the
form. Simply add a first page, calling attention to the fact that you are not
yet a US citizen because you arrived recently, but you expect to become
one in four years.’’ While I did not believe that my application would be
96 Wolf Prize in Agriculture
8 Karl Maramorosch
10 Karl Maramorosch
him that I was carrying out a serial passage of the ‘‘aster yellows virus’’ in
insect vectors (Maramorosch, 1952a). I was just finishing my last, 10th
passage and Dr. Black decided that he would wait with his publication
until mine would come out. Can you imagine that occurring today?
In 1952, I described the multiplication of the aster yellows pathogen
(Maramorosch, 1952a) (considered at that time to be a virus, and in 1968
recognized as a phytoplasma) and I entered the detailed description of the
serial passage of the aster yellows ‘‘virus’’ for the Cressy Morrison Prize
competition of the New York Academy of Sciences. My winning of this
prize started my intensive activities at the New York Academy, where I
became chair of the Microbiology Section, and later Recording Secretary
and Vice President. Work as a member of the committee responsible for
the organization of academy conferences gave me the experience in orga-
nizing later comparative virology and other national and international
conferences.
In 1952, I attended a New York Academy conference on virus
taxonomy. Among the invited participants were Dr. Kenneth M. Smith
from Cambridge and Sir Frederick C. Bawden (Fig. 4) Director of
the Rothamsted Experimental Station in Harpenden, Hearts, United
Kingdom. The two plant virologists were recognized as the world’s
leading authorities on plant viruses. I met both for the first time and
12 Karl Maramorosch
14 Karl Maramorosch
driving with Dr. Porter to take part in the June symposium and they took
me along, to see the beautiful location and to apply to Dr. Demerec
personally. With no written application and no formalities, the permis-
sion was granted and this stroke of good luck had a profound influence
on my career.
During the 1950s, I spent eight summers at the Cold Spring Harbor
Laboratories on Long Island, New York. Dr. Barbara McClintock permit-
ted me to use her greenhouses while she was working outdoors with corn
(Zea mays). Each year at the end of August, Dr. Alfred Hershey organized
a bacteriophage symposium for invited bacterial virologists. Although I
did not work on bacteriophages, I was permitted to attend these meetings,
where as yet unpublished findings were presented by the virologists.
Throughout the summers, Drs. Max Delbruck, Salvator Luria, and Ernst
Mayr were working in Cold Spring Harbor, lecturing, and socializing
with the small group of scientists. Dr. Luria was working on his textbook
on viruses and I was greatly impressed watching him dictate into a tape
recorder each morning, and then mailing the tape to his secretary in
Urbana for typing. When the typed version came back to Cold Spring
Harbor, Dr. Luria made small corrections and each chapter was ready for
publication. Few times he asked me for comments and when the book was
published, he donated a copy to me and I found that he acknowledged my
reviewing of a couple of chapters in his book.
One day Dr. Luria suggested that I should invite Japanese postdoc-
toral scientists to my laboratory and he added: ‘‘Get a good Japanese
postdoc, but never more than one. You will rapidly advance with your
work, but if you get more than one Japanese associate, you will no longer
have any time with your daughter and your wife, because you will try to
keep up with your Japanese postdocs and spend 7 days a week in the lab.’’
I remembered the first part of Luria’s suggestion and followed it when I
left Rockefeller University in 1961 and moved to the Boyce Thompson
Institute. But I did not follow the advice concerning the limitation of
invited Japanese postdocs. I soon found out how correct Luria was
when he told me never to get more than one Japanese coworker at a
time. When I got three Japanese associates at the same time, my own
working habit changed as I felt compelled to keep up with my Japanese
coworkers.
Thanks to Dr. McClintock’s generosity in Cold Spring Harbor where
she permitted me to use her greenhouses during the summer, I could
carry out an experiment in which I injected antibiotics into abdomens of
leafhoppers, exposed to presumptive viruses of aster yellows and corn
stunt. I used penicillin, streptomycin, and tetracycline, convinced that the
causative agents of the two plant diseases were viruses. Penicillin and
streptomycin injections did not prevent transmissions, but tetracycline-
injected leafhoppers failed to infect the exposed seedlings. Convinced that
Karl Maramorosch 105
16 Karl Maramorosch
tetracycline has no effect on viruses, I did not believe the results of the
tests, and assumed that the failed transmission was due to the heat in the
greenhouses. I did not repeat the experiment after I returned to the Rocke-
feller greenhouses and I published the results and my wrong conclusion
in the Transactions of the New York Academy (Maramorosch, 1954). Had I
repeated the tests, I would have found that not the summer heat but
tetracycline interfered with the presumptive viruses. Ten years later, my
Japanese colleagues in Tokyo discovered the phytoplasma nature of the
aster yellows disease, but I missed the boat.
did not. It would seem easy to stop the spread of the cadang-cadang
viroid by dipping the cutting tools of plantation workers in a calcium
chloride solution (Maramorosch, 1985, 1993). As far as I know, this has not
been implemented and more than 50 million palms have been destroyed
by cadang-cadang in the Philippines (Maramorosch, 2004). Losses are
partially alleviated by replanting with susceptible, but earlier maturing,
coconut palms.
18 Karl Maramorosch
20 Karl Maramorosch
During the past three decades, invertebrate cell culture became widely
used in biotechnology and basic research in virology. Use of baculoviruses
in insect cell cultures is gaining popularity for the production of recombi-
nant proteins, viral insecticides, and the production of vaccines. In vitro
techniques are indispensable for studies of insect virus expression systems.
Application of invertebrate cell culture and molecular biology is leading to
significant progress in the understanding of cellular and molecular interac-
tions between insect cells and viruses. Often unexpected results are
obtained as was the case with our M&M medium, developed for leafhopper
cell culture, and later found best suitable for mosquito cell cultivation and
the study of arboviruses in mosquito cells (Maramorosch, 1979b).
22 Karl Maramorosch
IX. BOOKS
In 1960 at the AAAS Annual Meeting, I stopped at the book exhibit of
Academic Press where I met Vice President, Kurt Jacoby. We spoke for
quite a while about his former work in Germany and the creation of
Academic Press in New York. I asked Mr. Jacoby whether symposium
papers on biological transmission of animal and plant disease agents could
be published by Academic Press. I was organizing a 2-day symposium on
this subject, to be held at the Annual Meeting of the Entomological Society
of America in Atlantic City, NY. Mr. Jacoby agreed and my first book, of
192 pages, ‘‘Biological Transmission of Disease Agents,’’ was published in
1962. As agreed, I received no royalties. Years later, I was told that all 1800
copies were sold when the book went out of print. The idea of publishing
the presentations came only after the conference participants had agreed
to be symposium speakers. I had considerable difficulty in persuading
some authors to submit manuscripts for publication. Foreign participants,
Dr. W. C. Willett from Kaduna, Nigeria, and Dr. D. Blascovic from Brati-
slava, Slovakia, were among the first to send their contributions. The
Rockefeller Foundation arranged the travel of these eminent participants
through a grant to the Entomological Society of America.
The success of my first book prompted me to again try Academic Press
for the publication of a more voluminous volume of 666 pages. The
treatise was based on a United States–Japan seminar, which I organized
in Tokyo together with Dr. Paul Oman. Mr. Jacoby was not interested this
time because as he explained, symposia were not selling well. Wiley
Interscience agreed to publish the book when I added several additional
authors who did not participate in the Tokyo meeting. I also used the title
of this second book, ‘‘Viruses, Vectors, and Vegetation’’ (1969) for the title
of my current autobiographical chapter.
During the following years several volumes on viruses, edited by me
alone or jointly with other virologists, were mainly published by Aca-
demic Press. In 1968, Springer published ‘‘Insect Viruses’’ (192 pp.). In
1971, ‘‘Comparative Virology,’’ edited by me and E. Kurstak, (Academic
Press, 584 pp.) was followed by ‘‘Viruses, Evolution, and Cancer’’ (813
pp., 1974). In 1975, with R. E. Shope, we edited ‘‘Invertebrate Immunity’’
(Academic Press, 365 pp.)
Viruses and virus diseases were included in ‘‘Tropical Diseases
of Legumes,’’ edited by Julio Bird and me in 1975. In 1977, I edited
112 Wolf Prize in Agriculture
24 Karl Maramorosch
X. INTERNATIONAL CONNECTIONS
26 Karl Maramorosch
ACKNOWLEDGMENTS
I want to express my special thanks to all my past graduate students and postdoctoral
associates who worked with me in the Unites States and abroad during the past years. To
them I express my sincere thanks and best wishes for their continuous successful research
and happy life.
116 Wolf Prize in Agriculture
REFERENCES
Bird, J., and Maramorosch, K. (1975). Viruses and virus diseases associated with whiteflies.
Adv. Virus Res. 22:55–110.
Black, L. M. (1953). Transmission of plant viruses by Cicadellids. Adv. Virus Res. 1:69–89.
Brakke, M. K., Maramorosch, K., and Black, L. M. (1953). Properties of the wound-tumor
virus. Phytopathology 43:387–390.
Corner, G. W. (1964). ‘‘A History of the Rockefeller Institute 1901–1953.’’ The Rockefeller
Institute Press, New York City.
Fukushi, T. (1935). Multiplication of virus in its insect vector. Proc. Imp. Acad. Japan
11:301–303.
Ginsberg, H. S. (1999). The life and times of adenoviruses. Adv. Virus Res. 54:1–13.
Goodman, R. M. (1977). Single-stranded DNA genome in a whitefly-transmitted virus.
Virology 83:171–179.
Granados, R. R., Hirumi, H., and Maramorosch, K. (1967). Electron microscopic evidence for
wound-tumor virus accumulation in various organs of an inefficient leafhopper vector,
Agalliopsis novella. J. Invertebr. Pathol. 9:147–159.
Granados, R. R., Ward, L., and Maramorosch, K. (1968). Insect viremia caused by a plant-
pathogenic virus: Electron microscopy of vector hemocytes. Virology 34:790–796.
Hirumi, H., and Maramorosch, K. (1963). Recovery of aster yellows virus from various
organs of the insect vector, Macrosteles fascifrons. Contrib. Boyce Thompson Inst. 22:165–173.
Hirumi, H., and Maramorosch, K. (1964). The in vitro cultivation of embryonic leafhopper
tissues. Exp. Cell Res. 36:625–631.
Hirumi, H., and Maramorosch, K. (1968). Electron microscopy of wound-tumor virus in
cultured embryonic cells of the leafhopper Macrosteles fascifrons. In ‘‘Second International
Colloquium on Invertebrate Tissue Culture’’ (C. Barigozzi, ed.), pp. 203–217. Lombardo
Acad. Sci. Lettere, Milano.
Hirumi, H., and Maramorosch, K. (1971). Cell culture of Hemiptera. In ‘‘Invertebrate Tissue
Culture’’ (C. Vago, ed.), Vol. 1, pp. 307–340. Academic Press, New York.
Hirumi, H., Burton, G. J., and Maramorosch, K. (1971). Preliminary studies on electron
microscopy of Friend murine leukemia virus in the midgut of experimentally infected
mosquitoes. J. Virol. 8:801–804.
Hirumi, H., Frankel, J. W., Prickett, C. O., and Maramorosch, K. (1974). Coexistance of
particles resembling herpes virus and Type-C virus in chicken feather follicle epithelium.
J. Natl. Cancer Inst. 52:303–306.
Koren, Z., Van Damme, O., Hirumi, H., and Maramorosch, K. (1971). The growth and
stimulation of mouse trophoblastic cells in culture by polyoma virus. Am. J. Obstet.
Gynecol. 111:846–850.
Maramorosch, K. (1950). Influence of temperature on incubation and transmission of the
wound-tumor virus. Phytopathology 40:1071–1093.
Maramorosch, K. (1951a). Handy insect-vector cage. J. NY Entomol. Soc. 59:49–50.
Maramorosch, K. (1951b). A simple needle for micro-injections. Nature. 167:734.
Maramorosch, K. (1952a). Multiplication of aster-yellows virus in its vector. Nature
169:194–195.
Maramorosch, K. (1952b). Toxicity of cellulose acetate sheets to plants and fish. Science
115:236.
Maramorosch, K. (1953). A versatile virus. Sci. Am. 188:78–86.
Maramorosch, K. (1954). Biological transmission of plant viruses by animal vectors. Trans.
NY Acad. Sci. 16:189–195.
Maramorosch, K. (1955). Multiplication of plant viruses in insect vectors. Adv. Virus Res.
3:221–249.
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28 Karl Maramorosch
Mitsuhashi, J., and Maramorosch, K. (1964). Inoculation of plant tissue cultures with aster
yellows virus. Virology 23:277–279.
Randles, J. W. (1975). Association of two ribonucleic acid species with cadang-cadang disease
of coconut palm. Phytopathology 65:163–167.
Shikata, E., and Maramorosch, K. (1965a). Plant tumor virus in arthropod host: Microcrystal
formation. Nature 288:507–508.
Shikata, E., and Maramorosch, K. (1965b). Electron microscopic evidence for the systemic
invasion of an insect host by a plant pathogenic virus. Virology 27:461–475.
Shikata, E., and Maramorosch, K. (1966a). An electron microscope study of plant neoplasia
induced by wound tumor virus. J. Natl. Cancer Inst. 36:97–116.
Shikata, E., and Maramorosch, K. (1966b). Electron microscopy of a pea enation mosaic virus
in plant cell nuclei. Virology 30:439–454.
Shikata, E., and Maramorosch, K. (1967a). Electron microscopy of wound tumor virus
assembly sites in insect vectors and plants. Virology 32:363–377.
Shikata, E., and Maramorosch, K. (1967b). Electron microscopy of the formation of wound
tumor virus in abdominally inoculated insect vectors. J. Virol. 1:1052–1073.
Shikata, E., and Maramorosch, K. (1969). Electron microscopy of insect-borne viruses in situ.
In ‘‘Viruses, Vectors, and Vegetation’’ (K. Maramorosch, ed.), pp. 393–415. Interscience,
John Wiley & Sons, New York.
Shikata, E., Maramorosch, K., and Granados, R. R. (1966). Electron microscopy of pea
enation mosaic virus in plants and aphid vectors. Virology 29:426–436.
Shikata, E., Orenski, S. W., Hirumi, H., Mitsuhashi, J., and Maramorosch, K. (1964). Electron
micrographs of wound-tumor virus in an animal host and in a plant tumor. Virology
23:441–444.
Storey, H. H. (1933). Investigations of the mechanism of the transmission of plant viruses by
insect vectors. I. Proc. R. Soc. Lond. Ser. B 113:463–485.
Streissle, G., and Maramorosch, K. (1963). Reo virus and wound-tumor virus: Serological
cross reactivity. Science 140:996–997.
Szybalski, W. (1999). Maintenance of human-fed live lice in the laboratory and production of
Weigl’s exanthematous typhus vaccine. In ‘‘Maintenance of Human, Animal, and Plant
Pathogen Vectors’’ (K. Maramorosch and F. Mahmood, eds.), pp. 161–179. Science Pub-
lishers, Enfield, NH, USA.
Tokumitsu, T., and Maramorosch, K. (1966). Survival of aphid cells in vitro. Exp. Cell Res.
44:652–655.
Tokumitsu, T., and Maramorosch, K. (1967). Cytoplasmic protrusions in insect cells during
mitosis in vitro. J. Cell Biol. 34:677–683.
Yamada, K., and Maramorosch, K. (1980). Propagation of Heliothis zea single embedded
baculovirus in a H. zea cell line. Abstracts 16th International Entomology Congress
Tokyo, Japan, p. 193.
Yamada, K., and Maramorosch, K. (1981). Plaque assay of Heliothis zea baculovirus employ-
ing a mixed agarose overlay. Arch. Virol 67:187–189.
Yamada, K., Sherman, K. E., and Maramorosch, K. (1981). In vivo infectivity of early and
late passaged Heliothis zea polyhedra produced in tissue culture. Appl. Entomol. Zool.
16:504–505.
BOOKS
K. Maramorosch (ed.) (1962). Biological Transmission of Disease Agents. Academic Press,
New York and London.
K. Maramorosch and H. Koprowski (eds.) (1967–1984). Methods in Virology, Vols. 1–8.
Academic Press, New York and London.
Karl Maramorosch 119
30 Karl Maramorosch
K. Maramorosch (ed.) (1968). Insect Viruses, Springer Verlag, New York, Berlin.
K. Maramorosch (ed.) (1969). Viruses, Vectors, and Vegetation. Wiley-Interscience,
New York, London, Toronto.
K. Maramorosch and E. Kurstak (eds.) (1971). Comparative Virology. Academic Press,
New York.
M. A. Lauffer, F. B. Bang, K. Maramorosch, and K. M. Smith (eds.) (1973–1981). Advances in
Virus Research, Vols. 22–26. Academic Press, New York.
E. Kurstak and K. Maramorosch (eds.) (1974). Viruses, Evolution, and Cancer. Academic
Press, New York, London.
K. Maramorosch and R. E. Shope (eds.) (1975). Invertebrate Immunity. Academic Press,
New York and London.
J. Bird and K. Maramorosch (eds.) (1975). Tropical Diseases of Legumes. Academic Press,
New York and London.
K. Maramorosch (ed.) (1976). Invertebrate Tissue Culture: Research Applications. Academic
Press, New York, London.
E. Kurstak and K. Maramorosch (eds.) (1976). Invertebrate Tissue Culture: Applications in
Medicine, Biology, and Agriculture. Academic Press, New York and London.
K. Maramorosch (ed.) (1977). The Atlas of Insect and Plant Viruses. Academic Press,
New York and London.
K. Maramorosch and H. Hirumi (eds.) (1979). Practical Tissue Culture Applications.
Academic Press, New York.
E. Kurstak, K. Maramorosch, and A. Dubendorfer (eds.) (1977). Invertebrate Systems In Vitro.
Elsevier-North Holland Biomedical Press.
K. F. Harris and K. Maramorosch (eds.) (1977). Aphids as Virus Vectors. Academic Press,
New York and London.
K. Maramorosch and K. F. Harris (eds.) (1979). Leafhopper Vectors and Plant Disease
Agents. Academic Press, New York.
K. F. Harris and K. Maramorosch (eds.) (1980). Vectors of Plant Pathogens. Academic Press,
New York.
M. A. Lauffer and K. Maramorosch (eds.) (1981–1984). Advances in Virus Research, Vols. 27
and 28. Academic Press, New York.
K. Maramorosch and K. F. Harris (eds.) (1981). Plant Diseases and Vectors: Ecology and
Epidemiology. Academic Press, New York.
K. F. Harris and K. Maramorosch (eds.) (1982). Pathogens, Vectors, and Plant Diseases:
Approaches to Control. Academic Press, New York.
J. J. McKelvey, Jr., B. F. Eldridge, and K. Maramorosch (eds.) (1980). Vectors of Disease
Agents: Interactions with Plants, Animals and Man. Praeger, New York.
E. Kurstak and K. Maramorosch (eds.) (1978). Viruses and Environment. Academic
Press, New York.
K. Maramorosch (ed.) (1981–1987). Advances in Cell Culture, Vols. 1–5. Academic Press,
New York.
K. Maramorosch and G. H. Sato (eds.) (1988–1989). Advances in Cell Culture, Vols. 6 and 7.
Academic Press, New York.
K. Maramorosch and J. Mitsuhashi (eds.) (1982). Invertebrate Cell Culture Applications.
Academic Press, New York.
K. Maramorosch and J. J. McKelvey, Jr. (eds.) (1985). Subviral Pathogens of Plants and
Animals: Viroids and Prions. Academic Press, New York.
K. Maramorosch, F. A. Murphy, and A. J. Shatkin (eds.) (1985–2007). Advances in Virus
Research Vols. 29–70. Academic Press, Elsevier, London, San Diego, Amsterdam etc.
K. Maramorosch and K. E. Sherman (eds.) (1985). Viral Insecticides for Biological Control.
Academic Press, Orlando, New York, London, Tokyo.
120 Wolf Prize in Agriculture
K. Maramorosch (ed.) (1987). Biotechnology in Insect Pathology and Cell Culture. Academic
Press, San Diego, CA.
E. Kurstak, Y. Kuroda, and K. Maramorosch (eds.) (1988). Invertebrate and Fish Tissue
Culture. Springer, Berlin and Tokyo.
K. Maramorosch (ed.) (1991). Viroids and Satellites: Molecular Parasites at the Frontier of
Life. CRC Press, Boca Raton, FL, USA.
K. Maramorosch (ed.) (1992). Plant Diseases of Viral, Viroid, Mycoplasma, and Uncertain
Etiology. Oxford and IBH, New Delhi, INDIA.
K. Maramorosch and A. H. McIntosh (eds.) (1994). Arthropod Cell Culture Systems.
CRC Press, Boca Raton, Ann Arbor, London, Tokyo.
K. Maramorosch and A. H. McIntosh (eds.) (1994). Insect Cell Biotechnology. CRC Press,
Boca Raton, FL.
K. Maramorosch and J. Mitsuhashi (eds.) (1997). Invertebrate Cell Culture: Novel Directions
and Biotechnology Applications. Science Publishers, Enfield, NH, USA.
K. Maramorosch and M. J. Loeb (eds.) (1997). Invertebrate Cell Culture: Looking Toward
the XXI Century. SIVB, Columbia, MD.
K. Maramorosch and F. Mahmood (eds.) (1999). Maintenance of Animal/Human and
Plant Pathogen Vectors. Science Publishers, Enfield, NH, USA.
John O. Almquist
Pennsylvania State University
University Park, Pennsylvania, USA
k
Professor Almquist’s early work on the addition of antibiotics to bull semen resulted
in substantial increases in breeding efficiency. This, coupled with his remarkable
achievements in developing methods for processing, freezing, and thawing of frozen
semen, significantly enhanced the practical utilization of artificial insemination in
the livestock industry. Many techniques developed by him for cattle have been
applied to other species, including the human male.
CURRICULUM VITAE
Date and Place of Birth - February 10, 1921, Hoidrege, Nebraska
ACADEMIC DEGREES
Cornell University, B.S., 1942
Purdue University, M.S., 1944
The Pennsylvania State University Ph.D., 1947
121
122 Wolf Prize in Agriculture
ESSENTIAL BIBLIOGRAPHY
Author or coauthor of 160 scientific publications, including 78 papers in refereed
journals, 3 bulletins, 3 book chapters and over 70 papers given at scientific meetings.
4. Most young dairy and beef bulls can ejaculate sufficient sperm for
progeny testing by l4 months of age.
5. The testes of mature bulls produce prodigious numbers of sperm —
about 7 billion per day in Holsteins.
6. Postpuberal increases in weekly sperm output are much more rapid in
Charolais and Holstein than in Angus and Hereford bulls.
7. Charolais bulls attain puberty at a significantly earlier age (41 wks)
than Angus and Hereford bulls (45 wks). This finding contradicts the
commonly held viewpoint that Charolais bulls mature rather slowly.
8. On a within herd basis, much valuable growth data can be gathered for
Charolais if the top 60% of bulls at 365 days of age are continued on
gain test to 550 days of age.
9. Bull calves should be fed ad libitum for achieving puberty at the earliest
possible age.
d. From a 10-year study of 18 Holstein bulls, he and his collaborator (Dr. R.G.
Thompson, Dept. of Pathology, University of Guelph, Ontario, Canada)
demonstrated that there was no difference in spinal bone lesions between
bulls ejaculated continuously six times per week from 1 to 9.5 years of age
and the controls ejaculated only once weekly to the same age. This report
helped to allay fears in the AI industry that high frequency of ejaculation
involving three false mounts preceding each ejaculation shortens the useful
reproductive life of a bull by increasing the number and severity of vertebral
body osteophytes.
e. Important findings of recent laboratory and fertility studies include the
following:
1. Developed new antibiotic combinations, lincomycin-spectinomycin and
penicillin-neomycin, which were shown to be satisfactory substitutes if
penicillin-streptomycin is not available. Each combination effectively
controlled bacterial growth in semen without reducing fertility
(1973–74).
2. Explored an area of male reproduction — physiology of ejaculation —
which had not been studied extensively despite its obvious importance
in animal and human reproduction. Three recent publications described
a new technique for making the first quantitative in viva pressure
measurements of contractions of the vas deferens and vesicular glands
in both anesthetized and conscious rabbits by implanting silicone tubing
sensors.
3. Published in 1973 the first two papers on new methods for freezing and
thawing semen packaged in Continental US plastic straws. Dr. Almquist’s
research showed that fast thawing was much mare important than rate
128 Wolf Prize in Agriculture
of freezing. Freezing rate over a rather wide range did not affect sperm
survival in US straws provided they were thawed rapidly (7 to 15
seconds). He and his graduate students also found that both acrosomal
maintenance and motility of spermatozoa were affected more by thawing
rate than by time of exposure to glycerol, a cryoprotective agent.
Optimum glycerol level for freezing semen in straws was 11%, or the
same as that reported earlier for freezing semen in glass ampules using
skimmilk extender.
4. Showed that semen in straws could be placed onto canes before freezing
and that loads of up to 7500 straws could be frozen in an automatic
forced vapor freezer. Results were equally as good as those with the
procedure presently used by the AI industry in which about 350 straws
are individually cooled and then placed into goblets and onto canes
while at 196°C. This automatic method provides a 20-fold increase in
the number of straws which can be frozen at one time, reduces labor
and improves product quality.
5. Conventionally, semen is warmed to 5°C and then used for insemination.
From studies with thawing baths ranging from 5°C to 135°C, he showed
that warming semen rapidly in straws directly to body temperature
while thawing was more desirable. By placing straws of frozen semen
into a 35°C water bath for 30 seconds, seminal temperature was raised
rapidly to near body temperature and fertility involving over 21,000
experimental inseminations was significantly higher than for semen
warmed to only 5°C.
f. Other important early research findings include the following:
1. Developed a safe, practical method of coloring semen to distinguish
semen from bulls of different breeds. Reported in 1946 and still used by
AI studies in most countries.
2. Established relationships between numbers and types of bacteria in bull
semen and fertility.
3. Developed a commonly used method of staining bull sperm to
differentiate living and dead cells.
4. Collaborated with Dr. R.J. Flipse to show that large quantities of corn
and grass silage could be fed safely to dairy breeding bulls and that
dairy bull calves mature later on low-energy rations.
5. Collaborated with Dr. B.B. Hale to conduct the first quantitative studies
on sexual behavior and semen production by dairy bulls.
6. Cooperated with Dr. T.Y. Tanabe to show that pregnancy interruption in
AI of dairy cattle is reduced by addition of antibiotics to bull semen and
deposition of semen in the mid-cervical region of the uterus.
John O. Almquist 129
2) The top breeding AI bulls sire many more offspring. Faster genetic
improvement has been achieved by identifying and using the best bulls
more efficiently. Fewer bulls are needed to breed the same number of
females. One Holstein-Friesian bull in the USA sired about 250,000 offspring
in his lifetime. Based on actual production records for his 27,754 tested
daughters, extensive use of this superior sire resulted in over 17.5 million
dollars more income to the owners of these daughters. The estimated value
of the extra milk produced by the approximately 100,000 tested and
untested daughters of this bull amounted to over 600 million dollars.
Breeding potential for top bulls is tremendous; recently two outstanding
bulls in a Pennsylvania Al stud using Dr. Almquist’s recommendations
were mated to 120,000 and 125,000 cows in one year. Such figures seemed
impossible a short while ago.
Such potent “bull power” was unknown until the prodigious production of
spermatozoa by bulls (6 to 9 billion per day) and means of obtaining them
were revealed through original research done by Almquist and coworkers
starting in 1951. They also were the first to provide quantitative data on the
sexual behavior of bulls. The new techniques developed in the 1950’s for
measuring the numbers of sperm which bulls are capable of producing, storing
and ejaculating have been adopted by reproductive researchers for determining
these parameters in many species, including man. This pioneering work on
male reproductive physiology has received international recognition and has
aided the surge in knowledge about the role of the male in the reproductive
process and in overcoming infertility in both man and domestic animals.
Dr. Almquist became convinced at an early age that the usefulness of AI
should be expanded beyond dairy cattle to increase production in other species
of economic importance for providing food and fiber to man. With characteristic
foresight, he initiated research on the reproductive capacity of beef bulls in
1962. Much to the surprise of many animal breeders, he demonstrated that
many beef bulls possess great reproductive potential for use in AI to increase
meat production. Prior to his research, it was commonly believed that beef
bulls should not be used for breeding before 18 months of age. He showed that
semen collected, frozen and used from beef bulls under 1 year of age yielded
economically satisfactory AI fertility rates, and thus, beef bulls could be sampled
in a progeny testing program at a much earlier age than previously believed
possible.
The original research by Dr. Almquist showing considerably higher AI
conception rates with antibiotic-treated semen provided a major impetus for
widespread acceptance and more rapid growth of cattle AI in Pennsylvania
and the USA. Antibiotics in semen aid in controlling spread of reproductive
diseases and significantly reduce the costly losses due to early death of embryos
in infected cows.
John O. Almquist 131
the cost of existing extenders and subsequent widespread use in the USA and
excellent levels of fertility.
Before frozen semen became practical, he combined two of his research
findings to enable dairymen for the first time to make selected matings to the
bull of their choice every day of the week. Using his milk-glycerol extender
and collecting semen more frequently than previously believed possible, he
demonstrated to the AI industry that semen from superior bulls could be
available every day.
Recognizing that use of AI was limited or nonexistent in many areas of the
world where refrigeration to preserve spermatozoa was not readily available or
was very costly, in 1962 he originated an extender which enables semen to be
stored at ambient room temperatures. The essential ingredient was skimmilk,
together with glucose, sodium citrate and citric acid.
With his keen desire to develop practical procedures for the entire AT
industry and his awareness of the need to cut production costs, in 1978
Dr. Almquist introduced procedures for processing bull semen for freezing
which make it possible for laboratory technicians to do their work faster and
more comfortably at room temperature rather than in a 5°C walk — in cold
room. Using This egg-yolk-glycerol extender, freshly collected semen is extended
completely and packaged in plastic straws at room temperature rather than at
50oC before being frozen. This new method has been tested successfully and
used under routine field conditions in Pennsylvania for 3 years.
In 1979, Dr. Almquist reported a practical method for thawing frozen bull
semen rapidly which significantly improved fertility by about 2 percentage
points. While this might appear to be a small increase, nevertheless, use of the
new procedure could save annually over 5 million dollars in the USA over 100
million dollars worldwide. Several AI organizations already have adopted this
new thawing method.
Conclusions:
The founding fathers of artificial insemination probably did not envision that it
would evolve into an extensive, daily agricultural practice affecting the lives of
people throughout the world. Ever increasing world population has and will
continue to mandate that animal agriculture must become more efficient and
productive. Hunger and malnutrition know no race or creed and no human is far
removed from its overtones.
It is the responsibility of scientists such as Dr. Almquist to utilize their mental
and physical resources to develop programs that will undergird the food production
potential of the world. The contribution that artificial insemination has made to
both quality and, quantity of available food may be one of the greatest advances
ever offered through animal agriculture.
John O. Almquist 133
Professor Lardy, with the late Professor Paul H. Phillips, made the basic discovery
in the early forties that a nutrient medium of egg yolk dispersed in a phosphate
buffer forms an ideal environment for the storage of spermatozoa of bull, ram,
stallion, and turkey. This breakthrough paved the way to the rapid expansion of
the practical use of artificial insemination in livestock breeding. Further, the ability
to store semen made it possible to initiate fundamental studies on the metabolism
of spermatozoa. The results of this outstanding research work enabled improvements
to semen storage technique leading to more efficient and widespread utilization of
semen.
CURRICULUM VITAE
Birthdate: August 19, 1917
EDUCATION:
B.S., Chemistry, South Dakota State University, Brookings, South Dakota, 1939.
M.S., Biochemistry, University of Wisconsin, Madison, Wisconsin, 1941.
Ph.D., Biochemistry, University of Wisconsin, Madison, Wisconsin, 1943.
135
136 Wolf Prize in Agriculture
PROFESSIONAL EXPERIENCE:
1944-45 - N.R.C. Post-Doctoral Fellow, University of Toronto.
1945-47 - Assistant Professor, Department of Biochemistry, University of
Wisconsin.
1947-50 - Associate Professor, Department of Biochemistry, University of
Wisconsin.
1950-88 - Professor, Department of Biochemistry, University of Wisconsin.
1950-present - Chairman, Section II, Institute for Enzyme Research, University of
Wisconsin.
1966-88 - Vilas Professor of Biological Sciences, University of Wisconsin.
1988 - Professor Emeritus.
MEMBERSHIP IN SOCIETIES:
American Chemical Society (Chairman, Biological Division, 1958)
American Society of Biological Chemists (President, 1964)
National Academy of Sciences
American Academy of Arts and Sciences
American Philosophical Society
Harvey Society
Biochemical Society (Great Britain)
American Diabetes Association
The Society for the Study of Reproduction
EDITORIAL BOARDS:
Archives of Biochemistry and Biophysics - 1957-1960.
Journal of Biological Chemistry - 1958-1964 and 1980-1985.
Biochemical Preparations - 1952-1968.
Methods of Biochemical Analysis - 1954-1971.
Biochemistry - 1962-1973 and 1975-1981.
FEBS Letters - 1970-1975.
PUBLICATIONS:
1. The Vitamin D content of some South Dakota roughages. Wallis, G.C. and
Lardy, H.A. (1938). Proc. South Dakota Academy of Science, 18, 46.
2. Manganese content of some South Dakota feeds. Moxon, A.L. and Lardy, H.A.
(1939). Proc. South Dakota Academy of Science, 19, 57.
3. The effect of selenium and arsenic at various ratios on the fermentation of
glucose by baker’s yeast. Lardy, H.A., and Moxon, A.L. (1939). Proc. South
Dakota Academy of Science, 19, 109.
4. Preservation of spermatozoa. Lardy, H.A., and Phillips, P.H. (1939). Am. Soc.
Animal Production, 32nd Annual Proceedings of American Society of Animal
Production, p. 219.
5. Increasing the rate of excretion of selenium from selenized animals by the
administration of bromobenzene. Moxon, A.L., Schaefer, A.E., Lardy, H.A.,
DuBois, K.P., and Olson, O.E. (1940). J. Biol. Chem. 132, 785.
138 Wolf Prize in Agriculture
6. A yolk-buffer pablum for the preservation of bull semen. Phillips, P.H. and
Lardy, H.A. (1940). J. Dairy Sci., 23, 399.
7. H ion concentration of various fluids of the genital tract of the cow. Lardy,
H.A., Pounden, W.D., and Phillips, P.H. (1940). Proc. Soc. Exp. Biol. Med., 44,
517.
8. Sperm stimulation in the bull through the subcutaneous administration of
ascorbic acid. Phillips, P.H. and Lardy, H.A. (1940). J. Dairy Sci., 23, 873.
9. The relationship of ascorbic acid to reproduction in the cow. Phillips, P.H.,
Lardy, H.A., Boyer, P.D., and Werner, G.M. (1941). J. Dairy Sci., 24, 153.
10. The effect of certain inhibitors and activators on sperm metabolism. Lardy,
H.A. and Phillips, P.H. (1941). J. Biol. Chem., 138, 195.
11. The interrelation of oxidative and glycolytic processes as sources of energy
for bull spermatozoa. Lardy, H.A., and Phillips, P.H. (1941). Am. J. Physiol.,
133, 602.
12. Phospholipids as a source of energy for motility of bull spermatozoa. Lardy,
H.A. and Phillips, P.H. (1941). Am. J. Physiol., 134, 542.
13. The effect of colored ions on the photo-inactivation of invertase. Lardy, H.A.
and Anderson, T.F. (1942). Science 95, 330.
14. The ascorbic acid content of the livers of selenized rats and chicks. Lardy,
H.A. and Moxon, A.L. (1942). Proc. South Dakota Acad. Sci., 22, 39.
15. The role of potassium in muscle phosphorylations. Boyer, P.D., Lardy, H.A.,
and Phillips, P.H. (1942). J. Biol. Chem., 146, 673.
16. Effect of pH and certain electrolytes on the metabolism of ejaculated
spermatozoa. Lardy, H.A. and Phillips, P.H. (1943). Am. J. Physiol., 138, 741.
17. Inhibition of sperm respiration and reversibility of the effects of metabolic
inhibitors. Lardy, H.A. and Phillips, P.H. (1943). J. Biol. Chem., 148, 333.
18. Inhibition of sperm glycolysis and reversibility of the effects of metabolic
inhibitors. Lardy, H.A. and Phillips, P.H. (1943). J. Biol. Chem., 148, 343.
19. The effect of thyroxine and dinitrophenol on sperm metabolism. Lardy, H.A.
and Phillips, P.H. (1943). J. Biol. Chem., 149, 177.
20. Further studies on the role of potassium and other ions in the phosphorylation
of the adenylic system. Boyer, P.D., Lardy, H.A., and Phillips, P.H. (1943).
J. Biol. Chem., 149, 529.
21. Acetate utilization for maintenance of motility of bull spermatozoa. Lardy,
H.A. and Phillips, P.H. (1944). Nature 153, 168.
22. Ineffectiveness of sodium succinate in control of duration of barbiturate
anesthesia. Lardy, H.A., Hansen, R.G., and Phillips, P.H. (1944). Proc. Soc.
Exp. Biol. Med., 55, 277.
23. Determination of citric acid in fermentation media and biological materials.
Perlman, D., Lardy, H.A., and Johnson, M.J. (1944). Ind. Eng. Chem., 16,
515.
Henry A. Lardy 139
42. Circular bath and shaking mechanism for manometric microapparatus. Lardy,
H.A., Gilson, W.E., Hipple, J., and Burris, R.H. (1948). Anal. Chem. 20, 1100.
43. A metabolic regulator in mammalian spermatozoa. Lardy, H.A., Ghosh, D.,
and Plaut, G.W.E. (1949). Science 109, 365.
44. Phosphorylation of hexoses by brain hexokinase. Wiebelhaus, V.D., and Lardy,
H.A. (1949). Arch. Biochem. 21, 321.
45. Comparative effectiveness of vitamin B12, whole liver substance and extracts
high in APA activity, as growth promoting materials for hyperthyroid animals.
Betheil, J.J. and Lardy, H.A. (1949). J. Nutrition 37, 495.
46. The net utilization of ammonium nitrogen by the growing rat. Lardy, H.A.
and Feldott, G. (1949). J. Biol. Chem. 179, 509.
47. A study of the metabolic activity of bull semen and spermatozoa in relation
to their fertilizing ability. Ghosh, D., Casida, L.E., and Lardy, H.A. (1949).
J. Animal Sci. 8, 265.
48. Metabolic functions of biotin. I. The role of biotin in bicarbonate utilization
by lactobacillus arabinosus studied with C14. Lardy, H.A., Potter, R.L., and
Burris, R.H. (1949). J. Biol. Chem. 179, 721.
49. Metabolic functions of biotin. II. The fixation of carbon dioxide by normal
and biotin-deficient rats. MacLeod, P.R., and Lardy, H.A. (1949). J. Biol.
Chem. 179, 733.
50. The oxalacetate decarboxylase of azotobacter vinelandii. Plaut, G.W.E. and
Lardy, H.A. (1949). J. Biol. Chem. 180, 13.
51. The synthesis of D-tagatose by biochemical oxidation and by an improved
chemical method. Totton, E.L., and Lardy, H.A. (1949). J. Am. Chem. Soc. 71,
3076.
52. The effect of the administration of various fatty acids on the blood ketone
levels of ruminants. Schultz, L.H., Smith, V.R., and Lardy, H.A. (1949).
J. Dairy Sci. XXXII, 817.
53. Metabolic functions of biotin. III. The synthesis of citrulline from ornithine
in liver tissue from normal and biotin-deficient rats. MacLeod, P.R., Grisolia,
S., Cohen, P.P., and Lardy, H.A. (1949). J. Biol. Chem. 180, 1003.
54. Phosphoric esters of biological importance. II. The synthesis of glucose-6-
phosphate from 1,2-isopropylidene-5,6-anhydro-D-glucofuranose. Lampson,
G.P., and Lardy, H.A. (1949). J. Biol. Chem. 181, 693.
55. Phosphoric esters of biological importance. III. The synthesis of propanediol
phosphate. Lampson, G.P., and Lardy, H.A. (1949). J. Biol. Chem. 181, 697.
56. Phosphoric ester of biological importance. IV. The synthesis and biological
activity of D-tagatose-6-phosphate. Totton, E.L., and Lardy, H.A. (1949).
J. Biol. Chem. 181, 701.
57. Relationship of vitamin B6 to the metabolism of D-amino acids. Armstrong,
K.L., Feldott, G., and Lardy, H.A. (1950). Proc. Soc. Exp. Biol. Med. 73, 159.
Henry A. Lardy 141
58. Orthophosphate uptake during the oxidation of fatty acids. Johnson, R.B. and
Lardy, H.A. (1950). J. Biol. Chem. 184, 235.
59. Systems for the separation of phosphoric esters by solvent distribution. Plaut,
G.W.E., Kuby, S.A., and Lardy, H.A. (1950). J. Biol. Chem. 184, 243.
60. 1,2-isopropylidene-D-glucitol, 1,2:5,6-diisopropylidene-D-glucitol and a new
synthesis of D,L-glyceraldehyde dimer. Pressman, B.C., Anderson, L., and
Lardy, H.A. (1950). J. Am. Chem. Soc. 72, 2404.
61. The relationship of folic acid to formate metabolism in the rat. Plaut, G.W.E.,
Betheil, J.J., and Lardy, H.A. (1950). J. Biol. Chem. 184, 795.
62. Stereochemical studies in the amino-desoxyinositol series. Meso-inosamine-
2 and scyllo-inosamine. Anderson, L. and Lardy, H.A. (1950). J. Am. Chem.
Soc. 72, 3141.
63. The net utilization of ammonium nitrogen by the growing rat. Lardy, H.A.
and Feldott, G. (1950). J. Biol. Chem. 186, 85.
64. Metabolism of ejaculated bull spermatozoa with particular reference to the
glycolysis of maltose. Plaut, G.W.E., and Lardy, H.A. (1950). Am. J. Physiol.
162, 598.
65. Incorporation of the carbons of acetone, formate, and carbonate into
acetoacetate. Plaut, G.W.E. and Lardy, H.A. (1950). J. Biol. Chem. 186, 705.
66. The mode of action of the antibiotic, usnic acid. Johnson, R.B., Feldott, G.,
and Lardy, H.A. (1950). Arch. Biochem. 28, 317.
67. The mechanism by which glyceraldehyde inhibits glycolysis. Lardy, H.A.,
Wiebelhaus, V.D., and Mann, K.M. (1950). J. Biol. Chem. 187, 325.
68. Phosphoric esters of biological importance. V. The synthesis of L-sorbose-1-
phosphate and L-sorbose-6-phosphate. Mann, K.M. and Lardy, H.A. (1950).
J. Biol. Chem. 187, 339.
69. Metabolic functions of the micro-catalytic B vitamins. Lardy, H.A. (1951).
Rec. Chem. Progress, Winter, p. 9.
70. The excretion of the citrovorum factor by hyperthyroid rats. Drysdale, G.R.,
Betheil, J.J., Lardy, H.A., and Baumann, C.A. (1951). Arch. Biochem. Biophys.
33, 1.
71. The nomenclature of the cyclohexitols and their derivatives. Fletcher, H.G.,
Jr., Anderson, L., and Lardy, H.A. (1951). J. Organ. Chem. 16, 1238.
72. Enzymatic incorporation of C14-bicarbonate into acetoacetate in the presence
of various substrates. Plaut, G.W.E., and Lardy, H.A. (1951). J. Biol. Chem.
192, 435.
73. Metabolic functions of biotin. IV. The role of carbamyl-L-glutamic acid in
the synthesis of citrulline by normal and biotin-deficient rats. Feldott, G.,
and Lardy, H.A. (1951). J. Biol. Chem. 192, 447.
74. 1,6-dibenzyl-3,4-isopropylidene-D-mannitol. Anderson, L., Lueptow, A.J., and
Lardy, H.A. (1951). J. Am. Chem. Soc. 73, 5002.
142 Wolf Prize in Agriculture
221. Paths of carbon in gluconeogenesis. Lardy, H.A., Veneziale, C., and Gabrielli,
F. (1969). FEBS Symposium 19, 55.
222. Site-specific inhibitors of gluconeogenesis. Lardy, H.A. In Colloq. der
Gesellschaft fur Biol. Chem., p. 374, Mosbach-Baden, Germany, April 14-16.
223. Inhibition by hydrazine of gluconeogenesis in the rat. Ray, P.D., Hanson, R.L.,
and Lardy, H.A. (1970). J. Biol. Chem. 245, 690.
224. Bongkrekic acid: An inhibitor of the adenine nucleotide translocase of
mitochondria. Henderson, P.J.F. and Lardy, H.A. (1970). J. Biol. Chem. 245,
1319.
225. Inhibition of phosphofructokinase by quinone methide and α-methylene
lactone tumor inhibitors. Hanson, R.L., Lardy, H.A., and Kupchan, S.M. (1970).
Science 168, 378.
226. Antibiotic inhibition of mitochondrial energy-transfer reactions. Henderson,
P.J.F. and Lardy, H.A. (1970). In Antimicrobial Agents and Chemotherapy-
1969, p. 18, Am. Soc. for Microbiology.
227. Gluconeogenesis. Lardy, H.A. (1970). In Nobel Symposium 13 (Cerasi, E. &
Luft, R., eds.), p. 199, Wiley and Sons.
228. Factors affecting the inhibition of adenine nucleotide translocase by bongkrekic
acid. Henderson, P.J.F., Lardy, H.A., and Dorschner, E. (1970). Biochemistry
9, 3453.
229. Gluconeogenesis from pyruvate in isolated perfused rat liver. Veneziale, C.M.,
Gabrielli, F., and Lardy, H.A. (1970). Biochemistry 9, 3960.
230. Mode of action of hypoglycemic agents. III. Studies on 5-methoxyindole-2-
carboxylic acid and quinaldic acid. Reed, J. and Lardy, H.A. (1970). J. Biol.
Chem. 245, 5297.
231. Effects of phosphodiesterase inhibitors and cyclic nucleotides on sperm
respiration and motility. Garbers, D.L., Lust, W.D., First, N.L., and Lardy, H.A.
(1971). Biochemistry 10, 1825.
232. Rat liver pyruvate carboxylase. I. Preparation, properties, and cation specificity.
McClure, W.R., Lardy, H.A., and Kneifel, H.P. (1971). J. Biol. Chem. 246,
3569.
233. Rat liver pyruvate carboxylase. II. Kinetic studies of the forward reaction.
McClure, W.R., Lardy, H.A., Wagner, M., and Cleland, W.W. (1971). J. Biol.
Chem. 246, 3579.
234. Rat liver pyruvate carboxylase. III. Isotopic exchange studies of the first
partial reaction. McClure, W.R., Lardy, H.A., and Cleland, W.W. (1971).
J. Biol. Chem. 246, 3584.
235. Rat liver pyruvate carboxylase. IV. Factors affecting the regulation in vivo.
McClure, W.R., and Lardy, H.A. (1971). J. Biol. Chem. 246, 3591.
236. Contaminant of malic acid: A malic enzyme inhibitor. Anderson, D.B.,
Ferguson, S.M.F., and Lardy, H.A. (1971). FEBS Letters 14, 283.
152 Wolf Prize in Agriculture
S.M., Ebel, R.E., and Lardy, H.A. (1975). Arch. Biochem. Biophys. 171,
656-661.
292. Regulation of intermediary carbohydrate metabolism. Clark, M.G. and
Lardy, H.A. (1975). In Biochemistry of Carbohydrates (Whelan, W.J., ed),
pp. 223-266, University Park Press, Baltimore, MD.
293. Ionophore A23187: The effect of H+ concentration on complex formation
with divalent and monovalent cations and the demonstration of K+ transport
in mitochondria mediated by A23187. Pfeiffer, D.R., and Lardy, H.A. (1976).
Biochemistry 15, 935-943.
294. Essential arginyl residues in mitochondrial adenosine triphosphatase.
Marcus, F., Schuster, S.M., and Lardy, H.A. (1976). J. Biol. Chem. 251,
1775-1780.
295. Effect of epinephrine on the concentration of cGMP in rat liver cells. Bosch,
A., Kneer, N., and Lardy, H.A. (1976). In Use of Isolated Liver Cells and
Kidney Tubules in Metabolic Studies (Tager, J.M., Söling, H.D. & Williamson,
J.R., eds.), pp. 285-290, North-Holland Publ. Co., Amsterdam, The
Netherlands.
296. Interaction of anions and divalent metal ions with phosphoenolpyruvate
carboxykinase. Bentle, L.A. and Lardy, H.A. (1976). J. Biol. Chem. 251,
2916-2921.
297. A protein factor required for activation of phosphoenolpyruvate carboxykinase
by ferrous ions. Bentle, L.A., Snoke, R.E., and Lardy, H.A. (1976). J. Biol.
Chem. 251, 2922-2928.
298. Some effects of ionophore A23187 on energy utilization and the distribution
of cations and anions in mitochondria. Pfeiffer, D.R., Hutson, S.M., Kauffman,
R.F., and Lardy, H.A. (1976). Biochemistry 15, 2690-2697.
299. Action of ionophore A23187 at the cellular level. Separation of effects at
the plasma and mitochondrial membranes. Babcock, D.F., First, N.L., and
Lardy, H.A. (1976). J. Biol. Chem. 251, 3881-3886.
300. Effect of cations and anions on the steady state kinetics of energy-dependent
Ca2+ transport in rat liver mitochondria. Hutson, S.M., Pfeiffer, D.R., and
Lardy, H.A. (1976). J. Biol. Chem. 251, 5251-5258.
301. Activation of bovine epididymal sperm respiration by caffeine. Its transient
nature and relationship to the utilization of acetyl carnitine. Milkowski, A.L.,
Babcock, D.F., and Lardy, H.A. (1976). Arch. Biochem. Biophys. 176,
250-256.
302. Effect of inosine 5'-(β,γ-imido) triphosphate and other nucleotides on beef
heart mitochondrial ATPase. Schuster, S.M., Gertschen, R.J., and Lardy, H.A.
(1976). J. Biol. Chem. 251, 6705-6710.
303. Studies on the kinetic mechanism of oxidative phosphorylation. Schuster,
S.M., Reinhart, G.D., and Lardy, H.A. (1977). J. Biol. Chem. 252, 427-432.
Henry A. Lardy 157
372. The structure of caltrin, the calcium transport structure of caltrin, the calcium
transport inhibitor of bovine inhibitor of bovine seminal plasma. Lewis, R.V.,
San Agustin, J., Kruggel, W., and Lardy, H.A. (1985). Proc. Natl. Acad. Sci.
USA 82, 6490-6491.
373. Catecholamine and vasopressin stimulation of gluconeogenesis from
dihydroxyacetone in the presence of atractyloside. Wernette-Hammond, M.E.,
and Lardy, H.A. (1985). J. Biol. Chem. 260, 12647-12652.
374. Multiple requirements for glycogen synthesis by hepatocytes isolated from
fasted rats. Chen, K. and Lardy, H.A. (1985). J. Biol. Chem. 260, 14683-
14688.
375. A 31P NMR study of the epididym is and epididymal sperm of the bull and
hamster. Smith, M.B., Babcock, D.F., and Lardy, H.A. (1985). Biol. of Reprod.
33, 1029-1041.
376. A half century biochemistry. Lardy, H.A. (1985). In Comprehensive
Biochemistry. Personal Recollections, II, Vol. 36 (Semenza, G., ed),
p. 297-325, Elsevier.
377. The effect of different types of physical exercise on glucose and citrulline
synthesis in isolated rat liver parenchymal cells. Bobyleva-Guarriero, V., and
Lardy, H.A. (1986). FEBS Letters 194, 56-59.
378. The role of malate in exercise-induced enhancement of mitochondrial
respiration. Bobyleva-Guarriero, V. and Lardy, H.A. (1986). Arch. Biochem.
Biophys. 245, 470-476.
379. The role of malate in hormone-induced enhancement of mitochondrial
respiration. Bobyleva-Guarriero, V., Wehbie, R.S., and Lardy, H.A. (1986).
Arch. Biochem. Biophys. 245, 477-482.
380. Methylisobutylxanthine blocks insulin antagonism of cAMP-stimulated
glycogenolysis at a site distinct from phosphodiesterase. Evidence favoring an
insulin-insensitive calcium release mechanism. Gabbay, R.A. and Lardy, H.A.
(1986). J. Biol. Chem. 261, 4002-4007.
381. Enzymatic adaptation to physical training under β-blockade in the rat:
Evidence of a β2-adrenergic mechanism in skeletal muscle. Ji, L.L., Lennon,
D.L.F., Kochan, R.G., Nagle, F.J., and Lardy, H.A. (1986). J. Clin. Invest. 78,
771-778.
382. Stereochemical course of thiophosphoryl transfer catalyzed by cytosolic
phosphoenolpyruvate carboxykinase. Konopka, J.M., Lardy, H.A., and
Frey, P.A. (1986). Biochemistry 25, 5571-5575.
383. Caltrin: A regulator of calcium transport by bovine spermatozoa,
Development, Growth andDifferentiation. Supplement, 1986. Lardy, H.A.,
San Agustin, J., Lewis, R., and Hughes, P. Abst. 48.
384. Chronic exercise training alters kinetic properties of rat skeletal muscle and
myocardial lactate dehydrogenase. Ji, L.L., Stratman, F.W., and Lardy, H.A.
(1986). FEBS 208, 297-300.
Henry A. Lardy 163
412. Bovine seminal plasma constituents modulate the activity of caltrin, the
calcium-transport regulating protein of bovine spermatozoa. San Agustin,
J.T. and Lardy, H.A. (1990). J. Biol. Chem. 265, 6860-6867.
413. Biochemical aspects of obesity. Lardy, H.A. and Shrago, E. (1990). Annu. Rev.
Biochem.59, 689-710.
414. Calcium transport in sperm mitochondria: Effect of substrates and phosphate.
Breitbart, H., Wehbie, R., and Lardy, H.A. (1990). Biochim. Biophys. Acta
1026, 57-63.
415. Regulation of calcium in bovine spermatozoa. Breitbart, H., Wehbie, R., and
Lardy, H.A. (1990). Biochim. Biophys. Acta 1027, 72-78.
416. Stimulation of Ca2+ uptake into epididymal bull spermatozoa by analogues
of amiloride. Breitbart, H., Cragoe, E.J. Jr., and Lardy, H.A. (1990). Eur. J.
Biochem. 192, 529-535.
417. Role of malate in the enhancement of mitochondrial respiration in vitro.
Bobyleva-Guarriero, V., Wehbie, R.S., Muscatello, U., and Lardy, H.A. (1991).
Biokhimiya 56, 542-551.
418. Induction of hepatic mitochondrial glycerophosphate dehydrogenase
in rats by dehydroepiandrosterone. Su, C-Y. and Lardy, H.A. (1991).
J. Biochemistry 110, 207-213.
419. Antioxidant enzyme response to selenium deficiency in rat myocardium.
Ji, L.L., Stratman, F.W., and Lardy, H.A. (1992). J. Am. College of Nutrition
11, 79-86.
420. Bovine spermatozoa incorporate 32Pi into ADP by an unknown pathway.
Cheetham, J. and Lardy, H.A. (1992). J. Biol. Chem. 267, 5186-5192.
421. Changes in liver structure and function after short-term and long-term
treatment of rats with dehydroepiandrosterone. Bellei, M., Battelli, D., Fornieri,
C., Mori, G., Muscatello, U., Lardy, H.A., and Bobyleva-Guarriero, V. (1992).
J. Nutrition 122, 967-976.
422. Functional properties of caltrin proteins from seminal vesicle of the
guinea pig. Coronel, C. and Lardy, H.A. (1992). Molecular Reprod. and
Development 33, 74-80.
423. Purification, structure and characterization of caltrin proteins from seminal
vesicle of the rat and mouse. Coronel, C.E., Winnica, D.E., Novella, M.L., and
Lardy, H.A. (1992). J. Biol. Chem. 267, 20909-20915.
424. Isolation and characterization of a 54-kilodalton precursor of caltrin, the
calcium transport inhibitor protein from seminal vesicles of the rat. Coronel,
C.E., Novella, M.L., Winnica, D.E., and Lardy, H.A. (1993). Biol. Repro. 48,
1326-1333.
425. Comparative studies of effects of dehydroepiandrosterone on rat and chicken
liver. Bobyleva, V., Kneer, N., Bellei, M., Battelli, D., Muscatello, U., and Lardy,
H.A. (1993). Comp. Biochem. Physiol. 105B, 643-647.
166 Wolf Prize in Agriculture
1910–1994
While working at Cornell University and the University of Illinois, Salisbury did
extensive research on semen extendors, number of sperm per insemination, and
insemination techniques. His contributions have been pivotal in the resolution of
problems limiting successful reproduction following artificial insemination. He is
known for his ability to stimulate thinking and interest in his field and for his bold
scientific approach to the problems facing the animal industry.
171
172 Wolf Prize in Agriculture
has been highly instrumental in leading and developing groups that have made
significant contributions to agricultural industry. His service to agriculture cannot
be measured by just his awn accomplishments but must include the influence of
all who have been associated with him.
ESSENTIAL BIOGRAPHY:
Glenn Wade Salisbury was born at Sheffield, Ohio, in 1910. He received the
B. S. degree in agriculture from Ohio State University in 1931 and the Ph.D. degree
from Cornell University in 1934. He served as graduate assistant, instructor, assistant
professor, associate professor, and professor at Cornell University from 1931 to
1947. He served as Professor and Head, Department of Dairy Science, University
of Illinois, fran 1947 to 1969 and then became Associate Dean, and Director of
the Illinois Agricultural Experiment Station until his retirement in 1978. He is now
Professor Emeritus. He has had an outstanding career in research, teaching and
public service. He has been a prolific writer with over 260 scientific and popular
articles published.
CITATION
To Professor Glenn W. Salisbury
In appreciation for 22 years of leadership as Head of the Department of Dairy
Science at the University of Illinois Urbana-Champaign Campus.
His research accomplishments and foreign travels have given him an
international reputation as an extraordinary scientist, scholar, and teacher. His
abilities and dedication to serve have resulted in his appointment to numerous
local, national, and international committees, as well as to his new position. These
services have brought recognition to the department and to the university. His
initiative and drive, and his broad experience and understanding have served as a
stimulus for conducting and coordinating basic and applied research, and extension
activities of the department. His example has been an inspiration to his colleagues.
He is warm and considerate on a professional and personal basis. We wish to
acknowledge his contributions to the department and consider it a privilege to
have had this association with him.
PROFESSORIAL STAFF
DEPARTMENT OF DAIRY SCIENCE
UNIVERSITY OF ILLINOIS
JUNE 11, 1969
Wendell L. Roelofs
New York State Agricultural Experiment Station,
College of Agriculture and Life Sciences,
Cornell University, Geneva, New York, USA
k
173
174 Wolf Prize in Agriculture
CURRICULUM VITAE
Education
1964 - Ph.D., Indiana University
1960 - B.S., Central College
Professional Experience
1962-1964 - NIH Predoctoral Fellow
1964 - NIH Postdoctoral Fellow, Massachusetts Institute of Technology
ACADEMIC RESPONSIBILITIES
Research Responsibilities
• Current Postdoctoral Associates (list names)
Paul Robbins
• Past Postdoctoral Associates
Bingye Xue 2003-2006
Grace Hao, 2000-2002
Satoshi Nojima, 2000-2003
Bruce Morris, North Dakota St. 2000-2001
Aijun Zhang, USDA, Beltsville, MD 1997-2000
Weitian Liu, Univ. South Dakato Medical School 1998-2002
Nancy Murray, Hobart and William Smith Colleges 2000-2001
Prior to 5 years ago from 1969:
Thomas Baker, Jan Kochansky, Jim Miller, Ring Carde´, Ashok Tamhankar
Lou Bjostad, Henry Arn, Jim Tette, Ada Hill, Walter Wolf, Ritzuo Nishida,
Boris Kovalev, Jia-Wei Du, Stephen Foster, Ralph Charlton, Stuart Krasnoff,
Russell Jurenka, Peter Ma,
• Other Current Research Professionals Supervised (list names)
Charles Linn, Senior Research Associate
176 Wolf Prize in Agriculture
Professional Honoraries
1985 Honorary Doctor of Science, Central College
1988 Honorary Doctor of Science, Hobart & William Smith Colleges, Geneva, NY
1988 Honorary Doctor of Science, Indiana University
1989 Honorary Doctor of Philosophy, University of Lund, Sweden
1989 Honorary Doctor of Science, Free University of Brussels, Belgium
Editorial Boards
Journal of Insect Physiology
Insect Biochemistry and Molecular Biology
Journal of Chemical Ecology
Committee Assignments
• International/National:
2000-03 Secretary, Class VI, National Academy of Sciences
1999 Chairman, Section B, National Entomology Society of America
1998 Vice Chairman, National Entomology Society of America
1997 Secretary, National Entomology Society of America
• University:
2005 Chairman, CALS Research and Extension Awards Committee
2003 Geneva Director Search Committee
2000-present Chairman, Facility Use Committee at Geneva
Wendell L. Roelofs 177
LIST OF PUBLICATIONS
1. Campaigne, E., D. R. Maulding, and W. L. Roelofs. 1964. Ring closure of
ylidenemalononitriles. III. Formation of six-membered rings and related
chemistry. J. Org. Chem. 29: 1543-1549.
2. Campaigne, E. and W. L. Roelofs. 1965. Ring closure of ylidenemalononitriles.
IV. Attempted cyclizations of saturated malononitrile derivatives. J. Org. Chem.
30: 396-400.
3. Campaigne, E. and W. L. Roelofs. 1965. Reductions of 2-carboxamidotetra-
hydroacenaphthenone derivatives. J. Org. Chem. 30: 2610-2614.
4. Campaigne, E., W. L. Roelofs, and R. F. Weddleton. 1966. Succinimido
[3,4-b]-3a,4,5,6-tetrahydroacenaphthen-10-ones. U. S. Pat. 3, 227,729,
Jan. 4.
178 Wolf Prize in Agriculture
81. Cardé, R. T., T. C. Baker, and W. L. Roelofs. 1975. Behavioral role of individual
components of a multichemical attractant system in the oriental fruit moth.
Nature 253: 348-349.
82. Roelofs, W. L. 1975. Manipulating sex pheromones for insect suppression.
Environ. Lett. 8: 41-59.
83. Liebherr, J. and W. L. Roelofs. 1975. Laboratory hybridization and mating
period studies using two pheromone strains of Ostrinia nubilalis. Ann.
Entomol. Soc. Amer. 68: 305-309.
84. Hill, A. and W. L. Roelofs. 1975. Sex pheromone components of the
omnivorous leafroller moth, Platynota stultana. J. Chem. Ecol. 1: 91-99.
85. Roelofs, W. L. 1975. Insect communication — chemical. In: “Insects, Science
and Society,” D. Pimentel [ed.] Academic Press, NY: 79-99.
86. Cardé, R. T., K. Trammel, and W. L. Roelofs. 1975. Disruption of sex attraction
of the redbanded leafroller (Argyrotaenia velutinana) with microencapsulated
pheromone components. Environ. Entomol. 4: 448-450.
87. Roelofs, W. L., J. P. Kochansky, R. T. Cardé, G. G. Kennedy, C. A. Henrick,
J. N. Labovitz, and V. L. Corbin. 1975. Sex pheromone of the potato tuberworm
moth, Phthorimaea operculella. Life Sciences 17: 699-706.
88. Roelofs, W. L., J. Kochansky, E. Anthon, R. Rice, and R. Cardé. 1975. Sex
pheromone of the peach twig borer moth (Anarsia lineatella). Environ.
Entomol. 4(6): 580-582.
89. Kochansky, J., J. Tette, E. F. Taschenberg, R. T. Cardé, K. E. Kaissling, and W. L.
Roelofs. 1975. Sex pheromone of the moth, Antheraea polyphemus. J. Insect
Physiol. 21: 1977-1983.
90. Cardé, R. T., T. C. Baker, and W. L. Roelofs. 1975. Ethological function of
components of a sex-attractant system for oriental fruit moth males,
Grapholitha molesta (Lepidoptera: Tortricidae). J. Chem. Ecol. 1: 475-491.
91. Roelofs, W. L., A. S. Hill, T. C. Baker, and R. T. Cardé. 1975. Novel attractant
components for males of the tobacco budworm moth. U. S. Pat. 3,917,711,
Nov. 4.
92. Roelofs, W. L., J. R. Miller, and T. C. Baker. 1976. Pheromones of lepidopterous
insects. In: “Perspectives of Forest Pest Management,” Anderson and Kaya
[eds.] Academic Press, NY.
93. Roelofs, W. L., A. S. Hill, and T. C. Baker. 1976. Novel attractant components
for males of the tobacco budworm moth. U. S. Pat. 3,952,093, Apr. 20.
94. Roelofs, W. L., R. Cardé, E. F. Taschenberg, and R. W. Weires, Jr. 1976.
Pheromone research for control of lepidopterous species in New York. In:
Beroza, M. ed., ACS Symp. Ser. 23: 75-87.
95. Miller, J. R., T. C. Baker, R. T. Cardé, and W. L. Roelofs. 1976. Reinvestigation
of oak leafroller sex pheromone components and the hypothesis that they
vary with diet. Science 192: 140-143.
184 Wolf Prize in Agriculture
96. Roelofs, W. L., A. Hill, A. Cardé, R. Cardé, H. Madsen, and J. Vakenti. 1976.
Sex pheromone of the European leafroller, Archips rosanus. Environ. Entomol.
5: 362-364.
97. Roelofs, W. L. 1976. The importance of defining lepidopteran pheromone
blends. NYS Agr. Exp. Sta., Geneva. Search Agriculture 6: 1-4.
98. Roelofs, W. L. 1976. Pheromones. In: “The Future for Insecticides: Needs and
Prospects,” R. L. Metcalf and J. J. McKelvey, Jr. [eds.] Wiley & Sons, Inc., NY,
pp. 445-466.
99. Roelofs, W. L., A. Cardé, A. Hill, and R. Cardé. 1976. Sex pheromone of the
threelined leafroller. Environ. Entomol. 5: 649-652.
100. Taschenberg, E. F. and W. L. Roelofs. 1976. Pheromone communication
disruption of the grape berry moth with microencapsulated and hollow
fiber systems. Environ. Entomol. 5: 688-691.
101. Baker, T. C., R. T. Cardé, and W. L. Roelofs. 1976. Behavioral responses of
male Argyrotaenia velutinana (Lepidoptera: Tortricidae) to components of
its sex pheromone. J. Chem. Ecol. 2: 333-352.
102. Comeau, A., R. T. Cardé, and W. L. Roelofs. 1976. Relationship of ambient
temperature to diel periodicities of sex attraction in six species of Lepidoptera.
Can. Entomol. 108: 415-418.
103. Roelofs, W. L. 1976. Pheromone chemistry: Overview and discussion. In:
“Chemical Control of Insect Behavior: Theory and Application,” J. J. McKelvey,
Jr. and H. Shorey [eds.] Wiley & Sons, NY: 287-297.
104. Baker, T. C. and W. L. Roelofs. 1976. Electroantennogram responses of the
male moth Argyrotaenia velutinana to mixtures of sex pheromone components
of the female. J. Insect Physiol. 22: 1357-1364.
105. Roelofs, W. L. and R. T. Cardé. 1976. Responses of Lepidoptera to synthetic
sex pheromone chemicals and their analogs. Ann. Rev. Entomol. 22:
377-399.
106. Roelofs, W. L. 1976. Sex pheromones, hormones and trapping. In: Nat’l
Acad. Sci. Report. Insect Control in China.
107. Arn, H., S. Rauscher, H-R. Buser, and W. L. Roelofs. 1976. Sex pheromone of
Eupoecilia ambiguella: cis-9-dodecenyl acetate as a major component.
Z. Naturforsch. 31: 499-503.
108. Cardé, R. T., T. C. Baker, and W. L. Roelofs. 1976. Sex attractant responses of
male oriental fruit moths to a range of component ratios-pheromone
polymorphism? Experientia 32: 1406-1407.
109. Doolittle, R. E., W. L. Roelofs, J. D. Solomon, R. T. Cardé, and M. Beroza. 1976.
(Z,E)-3,5-tetradecadien-1-ol acetate sex attractant for the carpenterworm
moth, Prionoxystus robiniae (Peck) (Lepidoptera: Cossidae). J. Chem. Ecol. 2:
399-410.
Wendell L. Roelofs 185
138. Fein, B. L., W. H. Reissig, and W. L. Roelofs. 1978. Attraction of apple maggot
Rhagoletis pomonella (Walsh) (Diptera: Tephritidae) females to apple volatiles
in wind tunnel bioassays. J.N.Y. Entomol. Soc. 4: 286.
139. Hill, A. S. and W. L. Roelofs. 1979. Sex pheromone components of the
obliquebanded leafroller, Choristoneura rosaceana. J. Chem. Ecol. 5: 3-11.
140. Gieselmann, M., D. J. Moreno, J. Fargerlund, H. Tashiro, and W. L. Roelofs.
1979. Identification of the sex pheromone of the yellow scale. J. Chem. Ecol.
5: 25-31.
141. Tashiro, H., M. J. Gieselmann, and W. L. Roelofs. 1979. Residual activity of a
California red scale synthetic pheromone component. Environ. Entomol. 8:
931-934.
142. Hill, A. S., R. W. Rings, S. R. Swier, and W. L. Roelofs. 1979. Sex
pheromone of the black cutworm moth, Agrotis ipsilon. J. Chem. Ecol. 5:
439-457.
143. Gieselmann, M. J., R. E. Rice, R. A. Jones, and W. L. Roelofs. 1979. Sex
pheromone of the San Jose scale. J. Chem. Ecol. 5: 891-900.
144. Roelofs, W. L. 1979. Pheromone perception in Lepidoptera. In:
“Neurotoxicology of Insecticides and Pheromones,” T. Narahashi, [ed.], Plenum
Press, NY, pp. 5-25.
145. Roelofs, W. L. 1979. Production and perception of lepidopterous pheromone
blends. In: “Chemical Ecology: Odour Communication in Animals,” F. J. Ritter
[ed.] Elsevier Press, NY, Pages 159-168, 427 pp.
146. Roelofs, W. L., A. S. Hill, and C. W. Berisford. 1979. Sex pheromone of
the subtropical pine tip moth, Rhyacionia subtropica. Environ. Entomol. 8:
894-895.
147. Roelofs, W. L. 1979. Electroantennograms. Chemtech. 9: 222-227.
148. Roelofs, W. L. 1979. Establishing efficacy of sex attractants and disruptants
for insect control. Entomol. Soc. Amer., 97 pp.
149. Roelofs, W. L. 1979. Pheromones and other chemical attractants: pheromone
trap specificity and potency. In: “Movement of Highly Mobile Insects: Concepts
and Methodology in Research,” R. L. Rabb and G. G. Kennedy [eds.] University
Graphica, North Carolina State University, Raleigh, NC, pp. 272-285.
150. Gieselmann, M. J., C. A. Henrick, R. J. Anderson, D. S. Moreno, and W. L.
Roelofs. 1980. Responses of male California red scale to sex pheromone
isomers. J. Insect Physiol. 26: 179-183.
151. Svirskaya, P. I., C. C. Leznoff, and W. L. Roelofs. 1980. A stereoselective
synthesis of a cis,cis conjugated dienol, a candidate pheromone. Syn. Comm.
10: 391-397.
152. Roelofs, W. L., A. J. Tamhankar, A. Comeau, A. S. Hill, and E. F. Taschenberg.
1980. Moth activity periods for uglynest caterpillar, Archips cerasivoranus
and identification of its sex pheromone. Ann. Entomol. Soc. Amer. 73:
631-634.
188 Wolf Prize in Agriculture
166. Baker, T. C., W. Meyer, and W. L. Roelofs. 1981. Sex pheromone dosage
and blend discrimination by Oriental fruit moth males. Ent. exp. appl. 30:
269-279.
167. Linn, C. E. and W. L. Roelofs. 1981. Modification of sex pheromone blend
discrimination in male Oriental fruit moths by pre-exposure to (E)-8-
dodecenyl acetate. Physiol. Entomol. 6: 421-429.
168. Suguro, T., W. L. Roelofs, and K. Mori. 1981. Stereoselective synthesis of
(+)-(E)-3,9-dimethyl-1-isopropyl-5,8-decadienyl acetate, the racemate of
the yellow scale pheromone, and its (Z)-isomer. Agric. Biol. Chem. 45:
2509-2514.
169. Bjostad, L. B., E. Gurevitz, S. Gothilf, and W. L. Roelofs. 1981. Sex attractant
for the honeydew moth, Cryptoblabes gridiella. Phytoparasi. 9: 95-99.
170. Roelofs, W. L. 1981. ESA Founders’ Memorial Award Lecture: Pheromones,
Plateaus and Platitudes. Bull. Entomol. Soc. Amer. 27: 3-7.
171. Hill, A. S. and W. L. Roelofs. 1981. Sex pheromone of the saltmarsh caterpillar
moth, Estigmene acrea. J. Chem. Ecol. 7: 655-668.
172. Clement, S. L., A. S. Hill, E. Levine, and W. L. Roelofs. 1981. Trap catches of
male Agrotis ipsilon with synthetic sex pheromone emitted for different
dispensers. Environ. Entomol. 10: 521-523.
173. Bjostad, L. B. and W. L. Roelofs. 1981. Sex pheromone biosynthesis from
radiolabeled fatty acids in the redbanded leafroller moth. J. Biol. Chem. 256:
7936-7940.
174. Anderson, R. J., M. J. Gieselmann, H. R. Chinn, K. G. Adams, C. A. Henrick,
R. E. Rice, and W. L. Roelofs. 1981. Synthesis of identification of a third
component of the San Jose scale sex pheromone. J. Chem. Ecol. 7: 695-706.
175. Baker, T. C., R. Nishida, and W. L. Roelofs. 1981. Close-range attraction of
female Oriental fruit moths to herbal scent of male hairpencils. Science 214:
1359-1361.
176. Hill, A. S., B. G. Kovalev, L. N. Nikolaeva, and W. L. Roelofs. 1982. Sex
pheromone of the fall webworm, Hyphantria cunea. J. Chem. Ecol. 8:
383-396.
177. Nishida, R., T. C. Baker, and W. L. Roelofs. 1982. Hairpencil pheromone
components of male Oriental fruit moths, Grapholitha molesta. J. Chem.
Ecol. 8: 947-959.
178. Fein, B. L., W. H. Reissig, and W. L. Roelofs. 1982. Identification of apple
volatiles attractive to apple maggot flies, Rhagoletis pomonella. J. Chem. Ecol.
8: 1473-1487.
179. Meyer, W. L., G. L. DeBarr, C. W. Berisford, L. R. Barber, and W. L. Roelofs.
1982. Identification of the sex pheromone of the webbing coneworm
moth, Dioryctria disclusa (Lepidoptera: Pyralidae). Environ. Entomol. 11:
986-988.
190 Wolf Prize in Agriculture
206. Linn, C. E., Jr. and W. L. Roelofs. 1985. Response specificity of male pink
bollworm moths to different blends and dosages of sex pheromone. J. Chem.
Ecol. 11: 1583-1590.
207. Reissig, W. H., B. H. Stanley, W. L. Roelofs, and M. R. Schwarz. 1985. Tests of
synthetic apple volatiles in traps as attractants for apple maggot flies (Diptera:
Tephritidae) in commercial apple orchards. Environ. Entomol. 14: 55-59.
208. Löfstedt, C. and W. L. Roelofs. 1985. Sex pheromone precursors in two
primitive New Zealand tortricid moth species. Insect Biochem. 15: 729-734.
209. Glass, E. H. and W. L. Roelofs. 1985. Argyrotaenia velutinana. In: “Handbook
of Insect Rearing,” Vol. 2, Elsevier Science Publishers, BV Amsterdam.
pp. 197-205.
210. Linn, C. E., Jr. and W. L. Roelofs. 1986. Neuropharmacological effects on
olfactory perception. In: “Mechanisms of Insect Olfaction.” T. L. Payne,
M. C. Birch, and C. E. J. Kennedy [Eds.] Oxford Univ. Press, pp. 209-215.
211. Linn, C. E., Jr., M. G. Campbell and W. L. Roelofs. 1986. Male moth sensitivity
to multicomponent pheromones: The critical role of the female released
blend in determining the functional role of components and the active space
of the pheromone. J. Chem. Ecol. 12: 659-668.
212. Foster, S. P., J. R. Clearwater, S. J. Muggleston and W. L. Roelofs. 1986.
Probable sibling species complexes within two described New Zealand
leafroller moths. Naturwissenschaften 73: 156-158.
213. Wolf, W. A. and W. L. Roelofs. 1986. Properties of the “11-desaturase enzyme
used in cabbage looper moth sex pheromone biosynthesis. Arch. Insect
Biochem. & Physiol. 3: 45-52.
214. Bjostad, L. B. and W. L. Roelofs. 1986. Sex pheromone biosynthesis in the
redbanded leafroller moth, studied by mass-labeling with stable isotopes and
analysis with mass spectrometry. J. Chem. Ecol. 12(2): 431-450.
215. Linn, C. E., Jr. and W. L. Roelofs. 1986. Modulatory effects of octopamine
and serotonin on thresholds for male sensitivity and periodicity of response
to sex pheromone in the cabbage looper, Trichoplusia ni. Arch. Insect Biochem.
Physiol. 3: 161-172.
216. Meyer, W. L., G. L. DeBarr, J. L. Hanula, B. Kovalev, R. S. Cameron, C. W.
Berisford, and W. L. Roelofs. 1986. Z-11-Hexadecenyl acetate, a sex
pheromone component for the southern pine coneworm, Dioryctria amatella
(Lepidoptera: Pyralidae). Environ. Entomol. 15: 316-320.
217. Roelofs, W. L., J. W. Du, C. E. Linn, Jr., T. J. Glover, and L. B. Bjostad. 1987.
The potential for genetic manipulation of the redbanded leafroller moth sex
pheromone gland. In: “Evolutionary Genetics of Invertebrate Behavior,”
M. D. Huettel [Ed.] Plenum Pub. Corp., New York, pp. 263-272.
218. Glover, T. J., X.-H. Tang, and W. L. Roelofs. 1987. Sex pheromone blend
discrimination by male moths from E and Z strains of European corn borer.
J. Chem. Ecol. 13: 143-151.
Wendell L. Roelofs 193
246. Sreng, I., T. J. Glover, and W. L. Roelofs. 1989. Canalization of the redbanded
leafroller moth sex pheromone blend. Arch. Insect. Biochem. Physiol. 10:
73-82.
247. Wolf, W. A. and W. L. Roelofs. 1989. Enzymes involved in the biosynthesis of
sex pheromones in moths. In: “Biocatalysis in Agricultural Biotechnology,”
J. R. Whitaker and P. E. Sonnet [Eds.], American Chemical Society Symposium
Series, pp. 323-331.
248. Jurenka, R. A. and W. L. Roelofs. 1989. Characterization of the
acetyltransferase used in pheromone biosynthesis in moths: specificity for
the Z isomer in Tortricidae. Insect Biochem. 19(7): 639-644.
249. Löfstedt, C., N. J. Vickers, W. L. Roelofs, and T. C. Baker. 1989. Diet related
courtship success in the Oriental fruit moth, Grapholita molesta (Tortricidae).
Oikas 55: 402-408.
250. Rule, G. S. and W. L. Roelofs. 1989. Biosynthesis of sex pheromone components
from linolenic acid in arctiid moths. Arch. Insect Biochem. Physiol. 12: 89-97.
251. Löfstedt, C., B. S. Hansson, W. L. Roelofs, and B. O. Bengtsson. 1989. The
linkage relationship between factors controlling female pheromone production
and male pheromone response in the European corn borer, Ostrinia nubilalis.
Genetics 123: 553-556.
252. Foster, S. P. and W. L. Roelofs. 1990. Biosynthesis of a monene and a conjugated
diene sex pheromone component of the light brown apple moth by
E11-desaturation. Experientia 46: 269-273.
253. Glover, T. J., M. G. Campbell, P. S. Robbins, and W. L. Roelofs. 1990. Sex-
linked control of sex pheromone behavioral responses in European corn
borer moths (Ostrinia nubilalis) confirmed with TPI marker gene. Arch.
Insect Biochem. Physiol. 15: 67-77.
254. Krasnoff, S. B. and W. L. Roelofs. 1990. Evolutionary trends in the male
pheromone systems of arctiid moths: Evidence from studies of courtship in
Phragmatobia fuliginosa and Pyrrharctia isabella (Lepidoptera: Arctiidae).
Zool. J. Linn Soc. 99: 319-338.
255. Dennehy, T. J., W. L. Roelofs, E. F. Taschenberg, and T. N. Taft. 1990. Mating
disruption for control of grape berry moth in New York vineyards. In:
“Behavior-Modifying Chemicals for Insect Management,” R. Ridgeway, R. M.
Silverstein, and M. Insco (Eds.), Marcel Dekker, Inc., New York. pp 223-240.
256. Tang, J. D., W. L. Roelofs, and D. C. Knipple. 1990. Construction of a cDNA
library from cabbage looper moth sex pheromone glands. In: “Molecular
Insect Science,” H. Hagedorn, J. Hildebrand, M. Kidwell, and J. Law, [Eds.],
Plenum, New York. [Abstract]
257. Jurenka, R. A., G. Fabrias, and W. L. Roelofs. 1991. Hormonal control of
female sex pheromone biosynthesis in the redbanded leafroller moth,
Argyrotaenia velutinana. Insect Biochem. 19: 639-644.
196 Wolf Prize in Agriculture
319. Facundo, H. T., Linn Jr., C.. E., Villani, M. G., and Roelofs, W. L. 1999.
Emergence, mating, and postmating behaviors of the Oriental beetle
(Coleoptera: Scarabaeidae). J. Insect Behav. 12: 175-192.
320. Liu, W., Ma, P. W. K., Marsella-Herrick, P., Rosenfield, C.-L., Knipple,
D. C., Roelofs, W. 1999. Cloning and functional expression of a cDNA
encoding a metabolic acyl-CoA “9-desaturase of the cabbage looper moth,
Trichoplusia ni. Insect Biochem. Molec. Biol. 29: 435-443.
321. Linn, Jr., C. E., Poole, K., Zhang, A., and Roelofs, W. 1999. Pheromone-blend
discrimination by European corn borer moths with inter-race and inter-sex
antennal transplants. J. Comp. Physiol. A 184: 273-278.
322. Zhang, A., Linn, Jr., C., Wright, S., Prokopy, R., Reissig, W., Roelofs, W. 1999.
Identification of a new blend of apple volatiles attractive to the apple maggot
Rhagoletis pomonella. J. Chem. Ecol. 25: 1221-1232.
323. Alm, Steven R., Villani, M. G., and Roelofs, W. 1999. Oriental beetles
(Coleoptera: Scarabaeidae): Current distribution in the United States and
optimization of monitoring traps. J. Econ. Entomol. 92: 931-935.
324. Ma, P. W. K., Garden, R., Niermann, J. T., O’Connor, M., Sweedler, J. V., and
Roelofs, W. L. 2000. Characterizing the Hez-PBAN gene products in neuronal
clusters with immunocytochemistry and MALDI MS. J. Insect Physiol. 46:
221-230.
325. Dalleerac, R., Labeur, C., Jallon, J.-M., Knipple D. C., Roelofs, W., Wicker-
Thomas, C. 2000. A ∆9 desaturase gene with a different substrate specificity
is responsible for the cuticular diene hydrocarbon polymorphism in
Drosophila melanogaster. Proc. Natl. Acad. Sci. USA 97: 9449-9454.
326. Rosenfield, C-L., You, K-M., Marsella-Herrick, P., Roelofs, W. L., and Knipple,
D. C. 2001. Structural and functional conservation and divergence among
acyl-CoA desaturase-encoding genes of two noctuid species, the corn
earworm, Helicoverpa zea, and the cabbage looper, Trichoplusia ni. Insect
Biochem. Molec. Biol. 31: 949-964.
327. Liu, W., Jiao, H., O’Connor, M. and Roelofs, W. L. 2002. Cloning and
expression of a moth desaturase that specifically produces a mixture of
Z/E11-tetradecenoic acids. Biochemistry 32: 1489-1495.
328. Liu, W., Jiao, H., Murray, N., O’Connor, M., and Roelofs, W. L. 2002. Gene
characterized for membrane desaturase that produces (E)-11 isomers
of mono- and diunsaturated fatty acids. Proc. Natl. Acad. Sci. USA 99:
620-624.
329. Hao, G., Liu, W., O’Connor, M., and Roelofs, W. L. 2002. Characterization
and functional expression of cDNAs encoding acyl-CoA Z9- and Z10-
desaturase of Planotortrix octo. Insect Biochem. Molec. Biol. 32: 961-966.
330. Ochieng, S. A., Robbins, P. S., Roelofs, W. L., and Baker, T. C. 2002. Pheromone
reception in the scarab beetle, Phyllophaga anxia (Coleoptera: Scarabaeidae).
Ann. Entomol. Soc. Amer. 95: 97-102.
202 Wolf Prize in Agriculture
331. Ma, W. K. P. and Roelofs, W. L. 2002. Sex pheromone gland of the female
European corn borer moth, Ostrinia nubilalis (Lepidoptera, Pyralidae):
ultrastructural and biochemical evidences. Zool. Sci. 19: 501-511.
332. Roelofs, W. L., Liu, W., Hao, G., Jiao, H., Rooney, A. P., and Linn, Jr., C. E.
2002. Evolution of moth sex pheromones via ancestral genes. Proc. Natl.
Acad. Sci. USA 99: 13621-13626.
333. Hao, G., O’Connor, M., Liu, W., Roelofs, W. L. 2002. Characterization of
Z/E11- and Z9-desaturases from the obliquebanded leafroller moth,
Choristoneura rosaceana J. Insect Sci. 2:26 online: insectscience.org/2.26.
334. Ochieng, S. A., Poole, K., Linn, Jr., C., Vickers, N., Roelofs, W. L.,
Baker, T. C. 2002. Unusual pheromone receptor neuron responses in
heliothine moth antennae derived from inter-species imaginal disc
transplantation. J. Comp. Physiol. A 189: 19-28.
335. El-Sayed, A.M., Delisle, J., De Lury, N., Gut, L. J., Judd, G. J. R., Legrand, S.,
Reissig, W. H., Roelofs, W. L., Unelius, C. R., and Trimble, R. M. 2003.
Geographic variation in pheromone chemistry, antennal electrophysiology,
and pheromone-mediated trap catch of North American populations of the
obliquebanded leafroller. Environ. Entomol. 32; 470-476.
336. Nojima, S., Sakata, T., Yoshimura, K., Robbins, P. S., Morris, B. D., and
Roelofs, W. L. 2003. Male-specific EAD active compounds produced by
female European chafer, Rizotrogus majalis (Razoumowshy) (Coleoptera:
Scarabaeidae, Melolonthinae). J. Chem. Ecol. 29: 503-507.
337. Nojima, S., Linn, Jr., C., Morris, B. D., Zhang, A. and Roelofs, W. L.
2003. Identification of host fruit volatiles from hawthorn (Crataegus spp.)
attractive to hawthorn-orign Rhagoletis pomonella flies. J. Chem. Ecol. 29:
321-336.
338. Linn, Jr., C., O’Connor, M., Roelofs, W. 2003. Silent genes and rare males: A
fresh look at pheromone blend response specificity in the European corn
borer moth, Ostrinia nubilalis. Journal of Insect Science, 3:15, Available
online: insectscience.org/3.15.
339. Zhang, A., Robbins, P. S., Averill, A. L., Weber, D. C., Linn, C. E., Jr.,
Roelofs, W. L., Villani, M. G. 2003. Identification of the female-produced
sex pheromone of the scarab beetle, Hoplia equina. J. Chem. Ecol. 29:
1635-1642.
340. Roelofs, W. L. and Rooney, A. P. 2003. Molecular genetics and evolution
of pheromone biosynthesis in Lepidoptera. Proc. Natl. Acad. Sci. USA 100:
9179-9184.
341. Linn, Jr., C., Feder, J. L., Nojima, S., Dambroski, H. R., Berlocher, S. H.,
Roelofs, W. L. 2003. Fruit odor discrimination and sympatric host race
formation in Rhagoletis. Proc. Natl. Acad. Sci. USA 100: 11490-11493.
Wendell L. Roelofs 203
342. Robbins, P. S., Crocker, R. L., Nojima, S., Morris, B. D., Roelofs, W. L.,
Villani, M. G. 2003. Methyl 2-(methylthio)benzoate: the unique sulfur-
containing sex pheromone of Phyllophaga crinita. Naturwissenschaften 90:
517-520.
343. Nojima, S., Robbins, P. S., Salsbury, G. A., Morris, B. D. Roelofs, W. L.,
Villani, M. G. 2003. L-Leucine methyl ester: the female-produced sex
pheromone of the scarab beetle, Phyllophaga lanceolata. J. Chem. Ecol. 29:
2439-2446.
344. Knipple, D. C. and W. L. Roelofs. 2003. Molecular biochemical investigations
of pheromone desaturases. In: “Insect Pheromone Biochemistry and Molecular
Biology,” G. J. Blomquist and R. G. Vogt [Eds.]. Elsevier Academic Press, New
York, pp. 81-106.
345. Nojima, S., C. Linn Jr., and W. Roelofs. 2003. Identification of host fruit
volatiles from flowering dogwood (Cornus florida) attractive to dogwood-
origin Rhagoletis pomonella flies. J. Chem. Ecol. 29: 2347-2357.
346. Linn, Jr., C. E., Dambroski, H. R., Feder, J. L., Berlocher, S. H., Nojima, S.,
Roelofs, W. L. 2004. Postzygotic isolating factor in sympatric speciation in
Rhagoletis flies: Reduced response of hybrids to parental host-fruit odors.
Proc. Natl. Aca. Sci. USA 10: 17753-17758.
347. Nojima, S., Kiemle, D. J., Webster, F. X., Roelofs, W. L. 2004. Submicro
scale NMR sample preparation for volatile chemicals. J. Chem. Ecol. 30:
2153-2161.
348. Rodriguez, S., Hao, G., Liu, W., Pina, B. Rooney, A., Camps, F., Roelofs, W.,
Fabrias, G. 2004. Expression and evolution of delta-9 and delta-11 desaturase
genes in the moth Spodooptera littoralis. Insect Biochem. Molec. Biol. 34:
1315-1328.
349. Liu, W., Rooney, A. P., Xue, B., Roelofs, W. L. 2004. Desaturases form the
spotted fireworm moth (Choristoneura parallela) shed light on the
evolutionary origins of novel moth sex pheromone desaturases. Gene 342:
303-311.
350. Nojima, S., Kiemle, D. J., Webster, F. X., Roelofs, W. L. 2004. Submicro
scale NMR sample preparation for volatile chemicals. J. Chem. Soc. 30:
2147-2155.
351. Linn, C., Jr., Nojima, S., Roelofs, W. 2005. Antagonist effects of non-host
fruit volatiles on discrimination of host fruit by Rhagoletis flies infesting
apple (Malus pumila), hawthorn (Crataegus spp.), and flowering dogwood
(Cornus florida). Entomol. Exp. Appl. 114: 97-105.
352. Nojima, S., Schal, C., Webster, F. X., Santangelo, R. G., Roelofs, W. L. 2005.
Identification of the sex pheromone of the German cockroach, Blattella
germanica. Science 307: 1104-1106.
204 Wolf Prize in Agriculture
353. Burand JP, Tan W, Kim W, Nojima S, Roelofs W. 2005. Infection with the
insect virus Hz-2v alters mating behavior and pheromone production in
female Helicoverpa zea moths. 6pp. Journal of Insect Science 5:6, Available
online: insectscience.org/5.6
354. Heath, Jeremy J. , Zhang, Aijun, Roelofs, Wendell L., Smith, Robert F. 2005.
Flight activity and further evidence for a female-produced sex pheromone of
the apple leaf midge, Dasineura mali , in Nova Scotia. Northeastern Naturalist
12 (1), 93-102.
355. Linn, Jr., C. E. Dambroski, Hattie, Nojima, Satoshi, Feder, Jeffrey L., Berlocher,
Stewart H., Roelofs, Wendell L. 2005. Variability in response specificity of
apple, hawthorn, and flowering dogwood-infesting Rhagoletis flies to host
fruit volatile blends: Implications for sympatric host shifts. Entomol. Exp.
Appl. 116: 55-64.
356. Dambroski, H. R., Linn, Jr., C. E., Berlocher, S. H., Forbes, A., Roelofs, W.,
Feder, J. L. 2005. The genetic basis for fruit odor discrimination in Rhagoletic
flies and its significance for sympatric host shifts. Evolution 59: 1953-1964.
357. Olsson, S. B., Linn, Jr., C. E., Roelofs, W. L. 2006. The chemosensory basis for
behavioral divergence involved in sympatric host shifts II: olfactory receptor
neuron sensitivity and temporal firing pattern to individual key host volatiles.
J. Comp. Physiol. A. 192: 289-300.
358. Olsson, S. B., Linn, Jr., C. E., Roelofs, W. L. 2006. The chemosensory basis for
behavioral divergence involved in sympatric host shifts. I. Characterizing
olfactory receptor neuron classes responding to key host volatiles. J. Comp.
Physiol. A. 192: 279-288.
359. Robbins, P. S., Zhang, A., Averill, A. L., Linn, Jr., C. E., Roelofs, W. L.,
Sylvia, M. M., Villani M. G. 2006. Sex pheromone of the cranberry root
grub Lichnanthe vulpine. J. Chem. Ecol. 32: 1663-1672.
360. Robbins, P.S. and 44 collaborators. 2006. Trapping Phyllophaga spp.
(Coleoptera: Scarabaeidae: Melolonthinae) in the United States and Canada
using sex attractants. http://www.insectscience.org/6.39/
361. Olsson, S. B., Linn, Jr., C. E., Michel, A., Dambroski, H. R., Berlocher, S. H.
Feder, F. L., Roelofs, W. L. 2006. Receptor expression and sympatric speciation:
unique olfactory receptor neuron responses in F1 hybrid Rhagoletis
populations. J. Exp. Biol. 209: 3729-3741.
362. Linn, C., Jr., Musto, C., Roelofs, W. 2007. More Rare Males in Ostrinia:
Response of Asian Corn Borer Moths to the Sex Pheromone of the European
Corn Borer. J. Chem. Ecol. 33: 199-212.
363. Domingue M. J., Roelofs, W. L., Linn Jr., C. E., Baker, T.C. 2006. Effects of
egg-to-adult development time and adult age on olfactory neuron response
to semiochemicals in European corn borers. J. Insect. Physiol. 52: 975-983.
Wendell L. Roelofs 205
364. Xue, B., Rooney, A. P., Kajikawa, M., Okada, N., Roelofs, W. L. 2007. Novel
sex pheromone desaturases in the genomes of corn borers generated
through gene duplication and retroposon fusion. Proc. Natl. Aca. Sci. USA
104: 4467-4472.
365. Linn, C. E., Jr., Domingue, M. J., Musto, C., Baker, T. C., Roelofs, W L. 2007.
Support for (Z)-11-hexadecanal as a pheromone antagonist in Ostrinia
nubilalis: flight tunnel and single sensillum studies with a New York
population. J. Chem. Ecol. 33: 909-921.
366. Domingue, M. J., Musto, C., Linn, C. E., Jr., Roelofs, W L., Baker, T. C. 2007.
Evidence of olfactory antagonistic imposition as a facilitator of evolutionary
shifts in pheromone blend usage in Ostrinia spp. (Lepidoptera: Crambidae).
J. Insect Physiol. 53: 488-49.
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Cornelis T. De Wit
Agricultural University
Wageningen, Netherlands
k
1924–1993
IN MEMORIAM
On October 10, 2008 it is the 40th anniversary of Kees de Wit’s inaugural address
(Theory and model, in Dutch) at Wageningen Agricultural University, as Professor
of Theoretical Production Ecology. Although it is 15 years since he passed away,
much regretted, his mark on Wageningen University is still present, through the
C.T. De Wit Graduate School for Production Ecology and Resource Conservation
and through the ‘PhD children and grandchildren’ that serve the University in
various positions. This year therefore seems an appropriate moment to reflect on
40 years Theory and Model at Wageningen University and to take stock of the
developments in the last 15 years.
Kees (Cornelis) De Wit grew up in a rural village in the eastern part of the
Netherlands, and spent most of World War II as a farm laborer. This aroused his
interest in the complexities of farming and in farmers and was to guide his
professional career. He completed his studies after the war with a Ph.D. thesis
(‘A physical theory on placement of fertilizers’, 1953). His subsequent employment
at the Ministry of National Planning of the Union of Birma laid the foundation for
his strong commitment to agriculture in developing countries.
After his return in 1956, De Wit was employed at the Institute for Biological
and Chemical Research on Field Crops and Herbage (IBS) and its successor, CABO
(Centre for Agrobiological Research), and in the next decade produced some of his
most influential papers: ‘Transpiration and crop yields’ (1958), a major re-
interpretation of crop water use data, based on a physical analysis of canopy
207
208 Wolf Prize in Agriculture
LIST OF PUBLICATIONS
Wit, C.T. de, 1953. A physical theory on placement of fertilizers. Agricultural
Research Reports 59.4, 71 pp.
Cornelis T. De Wit 209
Wit, C.T. de, 1958. Transpiration and crop yields. Agricultural Research Reports
64.6, Pudoc, Wageningen, 88 pp.
Wit, C.T. de, 1960. On competition. Agricultural Research Reports 66.8, Pudoc,
Wageningen, 82 pp.
Wit, C.T. de, 1965. Photosynthesis of leaf canopies. Agricultural Research Reports
633, Pudoc, Wageningen, 57 pp.
Wit, C.T. de, 1972. Food production: past, present and future. Stikstof 15, 65-80.
Wit, C.T. de, 1972. De moderne landbouw in het westen. Landbouwkundig
Tijdschrift 84(9), 310-312.
Wit, C.T. de & H. van Keulen, 1972. Simulation of transport processes in soils.
Simulation Monographs, Pudoc, Wageningen, 109 pp.
Goudriaan, J. & C.T. de Wit, 1973. A re-interpretation of Gause’s population
experiments by means of simulation. Journal of Animal Ecology 42,
521-530.
Wit, C.T. de & A.Th.J. Nooij, 1973. Over eten en over leven. Diësrede LH. Intermediair
9, nr 11, 24 pp.
Dekkers, W.A., J.M. Lange & C.T. de Wit, 1974. Energy production and use in
Dutch agriculture. Netherlands Journal of Agricultural Science 22, 107-118.
Wit, C.T. de, 1974. Early theoretical concepts in soil fertility. Netherlands Journal
of Agricultural Science 22, 319-324.
Wit, C.T. de & J. Goudriaan, 1974. Simulation of ecological processes. Simulation
Monographs, Pudoc, Wageningen, 159 pp.
Sinclair, T.R. & C.T. de Wit, 1975. Photosynthesis and nitrogen requirements for
seed production by various crops. Science 189, 565-567 (15 August).
Wit, C.T. de, 1975. Energetische aspecten van de plantaardige produktie. Nederlands
Tijdschrift. voor Natuurkunde 41(12), 166-168.
Wit, C.T. de, 1975. Agriculture’s uncertain claim on world energy resources. SPAN
18(1), 2-4.
Wit, C.T. de, 1975. Landbouw in Nederland. In: Nederland en de grenzen aan
de groei. Symposium Tussentijds Bestek. Aula 548, Utrecht/Antwerpen,
pp. 29-34.
Wit, C.T. de, 1975. Modellen als brug tussen proces en systeem. In: Produktiviteit
in biologische systemen. Pudoc, Wageningen, pp. 157-170.
Wit, C.T. de, 1975. Substitution of labour and energy in agriculture and options
for growth. Netherlands Journal of Agricultural Science 23, 145-162.
Wit, C.T. de, 1975. Over de bruikbaarheid van simulatiemodellen van ecologische
systemen. In: Verhandeling van de Faculteit Landbouwwetenschappen te Gent,
nr 13, XXIVe Post-Universitaire Onderwijsdag, 5 pp.
Arnold, G.W. & C.T. de Wit (Eds), 1976. Critical evaluation of systems analysis
in ecosystems research and management. Simulation Monographs, Pudoc,
Wageningen, 114 pp.
Keulen, H. van, C.T. de Wit & H. Lof, 1976. The use of simulation models for
productivity studies in arid regions. In: Ecological Studies. Analysis and
Synthesis, Vol. 19. Eds O.L. Lange, L. Kappen & E.D. Schulze. Springer Verlag,
Heidelberg, pp. 408-420.
Sinclair, T.R. & C.T. de Wit, 1976. Analysis of the carbon and nitrogen limitations
to soybean yield. Agronomy Journal 68, 319-324.
Cornelis T. De Wit 211
Wit, C.T. de, 1976. On the usefulness of simulation models of ecological systems
(some thoughts for further discussion). In: Simulation of continuous systems.
Proceedings of a seminar. August 25, 26, 27. SAHR/NRMG, pp. 126-130.
Wit, C.T. de & H.D.J. van Heemst, 1976. Aspects of agricultural resources. In:
Chemical Engineering in a changing world. Ed. W.T. Koetsier. Proceedings of
the Plenary Sessions of the First World Congress on Chemical Engineering.
Amsterdam, June 28 - July 1, 1976. Elsevier Scientific Publishing Company,
Amsterdam, pp. 125-145.
Laar, H.H. van, D. Kremer & C.T. de Wit, 1977. Maize. In: Crop photosynthesis:
methods and compilation of data obtained with a mobile field equipment. Eds
Th. Alberda et al. Agricultural Research Reports no. 865. Pudoc, Wageningen,
pp. 12-21.
Sinclair, T.R., J. Goudriaan & C.T. de Wit, 1977. Mesophyll resistance and CO2
compensation concentration in leaf photosynthesis models. Photosynthetica
11(1), 56-65.
Wit, C.T. de, 1977. Model studies on actual and potential herbage production in
arid regions. Proceedings of the Seminar: Evaluation and mapping of tropical
African rangelands. Bamako, Mali, 3-8 March 1975. ILCA, Addis Ababa,
pp. 329-331.
Wit, C.T. de, 1977. Selectieve ontwikkeling van de landbouw in een kader van
verantwoord energie- en omgevingsbeleid. In: Verslag van het Symposium
Landbouwkundig Onderzoek in Nederland. Pudoc, Wageningen, pp. 93-108.
Wit, C.T. de, 1977. Problemen van de voedselproduktie. De Ingenieur 89(17),
334-338.
Wit, C.T. de, 1977. Modelle der Ertragsbildung als Brücke zwischen Prozess und
System. In: Bio-physikalische Analyse pflanzlicher Systeme. Ed. K. Unger. VEB
Gustav Fischer Verlag, Jena, pp. 19-30.
Wit, C.T. de, 1977. Concluding remarks. In: Environmental Effects on Crop
Physiology. 5th Long Ashton Symp., 13-16 April 1975. Eds J.J. Landsberg &
C.V. Cutting, Academic Press, London, pp. 365-369.
Wit, C.T. de, 1978. Simulatie van levende systemen. Landbouwkundig Tijdschrift/
pt 90-8a, 237-240.
Wit, C.T. de, 1978. Een elementair model van gewasgroei. Landbouwkundig
Tijdschrift/pt 90-8a, 275-281.
Wit, C.T. de, 1978. Summative address: One. In: Plant Relations in pastures.
Ed. John R. Wilson, C.S.I.R.O., Australia, pp. 405-410.
Wit, C.T. de & J. Goudriaan, 1978. Simulation of ecological processes. Revised
version. Simulation Monographs, Simulation Monographs, Pudoc, Wageningen,
183 pp.
Wit, C.T. de et al., 1978. Simulation of assimilation, respiration and transpiration
of crops. Simulation Monographs, Pudoc, Wageningen, 148 pp.
212 Wolf Prize in Agriculture
Wit, C.T. de, 1979. The efficient use of labour, land and energy in agriculture.
Agricultural Systems, 279-287.
Wit, C.T. de & R. Rabbinge, 1979. Systems analysis and dynamic simulation. EPPO
bull. 9(3), 149-153.
Wit, C.T. de & H.D.J. van Heemst, 1979. Aspekte landwirtschaftlicher Produktion
quellen. In: Tropische Landwirtschaft. Forschung und Information, Band 25,
Colloquium Verlag, Berlin, pp. 28-43.
Wit, C.T. de, H.H. van Laar & H. van Keulen, 1979. Physiological potential of crop
production. In: Plant Breeding perspectives. Eds J. Sneep and A.J.T. Hendriksen,
Pudoc, Wageningen, pp. 47-82.
Keulen, H. van & C.T. de Wit, 1980. Landhoedanigheden en produktiemogelijkheden.
Landbouwkundig Tijdschrift/pt 92(2), 49-53
Keulen, H. van & C.T. de Wit, 1980. Simulation of plant production in arid regions:
a hierarchial approach. In: Arid-land ecosystems:structure, functioning and
management. Vol. 2, Cambridge University Press, Cambridge.
Wit, C.T. de, 1981. The energy spectrum: food and energy. Conference proceedings:
“Food security in a hungry world”, San Francisco, 4-6 March, University of
California, Davis, U.S.A., pp. 66-70.
Wit, C.T. de, 1981. Oude wijn in nieuwe zakken. Landbouwkundig Tijdschrift/pt
93(10), 257-262.
Wit, C.T. de, 1982. Simulation of living systems. In: F.W.T. Penning de Vries & H.H.
van Laar (Eds), Simulation of plant growth and crop production. Simulation
Monographs, Pudoc, Wageningen, pp. 3-8.
Wit, C.T. de, 1982. Coordination of models. In: F.W.T. Penning de Vries & H.H. van
Laar (Eds), Simulation of plant growth and crop production. Simulation
Monographs, Pudoc, Wageningen, pp. 26-31.
Wit, C.T. de & F.W.T. Penning de Vries, 1982. L’analyse des systemes de production
primaire. In: F.W.T. Penning de Vries & M.A. Djitèye (Eds), La productivité des
pâturages sahéliens. Pudoc, Wageningen, pp. 20-23.
Wit, C.T. de & F.W.T. Penning de Vries, 1982. La synthese et la simulation des
systemes de production primaire. In: F.W.T. Penning de Vries & M.A. Djitèye
(Eds), La productivité des pâturages sahéliens. Pudoc, Wageningen, pp. 23-27.
Wit, C.T. de, 1982. Des problemes analytiques et experimentaux. In: F.W.T. Penning
de Vries & M.A. Djitèye (Eds), La productivité des pâturages sahéliens. Pudoc,
Wageningen, pp. 27-32.
Wit, C.T. de & J.M. Krul, 1982. La production actuelle dans une situation d’équilibre.
In: F.W.T. Penning de Vries & M.A. Djitèye (Eds), La productivité des pâturages
sahéliens. Pudoc, Wageningen, pp. 275-281.
Keulen, H. van & C.T. de Wit, 1982. A hierarchical approach to agricultural
production modeling. In: Modeling Agricultural-Environmental Processes in
Crop Production. Eds G. Golubev & J. Shvytov. IIASA Collaborative proceedings
Series CP-82-S5, pp. 139-153.
Cornelis T. De Wit 213
Wit, C.T. de, H. Huisman & R. Rabbinge, 1987. Agriculture and its Environment:
Are There Other Ways? Agricultural Systems 23, 211-236.
Wit, C.T. de & H. van Keulen, 1987. Modelling production of field crops and its
requirements. Geoderma 40, 253-265.
Wolf, J., C.T. de Wit, B.H. Janssen & D.J. Lathwell, 1987. Modelling long-term
response to fertilizer phosphorus. I. The Model. Agronomy Journal 79,
445-451.
Wit, C.T. de, 1988. Duurzame landbouw: blijvende zorg. Verslag verenigde
vergadering van de Koninklijke Nederlandse Akademie van Wetenschappen
op 11 april 1988, Amsterdam, pp. 13-23.
Wit, C.T. de, 1988. Problemen van duurzaamheid van de landbouw. In: Naar
een duurzaam Nederland. Ed. M. Groen. SDU uitgeverij, ’s-Gravenhage,
pp. 93-97.
Wit, C.T. de, 1988. The agricultural environment in the European community.
Ecological Bulletins 39, 204-211 (Copenhagen).
Wit, C.T. de, 1988. Environmental impact of the common agricultural policy. In:
Environmental policy in a market economy. Eds Frank J. Dietz & Willem J.M.
Heijman. Selected papers from the Congress ‘Environmental policy in a
market economy’ Wageningen, Netherlands, 8-11 September 1987. Pudoc,
Wageningen, pp. 190-204.
Wit, C.T. de, 1988. Environmental impact of the CAP. European Review of
Agricultural Economics 15, 283-296.
Wit, C.T. de, 1988. Landbouw in Europees perspectief. De Europese gemeente, 23e
jaargang, nr 4, 32-37.
Wit, C.T. de, H. van Keulen, N.G. Seligman & I. Spharim, 1988. Application of
interactive multiple goal programming techniques for analysis and planning
of regional agricultural development. Agricultural Systems 26(3), 211-230.
Wit, C.T. de, 1989. Internationaal landbouwonderzoek voor ontwikkelingslanden
in perspectief. Afscheidsrede Landbouwuniversiteit Wageningen, 17 februari
1989, 16 pp.
Wit, C.T. de, 1989. Problemen van duurzaamheid in de landbouw. Landbouwkundig
Tijdschrift 101 (1), 18-20.
Wolf, J., C.T. de Wit & H. van Keulen, 1989. Modeling long-term crop response to
fertilizer and soil nitrogen. I. Model description and application. Plant and
Soil 120, 11-22.
Rabbinge, R. & C.T. de Wit, 1989. Theory of modelling and systems management.
In: Simulation and systems management in crop protection. Eds R. Rabbinge,
S.A. Ward & H.H.van Laar, Simulation Monographs 32, Pudoc, Wageningen,
the Netherlands, pp. 3-15.
Wit, C.T. de, 1990. Understanding and managing changes in agriculture. In: Systems
theory applied to agriculture and the food chain. Eds J.W.G. Jones & P.R.
Street, Elsevier Science Publishers, London and New York, pp. 235-249.
Cornelis T. De Wit 215
Wit, C.T. de, 1990. International agricultural research for developing countries. In:
Theoretical Production Ecology: reflections and prospects. Eds R. Rabbinge,
J. Goudriaan, H. van Keulen, F.W.T. Penning de Vries & H.H. van Laar,
Simulation Monographs 34, Pudoc, Wageningen, pp. 295-301.
Wit, C.T. de, 1990. Making predictions about farming. In: Scientific Europe, Research
and Technology in 20 Countries. Nature & Technology, Scientific Publishers
Ltd., Maastricht, Brussels, London, pp. 188-191.
Wit, C.T. de, 1991. On the efficiency of resource use in agriculture. In: Ziele und
Wege der Forschung im Pflanzenbau. Festschrift für Kord Baeumer zum 65.
Geburtstag. Editor Wolfgang Böhm, Triade-Verlag E. Claupein, Göttingen,
pp. 29-54.
Alberda, Th., H. van Keulen, N.G. Seligman & C.T. de Wit (Eds), 1992. Food from
dry lands. An integrated approach to planning af agricultural development.
Kluwer Academic Press, Dordrecht, Boston, London, 211 p.
Spharim, I., R. Spharim & C.T. de Wit, 1992. Modelling agricultural development
strategy. In: Food from dry lands. An integrated approach to planning af
agricultural development. Eds Th. Alberda, H. van Keulen, N.G. Seligman
& C.T. de Wit, Kluwer Academic Press, Dordrecht, Boston, London,
pp. 159-192.
Wit, C.T. de & N.G. Seligman, 1992. Introduction. In: Food from dry lands. An
integrated approach to planning af agricultural development. Eds Th. Alberda,
H. van Keulen, N.G. Seligman & C.T. de Wit, Kluwer Academic Press, Dordrecht,
Boston, London, pp. 1-5.
Wit, C.T. de, 1992. Summary. In: Food from dry lands. An integrated approach to
planning af agricultural development. Eds Th. Alberda, H. van Keulen, N.G.
Seligman & C.T. de Wit, Kluwer Academic Press, Dordrecht, Boston, London.
pp. 193-200.
Wit, C.T. de, 1992. Resource use efficiency in agriculture. Agricultural Systems 40,
125-151.
Wit, C.T. de, 1992. Over het efficiënte gebruik van hulpbronnen in de landbouw.
Spil (december, 1992) 5, 40-52.
Wit, C.T. de, 1993. Philosophy and terminology. In: On systems analysis and
simulation of ecological processes, with examples in CSMP and FORTRAN. Ed.
P.A. Leffelaar. Current Issues in Production Ecology Volume I, Kluwer Academic
Publishers, Dordrecht, pp. 3-9.
Wit, C.T. de & J. Goudriaan, 1993. The growth of yeast. In: On systems analysis
and simulation of ecological processes, with examples in CSMP and FORTRAN.
Ed. P.A. Leffelaar. Current Issues in Production Ecology Volume I, Kluwer
Academic Publishers, Dordrecht, pp. 41-50.
Wit, C.T. de, 1993. Tussen twee vuren. In: Intensivering of extensivering. Over de
toekomst van de Europese landbouw. Studium Generale, Landbouwuniversiteit
Wageningen, pp. 3-20.
216 Wolf Prize in Agriculture
Wit, C.T. de, 1994. Resource use analysis in agriculture: A struggle for
interdisciplinarity. In: The future of the land: Mobilising and integrating
knowledge for land use options. Eds L.O. Fresco, L. Stroosnijder, J. Bouma &
H. van Keulen, John Wiley & Sons, New York, pp. 41-55.
List of Ph.D. dissertations under partial or full guidance of Prof C.T. De Wit
Bergh, J.P. van den, 1968: An analysis of yields of grasses in mixed and pure
stands.
Penning de Vries, F.W.T., 1973: Substrate utilization and respiration in relation to
growth and maintenance in higher plants.
Janssen, J.G.M., 1974: System-ecological approaches to the microdistribution of
some winter annuals.
Fransz, H.G., 1974: The functional response to prey density in an acarine system
Pieters, G.A., 1974: The growth of sun and shade leaves of Populus euramericana
“Robusta” in relation to age, light intensity and temperature.
Egmond, F. van, 1975: The ionic balance of the sugar-beet plant.
Keulen, H. van, 1975: Simulation of water use and herbage growth in arid regions.
Challa, H., 1976: An analysis of the diurnal course of growth, carbon dioxide
exchange and carbohydrate reserve content of cucumber.
Rabbinge, R., 1976: Biological control of fruit-tree red spider mite.
Elderen, E. van, 1977: Heuristic strategy for scheduling farm operations.
Goudriaan, J., 1977: Crop micrometeorology: a simulation study.
Veen, J.A. van, 1977: The behaviour of nitrogen in soil. A computer simulation
model.
Bunnik, N.J.J., 1978: The multispectral reflectance of shortwave radiation by
agricultural crops in relation with their morphological and optical properties.
Spitters, C.J.T., 1979: Competition and its consequences for selection in barley
breeding.
Braakhekke, W.G., 1980: On coexistence: a causal approach to diversity and stability
in grassland vegetation.
Mutsaers, H.J.W., 1982: KUTUN, A morphogenetic model for cotton Gossypium
hirsutum L.
Sabelis, M.W., 1982: Biological control of two-spotted spider mites using phytoseiid
predators. Part I. Modelling the predator-prey interaction at the individual
level.
Chen Jialin, 1984: Mathematical analysis and simulation of crop micro-meteorology.
Bakker, Th.M., 1985: Eten van eigen bodem. Een modelstudie.
Lantinga, E.A., 1985: Productivity of grasslands under continuous and rotational
grazing.
Cissé, A.M., 1986: Dynamique de la strate herbacée des pâturages de la zone sud-
sahélienne.
Cornelis T. De Wit 217
Mohren, G.M.J., 1987: Simulation of forest growth, applied to Douglas fir stands
in the Netherlands.
Willigen, P. de, and M. van Noordwijk, 1987: Roots, plant production and nutrient
use efficiency.
Leffelaar, P.A., 1987: Dynamics of partial anaerobiosis, denitrification, and water
in soil: experiments and simulation.
Dulk, J.A. den, 1989: The interpretation of remote sensing, a feasibility study.
Kropff, M.J., 1989: Quantification of SO2 effects on physiological processes, plant
growth and crop production.
Klepper, O., 1989: A model of carbon flows in relation to macrobenthic food
supply in the Oosterschelde estuary (S.W. Netherlands).
Veeneklaas, F.R., 1990: Dovetailing technical and economic analysis.
Rappoldt, C., 1992: Diffusion in aggregated soil.
Miglietta, F., 1992: Simulation of wheat ontogenesis.
Ranganathan, Radha, 1993: Analysis of yield advantage in mixed cropping.
Nonhebel, Sanderine, 1993: The importance of weather data in crop growth
simulation models and assessment of climatic change effects.
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Don Kirkham
Iowa State University
Ames, Iowa, USA
k
1908–1998
219
220 Wolf Prize in Agriculture
Don Kirkham was born February 11, 1908 in Provo, Utah. He attended the public
schools in Salt Lake City and Berkeley, California. During high school and his first
year at the University of Utah he also studied music, graduating as a clarinetist
from the McCune School of Music, Salt Lake City, in 1926. As a youth he worked
summers on farmland owned by his father. He spent two and a half years in
Germany as a missionary for his church in 1927-30, and then continued his
education at Columbia University in New York City.
Columbia University awarded him the A. B. degree (with Honors in Physics) in
1933, the A. M. degree in 1934, and the Ph.D. degree, under S. L. Quimby, in
1938. His Ph.D. degree was received in Physics, his dissertation title being
“The variation of the initial susceptibility with temperature and the variation
of the magnetostriction and reversible susceptibility with temperature and
magnetization in nickel.”
From 1938 to 1940 he was instructor and assistant professor in mathematics
and physics at Utah State University. Here, through his colleague, Professor Willard
Gardner, he became interested in soil physics research and published several articles.
During the years of World War II (1940–46) he served as civilian scientist
with the United States Navy, working on the research physics of anti-mine warfare.
His naval work included designing, setting up, and operating the Navy’s anti-
magnetic mine program for all vessels using New York harbor. In 1946 he headed
a group of Navy physicists at the Bikini Atom Bomb Tests.
During the war years, in addition to his Navy work, he continued study in
theoretical soil physics problems and publication of articles. In 1946 he joined the
staff of Iowa State University as Associate Professor of Agronomy and Physics, and
was made Professor in 1949. In 1959 he was appointed Curtiss Distinguished
Professor of Agriculture, the Distinguished Professor Title being this institution’s
highest recognition for faculty excellence. He served as Director of the Iowa State
Water Resources Research Institute from 1964–73. Since 1973 he has continued
to carry on a full program of teaching and research. He was named Professor
Emeritus in 1978.
He was married and the father of three children.
Scientific Societies
International Water Resources Association
Netherlands Society of Agricultural Science
Don Kirkham 221
Honorary Societies
Sigma Xi, national honorary scientific research society
Gamma Sigma Delta, national honorary agricultural society
Phi Kappa Phi, national honorary scholastic society
theoretical streamlines with experimental streamlines. But this paper, like others
up to then, was for homogeneous soil. So Kirkham now analyzed the problem of
non-homogeneous soil and with a resulting two-part paper (31), (32), he received
the American Geophysical Union’s citation for the best paper in hydrology of the
year 1951.
These earlier papers were for ponded water, that is, completely saturated soil
and represented thus an extreme condition. He next considered the more common
condition of a curved water table resulting from successive recharges of water as
from irrigation, or as from successive rains. By first neglecting the hydraulic friction
in the soil in the water table arch region and then later correcting for this neglect,
he derived an explicit relation for the height of water table to be found between
drain lines at certain depths and spacings. He compared his theoretical results
with field data and found good agreement (64). In a paper with a graduate student
as senior author, the solution to this drainage problem was put in a graphical form
so that the various physical factors that entered into the problem could be considered
in the graph and the solution obtained graphically for the depth and spacing of
drains (98), (99), (99a). This same student in working for his Ph.D. degree
considered stratified soil. Two other papers resulted in which the ratio of hydraulic
conductivities of the layered soil could be taken into account. This long two-part
paper was published in the Irrigation and Drainage Division of the American
Society of Civil Engineers (173), (174).
All the foregoing work was for a simplified steady state condition of either
ponded water or steady recharge to the drain tubes. Kirkham next analyzed the
problem of a falling water table. He found equations which predicted where the
water table arch would be after a certain time when the steady recharge was
discontinued. He compared his theoretical results with actual field data that he
had obtained with other scientists in the Netherlands many years before and with
other field data. There was excellent agreement (130).
In all the above work involving tile drains, it was assumed that the water
entered the drain tube uniformly over its length. This would be true if there were
an envelope of coarse material around the drain tube. Actually drain pipes are
about 30 cm to 60 cm long and have about 1 to 6 mm spacing between each
individual drain pipe. At least this has been the condition up until recent times
when plastic tubes, perforated to let the water enter, are pulled into the soil. To see
just how much the cracks of individual tile lengths or the perforations in tubes
would control drainage flow if there was not a pervious envelope around them,
Kirkham authored or co-authored several articles — (28), (29), (197), (200). A
first one, in two parts, (28) and (29), was co-authored by one of his joint soil
physics-agricultural engineering Ph.D. candidates, who conceived the idea of using
plastic flexible tubing instead of clay pipe in soil. The two-part paper on the
influence of perforations on drainage flow to drain tubes received a citation of
Don Kirkham 223
the hydraulic conductivity of the soil about the hole, provided certain mathematical
formulas are found. Kirkham derived the formulas needed (19), (20), (66). More
detailed theory was later developed (181).
To solve a saturated soil-water movement problem, one wishes to know the
potential energy in the soil and also the flow paths or streamlines of the water.
Kirkham had found the potential energy and the streamlines for a number of
drainage problems, as has been noted. For some problems, as for axially symmetric
flow into an auger hole, he had not, however, determined the streamlines. One of
Kirkham’s Ph.D. candidates worked on the problem and two noteworthy papers
resulted. Streamline functions were developed which checked with auger–hole
flow (114), (138), and were later used in a number of well-flow problems.
Streamline functions and potential-energy functions, as Kirkham and his
students had so far developed them, could not solve water movement problems
where the flow domain was other than a relatively simple shape. Many seepage
domains are, however, not of simple shape and to determine flow for them, a
whole new mathematical method had to be developed. The needed new
mathematical method which he and his students developed is called the modified
Gram-Schmidt method (152), (195).
In the modified Gram-Schmidt method, Kirkham, with the help of graduate
students, developed equations in which needed parameters are not embedded in
complex expressions used by Gram-Schmidt. Kirkham’s expressions are tabulated
briefly in a paper in which W. L. Powers is senior author (152). The equations
which give the constants needed for this method of solving problems such as
piezometer problems are listed in more detail on pages 502-503 in Kirkham and
Power’s book (195). Numerous papers involving these functions have been
published. Those in which Kirkham is an author or co-author are (150), (152),
(157), (164), (180), (191), (185),(185a), (187) (189), (190), (191), (197), (200),
(201), (203), (204), (205), (206), (209), (212), (216), (217), (218), (221).
Agriculture is charged with polluting rivers and wells through runoff of
dissolved chemical fertilizers or pesticides or animal wastes. In recent years, a
number of pollution problems have been solved by Kirkham and his students. A
difficult one concerns the time required for a river that has been polluted to yield
polluted seepage flow to wells near the river’s side (209). Others concern the effect
of an improper seal around a well (217) and also the effect of a well casing (216)
and of a cover slab (218) in stopping pollution. Complicated aquifer shapes and
pollution are considered in (185) and (185a).
All the foregoing problems have dealt with water-saturated soil. This is a realistic
situation because water cannot move from soil into a drainage facility as a drain
tube, or a ditch, unless the soil at an atmospheric-pressure outflow point is saturated.
Nevertheless, water often moves in the soil in the unsaturated state. When Kirkham
came to Iowa State University, his first paper was on horizontal unsaturated flow
Don Kirkham 225
(14). In this paper he showed that the wetting front moved by a square root of
time law, but that, contrary to the statements of other workers, this movement was
not in accordance with constant-conductivity, heat-flow theory, even though the
wetting front movement corresponded to heat flow theory. This paper resulted in
British scientists suggesting that a diffusivity or variable conductivity function
needed to be used. The idea was followed through by other scientists and numerous
papers on the diffusion theory of capillary flow have now been published (195).
One on flow into drainage ditches where movement is in the unsaturated part of
the soil has generated many reprint requests (188). A much earlier paper on
movement in the capillary fringe of soil was published in two parts in Soil Science
(54), (55).
In irrigated areas salt moves to the soil surface by unsaturated flow and is
removed by saturated leaching flow. In a laboratory experiment, a layer of salt was
placed on a porous medium and leaching of the salt into simulated tile drains was
performed. The concentration of the drainage water was analyzed as the salt was
removed. A theoretical equation was derived for the salt concentration which
agreed well with the experimental data (183).
Although Kirkham’s papers are mainly on water movement in soil, he has
contributed to other aspects of soil physics. A paper on sediment transport by
rivers and canals gave a theoretical equation that checked experimental data (8).
Papers on heat flow are (65), (61), (171), (172), (208). Some papers on air and
oxygen movement are (13), (25), (120), (175), (223). Some on soil compaction
are (97), (101), (102). He and colleagues developed a neutron or subatomic particle
soil probe for measuring soil moisture content (36), (52), (86), (110). A portable
rainfall–simulator infiltrome was made (62) and patented (108). Some other soil
physics equipment was developed and patents obtained (197), (109), (110).
Kirkham’s research program has attracted and inspired students to do graduate
work at Iowa State University. Under his direction students have received 54 Ph.D.
degrees, 32 M.S. degrees; of these, 17 students have received both M.S. and Ph.D.
degrees. A list of his graduate students with their positions is found in Appendix P.
Through these students, his achievements on behalf of mankind for food production
and the protection of land against improper use are being promoted in countries
around the world. In his bibliography it is seen that a number of the papers are
with students as senior authors. Without the excellent and large input of capable
graduate students into the many papers on Kirkham’s list, the work could not have
been done.
The strong relationship between Kirkham and his students and Kirkham’ unique
professional contributions are described in the following paragraph quoted from
an article written by Dr. Dale Swartzendruber, Professor of Agronomy at the
University of Nebraska. (“In Recognition of Don Kirkham on His Seventieth
Birthday,” Soil Science, Vol. 125, No. 2, February 1978).
226 Wolf Prize in Agriculture
The wide application of Kirkham’s work is evident from the numerous national
and international journals in which his work is published. His papers have been
published in the Soil Science Society of America Proceedings (67 papers), Water
Resources Research (hydrology) (18 papers), Soil Science (16 papers), Transactions
American Geophysical Union (13 papers), Agronomy Journal (7 papers), Journal
of Geophysical Research (7 papers), Transactions of the State Agricultural College,
Ghent, Belgium (7 papers), Journal of Irrigation and Drainage of the American
Society of Civil Engineers (6 papers), Agricultural Engineering (4 papers), and in
many other publications.
Seven selected reprints of papers illustrating Kirkham’s pioneering and
fundamental work in solving difficult soil water flow problems in agriculture are
attached in Appendix E (7), (64), (98), (114), (130), (149), (152). In (7), see
especially p. 67, Fig. 5. In (64), see especially p. 904, Fig. 4 where the theoretical
curves fall quite closely on the field–data points. In (98), see especially the two
charts on pp. 513-514, Figs. 1 and 2. In (114), see espcially p. 159, Fig. 2 where
the theoretically derived streamlines (arrows) pass at right angles, as they should,
through the equipotential lines. In (130), see especially pp. 586, 588, 589,
Figs. 2–6; notice in Figs. 4, 5, and 6 that the theoretical curves of water table
height w versus days of fall t fit the field data “points.” In (149), p. 617, Fig. 8, see
that Dupuit-Forchheimer (D.F.) flow lines are not “horizontal,” as stated in the
literature, and that they agree quite well with actual streamlines; on p. 618, Fig. 9,
which is for tile drainage, the D.F. streamlines are the same as for ditch drainage,
Don Kirkham 227
Fig. 8, and do not agree well with exactly calculated streamlines. In (152), note
that Fig. 1 shows the problem of seepage of water in soil bedding, that Table 2
gives the mathematical functions developed to solve the problem, and that
Figs. 4–6 show the flow net solutions of the problem with the streamlines and
equipotential lines crossing at right angles as they should.
LIST OF PUBLICATIONS
1. Kirkham, Don. 1937. The variation of the initial susceptibility with
temperature, and the variation of the magneto striction and reversible
susceptibility with temperature and magnetization in nickel. Physical Review
52: 1162-1167.
2. Kirkham, Don. 1939. Abstract: Streamline flow of water from an artesian
basin into horizontal drains: Theory compared with experiment. Physical
Review 56: 852.
3. Kirkham, Don. 1939. Artificial drainage of land: Streamline experiments.
The artesian basin I. Trans. Amer. Geophys. Union, 20: 677-680.
4. Kirkham, Don. 1940. Artificial drainage of land: Streamline experiments.
The artesian basin II. Trans. Amer. Geophys. Union, 21: 587-593.
5. Kirkham, Don. 1940. Abstract: Solution of Laplace’s equation in application
to the artificial drainage of waterlogged land overlying an impervious layer.
Physical Review 57: 1058.
6. Kirkham, Don. 1941. Abstract: Conjugate potentials of a grid between
conducting plates. Physical Review 59: 111.
7. Kirkham, Don. 1940. Pressure and streamline distribution in waterlogged
land overlying an impervious layer. Soil Sci. Soc. Amer. Proc. 5: 65-68.
8. Kirkham, Don. 1942. Modification of a theory on the relation of suspended
to bed-material in rivers. Trans. Amer. Geophys. Union 23: 618-621.
9. Kirkham, Don. 1945. Artificial drainage of land: Streamline experiments.
The artesian basin III. Trans. Amer. Geophys. Union 26: 393-406.
10. Kirkham, Don. 1945. Proposed method for field measurement of permeability
of soil below the water table. Soil Sci. Soc. Amer. Proc. 10: 58-68.
11. Kirkham, Don. 1946. Discussion of C. E. Jacob’s article “Radial flow in a
leaky artesian aquifer.” Trans. Amer. Geophys. Union 26: 206-209.
12. Kirkham, Don. 1948. Abstract: A two-dimensional potential problem with
application to soil drainage. Physical Review 73: 1228.
13. J-1418. Kirkham, Don. 1946. Field method for determination of air
permeability of soil in its undisturbed state. Soil Sci. Soc. Amer. Proc. 11:
93-99.
14. J-1451. Kirkham, Don and C. L Feng. 1949. Some tests of the diffusion
theory, and laws of capillary flow, in soils. Soil Sci. 67: 29-40.
228 Wolf Prize in Agriculture
15. J-1472. Kirkham, Don. 1949. Flow of ponded water into drain tubes in soil
overlying an impervious layer. Trans. Amer. Geophys. Union 30: 369-385.
16. J-1503. Kirkham, Don. 1947. Reduction in seepage to soil under drains
resulting from their partial embedment in, or proximity to, an impervious
substratum. Soil Sci. Soc. Amer. Proc. 12: 54-59.
17. J-1504. Kirkham, Don. 1947. Studies of hillside seepage in the Iowan drift
area. Soil Sci. Soc. Amer. Proc. 12: 73-80.
18. J-1590. Hammond, L.C., and Don Kirkham. 1949. Growth curves of soybeans
and corn. Agronomy Journal 41: 23-29.
19. J-1624. van Bavel, C.H.M., and Don Kirkham. 1948. Field measurement of
soil permeability using auger holes. Soil Sci. Soc. Amer. Proc. 13: 90-96.
20. J-1625. Kirkham, Don, and C.H.M. Bavel. 1948. Theory of seepage into
auger holes. Soil Sci. Soc. Amer. Proc. 13: 75-82.
21. J-1644. Luthin, J.N., and Don Kirkham. 1949. A piezometer method
for measuring permeability of soil in situ below a water table. Soil Sci.
68: 349-358.
22. J-1650. Frevert, R.K. and Don Kirkham. 1948. A field method for measuring
the permeability of soil below a water table. Proc. Highway Res. Board
28: 433-442.
23. J-1673. Kirkham, Don. 1950. Seepage into ditches in the case of a plane
water table and an impervious substratum. Trans. Amer. Geophys. Union
31: 425-430.
24. J-1689. Kirkham, Don, and G.S. Taylor. 1949. Some tests of a four-electrode
probe for soil moisture measurement. Soil Sci. Soc. Amer. Proc. 14: 42-46.
25. J-1690. Evans, D.D., and Don Kirkham. 1949. Measurement of the air
permeability of soil in situ. Soil Sci. Soc. Amer. Proc. 14: 65-73.
26. J-1728. Kirkham, Don. 1950. Potential flow into circumferential openings in
drain tubes. Journal Applied Physics 21: 655-660.
27. Kirkham, Don. 1951. Abstract: Potential flow into circumferential openings
in drain tubes. Amer. Math. Monthly 58: 139.
28. J-1792. Kirkham, Don, and G.O. Schwab. 1951. The effect of circular
perforations on flow into subsurface drain tubes. Part I. Theory. Agric. Eng.
32: 211-214.
29. J-1793. Schwab, G.O. and Don Kirkham. 1951. The effect of circular
perforations on flow into subsurface drain tubes. Part II. Experiments and
results. Agric. Eng. 32: 270-274.
30. J-1823. Reeve, R.C. and Don Kirkham. 1951. Soil anisotropy and some
field methods for measuring permeability. Trans. Amer. Geophys. Union 32:
582-590.
31. J-1824. Kirkham, Don. 1951. Seepage into drain tubes in stratified soil. I —
Drains in the surface stratum. Trans. Amer. Geophys. Union 32: 422-433.
Don Kirkham 229
32. J-1827. Kirkham, Don. 1951. Seepage into drain tubes in stratified soil.
II — Drains below the surface stratum. Trans. Amer. Geophys. Union 32:
433-442.
33. J-1828. Kirkham, Don, and R.E. Gaskell. 1950. The falling water table in tile
and ditch drainage. Soil Sci. Soc. Amer. Proc. 15: 37-48.
34. J-1831. Evans, D.D., Don Kirkham, and R.K. Frevert. 1950. Infiltration and
permeability in soil overlying an impermeable layer. Soil Sci. Soc. Amer. Proc.
15: 50-54.
35. Kirkham, Don. 1951. Discussion of McNown and Hsu’s article “Effect of a
partial cutoff on seepage rates.” Trans. Amer. Geophys. Union 32: 283.
36. J-2007. Gardner, Wilford, and Don Kirkham. 1952. Determination of soil
moisture by neutron scattering. Soil Sci. 73: 391-401.
37. J-2071. Kirkham, Don, and J.W. de Zeeuw. 1952. Field measurements for
tests of soil drainage theory. Soil Sci. Soc. Amer. Proc. 16: 286-293.
38. J-2185. DeBoodt, M.F., and Don Kirkham. 1953. Anisotrophy and
measurement of air permeability of soil clods. Soil Sci. 76: 127-133.
39. J-2188. DeBoodt, M.F., Don Kirkham, and A.J. Englehorn. 1953. Fall vs.
spring plowing and soil physical conditions in a rotation experiment.
Agronomy Journal 45: 257-261.
40. Kirkham, Don. 1952. Abstract: On the zeros of Jn(xp)Ym(kxp)
Jm(kxp)Yn(xp). Amer. Math. Monthly 59: 501.
41. J-2205. Kirkham, Don, and J.R. Runkles. Evaluation of new soil conditioners.
Trans. Iowa State Horticultural Society 87: 41-46.
42. Kirkham, Don. 1953. Book Review of “Soil Physical Conditions and Plant
Growth”. SSSA Proc. 17: 175. See also: 1953. Agronomy Journal 44: 503.
43. J-2326. Swartzendruber, Dale, M.F. DeBoodt, and Don Kirkham. 1954.
Capillary intake rate of water and soil structure. Soil Sci. Soc. Amer. Proc 18:
1-7.
44. J-2337. Kirkham, Don, and W.V. Bartholomew. 1954. Equations for following
nutrient transformations in soil, utilizing tracer data. Soil Sci. Soc. Amer.
Proc. 18: 33-34.
45. J-2372. Kirkham, Don. 1954. Seepage of artesian and surface water into
drain tubes in stratified soil. Trans. Amer. Geophys. Union 35: 775-790.
46. J-2501. van Schilfgaarde, Jan, R.K Frevert, and Don Kirkham. 1954. A tile
drainage field laboratory. Agric. Eng. 35: 474-478.
47. Kirkham, Don, and W.V. Bartholomew. 1954. Abstract: Mathematical theory
for the utilization of tagged atoms in determining plant nutrient
transformation rates in soils. Amer. Math. Monthly 61: 514.
48. J-2505. Kirkham, Don. 1955. Measurement of the hydraulic conductivity of
soil in place. Symposium Amer. Soc. for Testing Materials, Special Tech. Publ.
No. 163, 80-97.
230 Wolf Prize in Agriculture
49. Rollins, R.L., M.G. Spangler, and Don Kirkham. 1954. Movement of soil
moisture under a thermal gradient. Highway Research Board Proc. 33:
492-508.
50. J-2605. Kirkham, Don, and W.V. Bartholomew. 1955. Equations for following
nutrient transformations in soil, utilizing tracer data: II. Soil Sci. Soc. Amer.
Proc. 19: 189-192.
51. J-2628. Maasland, Marinus, and Don Kirkham. 1955. Theory and
measurement of anisotropic air permeability in soil. Soil Sci. Soc. Amer. Proc.
19: 395-400.
52. J-2641. Stone, J.F., Don Kirkham, and A.A. Reed. 1955. Soil moisture
determination by a portable neutron scattering moisture meter. Soil Sci. Soc.
Amer. Proc. 19: 419-423.
53. van Schilfgaarde, Jan, Don Kirkham, and R.K. Frevert. 1956. Physical and
mathematical theories of tile and ditch drainage and their usefulness in
design. Agricultural Experiment Station, Iowa State College, Research Bulletin
No. 436: 667-706.
54. J-2773. Swartzendruber, Dale, and Don Kirkham. 1956. Capillary fringe
and flow of water in soil. Soil Sci. 81: 473-484.
55. J-2832. Swartzendruber, Dale and Don Kirkham. 1956. Capillary fringe and
flow of water in soil. II. Experimental results. Soil Sci. 82: 81-95.
56. Kirkham, Don. 1956. Book review of L.D. Baver,s “Soil Physics”. Soil Sci. Soc.
Amer. Proc. 20: 296; also in Agronomy Journal, 48:195. 1956.
57. J-2913. Kirkham, Don. 1956. Mathematical aspects of soil nitrogen studies.
Synposium on use of isotopes in agriculture sponsored by Argonne National
Laboratory. Government Printing Office, Washington 25, D.C. January, 1956.
58. Kirkham, Don. 1957. Theory of seepage of ponded water into drainage
facilities, p.139-181. In J.N. Luthin (ed.) Drainage of Agricultural Lands.
Agron. Monogr. 7, ASA and SSSA, Madison, Wisconsin.
59. J-2998. Kirkham, Don. 1958. Graphs and formulas for zeros of cross product
Bessel functions. Journal of Mathematics and Physics 36: 371-377.
60. J-3033. Schwab, G.O., Don Kirkham and H.P. Johnson. 1957. Effect of tile
spacing on crop yield and water table level in a planosol soil. Soil Sci.Soc.
Amer. Proc. 21: 448-452.
61. J-3052. Willis, W.O., W.E. Larson and Don Kirkham. 1957. Corn growth as
affected by soil temperature and mulch. Agronomy Journal 49: 323-328.
62. J-3073. Adams, J.E., Don Kirkham and D.R. Nielsen. 1957. A portable rainfall
simulator, infiltrometer and physical measurements of soil in place. Soil Sci.
Soc. Amer. Proc. 21: 473-477.
63. J-3106. Kirkham., Don. 1957. Potential and capacity of concentric coaxial
capped cylinders. Journal of Applied Physics 28: 724-731.
64. J-3179. Kirkham, Don. 1958. Seepage of steady rainfall through soil into
drains. Trans. Amer. Geophys. Union 39: 892-908.
Don Kirkham 231
65. J-3247. Jackson, Ray D. and Don Kirkham. 1958. Method of measurement
of the real thermal diffusivity of moist soil. Soil Sci. Soc. Amer. Proc. 22:
479-482.
66. J-3254. Kirkham., Don. 1958. Theory of seepage into an auger hole above
an impermeable layer. Soil Sci. Soc. Amer. Proc. 22: 204-208.
67. Nestroy, O. 1958. Report of two lectures delivered in German by Don Kirkham.
Wasserhaushalt und Pflanzenwachstum. Oesterreichische Wasserwirtschaft.
Hochschule fur Bodenkultur, Vienna, Austria, March 17 and 18.
68. Kirkham., Don. 1958. Advanced Soil Physics. A set of twenty lectures. Soil
Science Institute, Ghent, Belgium (mimeographed), 147 pages.
69. J-3255. Burrows, W.C., and Don Kirkham. 1958. Measurement of field
capacity with a neutron meter. Soil Sci. Soc. Amer. Proc. 22: 103-105.
70. J-3329. Adams, J.E., Don Kirkham and W.H. Scholtes. 1958. Soil erodibility
and other physical properties of some Iowa soils. Iowa Journal of Science 32:
485-540.
71. J-3331. Scholtes, W.H. and Don Kirkham. 1959. The use of radiocarbon for
dating soil strata. (Belgian Journal of) Pedologie 7: 316-323.
72. Nielsen, D.R., Don Kirkham and R.E. Phillips. 1959. Synthetic soil conditioners:
Soil aggregates and corn yields. Iowa Farm Science 13: 8-10.
73. J-3483. Nielsen, D.R., Don Kirkham and W.C. Burrows. 1958. Solids-water-
air space relations of some Iowa soils. Iowa State College Journal of Science
33: 111-116.
74. J-3566. Kirkham, Don, M.F. DeBoodt and L. De Leenheer. 1959. Air
permeability at the field capacity as related to soil structure and yields.
Mededelingen van de Landbouwhogeschool van de Staat te Gent 24:
377-391.
75. J-3567. Kirkham, Don, M.F. DeBoodt and L. De Leenheer. 1959. Modulus of
rupture determination on cylindrical core samples. Mededelingen van de
Landbouwhogeschool van de Staat te Gent 24: 369-376.
76. J-3568. Kirkham, Don, L. De Leenheer and M.F. DeBoodt. 1959. Physical
measurements and yields on some loam and clay soils in Belgium.
Mededelingen van de Landbouwhogeschool van de Staat te Gent 24:
324-334.
77. J-3598. Maasland, Marinus and Don Kirkham. 1959. Measurement of the
permeability of tri-axially anisotropic soil. Proc. Amer. Soc. Civil Engineers,
Soil Mechanics and Foundations Division, No. 2063: 25-34.
78. Kirkham, Don, M.F. DeBoodt and L. De Leenheer. 1959. Modulus of rupture
determination on undisturbed soil core samples. Soil Sci. 87: 141-144.
79. Kirkham, Don. 1959. Review of Drainage Investigations and Design, Adana
Plain Irrigation and Drainage Plan. Republic of Turkey Ministry of Public
Works and Tippetts-Abbett-McCarthy-Stratton Engineers, Adana, Turkey and
New York City, (mimeographed), 109 pages.
232 Wolf Prize in Agriculture
80. Kirkham, Don. 1959. Lorentz, Einstein and the discovery of relativity, a
letter to the Editor. American Scientist 47: 232A, 236A.
81. Kirkham, Don. 1959. Abstract: Method of solving a class of mixed boundary
value problems. The Am. Math. Monthly 66: 627.
82. J-3610. Nielsen, D.R., Don Kirkham and W.R. van Wijk. 1959. Measuring
water stored temporarily about the field moisture capacity. Soil Sci. Soc.
Amer. Proc. 23: 408-412.
83. J-3644. Phillips, R.E., C.R. Jensen and Don Kirkham. 1960. Use of radiation
equipment for plow-layer density and moisture. Soil Science 89: 2-7.
84. J-3670. Kirkham, Don. 1959. Exact theory of flow into a partially penetrating
well. Jour. Geophys. Res. 64: 1317-1327.
85. J-3713. Nielsen, D.R., Don Kirkham and E. R. Perrier. 1960. Soil capillary
conductivity: comparison of measured and calculated values. Soil Sci. Soc.
Amer. Proc. 24: 157-160.
86. J-3795. Stone, J.F., R.H. Shaw and Don Kirkham. 1960. Statistical parameters
and reproducibility of the neutron method of measuring soil moisture. Soil
Sci. Soc. Amer. Proc.24: 435-438.
87. J-3796. Kirkham, Don. 1960. Seepage into ditches from a plane water table
overlying a gravel substratum. J. Geophys. Res. 65: 1267-1272.
88. J-3803. DeBoodt, M.F., L. De Leenheer and Don Kirkham. 1961. Soil aggregate
stability indexes and crop yields. Soil Science 91: 138-146.
89. J-3830. Kunze, R.J. and Don Kirkham. 1961. Deuterium and the self-diffusion
coefficient of soil moisture. Soil Sci. Soc. Amer. Proc. 25: 9-12.
90. J-3874. Grover., Ben L. and Don Kirkham. 1961. A glassbead-glycerol
model for non-steady-state tile drainage. Soil Sci. Soc. Amer. Proc. 25:
91-94.
91. J-3878. Grover, Ben L., J.T. Ligon and Don Kirkham. 1960. Operational
characteristics of the laterals near the edge of a tile drainage system.
J. Geophys. Res. 65: 3733-3738.
92. Kirkham, Don. 1960. Book review of Dr. H. Franz’s treatise “Feldbodenkunde.
Soil Sci. Soc. Amer. Proc. 24.
93. J-3902. Nielsen, D.R., Don Kirkham and W.R. van Wijk. 1961. Diffusion
equation calculations of field soil water infiltration profiles. Soil Sci. Soc.
Amer. Proc. 24: 165-168.
94. J-3904. Kirkham, Don. 1961. An upper limit for the height of the water
table in drainage design formulas. Proc. Int. Soil Sci. Soc., Seventh Congress,
Madison, Wisconsin, August, 1960. Commission VI, Vol 1: 486-492.
95. J-3919. Bartholomew, W.V. and Don Kirkham. 1961. Mathematical
descriptions and interpretations of culture induced soil nitrogen changes.
Proc. Int. Soil Sci. Soc., Seventh Congress, Madison, Wisconsin, August, 1960.
Commission III. Vol. 2: 471-477.
Don Kirkham 233
112. J-4214. Kunze, R.J. and Don Kirkham. 1962. Simplified accounting for
membrane impedance in capillary conductivity determinations. Soil Sci. Soc.
Amer. Proc. 26: 421-426.
113. J-4233. Kirkham, Don and R.J. Kunze. 1962. Isotope methods and uses in
soil physics research. Advances in Agronomy Vol. 14: 325-358.
114. J-4293. Zaslavsky, D. and Don Kirkham. 1964. The streamline function
for axially symmetric groundwater movenent. Soil Sci. Soc. Amer. Proc. 28:
150-164.
115. J-4370. Ligon, J.T., H.P. Johnson and Don Kirkham. 1963 Glass bead-glycerol
model for studying the falling water table between open ditch drains.
Agricultural Engineering 6: 61-64.
116. J-4373. Benoit, G.R. and Don Kirkham. 1963. The effect of soil surface
conditions on evaporation of soil water. Soil Sci. Soc. Amer. Proc. 27: 495-498.
117. J-4377. Ligon, J.T., H.P. Johnson and Don Kirkham. 1962. Unsteady-state
drainage of fluid from a vertical column of porous material. J. Geophys. Res.
67: 5199-5204.
118. J-4446. Kirkham, Don.1963. Soil physics research in some overdeveloped
and underdeveloped countries. Food for Peace: Amer. Soc. Agronomy Special
Publication No.l, pp. 71-76.
119. J-4475. Ligon, J.T., Don Kirkham and H.P. Johnson. 1964. The falling water
table between open ditch drains. Soil Sci. 97: 113-118.
120. J-4476. Jensen, C.R. and Don Kirkham. 1963. Labeled oxygen: increased
diffusion rate through soil containing growing corn roots. Science 141:
735-736.
121. J-4477. Jensen, C.R. and Don Kirkham. 1963. Oxygen-18 as a tracer in soil
aeration studies. Soil Sci. Soc. Amer. Proc. 27: 499-501.
122. J-4498. Kunze, R.J. and Don Kirkham. 1964. Capillary diffusion and
selfdiffusion of soil water. Soil Sci. 97: 145-151.
123. Radke, J.K., R.H. Shaw and Don Kirkham. 1963. Planting ridge, plastic mulch,
speed corn growth. Crops and Soils, p. 24, vol. 15, No. 8.
124. J-4586. Flannery, R.D. and Don Kirkham. 1964. A soil core water permeameter
for field use. Soil Sci. 97: 233-240.
125. J-4643. Kunze, R.J. and Don Kirkham. 1965. Models and equations for
determining DOH exchange and enrichment in plants. Agronomy Journal
57: 279-282.
126. J-4740. Kirkham, Don. 1964. Recent trends in soil science research in
the United States. Mededeligen van de Landbouwhogeschool en de
Opzoekingsstations van de Staat Gent, Deel XXIX, No. 1 pp. 1-19.
127. J-4748. Kirkham, Don. 1964. Some physical processes causing movement of
ions and other matter through soil. Mededeligen van de Landbouwhogeschool
en de Opzoekingsstations van de Staat Gent, Deel XXIX No. 1. pp. 21-42.
Don Kirkham 235
128. Presentation of Don Kirkham for honorary doctors degree, Ghent, Belgium.
Mededeligen van de Landbouwhogeschool en de Opzoekingsstations van de
Staat Gent, Deel XXVIII Nr. 3.
129. Grover, Ben L. and Don Kirkham. 1964. Solving tile drainage problems by
using model data. Agricultural Experiment Station, Iowa State University
Research Bulletin No. 523.
130. J-4791. Kirkham, Don. 1964. Exact theory for the shape of the free
water surface about a well in a semiconfined aquifer. J. Geophysical Res. 69:
2537-2549.
131. J-4801. Kirkham, Don. 1964. Physical artifices and formulas for
approximating water table fall in tile-drained land. Soil Sci. Soc. Amer. Proc.
28: 585-590.
132. J-4930. Kirkham, Don and W.L. Powers. 1964. An exact theory of seepage of
steady rainfall into tile and ditch drained land of finite depth. Trans. Intern.
Congr. Soil Sci. 9th (Bucharest, Aug. 29 - Sept. 10).
133. Kirkham, Don. 1963. A sum formula. Problem 5156. Am. Math. Monthly
70: 1107. See also 71: 1141-1142 and 72: 796-797.
134. J-5014. Kirkham, Don. 1964. Some recent land drainage investigations
at Iowa State University of Science and Technology, Ames, Iowa. U.S.A.
Proc. Sodic-soils symposium, Aug. 9-16, Hungarian Academy of Science
Suppletr)ent TOM. 14: 229-234.
135. J-5032. Toksöz, Sadik, Don Kirkham and E. Robert Baumann. 1965. Two-
dimensional infiltration and wetting fronts. Proc. Am. Soc. of Civ. Eng. 91:
65-79.
136. J-5048. Kirkham, Don. 1965. Seepage of leaching water into drainage ditches
of unequal water level heights. J. of Hydrology 3: 207-224.
137. J-5060. Kirkham, Don. 1966. Steady-state theories for drainage. J. Irrigation
and Drainage Division Proc. Am. Soc. Civil Engr. 92-19-39.
138. Kirkham, Don. 1967. Closure of discussion on “Steady-state theories for
drainage”. J. Irrigation and Drainage Division. IR 2, pp. 69-74. (Refers to
IR3, pp. 81-82 and IR4, pp. 75-88).
139. Kirkham, Don and W.L. Powers. 1965. Mathematical Soil Physics. (A set of
Advanced Soil Physics class notes).
140. J-5131. Zaslavsky, Dan and Don Kirkham. 1965. Streamline functions for
potential flow in axial symmetry. Amer. J. Physics. 33: 677-679.
141. J-5182. Corey, J.C. and Don Kirkham. 1965. Miscible displacement of
N-15 tagged nitrate and tritiated water in water-saturated and water-
unsaturated soil. Proceedings of the International Atomic Energy Agency
(IAEA) Symposium on the “Use of Isotopes and Radiation in Soil-Plant
Nutrition Studies” at Ankara, Turkey, June 28. Published by IAEA, Vienna
(Austria), pp. 157-170.
236 Wolf Prize in Agriculture
142. J-5196. Rogowski, A.S., W.D. Shrader, Don Kirkham and H.P. Johnson. 1965.
Tile drainage experimentation on Webster soils: Results of years 1954-1963.
Iowa State Journal of Science 41: 25-40. 1965.
143. J-5254. Asseed, Mohamed and Don Kirkham. 1966. Depth of barrier
and water table fall in a tile drainage model. Soil Sci. Soc. Amer. Proc. 30:
292-298.
144. J-5277. Hinesly, T.D. and Don Kirkham. 1966. Theory and flow nets for rain
and artesian water seeping into soil drains. Water Resources Research 2:
497-511.
145. Kirkham, Don and Floyd Andre. 1966. A study of some aspects of higher
education in Argentina (section on agriculture). International Institute of
Education, United Nations Plaza, New York.
146. J-5310. Kirkham, Don. 1965. Saturated conductivity as a characterizer of
soil for drainage design, in “Drainage for Efficient Crop Production,
Conference Proceedings, December, 1965, pp. 24-31”, published by Am.
Soc. of Agric. Engineers, St. Joseph, Michigan.
147. J-5312. Kirkham, Don. 1967. Physical model for Dupuit-Forchheimer
drainage theory. International Soil Water Symposium, June 5-11, Prague,
Czechoslovakia. Czechoslovak National Committee of the International
Commission on Irrigation and Drainage. 385-395.
148. J-5381. Corey, J.C., Don Kirkham and Donald R. Nielsen. 1967. The movement
of chloride and nitrate through certain Iowa soils. Iowa Academy of Science
74: 130-141.
149. Kirkham, Don and W.L. Powers. 1966. Factors influencing hydraulic
conductivities of soils, etc., in “Environmental Biology”, published by
Federation of Amer. Societies for Experimental Biology, 9650 Rockville Pike,
Bethesda, Maryland, pp. 462-463.
150. J-5433. Corey, J.C., D.R. Nielsen, J.C. Picken, Jr. and Don Kirkham. 1967.
Miscible displacement through gamma radiation-sterilized soil columns.
J. of Environmental Science 1: 144-147.
151. J-5469. Kirkham, Don. 1967. Explanation of paradoxes in Dupuit-
Forchheimer seepage theory. Water Resources Research 3: 609-622.
152. J-5469. Kirkham, Don. 1967. Explanation of paradoxes in Dupuit-
Forchheimer seepage theory. Reprinted from Engr. News 12: 22-39. Quarterly
J. of West Pakistan Engr. Congreso, Lahore, West Pakistan.
153. J-5520. Powers, W.L., Don Kirkham and Gretchen Snowden. 1967. Seepage
of steady rainfall through soil into ditches of unequal water level heights.
Soil Sci. Soc. Amer. Proc. 31: 201-312.
154. Kirkham, Don. 1966. The use of isotopes in soil physics research. Abstract
llth Pacific Science Congress, Tokyo, Japan. Symposium 34. 1966.
Don Kirkham 237
155. J-5552. Powers, W.L., Don Kirkham and G. Snowden. 1967. Orthonormal
function tables and the seepage of steady rain through soil bedding.
J. Geophysical Res. 72: 6225-6237.
156. J-5572. Corey, J.C., D.R. Nielsen and Don Kirkham. 1967. Miscible
displacement of nitrate through soil columns. Soil Sci. Soc. Amer. Proc. 31:
497-501.
157. J-5593. Fritton, D.D., Don Kirkham and R.H. Shaw. 1967. Soil water and
chloride redistribution under various evaporation potentials. Soil Sci. Soc.
Amer. Proc. 31: 599-603.
158. Kirkham, Don ( Joint Editor). 1966. Plant environment and efficient water
use. American Society of Agronomy (ASA) and Soil Science Society of America
(SSSA). 295 pages.
159. J-5651. Kirkham, Don, D.E. Rolston and D.D. Fritton. 1967. Gamma-radiation
detection of water content in two-dimensional evaporation prevention
experiments. In “Isotope and Radiation Techniques in Soil Physics
and Irrigation Studies”, Istanbul, Turkey, June 12-16, 1967. Published by
International Atomic Energy Agency, Vienna (Austria), 1: 3-16. 1967.
160. J-5770. Kirkham, Don and Helmy Bakr. 1969. Some recent research on land
drainage. Proceedings Int. Land Reclamation Symposium, Yerevan, Armenia,
U.S.S.R. May 25-31.
161. Similar to J-5770 except published 1971 in Russian in The Transactions of
the Research Institute and Soil Science and Agrochemistry 6: 693-704,
Yerevan, Armenia.
162. J-5776. Rogowski, A.S., W.C. Moldenhauer and Don Kirkham. 1968.
Rupture parameters of soil aggregates. Soil Sci. Soc. Amer. Proc. 32:
720-724.
163. J-5796. Mansell, Robert S., Donald R. Nielsen and Don Kirkham. 1968. A
method for simultaneous control of aeration and unsaturated water movement
in laboratory soil columns. Soil Science 106: 114-121.
164. J-5807. Warrick, A.W. and Don Kirkham. 1968. Determination of equivalent
radii for half-tube and whole-tube drains in contact with an impermeable
barrier. Soil Sci. Soc. Amer. Proc. 32: 449-451.
165. J-5812. Kirkham, Don. 1968. Reply of the author to a note by E. C. Childs
and E. G. Youngs on the paper “Explanation of paradoxes in Dupuit-
Forchheimer seepage theory.” Water Resources Research 4:221-222.
166. J-5850. Kirkham, Don. 1969. Use of nuclear energy in water studies for
increasing agricultural productivity. Article in Proceedings of Conference,
“Application of Nuclear Energy to Increased Agricultural Productivity,”
Santiago, Chile, 9-12 January, 1968, of Interamerican Commission of Nuclear
Energy of Pan American Union, pp. 11-35, Washington, D.C.
238 Wolf Prize in Agriculture
167. Asseed, Mohamed and Don Kirkham. 1968. Advance of irrigation water
on the soil surface in relation to soil infiltration rate: A mathematical and
laboratory model study. Iowa Ag. & Home Ec. Expt. Station Res. Bul. 565.
168. Fritton, Daniel D. (single author, part of his Ph.D. thesis under Don Kirkham).
1969. Resolving time, mass absorption coefficient and water content with
gamma-ray attenuation. Soil Sci. Soc. Amer. Proc. 33: 651-655.
169. Kirkham, Don. Technique for solving mixed boundary value problems. (Being
reworked).
170. J-6099. Warrick, A.W. and Don Kirkham. 1969. Two-dimensional
seepage of ponded water to full ditch drains. Water Resources Research 5:
685-693.
171. Kirkham, Don. 1968. Book Review of “Physical Principles of Water Percolation
and Seepage”, UNESCO, Paris, by Bear, J. D., D. Zaslavsky and S. Irmay. Trans.
Amer. Geophys. Union. 49: 561. 1968.
172. J-6101. Rolston, Dennis E., D.R. Nielsen and Don Kirkham. 1969. Miscible
displacement of gases through soil columns. Soil Sci. Soc. Amer. Proc. 33:
488-492.
173. J-6240. Kirkham, Don. 1970. The importance of water resources research.
Symposium “Water Resources of Iowa”, Iowa Academy of Science, April
1969; published by Iowa Academy of Science, pp. 159-175. 1970.
174. J-6241. Srinilta, Sam-Arng, D.R. Nielsen and Don Kirkham. 1969. Steady
flow of water through a two-layer soil. Water Resources Research 5:
1053-1063.
175. J-6250. Selim, H., Don Kirkham and M. Amemiya. 1970. A comparison of
two methods for determining soil water diffusivity. Soil Sci. Soc. Amer. Proc.
34: 14-18.
176. J-6272. Fritton, D.D., Don Kirkham and R. H. Shaw. 1970. Soil-water
evaporation, isothermal diffusion, and heat and moisture transfer. Soil Sci.
Soc. Amer. Proc. 34: 183-189.
177. J-6279. Selim, H.M., and Don Kirkham. 1970. Soil temperature and water
content changes under drying as influenced by cracks, a laboratory
experiment. Soil Sci. Soc. Amer. Proc. 34: 565-569.
178. J-6302. Toksöz, Sadik, and Don Kirkham. 1971. Steady drainage of layered
soils: I. Theory. Journal of the Irrigation and Drainage Division, Proceedings
of the American Society of Civil Engineers 97: IR1: 1-18.
179. J-6335. Toksöz, Sadik, and Don Kirkham. 1971. Steady drainage of layered
soils: II. Nomographs. Journal of the Irrigation and Drainage Division,
Proceedings of the American Society of Civil Engineers 97: IR1: 19-37.
180. J-6402. Mansell, R.S., Don Kirkham and D.R. Nielsen. 1970. Nitrate and
detergent recovery in aerated soil columns. Soil Sci. Soc. Amer. Proc. 34:
883-889.
Don Kirkham 239
181. Rechard, Paul, and Don Kirkham. 1970. The university’s role in National
Water Policy, Discussion Report, Group 3 “Should the universities only be in
the business of extending methodology and procedures, or should they assist
in their application?” Proceedings of a Conference of the University Council
of Water Resources, Blacksburg, Virginia, July 27-29 p. 63.
182. J-6602. Khan, M.Y., Don Kirkham, and Sadik Toksöz. 1971. Steady
state flow around a well in a two-layered aquifer. Water Resour. Res. 7:
155-165.
183. J-6642. Khan, M.Y., Don Kirkham. 1971. Spacing of drainage wells in a
layered aquifer. Water Resour. Res. 7: 166-183.
184. J-6691. Yoo, Sun-Ho, and Don Kirkham. 1971. Flow cell system for miscible
displacement experiments. Water Resour. Res. 7: 211-213. U.S. AEC Contract
AT-(11-1)-1269. Report C00-1269-23.
185. J-6721. van der Ploeg, R.R., Don Kirkham, and C.W. Boast. 1971. Steady-
state well flow theory for a confined elliptical aquifer. Water Resour. Res. 7:
942-954.
186. J-6752. Boast, C.W., and Don Kirkham. 1971. Auger hole seepage theory.
Soil Sci. Soc. Amer. Proc. 35: 365-373.
187. J-6777. Kirkham, Don, and J.C. Corey. 1973. Recent progress in the design
of radiation equipment and its practical implications. Proceedings of the
International Atomic Energy Agency’s Symposium on “Soil-Moisture and
Irrigation Studies II” at Vienna, Austria, in Nov., 1970, published by Int.
Atomic Energy Agency, Vienna, Austria, pp. 17-39.
188. J-6886. Mulqueen, John, and Don Kirkham. 1972. Leaching of sodium
chloride into tile drains in a sand-tank model. Soil Sci. Soc. Amer. Proc. 36:
3-9.
189. Boersma, L.L., Don Kirkham, E.B. Norum, R. Ziemer, J.C. Guitjens, J. Davidson,
and J.N. Luthin. 1971. Soil Moisture. Trans. Amer. Geophys. Union 57:
279-285.
190. J-6957. Kirkham, Don, and R.R. van der Ploeg. 1974. Ground-water flow
patterns in confined aquifers and pollution. J. Amer. Water Works Assn.
66(3): 192-197.
191. J-6963. Selim, H.M., and Don Kirkham. 1973.Unsteady two-dimensional
flow of water in unsaturated soils above an impervious barrier. Soil Sci. Soc.
Amer. Proc. 37: 489-495.
192. J-6973. van der Ploeg, R.R., Don Kirkham, and L.V.A. Sendlein. 1973.
Groundwater flow patterns of the Ames aquifer. Proc. Iowa Acad. of Sci.
80(2): 103-110.
193. J-6994. Selim, H.M., and Don Kirkham. 1974. Unsteady state two-dimensional
water content distribution and wetting fronts in soils. Geoderma 11:
259-274.
240 Wolf Prize in Agriculture
194. J-6995. Selim, M.S., and Don Kirkham. 1972. Seepage through soil bedding
or a hillside due to a steady rainfall: I. Soil surface of constant slope. Soil Sci.
Soc. Amer. Proc. 36: 402-407.
195. J-7001. Selim, M.S., and Don Kirkham. 1972. Seepage through soil bedding
or a hillside due to a steady rainfall: II. Soil surface of arbitrary shape. Soil
Sci. Soc. Amer. Proc. 36: 407-412.
196. Kirkham, Don. 1973. Book Review of “Soil Physics (fourth edition)”, John
Wiley, New York, by Baver, L.D., Gardner, Walter H., and Gardner, Wilford
R. Trans. Amer. Geophys. Union (EOS) 54(4): 185.
197. J-7031. Selim, H.M., M. Sami Selim, and Don Kirkham. 1975. Mathematical
analysis of steady saturated flow through a multilayered soil with a sloping
surface. Soil Sci. Soc. Amer. Proc. 39 (May-June): 445-453.
198. J-7086. Kirkham, Don. 1971. Isotopes and radiation in relation to soil
moisture, irrigation, and efficient water use. In Proccedings, International
Symposium on Use of Isotopes and Radiation in Agriculture and Animal
Husbandry Research. New Delhi, India. Pp. 380-395.
199. J-7106. Timmons, J.F., Don Kirkham, and others. 1972. Soil Science in
relation to water resources development: V. Economics evaluation of water
resources data. Water Committee Paper (S878), Soil Sci. Soc. Amer. Proc. 36:
186-194.
200. J-7171. Khan, M.Y., and Don Kirkham. 1972. Reply. Water Resour. Res. 8:
1124-1127.
201. Kirkham, Don, and W.L. Powers. 1972. Advanced Soil Physics, 534 pp.
Wiley-Interscience, New York.
202. J-7343. Gabriels, D.M., W.C. Moldenhauer, and Don Kirkham. 1973.
Infiltration, hydraulic conductivity, and resistance to ater-drop impact of
clod beds as affected by chemical treatment. Soil Sci. Soc. Amer. Proc. 37:
634-637.
203. J-7448. Kirkham, Don, and M. Sami Selim. 1973. Theory of a gravel envelope
in drainage design. Soil Sci. Soc. Amer. Proc. 37: 517-521.
204. J-7590. Kirkham, Don. 1973. Soil physics and soil fertility. In Proceedings
International Soil Fertility Week. Agronomical Research Center, Gembloux,
Belgium, Sept. 2-7, pp.60-88. Also in Bulletin des Recherches Agronomiques
de Gembloux Faculte des Sciences Agronomique de l’Etat, n.s. Vol. 8(2):
60-88.
205. J-7652. Selim, M.S., and Don Kirkham. 1973. Reply. Seepage through soil
bedding or a hillside due to a steady rainfall. Soil Sci. Soc. Amer. Proc. 37:
817.
206. J-7665. Selim, M.S., and Don Kirkham. 1974. Screen theory for wells and
soil drain pipes. Water Resour. Res. 10: 1019-1030.
Don Kirkham 241
207. J-7691. Kirkham, Don, and R.R. van der Ploeg. 1973. Potential and flow fields
for multiple groundwater wells in a confined aquifer. Proceedings Symposium
on Development of Groundwater Resources, College of Engineering, University
of Madras, Madras, India, Vol. 2, part IV, pp. 153-164. (Nov. 26-29, 1973,
Congress dates).
208. Kirkham, Don. 1973. Iowa. (An annual report of the Iowa State Water
Resources Research Institute for 1973). In 1973 Annual Report, Cooperative
Water Resources Research and Training. Office of Water Resources Research,
U.S. Department of the Interior. Pp. 82-83. (Earlier Iowa Reports are in
earlier volumes 1964 to 1972).
209. J-7772. Kirkham, Don, Sadik Toksöz, and R.R. van der Ploeg. 1974. Steady
flow to drains and wells, p.203-243. In Jan van Schilfgaarde (ed.) Drainage
for Agriculture. Agron. Monogr. 17, ASA and SSSA, Madison, Wisconsin.
210. J-7804. Powell, N.L., and Don Kirkham. 1974. Flow patterns of steady rainfall
seeping through bedded land or a hillside with a barrier at great depth.
J. Hydrol. 23: 203-217.
211. J-7823. Powell, N.L., and Don Kirkham. 1976. Tile drainage in a bedded soil
or draw. Soil Sci. Soc. Amer. J. 40: 625-630.
212. J-8169. Khan, M.Y., Don Kirkham, and R.L. Handy. 1976. Shapes
of steady state perched groundwater mounds. Water Resour. Res. 12(3):
429-436.
213. J-8169a. Khan, M.Y., Don Kirkham, and R.L. Handy. 1977. Reply. Water
Resour. Res. 13(2): 503.
214. J-8304. Rogowski, A.S., and Don Kirkham. 1976. Strength of soil aggregates:
influence of size, density, and clay and organic matter content. Proc. Int.
Symp. On Soil Conditioners. Ghent University, Belgium, 8-12 December,
1975. Also (with Discussion) in Med. Fac. Landbouw. Rijksuniv. Gent 41/1-
1976, pp. 85-100.
215. J-8428. Affleck, S.B., Don Kirkham, and W.F. Buchele. 1976. Seedbed
preparation for optimum temperature, moisture, aeration and mechanical
impedance. Proceedings of the 7th Conference of the International Soil Tillage
Research Organization, ISTRO. Report No. 45, 1976, of the Division of Soil
Management, Agricultural College of Sweden, S-750 07. Uppsala.
216. J-8521. Kirkham, Don, and S.B. Affleck. 1977. Solute travel times to wells.
Ground Water 15: 231-242.
217. J-8611. Najmaii, M., Don Kirkham, and M.D. Dougal. 1978. Tube drainage
in stratified soil above an aquifer. Journal of the Irrigation and Drainage
Division, ASCE (Proceedings paper 13829) 104 (No. IR2): 209-228.
218. J-8716. Cushman, John, and Don Kirkham. 1978. Solute travel times to
wells in single or multiple layered aquifers. J. Hydrol. 37: 169-174.
242 Wolf Prize in Agriculture
219. J-8723. Kirkham, Don, and Lyle Prunty. 1977. Upslope recharge, downslope
waterlogging and interceptor drains. The Third National Drainage Symposium
Proceedings, American Society of Agricultural Engineers. P.O. Box 410, St.
Joseph, Michigan 49085.
220. J-8824. Mousavi, S.F., and Don Kirkham. 1978. Porous media tests of
groundwater mounds. Soil Science 125: 160-164.
221. J-8835. Kirkham, Don. 1976. Soil Physics and Plant Growth. In Anuario del
Centro de Edafologia y Biologia Aplicada de Salamanca.
222. Kirkham, Don. 1978. Water, our prime resource. Commencement Address,
Iowa State University, February 25, 1978. (Unpublished).
223. J-8853. Cushman, John, and Don Kirkham. 1978. A two-dimensional
linearized view of one dimensional unsaturated-saturated flow. Water Resour.
Res. 14: 319-323.
224. J-8865. Kirkham, Don, and M. Olga Sotres. 1978. Casing depths and solute
travel times to wells. Water Resour. Res. 14: 237-243.
225. Kirkham, Don. 1979. Some concepts and misconcepts of drainage theory.
Virgil Overholt Drainage Hall of Fame Award talk, at Ohio State University,
March 9, 1979, Columbia, Ohio. (Unpublished).
226. J-8881. Battikhi, Anwar, and Don Kirkham. 1979. Casing and leak depths,
and solute travel times to wells. Water Resources Bulletin 15(4): 1004-1015.
227. J-9111. Kirkham, Don, and Olga Sotres. 1979. Influence of cover slab
diameter on solute travel time to wells. Water Resour. Res.15(4): 941-948.
228. J-9126. Prunty, Lyle, and Don Kirkham. 1979. A drainage model for a
reclaimed surface mine. Soil Sci. Soc. Amer. J. 43: 28-34.
229. J-9339. Cushman, John H., Don Kirkham, and Roy F. Keller. 1979. A Galerkin
in time, linearized finite element model of 2-dimensional unsaturated porous
media drainage. Soil Sci. Soc. Amer. J. 43: 638-641.
230. J-9369. Prunty, Lyle, and Don Kirkham. 1980. Seepage versus terrace density
in reclaimed mineland soil. Journal of Environ. Qual. 9: 273-278.
231. J-9518. Henning, Stanley J., Don Kirkham, and Stephen B. Affleck. 1979.
Proceedings of the 8th Conference of the International Soil Tillage Research
Organization, ISTRO, Bundesrepublik Deutschland. Stuttgart, Hohenheim,
Germany. Sept. 8-15, 1979, Vol. 1: 103-108.
232. No J-no.. Kirkham, Don. 1980. Soil Physics. In Encyclopedia of Physics,
pp. 927-928. Addison-Wesley Publishing Co., Reading, Mass.
233. J-9924. Kanwar, R.S., J.L. Baker, H.P. Johnson, and Don Kirkham. 1980.
Nitrate movement with zero-order denitrification in a soil profile. Soil Sci.
Soc. Amer. J. 44: 898-902.
234. J-10055. Kanwar, R.S., H.P. Johnson, and Don Kirkham. 1982. Transport of
nitrate and gaseous denitrification in soil columns during leaching. J. Hydrol.
55: 171-184.
Don Kirkham 243
235. J-10221. Troeh, Frederick R., Jalal D. Jabro, and Don Kirkham. 1982. Gaseous
diffusion equations for porous materials. Geoderma 27: 239-243.
236. Brandyk, Tomasz, and Don Kirkham. 1982. Examination of diffusivity
theory for muck and sands. Annals of Warsaw Agricultural University, Land
Reclamation 19: 3-8.
237. J-10676. Kirkham, Don. 1982. Free surface potential theory for a gravity
well. In Papers International Symposium. Polders of the World, Vol. II,
pp. 13-23. International Institute for Land Reclamation and Improvement,
Wageningen, The Netherlands.
238. Mousavi, S.F., and Don Kirkham. 1982. Mineland seepage and its control.,
Iran Agricultural Research 1 (no. 2): 164-179.
239. Powers, W.L., and Don Kirkham. 1984. Advanced Soil Physics, revised edition,
539 pp., Robert E. Krieger Publishing Company, Inc., Malabar, Florida 32950.
240. J-11127. Modaihsh, Abdullah Saad, Robert Horton, and Don Kirkham.
1985. Soil water evaporation suppression by sand mulches. Soil Science 137:
357-361.
241. On my lecture tour of the People’s Republic of China 1985, I was presented
two volumes of translations into Chinese of sections from Kirkham and
Powers Advanced Soil Physics, Vol. I, October, 1980, dealt with Unsaturated
Water Flow; Vol. II, 1982, dealt with Saturated Water Flow. The publisher
was Tsinghua University, Department of Hydraulics, Water Resources,
Irrigationand Drainage Division. Beijing, People’s Republic of China. I did
not know until 1985 about these published translations.
242. J-12554. Kirkham, Don, and Robert Horton. 1986. Subirrigation with drainage
for drought-prone areas. Proceedings, International Conference of Water
Resources Needs and Planning in Drought-Prone Areas, Khartoum,
Sudan, Dec. 6-12. Published by Khartoum University Press. Designated a
“distinguished” paper, pp. 957-979.
243. J-12603. Walczak, R.T., R.R. van der Ploeg, and Don Kirkham. 1988. An
algorithm for the calculation of drain spacings for layered soils. Soil Sci. Soc.
Amer. J. 52: 336-340.
244. J-13651. Kirkham, Don, and Robert Horton. 1989. Groundwater management
by a dual-pipe subirrigation system. Proceedings of the Benidorm (Spain)
Symposium Oct. 1989 of the International Association of Hydrological
Sciences (IAHS) Publ. 188, pp. 53-56.
245. Kirkham, Don. 1991. Article: Soil Physics. P. 1124-1126. In Rita G. Lerner
and George L. Trigg (eds.) Encyclopedia of Physics. VCH Publishers, Inc.,
New York.
246. J-13907. Kirkham, Don, and Robert Horton. 1992. The stream function of
potential theory for a dual-pipe subirrigation-drainage system. Water Resour.
Res. 28(2): 373-387.
244 Wolf Prize in Agriculture
247. J-13655. Kirkham, Don, and Robert Horton. 1993. Modeling water flow
from subirrigation with drainage. Soil Sci. Soc. Amer. J. 57: 1451-1457.
248. Kirkham, Don, R.R. van der Ploeg, and R. Horton. 1997. Potential theory
for dual-depth subsurface drainage of ponded land. Water Resour. Res. 33:
1643-1654.
249. van der Ploeg, R.R., Maria Marquardt, and Don Kirkham. 1997. On the
history of the ellipse equation for soil drainage. Soil Sci. Soc. Am. J. 61:
1604-1606.
250. van der Ploeg, R. R., R. Horton and Don Kirkham. 1999. Steady flow to
drains and wells. p.213-263. In: R.W. Skaggs and J. van Schilfgaarde (eds.)
Agricultural Drainage. Agronomy Monograph No. 38, ASA/CSSA/SSSA,
Madison (WI), USA
Robert H. Burris
University of Wisconsin
Madison, Wisconsin, USA
k
SCIENTIFIC BACKGROUND
In the air above every hectare of land there are about 78 tons of elemental
nitrogen. The latter is chemically inert and, under ordinary conditions, does not
react with other elements. Hence most crop plants are likely to be starving in this
sea of nitrogen. Only when the nitrogen in the air is combined with other elements
it is possible for plants to use this nitrogen in their growth processes.
The mere fact that some bacteria are able to use the elemental form of nitrogen
that exists in the atmosphere has been known for nearly 100 years since its
discovery by the Russian biochemist S. N. Vinogradsky. Thus, these nitrogen-fixing
bacteria can grow in the absence of combined nitrogen and, at the same time,
produce nitrogenous substances in the soil that may be used later by crop plants.
However, during the first fifty years since the discovery of biological nitrogen
fixation by soil-inhabiting bacteria, very little information was available as regards
the biochemical mechanisms involved. This was quite surprising in view of the
paramount importance of this process in the nitrogen budget of our globe.
The state of the knowledge regarding this wonder of nature has changed
dramatically in the late 1930’s and early 1940’s when Professor Burris investigations
on the basic chemistry and physiology of the microorganisms that fix nitrogen,
actually opened up an entire new vista of basic research on biological nitrogen
245
246 Wolf Prize in Agriculture
fixation. He was the first to use a radioactive isotope of nitrogen (15N) in this
study, which led to the discovery of ammonia to be the key intermediate in this
biochemical process.
For over 45 years, following these pioneering efforts, Professor Burris and his
colleagues have purified and studied in great detail and depth the complex of
enzymes involved, and the metabolic pathways followed in this natural phenomenon.
At the same time they developed novel techniques for reliable, quantitative assay of
the nitrogen fixing activity of microorganisms.
The scope of his research in this particular area has not been limited to the
well recognized nitrogen-fixing bacteria that live in symbiosis with leguminous,
plants in nodules formed on their roots. His studies also embraced the free-living
microorganisms that are able to fix atmospheric nitrogen. Mention should be
made in this connection of Prof. Burris pioneering work on Azospirillum — a
free-living nitrogen-fixing genus of bacteria associated with plant roots, in particular
cereal plants.
One may sum up Burris’ contributions to fundamental biochemical research
on nitrogen fixation by concluding that his very careful, thorough investigations
have been in the heart of the efforts to elucidate the mechanism of this life-
supporting process. He has thus rightly earned the title of ‘world doyen of nitrogen
fixation’.
Because of the pressing food needs of the world on one hand, and the rising
cost of chemical nitrogen fixation for use in mineral fertilizers on the other, the
subject of biological nitrogen fixation is currently receiving greatly increased
attention throughout the world.
The lifelong fundamental studies of Prof. Burris provided the scientific
background for the now worldwide practice of inoculating legume crops with
industrially manufactured pre-cultured strains of nitrogen-fixing bacteria. The
latter are capable of supplying essentially all the nitrogen requirements of the
plan. Furthermore, the highly sensitive techniques developed by Prof. Burris for
measuring this biological activity, proved most instrumental in the selection of
more effective strains of bacteria for this purpose.
More recently, a new science-based biotechnology industry has been established
for the production of cultures of the aforementioned Azospirillum. These are
intended for inoculation of graminaceous crops in order to replace mineral nitrogen
fertilizers. Enhanced yields of these important staple food crops of the world can
thus be achieved at a remarkably reduced cost.
CURRICULUM VITAE
Date and Place of Birth: April 13, 1914, Brookings, South Dakota
Robert H. Burris 247
EDUCATION:
South Dakota State College, B.S. 1936 (Chemistry)
University of Wisconsin - Madison, M.S. 1938 (Bacteriology)
University of Wisconsin - Madison, Ph.D. 1940 (Bacteriology)
POSITIONS:
National Research Council Postdoctoral Fellow, Columbia University, 1940-41
Postdoctoral Appointment in ~Bacteriology, University of Wisconsin, 1941-44
Assistant Professor of Biochemistry, University of Wisconsin, 1944-46
Associate Professor of Biochemistry, University of Wisconsin, 1946-51
Professor of Biochemistry, University of Wisconsin, 1951-date
Guggenheim Fellow, University of Helsinki (Finland) and Cambridge University
(England), 1954
Chairman, Department of Biochemistry, University of Wisconsin, 1958-70
W. H. Peterson Professor of Biochemistry, 1976-date
PROFESSIONAL SOCIETIES:
National Academy of Sciences (Chairman, Section of Botany 1971-74; Executive
Committee, Assembly of Life Sciences 1973-78; Chairman, Division of
Biological Sciences 1977-78)
American Academy of Arts and Sciences
American Association of the Advancement of Science (Fellow)
American Society of Biological Chemists
American Chemical Society
American Society of Plant Physiologists (President, 1960)
American Society of Microbiologists
Biochemical Society (England)
American Philosophical Society
Japanese Society of Plant Physiologists
Sigma Xi
Gamma Alpha
Phi Sigma
1924–1999
Sir Ralph Riley led a research group, which increased precision of plant breeding
by experimentally describing the chromosomal architecture of wheat and related
species, and by comparing genetic activities of corresponding chromosomes. During
investigations which Dr. Riley started in 1958, he and his colleagues described the
genetic systems by which pairing of wheat chromosomes at meiosis is limited to
those which are fully homologous, and by which pairing between distantly related
chromosomes is precluded. This basic knowledge allowed Dr. Riley to pair and
recombine chromosomes in a way that is normally illegitimate.
CURRICULUM VITAE
Born 23 October 1924;
married 1949 Joan Elizabeth Norrington.
Education:
Audenshaw Grammar School; University of Sheffield.
251
252 Wolf Prize in Agriculture
Services:
1943-47 - Army; 1944, commissioned.
Served in 6th King’s Own Scottish Borders (15th Division) and 1st
South Lancashire Regiment (1st Division).
Final rank - Captain.
University:
1947-50 - University of Sheffield, B.Sc. 1st Class Honours (Botany)
Degrees:
1955 - Ph.D., University of Sheffield
1964 - D.Sc., University of Sheffield
1967 - M.A., University of Cambridge (Statute B, 111, 6).
Career:
1950-51 - Research student, Department of Botany, University of Sheffield, on
DSIR Studentship.
1951-52 - Demonstrator, Department of Botany, University of Sheffield
1952-78 - Staff of the Plant Breeding Institute, Cambridge:
from 1955-71 - Head of Cytogenetics Department
from 1971-78 - Director of the Institute
Since 1978 - Secretary to the Agricultural Research Council, London.
Distinctions:
1965 - Fellow of the Institute of Biology,
1967 - Fellow of The Royal Society
1967 - Fellow of Wolfson College, Cambridge
1969 - William Bate Hardy Prize of the Cambridge Philosophical Society
1973 - Sir Henry Tizard Memorial Lecturer
1973-75 - Member of the Council of The Royal Society
1973-75 - President of the Genetical Society
1975 - Foreign Fellow Indian National Science Academy
1976 - Woodhull Lecturer, Royal Institutions
1977 - Nilsson-Ehle Lecturer, Swedish Mendelian Society
Social Appointments:
1970-78 - Special Professor of Botany in the University of Nottingham.
Sir Ralph Riley 253
Editorial Activities:
Geneitical Research
Caryologia
Wheat Information Service
Theoretical and Applied Genetics
Current Advances in Plant Science
Family Circumstances: Married with two daughters
ESSENTIAL BIOGRAPHY
Appointments:
Plant Breeding Institute, Cambridge, from 1952 to 1978, head of cytogenetics
department 1954-72, director 1971-78; special professor of botany, University of
Nottingham, 1970-78; secretary, Agricultural Research Council since 1978. Fellow
of Wolfson College, Cambridge.
Offices held:
President, Genetical Society, 1973-75; Secretary, International Genetics Federation,
1973-78.
Relevant publications
R. Riley and V. Chapman, Genetic control of the cytologically diploid behaviour of
hexaploid wheat. Nature Lond. 182: 713-715, 1958.
R. Riley, Diploidization of polyploid wheat. Heredity, 15: 407-429, 1960.
R. Riley, V. Chapman and R. Johnson, The incorporation of alien disease resistance
in wheat by genetic intergerence with the regulation of meiotic chromosome
synapsis. Genet. Res., Camb. 12: 199-219, 1968.
SCIENTIFIC CONTRIBUTIONS
A principal achievement of Dr. R. Riley has been to device and use a form of
genetic engineering by which useful genetic variation from related species of wild
grass can be incorporated in cultivated wheat. Relevant findings arose from
cytogenetic investigations which commenced in 1952 at the Plant Breeding Institute,
Cambridge, which is supported financially by the Agricultural Research Council,
London. The research of Riley’s group had the general purpose of increasing the
precision of plant breeding and improving its rationale. Components of the work
included the experimental description of the chromosomal architecture of wheat
and related species and the comparison of the genetic activities of corresponding
chromosomes of wheat and of its relatives.
During these investigations, starting in 1958 Riley and his colleagues discovered
and described the genetic systems by which the pairing of chromosomes at meiosis
in wheat is limited to those which are fully homologous and by which pairing
Sir Ralph Riley 255
1910–1999
Ernest R. Sears synthesized for the first time in 1946 a hexaploid wheat. In 1950,
he completed work, which established the monosomic series in wheat, and later
created a mullisomic series, a trisomic series, and a telesomic series of chromosomes
in wheat. These have greatly enhanced development and breeding of modern
wheat varieties.
CURRICULUM VITAE
EDUCATION:
B.S. Oregon State College 1932
Ph.D. Harvard University 1936
U.S.D.A. Geneticist:
1936- Professor of Genetics, University of Missouri, Columbia
257
258 Wolf Prize in Agriculture
E.R. Sears led the pioneering effort in transferring rust resistance from aegilops to
common wheat by a unique procedure of hybridization and X-ray-induced
translocations. According to the estimates of the Texas Research Foundation of
Renner, Texas, the resistance factor that E R Sears introduced into the hexaploid
wheat can save from destruction 100 million bushels of wheat annually in
the United States alone. For this achievement the Texas awarded Dr. Sears
with the $10,000 Hoblitzelle National Award in the Agricultural Sciences
in 1958.
All of the scientific recognitions of E. R. Sears cannot be listed here; only the
major ones are mentioned:
There can be no question in the mind of anyone who is familiar with the
recent status of agriculture and genetics that Dr. E. R. Sears has made
one of the most serious impacts on the cultural and economic history of
mankind.
E. R. Sears has excelled, not only as a brilliant scientist and as an effective
teacher, but also as a great humanitarian. He is one of the most modest and
compassionate of men with the highest ethical standards.
Ernest R. Sears 259
A very quick comparison indicates that yield per area units is variable among these
species. These figures also indicate where the greatest possibilities for genetic
improvement exist. In order to realize the biological potentials of plant breeding,
scientifically well-founded and practical methods are required.
E. R. Sears, working for over forty years as a U.S.D.A. geneticist at the Missouri
Agricultural Experiment Station, has made far greater contributions to the scientific
development of wheat breeding than any other basic scientist, past or present.
In 1946 E. R. Sears (with E. S. McFadden) synthesized for the first time a
hexaploid wheat, thus laying down the foundations of experimental evolutionary
work in the species.
In the 1950’s Dr. Sears completed the monosomic series of wheat. This material,
available to research workers around the world, has remained the most important
tool of basic studies on this plant. Thus Dr. Sears is truly the founder of genetic
analysis at the chromosomal level in the allopolyploids.
E. R. Sears’ basic work on monosomics led him to the solution of the problem
of chromosome substitution. In the last few years the chromosome substitution
method has become the most powerful method of breeding hexaploid wheat, as
well as tobacco, cotton, oats, etc. It is now possible to introduce single isolated
chromosomes into an agronomically useful variety from other valuable strains.
Actually, practical techniques are available for the transfer of scientifically selected
desirable gene complexes, rather than entire chromosomes, from one variety to
another without losing any of the useful traits of the recipient. Never before in the
history of plant breeding, has it been possible to synthesize a favorable genetic
constitution in such a predictable manner.
This synopsis does not permit an adequate expositions of the theory or practice
of the development of substitution lines. It may suffice to mention that in 1971 the
European Wheat Aneuploid Cooperation was formed with the participation of
practically all European nations (including West and East) for the sole purpose of
implementing these scientific discoveries based upon E.R. Sears’ genetic research
on wheat.
260 Wolf Prize in Agriculture
In the late 1950’s E. R. Sears and one of his students, M. Okamoto, discovered
that the pairing of the wheat chromosomes is controlled primarily by the Ph gene
of the long arm of chromosome 5B. The recognition of this basic biological principle
opened up a most rewarding procedure for introducing useful genes into wheat
from the immense pool of wild relatives of wheat. Within a few years after this
discovery numerous disease-resistant varieties of wheat were developed and
introduced into commercial production in the United States as well as in several
other countries. These first success stories are just the beginning of a long and
comprehensive reshaping of the present varieties of wheat.
Primarily through the initiative and analytical endeavors of Aaron Aaronsohn,
the late Director of the Jewish Agricultural Experiment Station of Haifa, an
invaluable treasure chest of wild wheats exists now containing genes for large
kernel size, high amount and better quality of protein, disease resistance, etc. All
these can now be selectively and systematically incorporated into the best commercial
varieties of the world by the application of the genetic principles worked out by
E. R. Sears.
In such a brief summary it is not possible to point out — and even more
difficult to explain — the majority of the numerous contributions of Sears which
are detailed in his nearly one hundred scholarly papers and book chapters.
Theodor O. Diener
Plant Protection Institute
USDA, Beltsville, Maryland, USA
k
Dr. Diener’s fundamental and applied studies have firmly established the existence
of viroids, a new group of subviral pathogens.
Dr. Diener discovered that the pathogen causing potato spindle tuber disease is
not a virus, as previously believed, but a much smaller, free RNA molecule, which
he named viroid. The discovery by Diener and his colleagues of other viroids
affecting various cultivated plants such as chrysanthemum stunt, coconut cadang-
cadang and planta macho viroid of tomato soon followed; so that today viroids are
considered to be an important group of disease agents affecting potatoes, tomatoes,
citrus, avocado, etc. His pioneering studies on isolation and purification showed
inequivocally that viroids have a unique molecular structure, different from any
other pathogen. Dr. Diener found that viroids replicate without helper virus; that
viroid-complementary DNA, produced by insertion into bacteria, are themselves
infectious and that the RNA transcribed in potato cells is identical with the viroid.
Cloned viroid-specific cDNAs were also developed by him for novel diagnostic
tests.
Dr. Diener’s work has direct application to the control of viroid diseases of
many crops, as well as to the elucidation of the unique biological and molecular
properties of these small disease agents.
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264 Wolf Prize in Agriculture
CURRICULUM VITAE
Date and place of birth: 28 February 1921, Zurich, Switzerland
EDUCATION AND POSITIONS HELD:
1942-46 - Swiss Federal Institute of Technology, Dipl. sc. nat. ETH, 1946.
1946-48 - Research Assistant, Department of Botany, Swiss Federal Institute of
Technology, Dr. sc. nat. ETH, 1948.
1948-49 - Plant Pathologist, Swiss Federal Horticultural Experiment Station,
Waedenswil.
1950 - Assistant Professor of Plant Pathology, Rhode Island State University,
Kingston.
1950-59 - Assistant to Associate Plant Pathologist, Washington State University,
Prosser, Washington.
Since 1959 - Research Plant Pathologist, U.S. Department of Agriculture, Beltsville,
Maryland.
1) How can viroids replicate autonomously in susceptible cells even though they
contain only very limited genetic information and are not translated into viroid-
specific proteins?
2) What are the molecular mechanisms by which virolds cause diseases?
3) What did viroids originate from?
Dr. Diener, sometimes in collaboration with colleagues, greatly contributed toward
answers to these and other questions. His demonstration that viroid replication is
sensitive to actinoinycin D and, more recently, the finding in Dr. Diener’s laboratory
of replication intermediates much larger than the viroid itself are important
contributions toward the elucidation of viroid replication and have suggested a
model involving a rolling-circle type mechanism.
Molecular Probes
Dr. Diener’s laboratory was first in applying modern methods of recombinant
DNA technology to the viroid problem. Synthesis of viroid-complementary DNAs
(cDNAs) has yielded valuable probes with which to study viroid-specific molecular
events in infected cells. By insertion into bacteria, such DNAs can be produced in
any desired amounts. Of great importance is the recent demonstration, first made
in Dr. Diener’s laboratory and now confirmed by other investigators, that artificially
produced cDNAS representing the complete structure of PSTV are infectious
(exhibit #10). Because the DNA mirrors the structure of PSTV, the RNA transcribed
in susceptible cells from the DNA is identical with PSTV. Thus, plants into which
the DNA has been introduced, produce PSTV and display the characteristic symptoms
of PSTV infection. Recombinant DNA technology makes it now possible to modify
the recombinant DNA in various ways, to introduce precisely known nucleotide
exchanges, insertions, or deletions, or to produce chimeric viroids. Because each
DNA mutation is faithfully and predictably reflected in the transcribed RNA, these
experiments are apt to identify specific regions of the viroid molecule involved in
symptom formation, replication efficiency, or host specificity. Today, Dr. Diener’s
laboratory is in the forefront of these important investigations which, undoubtedly,
will permit us to correlate viroid structure with biological function and result in
the elucidation of the molecular mechanism of viroid pathogenesis. Most types of
symptoms of viroid infection have their counterpart in plants infected by
conventional viruses, and vice versa, indicating that viroids and viruses may, in
many cases, affect the same or similar plant metabolic systems. Thus, it is likely
that results obtained with viroids will explain mechanisms of virus, as well as
viroid, pathogenesis.
Viroid Origin
With the discovery of split genes and RNA splicing, it has been suggested (by
Dr. Diener and others) that viroids might have originated by circularization of
268 Wolf Prize in Agriculture
intervening sequences (introns) that are spliced out of precursor RNAs. The
identification, by Dr. Diener, of a region in the nucleotide sequence of PSTV that is
homologous with the 5’ end of small nuclear RNA Ui (which is involved in the
splicing of nuclear mRNA precursors) lent credence to the possibility that viroids
represent “escaped introns”. Recently, again in Dr. Diener’s laboratory, striking
sequence similarities have been discovered between viroids and class I introns,
further supporting a possible intron origin of viroids.
Epilogue
Dr. Diener has now been active in plant pathological research for 35 years and has
published some 180 scientific papers, including many book chapters and one
book. He has long been a recognized authority in plant pathology and particularly
in plant virology. With his discovery of the viroid, Dr. Diener’s international
Theodor O. Diener 269
recognition and stature have been extended to other areas of science, such as
animal virology, molecular biology, veterinary and human medicine, and
microbiology. He has received numerous invitations to lecture at prominent research
centers, medical and veterinary schools, and institutes of molecular biology, in the
U.S.A. and abroad. He has received many awards, and has been elected as a member
of several honorary societies, such as the U. S. National Academy of Sciences, the
American Academy of Arts and Sciences, and the German Academy of Natural
Scientists, Leopoldina.
Dr. Diener was actively engaged in research with his small research unit, to
remain in the forefront of fundamental research on viroids and viroid diseases.
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Ernest John Christopher Polge
Biotechnology Cambridge Ltd., Cambridge
United Kingdom
k
1926–2006
Dr. Ernest John Christopher Polge is renowned for his pioneering work on the
cryopreservation of living cells, such as sperm and in embryos. His work in this
area promoted the development of the science of cryobiology. During the last 30
years he has also contributed significantly to our understanding of embryonic
development, in vitro fertilization and embryo transplants. In addition, he led a
group to international recognition for pioneering techniques in cloning livestock
animals by embryo division and nuclear transfer, and initiated transgenic research
by insertion of gene constructs into pronuclei of recently fertilized eggs.
Throughout his career he has been concerned with the transfer and application
of science for the improvement of livestock production.
IN MEMORIAN1
Christopher Polge, who died on August 17 aged 80, attained scientific eminence at
a remarkably early age: his discovery, whilst still a doctoral student, of how to
preserve living cells and tissues at very low temperatures solved a long-standing
and intractable problem in biology.
This breakthrough not only formed the basis for the new science of cryobiology
but has also had profound and continuing practical implications for agriculture
and medicine.
1Telegraph, UK — 11/09/2006.
271
272 Wolf Prize in Agriculture
The key to cell survival during the freezing process was the discovery, by Polge
and his colleague Dr Audrey Smith, of a class of chemicals now known as
cryoprotective agents. Although the potential importance of cell preservation by
freezing had long been recognized, experiments in freezing living cells or tissues
had invariably resulted in their death.
It rapidly became clear from the work of Polge and Smith that many of the cell
types in the body had highly specific requirements for freezing and thawing. Polge
chose to focus on the preservation of sperm and eggs in mammals, and in 1950
produced the first chicks from eggs fertilized with frozen sperm.
Although these chicks were the first vertebrates to be produced in this way,
much greater acclaim followed Polge’s report two years later of high pregnancy
rates in cattle using sperm that had been frozen for periods in excess of a year.
These reports had far-reaching consequences for the future of artificial insemination
and genetic improvement in livestock.
Within 10 years virtually every cattle-breeding centre in the world had converted
to the use of frozen semen; Polge gave assistance and advice to these centres,
travelling throughout Britain and across the world, particularly in North and
South America.
In the early 1970s pig semen was successfully frozen in Polge’s laboratory, and
he finally felt that he had fulfilled his objective of developing methods for the
cryopreservation of semen from all the major species of livestock.
Ernest John Christopher Polge was born on August 16, 1926, the son of a
Buckinghamshire farmer. He was educated at Bootham School, York, then read
Agriculture at the University of Reading. After working briefly as an agricultural
economist he decided to follow his vocation as a research scientist.
Polge studied for his doctorate at the Division of Experimental Biology at the
National Institute for Medical Research at Mill Hill, London, before moving to the
Animal Research Station at Cambridge under Sir John Hammond.
Hammond’s method was to carry out rigorous fundamental science with a
view to embracing a practical outcome. This approach appealed to Polge, and was
to guide his research throughout his lifelong association with the Research Station,
which he was to lead for the last eight years of his career.
After his initial successes in the cryobiological field, Polge realised that the
freezing of embryos for use in embryo transfer programs offered the next practical
means of contributing to the improvement of livestock. He recruited Ian Wilmut
(later to lead the team which cloned Dolly the Sheep) to work specifically on the
problem of the low temperature preservation of embryos.
A series of advances by this team resulted in the birth, in 1973, of Frosty, the
first calf from a cryopreserved embryo. At that time it was not foreseen that
embryo freezing would find its greatest use in assisted human reproduction; rather
the focus was on the genetic improvement of livestock.
Ernest John Christopher Polge 273
BIOGRAPHICAL DATA
Date and place of birth: 16 August 1926 - Jordans, Buckinghamshire
1986 he has continued at the Animal Research Station site as Research Director of
a new Biotechnology Company — Animal Biotechnology Cambridge Ltd — where
his expertise is being utilized to develop commercial applications of much of the
breeding technology developed under his earlier guidance.
Throughout his career Dr Polge has been concerned with the application of
science to the practice of animal breeding. He does not live in an ‘ivory tower’, but
is as much concerned with the practical problems of farming as with basic science.
This philosophy has led to close liaisons with the industry in matters concerned
with animal breeding and has often enabled applications derived from basic research
to be applied quickly and effectively as is evidenced by new developments in
artifical insemination and embryo transplantation. He has undertaken numerous
lecture tours in the United Kingdom and abroad. He is the United Kingdom
representative on the EEC committee concerned with the physiology of reproduction
in farm animals. He is a consultant to the Food and Agriculture Organization on
the conservation of genetic resources, a member of the Genome Specialist Group
of the International Union for the Conservation of Nature and advises the Technical
Committee of the Rare Breeds Survival Trust. He has previously been Secretary
and Chairman of the Society for the Study of Fertility, Chairman of the Society for
Low Temperature Biology and is presently Chairman of the Journals of Reproduction
and Fertility Ltd. He is also a member of the Society for Endocrinology, Society for
the Study of Animal Breeding, Society for Cryobiology, Research Defence Society
and the Royal Institution.
Fertilization in the pig and horse. J. Repro. Fert. 514, 46l-470 (1978).
Embryo transplantation in the large domestic species: applications and perspectives
in the light of recent experiments with eggs and embryos. J. R. Agric. Soc.
141, 115-126 (1980). (With S.M. Willadsen).
Embryo transfer and preservation. In Control of Pig Reproduction (ed. D.J.A. Cole
& G.E. Foxoroft), pp. 277-291. London: Butterworths (1982).
Analysis of slow-warming injury of mouse embryos by cryomicroscopical and
physiochemical methods. Cryobiology 21, 106-121 (1984). (With W.F. Rall &
D.S. Reid).
How does embryo manipulation fit into present and future pig reproduction.
J. Repro. Fert. S33, 93-100 (1985).
Charles Thibault
Universite de Paris VI, Paris, France
k
1919–2003
BIOGRAPHICAL DATA
Date and place of birth: July 14, 1919, Paris.
277
278 Wolf Prize in Agriculture
Professor Emeritus
1950-1978 - Member of the CNRS Scientific Committees (Biology, Physiology)
1954-1958 - Dean of the National Center of Zootechnical Research, INRA
1960-1964 - Adviser of the Prime Minister for Biology
1976-1979 - President of Ph.D. Grant Committee (Biology, Agriculture, Medicine)
1979-1981 - President of the CNRS
Since 1986 - President of the French Society for the Study of Fertility
Prix scientifiques
1950 Prix Foulon (Zoologie).
1955 Prix Serres (Agriculture).
1962 Prix Foulon (Economie rurale).
1972 Prix do la Société française do Gynécologie (Annie. Jean d’ALSACE).
1980 Marschall Medal.
1983 Prix do Biologie do la Ville do Paris.
1987 Prix do l’Association pour I’Etude do. I’Endocrinologie et do Is Fertilité.
Decorations
1975 Officier de l’Ordre du Mérite (1967) et do la Legion d’Honneur.
1976 Commandeur des Palmes Académiques (1974) et du Mérite Agricole.
events up to the first cleavage (39, 47, 59, 76), What is more, he obtained, with
his group, the first true in vitro fertilization in 1954 (45, 47, 53). The birth of
young rabbits in 1959 in USA (CHANG) and in Jouy en Josas in 1961 (61) has
testified to the “normality” of the experimental procedure. Later on, in vitro
fertilization has been extended to domestic mammals in England, in USA, in Canada
and in our Institute. Trials on practical application are now in progres in France.
He used the in vitro fertilization technique as a tool for the study of sperm
capacitation, gamete ageing and the consequence of triploidy on subsequent
development (65, 66, 67, 76, 84, 85, 91, 99, 102). Triploid embryos (rabbit) or
parthenogenetically activated rabbit or sheep eggs are able to cleave but in no case
develop beyond implantation (10, 14, 23, 34, 36).
He studied the conditions of culture of rabbit, sheep, goat and bovine eggs
fertilized either in vivo or in vitro and was the first to observe that sheep, goat and
bovine egg development stops in culture after few cleavages whatever the medium
(43, 74) showing that fallopian tubes play a role in egg development in these
species but, the nature of the factors involved so far remains unknown.
-Other topics. He published papers on sperm survival in vitro (46, 48) but, above
all has been interested in sperm ascent in the female tract in relation to insemination
practice. He showed that insemination stress is able to postpone sperm ascent in
the ewe and that ocytocin stimulates sperm transport in unstressed ewes (75); as
ocytocin is released by cervical stimulation this hormone probably plays a
physiological role in female tract sperm transport after natural as well as artificial
insemination. He demonstrated that in the cow, after mating, spermatozoa are
present in the ampullary portion of the oviduct only 8 hours after natural mating
and that in lower isthmus and uterotubal junction polynuclear cells and
inacrophages are normally absent. This region, and not the cervix as generally
assessed, is the reservoir from which the fertilizing sperm moves to the ampulla
(95, 98, 101, 104, 135). This conclusion fits well with the more recent observations
in rabbit, sheep, hamster and bats.
We must also mention his enlightening participation in the studies of the role
of the photoperiodism on sexual activity and mainly on spermatogenesis (50, 52,
54, 69, 73). He gave a general survey on the importance of photoperiodism in the
reproduction of domestic and wild mammals, mainly based on the results of the
Department of Physiology in Jouy en Josas (73) and this paper has remained a
reference.
During the past twenty years he has been Professor of Reproductive Physiology
in Paris VI University. He has trained many hundred students, Biologists,
Veterinarians, Agronomists and Physicians. Nany of them are now in applied or
basic research teams in medicine, agriculture and biology. Five of them are university
professors. He has been a jury member at more than 300 PhD. He has published
three books and contributed to many others.
282 Wolf Prize in Agriculture
Narrative curriculum vitae for Professor P.M.Biggs CBE, DSc, Ph.D, CBiol,
FIBiol, FRCPath, FRCVS, FMed Sci, FRS.
After schooling in England and the USA Professor Biggs joined the Royal Air Force
in 1944 and spent the first six months of his service at the Queen’s University,
Belfast reading physics and engineering. On demobilisation in 1948 he entered the
Royal Veterinary College. London and graduated in veterinary science and as a
veterinary surgeon in 1953. He then took a PhD on the subject of lymphoid tissues
in the domestic chicken at Bristol University where he subsequently joined the
staff of the Veterinary School in 1955 as a lecturer in Clinical Pathology during
which time he started studies on the avian leucosis complex. In 1959 he moved to
the Houghton Poultry Research Station to form and Head a unit to study the
economically important lymphoid tumour conditions of the domestic fowl. He
built and led a group that established that there were two important lymphoid
tumour conditions of the fowl. One that had been shown in the USA to be caused
by what is now known as a retrovirus, now called lymphoid leucosis, and a second,
which he termed Marek’s disease after the Hungarian veterinary pathologist who
first described the disease, which was shown by the group to be caused by an
highly cell associated herpesvirus. Subsequently they developed an effective vaccine
285
286 Wolf Prize in Agriculture
against this serious tumour condition. This was the first vaccine effective against a
tumour in any species. Professor Biggs also contributed with his colleagues to
knowledge of the genetic control of resistance and susceptibility to infection with
retroviral tumour viruses of the fowl. He also described a novel disease in turkeys
which he named lymphoproliferative disease which he and colleagues showed was
caused by a retrovirus. Professor Biggs has published over 130 papers mainly on
viruses and poultry disease.
In 1971 Professor Biggs was appointed Deputy Director of Houghton Poultry
Research Station and its Director in 1974. During his Directorship he encouraged
the use of molecular biological techniques for the studies of avian disease pathogens
and appointed a number of molecular biologists to the staff of the Station. His
described a novel lymphoproliferative disease in turkeys and he and his research
group defined its retroviral cause.
On the re-organisation of the AFRC Institutes in 1986 he was appointed AFRC
Director of Animal Health charged with the remit of forming an Institute for
Animal Health comprising the former Animal Virus Research Institute, Pribright,
Houghton Poultry Research Station, Institute for Research on Animal Diseases,
Compton and the Neuropathogenesis unit in Edinburgh. He retired from this post
in 1988.
Between 1988 and 1994 he was an Andrew D. White Professor-at-Large at
Cornell University, Ithaca, N.Y. USA. and has been a Visiting Professor in Veterinary
Microbiology at the Royal Veterinary College, London University since 1982.
Professor Biggs is a Fellow of the Royal College of Veterinary Surgeons, founding
Fellow of the Academy of Medical Science, Fellow of the Royal College of Pathologists
and has been a Fellow of the Institute of Biologists since 1973 and was its President
from 1990 to 1992. He has been President of the British Veterinary Poultry
Association, the World Veterinary Poultry Association, The International Association
for Comparative Research on Leukaemia and Related Diseases and a Vice President
of the British Veterinary Association.
He has been a member of a number of Government and other Committees and
Working Groups including the Ashby Working Party on Genetic Manipulation in
Micro-organisms, the AFRC/ADAS Working Party on Animal Disease Research in
the United Kingdom, the Virus Working Group of the Advisory Committee on
Genetic Manipulation, Advisory Committee on Dangerous Pathogens chairing its
Working Group on the handling of Transmissible Spongiform Encephalopathies,
the Veterinary Products Committee, the Royal College of veterinary Surgeons’
Committee of Enquiry into Veterinary Research. He also chaired Visiting Groups to
the Ministry of Agriculture, Fisheries and Food Poultry Unit at the Experimental
Husbandry Farm, Gleadthorpe, the Pig Unit at the Experimental Husbandry Farm,
Terrington and the Central Veterinary Laboratory, Weybridge.
Peter M. Biggs 287
LIST OF PUBLICATIONS
Scientific papers
KING, A.S. and BIGGS, P.M. (l955) Experimental observations on the respiration
through the humerus of Gallus domesticus. J. Anat. (Lond.), 89: 567-568.
BIGGS, P.M. (1956) Lymphoid haemopoietic tissue. Vet. Rec., 68: 525-526.
BIGGS, P.M. (1957) The association of lymphoid tissue with the lymph vessels in
the domestic chicken (Gallus domesticus). Acta Anat., 29: 36-47.
KING, A.S. and BIGGS, P.M. (1957) General anaesthesia in Gallus domesticus for
non-survival laboratory experiments. Poult. Sci., 36: 490-495.
BIGGS, P.M. and KING, A.S. (1957) A new experimental approach to the problem
of the air pathway within the avian lung. J. Physiol., 138: 282-299.
BIGGS, P.M. and PAYNE, L.N. (1959) Cytological identification of proliferating
donor cells in chick embryos injected with adult chlcken blood. Nature, (Lond.),
184: 1594.
288 Wolf Prize in Agriculture
BIGGS, P.M. and PAYNE, L.N. (1961) Pathological changes following the inoculation
of chick embryos with adult cells. 1. Spleen cells. Immunology, 4: 24-37.
BIGGS, P.M. and PAYNE, L.N. (1961) Pathological changes following the inoculation
of chick embryos with adult cells. 2. Blood cells. Immunology, 4: 38-48.
BIGGS, P.M. (1962) Some observations on the properties of cells from the lesions
of Marek’s disease and lymphoid leukosis. Proceedings of the 13th Symposium
of the Colston Research Society on Animal Health and Production, pp. 83-99.
Butterworth, London.
BIGGS, P.M. and PAYNE, L.N. (1963) Transmission experiments with Marek’s disease
(fowl paralysis). Vet. Rec., 75: 177-179.
BIGGS, P.M. and PAYNE, L.N. (l963) Some properties of the agent of Marek’s
disease (fowl paralysis, neuro-lymphomatosis). Proceedings of the International
Symposium on Comparative Leukemia, IV, 2. Hannover.
PAYNE, L.N. and BIGGS, P.M. (1964) Differences between highly inbred lines of
chickens in the response to Rous sarcoma virus of the chorio-allantoic
membrane and of embryonic cells in tissue culture. Virology, 24: 610-616.
PAYNE, L.N. and BIGGS, P.M. (1964) A difference in susceptibility to lymphoid
leukosis virus and Rous sarcoma virus between cells from two inbred lines.
Nature (Lond.), 203: 1306-1307.
PAYNE, L.N. and BIGGS, P.M. (1964) Transmission experiments with Marek’s disease
(fowl paralysis) and lymphoid leukosis. World’s Poultry Science Journal,
20: 284.
Biggs, P.M. and Payne, L.N. (1964) The relationship of Marek’s disease (neuro-
lymphomatosis) to lymphoid leucosis. National Cancer Institute Monograph,
17: 83-98
BIGGS, P.M., PURCHASE, H.G., BEE, B.R. and DALTON, P.J. (1965) Preliminary
report on acute Marek’s disease (fowl paralysis) in Great Britain. Vet. Rec.,
77: 1339-1340.
PAYNE, L.N. and BIGGS, P.M. (1965) Effect of lymphoid leucosis virus on the
response of the chorio-allantoic membrane to Rous sarcoma virus. Virology,
27. 621-622
PAYNE, L.N., BIGGS, P.M., CHUBB, R.C. and BOWDEN R.S.T.(1966) Contamination
of egg-adapted canine distemper vaccine by avian leucosis virus. Vet. Rec.,
78: 45-49
HARRIS, R.J.C., DOUGHERTY, R.M., BIGGS, P.M., PAYNE, L.N., GOFFE, A.P.,
CHURCHILL, A.E. and MORTIMER, R. (1966) Contaminant viruses in two live
virus vaccines produced in chick cells. J. Hyg. Camb., 64; 1-7.
PAYNE, L.N. and BIGGS, P.M. (1966) Genetic basis of cellular susceptibility to
the Schmidt-Ruppin and Harris strains of Rous sarcoma virus. Virology, 29:
190-198.
OWEN, J.J.T., MOORE, M.A.S. and BIGGS, P.M. (1966) Chromosome studies in
Marek’s disease. J. natn. Cancer Inst., 37: 199-209.
Peter M. Biggs 289
KENZY, S.G. and B1GGS, P.M. (1967) Excretion of the Marek’s disease agent by
infected chickens. Vet. Rec., 80: 565-568.
BIGGS, P.M. and PAYNE, L.N. (1967) Studies on Marek’s disease. I. Experimental
transmission. J. natn. Cancer Inst., 39: 267-280.
PAYNE, L.N. and BIGGS, P.M. (l967) Studies on Marek’s disease. II. Pathogenesis.
J. natn. Cancer Inst., 39: 281-302.
CHURCHILL, A.E. and BIGGS, P.M. (1967) Agent of Marek’s: disease in tissue
culture. Nature (Lond), 215: 528-530.
PURCHASE. H.G. and BIGGS, P.M. (1967) Characterization of five isolates of Marek’s
disease. Res. vet. Sci., 8: 440-449.
BIGGS, P.M., THORPE, R.J. and PAYNE, L.N. (1968) Studies on genetic resistance to
Marek’s disease in the domestic chicken. Brit. Poult. Sci., 9: 37-52.
PURCHASE, H.G., CHUBB, R.C. and BIGGS, P.M. (1968) Effect of lymphoid leucosis
and Marek’s disease on the immunological responsiveness of the chicken.
J. natn. Cancer Inst., 40: 583-592.
CHUBB. R.C. and BIGGS. P.M. (1968) Neutralization of Rous sarcoma virus. J. gen.
Virol., 3: 87-96.
LONG, P.L., KENZY, S.G. and BIGGS, P.M. (1968) Relationship between Marek’s
disease and coccidiosis. I. Attempted transmission of Marek’s disease by avian
coccidia. Vet. Rec., 83: 260-262.
BlGGS, P.M., LONG, P.L., KENZY, S.G. and ROOTES, D.G. (1968) Relationship between
Marek’s disease and coccidiosis. II. The effect of Marek’s disease on the
susceptibility of chickens to coccidial infection. Vet. Rec., 83: 284-289.
EPSTEIN, M.A., ACHONG, B.G., CHURCHILL, A.E. and BIGGS, P.M. (1968) Structure
and development of a herpes-type virus of Marek’s disease. J. natn Cancer
Inst., 41: 805-820.
CHURCHILL, A.E. and BIGGS, P.M. (1968) A herpes-type virus isolated in cell
culture from the tumours of chickens with Mareik’s disease. II. Studies in
vivo. J. natn. Cancer Inst., 41: 951-956.
BIGGS, P.M., CHURCHILL, A.E., ROOTES, D.G. and CHUBB, R.C. (1968) The etiology
of Marek’s disease - an oncogenic herpes-type virus. In: Perspectives in
Virology, vol. 6, pp. 211-230. Edit. Pollard, M., Academic Press, New York
and London.
BIGGS, P.M., LONG, P.L., KENZY, S.G. and ROOTES. D.G. (1969) Investigations into
the association between Marek’s disease and coccidiosis. Proceedings of
International Symposium on Coccidiosis. Brno. Czechoslovakia. Acta.
veterinaria (Brno). 38: 65-75.
KENZY, S.G., LONG. P.L. and BIGGS, P.M. (1970) Relationship between Marek’s
disease and coccidiosis. III. Effects of infection with Eimeria mivati on Marek’s
disease in the chicken. Vet. Rec., 86: 100-104.
PAYNE, L.N. and BIGGS, P.M. (1970) Genetic resistance of fowl to MH2
reticuloendothelioma virus. J. gen.Virol., 7: 177-185.
290 Wolf Prize in Agriculture
BIGGS, P.M. PAYNE, L.N. MILNE, B.S. CHURCHI.LL. A.E. CHUBB. R.C. POWELL,
D.G. and HARRIS. A.H. (1970) Field trials with an attenuated cell associated
vaccine for Marek’s disease. Vet. Rec., 87: 704-709.
BIGGS, P.M. and MILNE, B.S. (1971) Use of the embryonating egg in studies on
Marek’s disease. Am, J. vet. Res., 32: 1795-1809.
B1GGS, P.M. and MILNE, B.S. (1972) Biological properties of a number of Marek’s
disease virus isolates. In: Oncogenesis and Herpesviruses, pp. 88-94. Edit.
Biggs, P.M., deThé, G. and PAYNE, L.N.. IARC scientific publication No.2.
International Agency for Research on Cancer, Lyon.
FRAZIER, J. A. and BIGGS, P.M. (1972) Marek’s disease herpesvirus particles in
tissues from chickens free of precipitating antibodies. J. natn. Cancer Inst.,
48: 15l9-l523.
BIGGS, P.M., POWELL, D.G., CHURCHILL, A.E. and CHUBB, R.C. (1972) The
epizootiology of Marek’s disease. 1. Incidence of antibody, viraemia and
Marek’s disease in six flocks. Avian Path., 1: 5-25.
PHILLIPS., P.A. and BIGGS, P.M. (1972) Course of infection in tissues of susceptible
chickens after exposure to strains of Marek’s disease virus and turkey
herpesvirus. J. natn. Cancer Inst., 49: 1367-1373.
WEISS, R.A. and BIGGS, P.M. (1972) Leukosis and Marek’s disease viruses of
feral red jungle fowl and domestic fowl in Malaya. J. natn. Cancer Inst., 49:
1713-1725.
ROSS, L.J.N., FRAZIER, J.A. and BIGGS, P.M. (1972) An antigen common to some
avian and mammalian herpesviruses. Oncogenesis and Herpesviruses.
pp. 480-484. Edit. Biggs, P.M., de Thé, G. and Payne, L.N., IARC scientific
publication No.2. International Agency for Research on Cancer, Lyon.
BIGGS, P.M., JACKSON, C.A.W., BELL, R.A., LANCASTER, F.M. and MILNE, B.S.
(1972) A vaccination study with attenuated Marek’s disease virus. In:
Qncogenesis and Herpesviruses, pp. 139-46. Edit. Biggs, P.M., de Thé, G. and
Payne, L.N., IARC scientific publication No.2. International Agency for Research
on Cancer, Lyon.
JACKSON, C.A.W., BIGGS, P.M., BELL, R.A., LANCASTER., F.M. and MILNE, B.S.
(1972) Vaccination against Marek’s disease with attenuated Marek’s disease
virus in a commercial egg laying flock. In: Proceedings 1972 Australasian
Poultry Science Convention, Auckland, New Zealand, pp. 229-240.
BIGGS, P.M., MILNE, B.S., GRAF, T. and BAUER, H. (1973) Oncogenicity of
non-transforming mutants of avian sarcoma viruses. J. gen. Virol., 18:
399-403.
ROSS, L.J.N., BIGGS, P.M. and NEWTON, ALISON A. (1973) Purification and
properties of the ‘A’ antigen associated with Marek’s disease virus infections.
J. gen. Virol., 18: 291-304.
PANI, P.K. and BIGGS, P.M. (1973) Genetic control of susceptibility to an A subgroup
sarcoma virus in commercial chickens. Avian Path., 2: 27-41.
Peter M. Biggs 291
BIGGS. P.M., SHILLETO. R.F.W. and MILNE, B.S. (1980) A quantitative sequential
study of viraemia and neutralizing antibody to HVT and MDV in a commercial
flock vaccinated with HVT. Avian Path., 9: 511-523.
McDOUGALL, J.S., SHILLETO, R.W. and BIGGS, P.M. (1980) Experimental infection
and, vertical transmission of reticuloendotheliosis virus in the turkey. Avian
Path., 9: 445-454.
McDOUGALL, J.S., SHILLETO, R.W. and BIGGS, P.M. (1981) Further studies on
vertical transmission of reticuloendotheliosis virus in turkeys. Avian Path.,
10: 163-169.
BIGGS, P.M., SHILLETO, R.W., LAWN, A.M. and COOPER, D.M. (1982) Idiopathic
polyneuritis in SPF chickens. Avian Path., 11: 163-178.
ROSS, L.J.N., MILNE, B.S. and BIGGS, P.M. (1983) Restriction endonuclease analysis
of Marek’s disease virus DNA and homology between strains. J. gen. Virol.,
64: 2785-2790.
BIGGS, P.M. (1975) Marek’s disease virus, a model for oncogenic herpesviruses -
chemical and antigenic properties. Proceedings of Xlth International Cancer
Congress, Florence. Excerpta Medica, Amsterdam. Vo. 1: 159-165.
BIGGS, P.M. (1976) Criteria for the differential diagnosis of avian lymphoid leukosis
and Marek’s disease. In: Differential diagnosis of avian lymphoid Leukosis
and Marek’s disease, pp. 67-85, Edit. Payne, L.N. CEC Publication, EURm
5494e, Luxembourg, 1976.
BIGGS, P.M. (1977) Marek’s disease and leukosis. In: Poultry Health, pp. 65-81.
Edit. Gordon, R.F. Bailliere Tindall, London.
BIGGS. P.M. (1978) The veterinary profession and an intensive animal industry.
(Wooldridge Memorial Lecture). Vet. Rec., 103: 251-255.
BIGGS, P.M. (1979) Marek’s disease now and then. In: Advances in Comparative
Leukemia Research, pp. 353-358. Proceedings of the IXth International
Symposium on Comparative Research on Leukemia and Related Diseases,
Pitsunda, U.S.S.R. Edit. Yohn, D.S., Lapin, B.A. and Blakeslee, J.R. New York,
Elsevier, 1980.
BIGGS, P.M. (1980) The biology of avian leukosis. In: Leukemias, Lymphomas, and
Pappillomas: Comparative Aspects, pp. 233-246. Edit. Bachmann, P.A. London,
Taylor and Francis.
BIGGS, P.M. (1982) The world of poultry disease. Avian Path. 11: 281-300.
ALLAN, W.H., ALEXANDER, D.J., BIGGS, P.M., GORDON, R.F., JORDAN, F.T.W.
and McFERRAN, J.B. (1982) Viral diseases. In: Poultry Diseases, 2nd ed.,
pp. 76-159. Edit. Gordon, R.F. and Jordan, F.T.W. London, Baillière Tindall.
BIGGS, P.M. (1982) Vaccination against Marek’s disease in layers and breeders. In:
Marek’s vaccination. Present situation and prospects for the future, pp.39-52.
Proceedings of the 3rd scientific meeting, IVAZ Laboratori Biologici, Padova,
Italy.
BIGGS, P.M. (1982) The epizootiology of avian herpesviruses in veterinary medicine.
Devl. Biol. Stand., 52: 3-11.
BIGGS, P.M. (1983) Disease prevention and control in poultry production. In:
Disease Prevention and Control in poultry Production. pp. 313-319. Sydney
Post-graduate Committee in Veterinary Science.
ALEXANDER. D., ALLAN. W.H., BIGGS. P.M.. BRACEWELL. C.D., DARBYSHI.RE, J.R.,
DAWSON, P.S., HARRIS. A.H., JORDAN, F.T.W., MACPHERSON, I., McFERRAN,
J.B., RANDALL, C.J., STUART, J.C., SWARBRICK, O. and WILDING. G.P. (1983)
A standard technique for haemagglutination inhibition tests for antibodies to
avian infectious bronchitis virus. Vet. Rec., 113: 64.
BIGGS, P.M. (1983) Vaccination. In: The Control of Infectious Diseases in Farm
Animals, pp. 21-27. BVA Trust Project: “Future of Animal Health Control”.
London, British Veterinary Association”.
Peter M. Biggs 295
BIGGS, P.M. (1984) Contagious diseases: an economic hazard to the poultry industry.
siipikarja, 66: 246-252.
BIGGS, P.M. (1984) Lympoproliferative disease of turkeys. In: Diseases of Poultry,
8th ed., pp. 418-419. Edit. Hofstad, M.S., Barnes, H.J., Calnek, B.W., Reid,
W.M. and Yoder, H.W. Iowa State University Press. Ames, Iowa, U.S.A.
BIGGS, P.M. (1985) Summing up of the 2nd International Symposium Marek’s
disease. Proceedings of the Internationa1 Symposium on Marek’s Disease,
1984, pp. 590-598. Cornell University, New York, U.S.A. Edit. Calnek, B.W.
and Spencer, J.L. Pennsylvania, American Association of Avian Pathologists,
Inc.
BIGGS, P.M. (1985) Infectious animal disease and its control. In: Philosophical
Transactions of the Royal Society, 30: 259-274.
BIGGS, P.M. (1985) Spread of Marek’s disease. In: Marek’s disease. Scientific Basis
and Methods of Control, pp. 329-340. Edit. Payne, L.N. Boston/Dordrecht/
Lancaster, Martinus Nijhoff Publishing.
ROSS, L.J.N. and BIGGS. P.M. (1986) Vaccination against Marek’s disease. In:
Leukaemia and Lymphoma Research. 3. Vaccine intervention against virus
induced tumours. Eds. Goldman. J.M. and Epstein, M.A. Macmillan Press.
BIGGS, P.M. (1988) Disease and its control in poultry production. First International
Poultry and Poultry Diseases Symposium, Manisa, Turkey.
BIGGS, P.M. (1988) Summing up of the 3rd International Symposium on Marek’s
disease. In: Advances in Marek’s disease research, 1998. Edits. Kato, S.,
Horuichi, T., Mikami, T. and Hirai, K. Japnese Association on Marek’s disease.
BIGGS, P.M. (1988) The place of molecular biology in veterinary research and
teaching. Vet. Rec., 123: 176-179.
BIGGS, P.M. (1989) La place de la biologie moléculaire dans la recherche et
l’enseignment vétérinaiares. Ann. Méd. Vét., 133: 185-189.
BIGGS, P.M. (1990) Use and desired properties of poultry vaccines. In: Genetic
engineering of animals. Edit. Hansel, H. and Weir, Barbara J. J. Reprod. Fert.
Suppl, 41, pp 149-152.
BIGGS, P.M. (1990) Gordon Memorial Lecture. Vaccines and vaccination – past,
present, and future. Brit. Poult. Sci. 31: 3-32.
BIGGS, P.M. (1990) Why biological control? In: Biological control of pests and
diseases. Edit. McCrachen, A.R. and Mercer, P.C.. Agriculture and Food Science
Centre, Belfast. 1990.
BIGGS, P.M. (1991) Lymphoproliferative disease of turkeys. In: Diseases of Poultry,
9th Edn. Edit. Calnek, B.W. pp. 456-459. Iowa State University Press, Ames,
Iowa, U.S.A.
BIGGS, P.M. (1992) Society’s changing view of science and the effect on scientific
research. Biologist, 39: 197-200.
296 Wolf Prize in Agriculture
BIGGS, P.M. (1993) Factors affecting Livestock production in the 21st century.
In: Animal Health and production for the 21st century. Edit. Beh, K.J.
pp. 148-154. CSIRO, Australia.
BIGGS, P.M. (1993) Developments in Vaccines. In: The advancement of veterinary
science. Vol. 4. Veterinary Science – Growth points and comparative medicine.
Edit. Michell, A.R. pp. 13-127. C.A.B. International.
BIGGS, P.M. (1997) Lympoproliferative disease of turkeys. In: Diseases of poultry,
10th Edn. Edit. Calnek, B.W. pp.485-489. Iowa State University Press. Ames,
Iowa, U.S.A.
BIGGS, P.M. (1997) The Leeuwenhoek Lecture 1997. Marek’s disease herpesvirus:
oncogenesis and prevention. Phil. Trans, R. Soc. London B 352: 1951-1962.
BIGGS, P.M. (2001) The history and biology of Marek’s disease virus. In: Marek’s
disease. Edit.Hirai, K. Springer-Verlag, Berlin.
BIGGS, P.M. (2002) Response to Whitby, Millett and Dando. Medicine, Conflict
and Survival. 18: 157-160.
BIGGS, P.M. (2004) Marek’s disease: long and difficult beginnings. In: Marek’
disease – an evolving problem. Edit. Davison, F. and Nair, V. Elsevier Academic
Press, London.
BIGGS, P.M. (2006) 50 years of Poultry Research. In 50th Anniversary of the
British Veterinary Poultry Association. British Veterinary Poultry Association.
BIGGS, P.M. (2006) The World Veterinary Poultry Association: the Beginnings and
first 25 years. Avian Path., 35: 417-428.
Books edited
Resistance and immunity to Marek’s disease. Edit. Biggs, P.M., C.E.C. Luxemburg.
Oncogenesis and Herpesviruses, pp. 88-94. Edit. Biggs, P.M., deThé, G. and PAYNE,
L.N.. IARC scientific publication No.2. International Agency for Research on Cancer,
Lyon.
Abstracts
BIGGS, P.M. (1966) Avian Leukosis. Paper presented at Symposium on “Tumour
viruses and other non-cytocidal agents”. Virus group of the Society for General
Microbiology, London. J. gen. Microbiol., 42: No.3, p. xv.
BIGGS, P.M. and ROOTES, D.G. (1967) Some properties of the infectious agent of
Marek’s disease (neurolymphomotosis) (Abs) J. Am. Vet. med. Ass., 150: 1341.
PURCHASE, H.G., CHUBB, R.C. and BIGGS, P.M. (1967) Immunologic disturbances
in birds infected with lymphoid leukosis and Marek’s disease. (Abs.) J. Am.
vet. med. Ass., 150: 1341-1342.
BIGGS, P.M. (1975) Epidemiology and control of Marek’s disease. (Abs) 3rd
International Congress for Virology, Madrid, p. 168.
BIGGS, P.M. (1977) Vertical transmission of avian RNA tumour Viruses and its
significance in oncogenesis. Proceedings of the Society for General
Microbiology, 4: 62.
BIGGS, P.M., McDOUGALL, J.S., MILNE, B.S. and SHILLETO, R.W. (1977) Leukosis
in turkeys, p. 13 (abstract). VIth International Congress of the World Veterinary
Poultry Association, Atlanta, Georgia.
Popular Articles
BIGGS, P.M. (1966) Avian leucosis and Marek’s disease. Poultry Digest, 25: 68-72.
BIGGS, P.M. (1977) Animal Health: the next ten years. 4. Poultry Span, Vol. 20,
No. 3: 106-107.
BIGGS, P.M. (1988) Perspectives. Turkeys, Vol. 36, No. 4, p. 1.
298 Wolf Prize in Agriculture
This article is reprinted with permission from Avian Pathology (published by Taylor & Francis,
Informa Ltd., London, UK).
Peter M. Biggs 299
300 Wolf Prize in Agriculture
Peter M. Biggs 301
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Michael Elliott
AFRC Institute of Arable Crops Research
Rothamsted, United Kingdom
k
1924–2007
Dr. Michael Elliot has demonstrated scientific vision in the field of crop protection
against insects. In the 1940’s he recognized that the naturally occurring insecticidal
compounds of plant in the Composite (Sunflower family) Chrysanthemum
cinevariaefolium known as pyrethroids had important crop protective potential
but suffered from serious disadvantages of lack of potency, stability and selectivity.
He, therefore, embarked on an imaginative program for the development of synthetic
analogues of the naturally occurring compounds and produced two, resmethrin
(1962) and bioresmethrin (1985) which had greatly enhanced potency and insect
specificity, combined with low mammalian toxicity. These compounds were
particularly effective for use in domestic and stored products, but were
photodegradable and therefore relatively ineffective under the field conditions. By
1973 Dr. Elliott had produced further analogues which combined crop surface
stability and insecticidal potency with exceptionally low mammalian toxicity; they
also had the advantage over the organochlorine insecticides of being readily
biodegradable in the soil without the formation of toxic residues. These compounds
have proved to be of enormous economic Importance in the protection of a very
wide variety of herbaceous and tree crops. At the fundamental level Dr. Elliott’s
work has made possible the establishment of relationships between chemical
303
304 Wolf Prize in Agriculture
CURRICULUM VITAE
BRIEF DETAILS OF CAREER
more stable in light and air, rendering them suitable for use in a wide range of
agricultural situations. In addition, they were highly active (deltamethrin is one of
the most potent insecticides known) and had the low mammalian toxicity generally
associated with pyrethroids. The first indications of all these favourable properties
were shown in experiments at Rothamsted (insecticidal activity and photostability)
and at the MRC unit at Carshalton and the University of California at Berkeley
(mammalian toxicity and metabolism), all initiated by Dr. Elliott at exactly the time
they were needed.
The natural pyrethrins are a mixture of six complex molecules, each containing
over fifty atoms arranged in structures with three chiral centres, and three
opportunities for additional isomerism. Despite this daunting complexity, Elliott
was able to establish which parts of the molecule could be replaced, and to
determine the stereochemical features that control the shape of active analogues.
The culmination of this research was deltamethrin, a single crystalline isomer (of
the eight possible), in which well over half of the structural features of the natural
pyrethrin molecule have been replaced by new groups. These pyrethroids,
particularly deltamethrin, have substantially enhanced compatibility with integrated
pest management (IPM) over the earlier organochlorines, organophosphates and
carbamates, and include safety to pollinators under prescribed conditions. The
advances are summarised in the following table:
Aldicarb 10 10 1 weeks
(carbamate)
Dimethoate 15 550 37 weeks
(organophosphate)
Pyrethrins 0.2 900 4,500 hours
Permethrin ~0.2 ~2,000 ~10,000 weeks
Cypermethrin ~0.06 ~500 ~8,000 weeks
Deltamethrin ~0.008 ~100 ~13,000 weeks
The research led by Dr. Elliott and the discovery of exceptionally effective
synthetic pyrethroids has been described as “a classic example of applied science”.
His modesty made him reluctant to accept another phrase frequently applied to
him: “The father of modern pyrethroids”. The international significance of the
work has been recognised in several important ways, the most prestigious being
the award to Rothamsted of the UNESCO Science Prize for 1978.
Elliott eagerly accepted the responsibilities that went with his success — to
respond to requests for information, to write reviews and to present lectures.
308 Wolf Prize in Agriculture
Statement of Impact
Of a million insect species, approximately 10,000 can be regarded as pests, and
about 30% of all the food we grow feeds insects rather than people.1 Despite the
success of alternative control methods, the protection of world food production
continues to rely very heavily on insecticides. Consequently, much effort has been
expended towards developing safer and more effective insecticides. Insects, being
animals, have many physiological processes in common with ourselves, and it is
therefore technically extremely demanding to produce insecticides that are also
benign to human beings and other warm-blooded animals. The immense impact
of the pyrethroids was assured by their being so highly insecticidal and yet rapidly
detoxified by mammals. With the discovery of the field-stable pyrethroids, e.g.
permethrin, and subsequently those optically resolved such as deltamethrin,
commercial development followed rapidly. The compounds have proved to be
suitable for lepidopterous pests worldwide and also for a multiplicity of other uses
in agriculture, animal husbandry and horticulture, generally at extremely low dose
rates, down to 10 g/ha. They were swiftly accepted because, unlike DDT and other
organochlorines, they are readily metabolised to harmless products in biotic media
such as soil. The research provided a new group of insecticides which immediately
rivalled the organophosphates as leading products for commercial crop protection.
This was achieved at a time when new insecticides were urgently needed to replace
compounds that had properties no longer considered suitable, or to which insects
had become resistant.
Once the discoveries had been announced, they were eagerly taken up by
industry, initially by eight separate firms. Even today, much of Elliott’s chemistry
remains the basis for the modern pyrethroid industry. For example, deltamethrin is
produced, stereochemically pure, on an enormous scale (hundreds of tons)
essentially by the originally published route (Hoechst Roussel), and Shell (now
Cyanamid) have responded to Elliott’s constant stress on the importance of
stereochemical purity to reduce environmental loading by developing a two-isomer
form of cypermethrin, again using similar chemistry. The immense impact of
Elliott’s work lies, therefore, not only in providing safer food protectants but also
in opening up the opportunity for discovering new and more acceptable insecticides
in a novel chemical class.
The new class of agricultural insecticides that resulted from Elliott’s work
remains important more than 20 years after their introduction. Pyrethroids currently
(1997) represent 19% of the total world sales of insecticide ($1,530M of $8070M).2
Michael Elliott 309
Of the sales of pyrethroids, about 70% are used on food crops. Although generic
products have subsequently been developed, the Elliott compounds still comprise
about half of the pyrethroid market and continue to be used extensively in
developing agricultural regions.*
Rarely in the field of agricultural pest control can one contribution have had
such a significant and lasting influence on food production. Dr. Elliott’s research,
combined with that of John E. Casida, undoubtedly constitutes a major contribution
to the efficiency and safety of food production worldwide, whilst minimising
undesirable effects on the environment. The award of the Wolf Foundation Prize
in Agriculture is a fitting tribute to the outstanding scientific integrity he has
shown so clearly throughout the whole of his career and his single-minded
determination to help in the never-ending quest for the increased availability of
healthy food.
References
1. Elliott, M. (1990). The Contribution of Pyrethroid Insecticides to Human Welfare.
L’actualité chimique, 57-70.
2. Wood Mackenzie report, 1998, 52-53. (Confidential - Not for general
dissemination).
LIST OF PUBLICATIONS
CROMBIE, L., ELLIOTT, M and HARPER, S.H. 1948. Total synthesis of some
pyrethrins. Nature 162, 222.
CROMBIE, L., ELLIOTT, M. and HARPER, S.H. 1950. Experiments on the synthesis
of the pyrethrins. III - Synthesis of dihydrocinerin-I and tetrahydropyrethrin-
I; a study of the action of N-bromosuccinimide on 3-methyl-1-n-alkyl (and
alkenyl)- cyclopent-3-en-1-ones. Journal of the Chemical Society, 971.
ELLIOTT, M., NEEDHAM, P.H. and POTTER, C. 1950. The insecticidal activity
of substances related to the pyrethrins. I - The toxicities of two synthetic
pyrethrin-like esters relative to that of the natural pyrethrins and the
significance of the results in the bioassay of closely related compounds. The
Annals of Applied Biology 37, 490.
ELLIOTT, M. 1951. The insecticidal activity of the pyrethrins and related compounds.
Pyrethrum Post 2(3), 18.
ELLIOTT, M. 1954. Allethrin. Journal of the Science of Food and Agriculture 11,
505.
ELLIOTT, M. 1956. The pyrethrins and related compounds. I - The preparation of
cyclopentenones from the products of Stobbe condensations with aliphatic
ketones. Journal of the Chemical Society, 2231.
310 Wolf Prize in Agriculture
POTTER, C., ELLIOTT, M and WARD, J. 1956. The pyrethrins - a British viewpoint.
Pyrethrum Post 4(1), 15.
ELLIOTT, M. 1958. Isolation and purification of (+)-pyrethrolone from pyrethrum
extract: reconstitution of pyrethrins I and II. Chemistry and Industry, 685.
ELLIOTT, M., OLEJNICZAK, J.S. and GARNER, J.J. 1959. Laboratory scale molecular
distillation of the pyrethrins. Pyrethrum Post 5(2), 8.
ELLIOTT, M. 1960. The structures of the enols of pyrethrolone. Proceedings of the
Chemical Society 406.
ELLIOTT, M. 1960. Pyrethrolone and related compounds. Chemistry and Industry,
1142.
ELLIOTT, M. 1961. Pyrethrolone. Japanese Journal of Pharmacy and Chemistry
33(12), 22.
CROMBIE, L. and ELLIOTT, M. 1961. Chemistry of the natural pyrethrins.
Fortschritte der Chemie organischer Naturstoffe 19, 120.
ELLIOTT, M. and JEFFS, K.A. 1961. A synthesis of calythrone. Proceedings of the
Chemical Society, 374.
ELLIOTT, M. 1961. The pyrethrins and related compounds. II - Infra-red spectra
of the pyrethrins and of other constituents of pyrethrum extract. Journal of
Applied Chemistry II, 19.
ELLIOTT, M. 1961. La distillation moleculaire des pyrethrines. Feuilles d’information
du Cofap 5, 6.
SAWICKI, R.M., ELLIOTT, M., GOWER, J.C., SNAREY, M. and THAIN, E.M. 1962.
Insecticidal activity of pyrethrum extract and its four insecticidal constituents
against house flies. I - Preparation and relative toxicity of the pure constituents;
statistical analysis of the action of mixtures of these components. Journal of
the Science of Food and Agriculture 13, 172.
ELLIOTT, M. 1964. The pyrethrins and related compounds.III - Thermal
isomerization of cis-pyrethrolone and its derivatives. Journal of the Chemical
Society, 888.
ELLIOTT, M. 1964. The pyrethrins and related compounds. V - Purification of (+)-
pyrethrolone as the monohydrate, and the nature of ‘Pyrethrolone-C’. Journal
of the Chemical Society, 5225.
ELLIOTT, M. 1965. The pyrethrins and related compounds. VI - The structures of
the ‘enols’ of pyrethrolone. Journal of the Chemical Society, 3097.
SAWICKI, R.M. and ELLIOTT, M. 1965. Insecticidal activity of pyrethrum extract
and its four insecticidal constituents against house flies. VI - Relative toxicity
of pyrethrin I and pyrethrin II against four strains of house flies. Journal of
the Science of Food and Agriculture 16, 85.
ELLIOTT, M., JANES, N.F., JEFFS, K.A. NEEDHAM, P.H. and SAWICKI, R.M.
1965. New pyrethrin-like esters with high insecticidal activity. Nature 207,
938.
Michael Elliott 311
ELLIOTT, M., JANES, N.F., JEFFS, K.A., NEEDHAM, P.H., SAWICKI, R.M. and
STEVENSON, J.H. 1965. New insecticidal esters of chrysanthemic acid.
Proceedings of the 3rd British Insecticide and Fungicide Conference 432.
CORRAL, C. and ELLIOTT, M. 1965. The pyrethrins and related compounds. VII -
New pyrethrin-like compounds with ester and ketonic groups in the alcoholic
side chain. Journal of the Science of Food and Agriculture 16, 514.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H., PEARSON, B.C.and
STEVENSON, J.H. 1967. New synthetic insecticidal compounds related to the
pyrethrins. Proceedings of the 4th British Insecticide and Fungicide Conference
437.
ELLIOTT, M. 1967. Synthesizing pyrethrin-like insecticides. Science Journal
(3 March).
ELLIOTT, M., JANES, N.F. and JEFFS, K.A. 1967. 2,4-Dimethylphenylacetic acid
from cyanodihydrocarvone. Chemistry and Industry, 1175.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H. and PEARSON, B.C.
1967. 5-Benzyl-3-furylmethyl chrysanthemate: a new potent insecticide.
Nature 213, 493.
ELLIOTT, M., HARPER, S.H. and KAZI, M.A. 1967. Experiments on the synthesis
of the pyrethrins XIV -Rethrins and the cyclopentadienone related to
3-methylcyclopent-2-enone. (The pyrethrins and related compounds.
VIII-) Journal of the Science of Food and Agriculture 18, 167.
ELLIOTT, M., JANES, N.F. and PEARSON, B.C. 1967. The pyrethrins and related
compounds. IX - Alkenylbenzyl and benylbenzyl chrysanthemates. Journal of
the Science of Food and Agriculture 18, 325.
ELLIOTT, M., and JANES, N.F. 1967. Synthesis of 4-allyl- and 4-benzyl-2,6-
dimethylbromobenzene from 2,6-xylidine. Journal of the Chemical Society
(C), 1780.
ELLIOTT, M., 1968. The search for safer insecticides. Article No. 0911/8, No.7 in
Features Newsletter No.35, 1968.
ELLIOTT, M. and JANES, NF. 1969. Pyrethrin II and related esters obtained by
reconstitution. Chemistry and Industry, 270
BRAMWELL, A.F., CROMBIE, L., HEMESLEY, P., PATTENDEN, G., ELLIOTT, M. and
JANES, N.F. 1969. Nuclear magnetic resonance spectra of the natural pyrethrins
and related compounds. Tetrahedron 26, 1727.
ELLIOTT, M., JANES, N.F., JEFFS, K.A., CROMBIE, L., GOULD, R. and PATTENDEN,
G. 1969, Oxidative dimerisation of 2-alkenylcyclopent-2-ene-1, 4-diones by
manganese dioxide. Tetrahedron Letters, 373.
ELLIOTT, M., NEEDHAM, P.H. and POTTER, C. 1969. Insecticidal activity of the
pyrethrins and related compounds. II - Relative toxicity of esters from optical
and geometrical isomers of chrysanthemic, pyrethric and related acids and
optical isomers of cinerolone and allethrolone. Journal of the Science of Food
and Agriculture 20, 561.
312 Wolf Prize in Agriculture
ELLIOTT, M., JANES, N.F. and JEFFS, K.A. 1969. Condensation of ketones with
dimethyl dimethylmaleate. A synthesis of calythrone. Journal of the Chemical
Society (C), 1845.
ELLIOTT, M., 1969. Structural requirements for pyrethrin-like activity. Chemistry
and Industry, 776.
ELLIOTT, M., KIMMEL, E.C. and CASIDA, J.E. 1969. 3H-Pyrethrin I and -pyrethrin
II: Preparation and use in metabolism studies. Pyrethrum Post 10(2), 1.
ELLIOTT, M., JANES, N.F. and JEFFS, K.A. 1970. The pyrethrins and related
compounds. X - The methylbenzyl chrysanthemates. Pesticide Science 1, 49.
ELLIOTT, M., FORD, M.G. and JANES, N.F. 1970. Insecticidal activity of the pyrethrins
and related compounds. III - Penetration of pyrethroid insecticides into mustard
beetles (Phaedon cochleariae). Pesticide Science 1, 220.
CROMBIE, L., ELLIS, J.A., GOULD, R., PATTENDEN, G., ELLIOTT, M., JANES, N.F.
and JEFFS, K.A. 1971. Oxidative dimerizations of natural rethrolones and
related compounds with manganese dioxide. Journal of the Chemical Society
(C),9.
BARLOW, F., ELLIOTT, M., FARNHAM, A.W., HADAWAY, A.D., JANES, N.F.,
NEEDHAM, P.H. and WICKHAM, J.C. 1971. Insecticidal activity of the
pyrethrins and related compounds. IV - Essential features for insecticidal
toxicity in chrysanthemates and related cyclopropane esters. Pesticide Science
2, 115.
ELLIOTT, M., JANES, N.F. and PAYNE, M.C. 1971. The pyrethrins and related
compounds. XI - Synthesis of insecticidal esters of 4-hydroxycyclopent-2-
enones (nor-rethrins). Journal of the Chemistry Society (C), 2548.
ELLIOTT, M., JANES, N.F. and PEARSON, B.C. 1971. The pyrethrins and related
compounds. XII - Synthesis of 5-substituted 3-furoates and 3-thenoates,
intermediates for synthesis of insecticidal esters. Journal of the Chemical
Society (C), 2551.
ELLIOTT, M., JANES, N.F., KIMMEL, E.C. and CASIDA, J.E. 1971. Mammalian
metabolites of pyrethroids. Proceedings of the Second International Congress
of Pesticide Chemistry, February 1971.
ELLIOTT, M., JANES, N.F. and PEARSON, B.C. 1971. The pyrethrins and related
compounds. XIII - Methyl-, alkenyl- and benzyl-substituted furfuryl and
furylmethyl chrysanthemates. Pesticide Science 2, 243.
CASIDA, J.E., KIMMEL, E.C. ELLIOTT, M. and JANES, N.F. 1971. Oxidative metabolism
of pyrethrins in mammals. Nature 230, 327.
ELLIOTT, M. 1971. The relationship between the structure and the activity of
pyrethroids. Bulletin of the World Health Organization 44, 315.
ELLIOTT, M. and CASIDA, J.E. 1972. The pyrethrins and related compounds. XIV -
Optically pure pyrethroids labelled with deuterium and tritium in the
methylcyclopentenonyl ring. Journal of Agricultural and Food Chemistry
20, 295.
Michael Elliott 313
ELLIOTT, M., JANES, N.F., KIMMEL, E.C. and CASIDA, J.E. 1972. The pyrethrins
and related compounds. XV - Metabolic fate of pyrethrin I, pyrethrin II, and
allethrin administered orally to rates. Journal of Agricultural and Food
Chemistry 20, 300.
ELLIOTT, M., GAUGHAN, L.C. and CASIDA, J.E. 1972. The pyrethrins and
related compounds XVI - Preparation of tritium-labelled tetramethyl-
cyclopropanecarboxylic acid and insecticidal esters from it. Journal of
Agricultural and Food Chemistry 20, 731.
ELLIOTT, M., FARNHAM, A.W., FORD, M.G., JANES, N.F. and NEEDHAM, P.H. 1972.
Insecticidal activity of the pyrethrins and related compounds. V - Toxicity of
the methylbenzyl chrysanthemates to houseflies (Musca domestica) and
mustard beetles (Phaedon cochleariae). Pesticide Science 3, 25.
ELLIOTT, M., JANES, N.F. and SPANNER, J.A. 1973. The pyrethrins and related
compounds. XVII - Preparation of the insecticide bioresmethrin (5-benzyl-3-
furylmethyl(+)-trans-chrysanthemate) and related compounds labelled with
deuterium or tritum in the furan ring. Pesticide Science 4, 677.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H. and PULMAN D.A.
1973. Potent pyrethroid insecticides from modified cyclopropane acids. Nature
244, 456.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H., PULMAN, D.A. and
STEVENSON, J.H. 1973. A photostable pyrethroid. Nature 246, 169.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H., PULMAN, D.A.and
STEVENSON, J.H. 1973, NRDC 143, a more stable pyrethroid. Proceedings of
the 7th British Insecticide and Fungicide Conference, 721.
ELLIOTT, M. and JANES, N.F. 1973. Chemistry of the natural pyrethrins. Pyrethrum,
the Natural Insecticide. (Edited by J.E. Casida). Academic Press, p.55.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H. and PULMAN, D.A.
1974. Insecticidally active conformation of pyrethroids. ACS Symposium Series
No.2 80-91.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H. and PULMAN, D.A.
1974. Synthetic insecticide with a new order of activity. Nature 248,
710-711.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F. and NEEDHAM, P.H. 1974. Insecticidal
activity of the pyrethrins and related compounds. VI - Methyl-, alkenyl-,
benzyl-furfuryl and 3-furylmethyl chrysanthemates. Pesticide Science 5,
491-496.
BRIGGS, G.G., ELLIOTT, M., FARNHAM, A.W. and JANES, N.F. 1974. Structural
aspects of the knockdown of pyrethroids. Pesticide Science 5, 643-649.
ELLIOTT, M., JANES, N.F. and PULMAN, D.A. 1974. The pyrethrins and related
compounds. XVIII - Insecticidal 2,2-dimethylcyclopropane carboxylates with
new unsaturated substituents at position 3 of the cyclopropane ring. Journal
of the Chemical Society, Perkin Transactions 1, 2470-2474.
314 Wolf Prize in Agriculture
BURT, P.E., ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H.
and PULMAN, D.A. 1974. The pyrethrins and related compounds. XIX -
Geometrical and optical isomers of 2,2-dimethyl-3(2,2-dichlorovinyl)-
cyclopropanecarboxylic acid and insecticidal esters with 5-benzyl-furylmethyl
and 3-phenoxybenzyl alcohols. Pesticide Science 5, 791-799.
JANES, N.F. and ELLIOTT, M. 1975. Utilisation de la RMN en chimie des
pyrethrinoids. Feuille d’information du CILDA, No. 6, 5-14.
ELLIOTT, M and JANES N.F. 1075. Quelques aspects du developpement des
pyrethrinoides de synthese. Feuille d’information du CILDA, No.6, 15-26.
ELLIOTT, M., JANES, N.F. and GRAHAM-BRYCE, I.J. 1975. Retrospect on the discovery
of a new insecticide. Proceedings of 8th British Insecticide and Fungicide
Conference, 373-379.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H and PULMAN, D.A.
1975. Insecticidal activity of the pyrethrins and related compounds. VII -
Insecticidal dihalovinyl analogues of cis and trans chrysanthemates. Pesticide
Science 6, 537-542.
ELLIOTT, M., JANES, N.F., PULMAN, D.A., GAUGHAN, L.C., UNAI, T. and CASIDA,
J.E. 1976. Radiosynthesis and metabolism in rats of the 1R isomers of the
insecticide permethrin. Journal of Agricultural and Food Chemistry 24,
270-276.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H. and PULMAN, D.A.
1976. Insecticidal activity of the pyrethrins and related compounds.
IX - 5-Benzyl-3-furylmethyl-2,2-dimethylcyclopropane carboxylate with non-
ethylenic substituents at position 3 on the cyclopropane ring. Pesticide Science
7, 492-498.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H. and PULMAN, D.A.
1976. Insecticidal activity of the pyrethrins and related compounds. X -
5-Benzyl-3-furylmethyl-2,2-dimethylcyclopropanecarboxylate with ethylenic
substituents at position 3 on the cyclopropane ring. Pesticide Science 7,
499-502.
ELLIOTT, M. 1976. Chemistry, biochemistry and insecticidal action of synthetic
pyrethroids. Pesticide Science 7, 223-224.
BRIGGS, G.G., ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H.,
PULMAN, D.A. and YOUNG, S.R. 1976. Insecticidal activity of the pyrethrins
and related compounds. VII - Relation of polarity with activity in pyrethroids.
Pesticide Science 7, 236-240.
ELLIOTT, M. 1976. Properties and application of pyrethroids. Environmental Health
Perspectives 14, 3-13.
BURT, P.E., ELLIOTT, M., FARNHAM, A.W., JANES, N.F., NEEDHAM, P.H and
STEVENSON, J.H. 1977. Evaluation of pyrethroids for insect control in ‘Crop
Protection Agents’, Academic Press, 393-411.
Michael Elliott 315
ELLIOTT, M., FARNHAM, A.W., JANES, N.F. and KHAMBAY, B.P.S. 1982. Insecticidal
activity of the pyrethrins and related compounds. Part XIII - α-Substituted 3-
phenoxybenzyl esters. Pesticide Science 13, 407-414.
ELLIOTT, M. 1982. University research and industrial development. Proceedings of
a symposium, Research and Industry: a community? Paris: l’Ecole Nationale
Superieure Chimie de Paris.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., KHAMBAY, B.P.S. and PULMAN, D.A.
1983. The pyrethrins and related compounds. Part XXVI - Analogues of
fenvalerate from acyclic substituted 3-methylbutyric and related acids. Pesticide
Science 14, 1-8.
ELLIOTT, M., JANES, N.F. and KHAMBAY, B.P.S. 1983. The pyrethrins and related
compounds. Part XXVII - Esters of pulegenic acid and related acids. Pesticide
Science 14, 9-16.
ELLIOTT, M., JANES, N.F. and KHAMBAY, B.P.S. 1983. Insecticidal activity of the
pyrethrins and related compounds. Part XIII - Comparison of the effects of
a-substituents in different esters. Pesticide Science 14, 182-190.
BRIGGS, G.G., ELLIOTT, M. and JANES, N.F. 1983. Present status and future prospects
for synthetic pyrethroids. IUPAC Pesticide Chemistry. Human Welfare and the
Environment. Ed. J Miyamoto et al., Pergamon Press, Oxford and New York,
157164.
ELLIOTT, M. 1983. Developments in the chemistry and action of pyrethroids.
Natural products for innovative pest management. Ed. D.L. Whitehead and
W.S. Bowers. Pergamon Press, Oxford and New York, 127-150.
ELLIOTT, M., JANES, N.F., STEVENSON, J.H. and WALTERS, J.H.H. 1983. Insecticidal
activity of the pyrethrins and related compounds. Part XIV - Selectivity of
pyrethroid insecticides between Ephestia kuehniella and its parasite Venturia
canescens. Pesticide Science 14, 423-426.
ELLIOTT, M and JANES, N.F. 1983. Evolution of pyrethroid insecticides for
crop protection. Proceedings of the 10th International Congress of Plant
Protection Brighton, 1983. Plant Protection for Human Welfare. Volume 1,
216-223.
LAUFER, J., ROCHE, M., PELHATE, M., ELLIOTT, M., JANES, N.F. and SATELLE, D.B.
1984. Pyrethroid insecticides: action of deltamethrin and related compounds
on insect axonal sodium channels. Journal of Insect Physiology, Volume 30,
No. 5, 341-349.
ELLIOTT, M., JANES, N.F. and KHAMBAY, B.P.S. 1984. Relative insecticidal
activity of various 3-polyhaloethyl- 2,2-dimethylcyclopropane carboxylates.
Proceedings of the 1984 British Crop Protection Conference, Volume 3,
849-852.
ELLIOTT, M. 1985. A survey of the development and environmental chemistry of
pyrethroids, Pesticide Science 16, 192-193.
Michael Elliott 317
ELLIOTT, M. 1985. Lipophilic insect control agents. In: Recent Advances in the
Chemistry of Insect Control, R.S.C. Special Publication No. 53; Ed. JANES,
N.F., 73-102.
ELLIOTT, M. FARNHAM, A.W., JANES, N.F., JOHNSON, D.M., PULMAN, D.A. and
SAWICKI R.M. 1986. Insecticidal amides with selective potency against a
resistant (super-kdr) strain of houseflies (Musca domestica L.). Agricultural
and Biological Chemistry 50(5), 1347-1349.
BAYDAR, A.E., ELLIOTT, M., JANES, N.F. and KHAMBAY, B.P.S. 1985. Structure-
activity relationships in non-ester pyrethroids. 190th American Chemical
Society National Meeting, Chicago, 1985, Abstract No.18.
BRIGGS, G.G., ELLIOTT, M., JANES, N.F. and KHAMBAY, B.P.S. 1985. The
influence of fluorine substituents on the biological activity of pyrethroids.
190th American Chemical Society National Meeting, Chicago, 1985, Abstract
No.90.
ELLIOTT, M. 1985. A survey of the development and environmental chemistry of
pyrethroids. Pesticide Science 16, 192-193.
ELLIOTT, M. and JANES, N.F. 1984. L’evoluzione degli insetticidi piretroidi in
agricoltura. Informatore Fitopatologica (9), 11-14.
ELLIOTT, M., ELLIOTT, R.L., JANES, N.F., KHAMBAY, B.P.S. and PULMAN, D.A. 1986.
The pyrethrins and related compounds. Part XXVIII. Alkenyl- and Alkynyl-
substituted Benzyl Esters. Pesticide Science 7, 691-700.
ELLIOTT, M., ELLIOTT, R.L., JANES, N.F., KHAMBAY, B.P.S. and PULMAN, D.A. 1986.
The pyrethrins and related compounds. Part XXIX. Haloallylbenzyl Esters.
Pesticide Science 17, 701-707.
ELLIOTT, M., ELLIOTT, R.L., JANES, N.F., KHAMBAY, B.P.S. and PULMAN, D.A. 1986.
The pyrethrins and related compounds. Part XXX. Esters from Acids with
Mono-Halovinyl Side Chains. Pesticide Science 17, 708-714.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. KHAMBAY, B.P.S. and
SAWICKI R.M. 1987. Selectivity and resistance to non-ester pyrethroids and
N-akylamides in houseflies (Musca domestica L.) In: Combating resistance to
xenobiotics. Biological and chemical approaches. Eds. M.G. Ford, D.W.
Hollomon, B.P.S. Khambay and R.M. Sawicki. Chichester: Ellis Horwood,
pp. 306-313.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. and PULMAN, D.A.
1987. Synthesis and insecticidal activity of lipophilic amides. Part 1:
Introductory survey, and discovery of an active synthetic compound. Pesticide
Science 18, 191-201.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. and PULMAN, D.A.
1987. Synthesis and insecticidal activity of lipophilic amides. Part 2: Effect of
heteroatom replacements and of introducing methyl groups in the pentadiene
system. Pesticide Science 18, 203-209.
318 Wolf Prize in Agriculture
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. and PULMAN, D.A.
1987. Synthesis and insecticidal activity of lipophilic amides. Part 3: Influence
of chain length and terminal group in N-(2- methylpropyl)-2,4-dienamides.
Pesticide Science 18, 211-221.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M and PULMAN D.A.
1987. Synthesis and insecticidal activity of lipophilic amides. Part 4: The
effect of substituents on the phenyl group of 6-phenylhexa-2,4-dienamides.
Pesticide Science 18, 223-228.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. and PULMAN. D.A.
1987. Synthesis and insecticidal activity of lipophilic amides. Part 5: Influence
on activity of varying the substituent on nitrogen. Pesticide Science 18,
229-238.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. and PULMAN, D.A.
1987. Synthesis and insecticidal activity of lipophilic amides. Part 6: 6-
(Disubstituted-phenyl)hexa-2,4-dienamides. Pesticide Science 18, 239-244.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F. and KHAMBAY, B.P.S. 1988. The
pyrethrins and related compounds. Part XXXI: Alkoxyimino- substituted esters.
Pesticide Science 22, 231-249.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F. and KHAMBAY, B.P.S. 1988. The
pyrethrins and related compounds. Part XXXIII: Effective combinations in the
gemdimethyl-central group region of non-ester pyrethroids. Pesticide Science
23, 231-246.
BAYDAR, A.E., ELLIOTT, M., FARNHAM, A.W., JANES, N.F. and KHAMBAY, B.P.S.
1988. The pyrethrins and related compounds. Part XXXIV: Optimisation of
insecticidal activity in non-esters. Pesticide Science 23, 247-257.
ELLIOTT, M., FARNHAM, A.W., JANES, N.F., JOHNSON, D.M. and PULMAN, D.A.
1989. Synthesis and insecticidal activity of lipophilic amides. Part 7: Alternative
aromatic groups for phenyl in 6-phenylhexa-2,4- dienamides. Pesticide Science
26, 199-208.
ELLIOTT, M. 1989. The pyrethroids: Early Discovery, Recent Advances and the
Future. Pesticide Science 17, 337-351.
ELLIOTT, M. 1990. The Contribution of Pyrethroid Insecticides to Human Welfare.
L’actualité chimique, 57-70.
ELLIOTT, M. 1990. Safer Crop Protection - A Citation Classic Commentary. Current
Contents (Agriculture, Biology and Environmental Sciences) 21(36), 20.
ELLIOTT, M., PULMAN, D.A., LARKIN, J.P. and CASIDA, J.E. 1992. Insecticidal 1,3-
dithianes. J. Agricultural Food Chemistry 40, 147-151.
ELLIOTT, M. 1992. Pyrethroids at Rothamsted: A Case History. Proceedings of the
Conference: Technology Transfer and Implementation, London July 1992.
ELLIOTT, M. 1995. Chemicals in Insect Control. in ‘Pyrethrum Flowers: Production,
Chemistry, Toxicology and Uses. Eds. J.E. Casida and G.B. Quistad, Oxford
University Press, New York and Oxford, pp. 3-26.
Michael Elliott 319
1935–2003
Professor Jozef Stefaan Schell played a key role in the development of Agrobacterium
and its Ti plasmid as a vector for inserting genes into plants.
His subsequent work has moved to the use of transgenic plants to bring a new
dimension into the study of classical problems in plant physiology, particularly in
relation to hormone receptors and the mechanisms of hormone action.
Another particularly notable feature of Professor Schell’s work has been his
orchestration of collaborative efforts to spread the knowledge and use of these
techniques around the world.
The impact of his pioneering contribution and leadership is now beginning to
appear as companies are developing transgenic plants for pest and disease resistance
and for effecting a range of compositional changes of potential industrial
importance.
Ti-plasmid-derived genetic constructions are the only proven vehicles for the
introduction of foreign DNA into plants. The newly acquired DNA integrates into
the plant’s genome and becomes a stable constituent of the genetic make-up of the
transformed plant.
321
322 Wolf Prize in Agriculture
Professor Schell’s studies and research work of over many years led to the
elucidation of the molecular basis of crown gall formation on plants by
Agrobacterium tumefaciens and the mechanism of DNA transfer to higher plants,
and formed the basis for the development of sophisticated vectors and methods for
the introduction of novel or modified genes into plants. These developments now
allow the stable introduction of desirable genetic traits into plants, also those of
agricultural interest. Furthermore, the techniques and vector systems originating
from this work are being used to study gene expression in higher plants, in response
to hormones, heat shock, pathogens, light and symbiotically nitrogen fixing bacteria
and in a tissue specific manner. This led to the identification of tissue specific
promoters which can be used to direct the expression of newly introduced genes
in the desired tissue (organ) of the plant.
The work of Schell and his coworkers, therefore, represents a major contribution
not only to a scientific (molecular) understanding of gene transfer to and gene
expression in higher plants, but also offers major new tools for modern plant
breeding.
CURRICULUM VITAE
Personal data
Place of birth Antwerp (Belgium)
Date of birth July 20th, 1935
Citizenship Belgian
Marital status Married
Children 2
Education
October 1953-July 1957 - State University Gent (Belgium), Licentiate Zoology
September 1957-November 1960 - State University Gent (Belgium), Laboratory of
Microbiology (Dir.: Prof. J. De Ley). Field: Ph.D. in Microbiology (Comparative
Biochemistry)
April 1959-September 1959 - State University Utrecht (The Netherlands), Laboratory
of Microbiology (Dir.: Prof. K.C. Winkler). Field: Part of Ph.D. training in
Microbiology
September 1962-July 1963 - Hammersmith Hospital, London (U.K.), Microbial
Genetics Research Unit of the Medicinal Research Council (Dir.: Prof. W.
Hayes). Field: Post-doctoral fellow in Microbial Genetics
September 1967-October 1967 - National Institutes of Health (Dir.: Dr. A.
Weissbach), Bethesda, Maryland (USA). Field: Isolation of circular DNA species
from bacteria
Jozef Stefaan Schell 323
Positions held
1957-1960 Bursar of the “Instituut tot Aanmoediging van het
Wetenschappelijk Onderzoek in Nijverheid en Landbouw”
(I.W.O.N.L. - Belgium)
1960-1961 “Aspirant” of the “Nationaal Fonds voor Wettenschappelijk
Onderzoek” (N.F.W.O. - Belgium)
1961-1967 “Werkleider” of the Laboratory of Microbiology State
University of Gent (Belgium)
1967-1970 Associate Professor (Docent) and Director of the Laboratory
of General Genetics, State University of Gent (Belgium)
1970-1978 Full Professor (Gewoon Hoogleraar) and Director of the
Laboratory of General Genetics, State University Gent (Belgium)
1972-1978 Extraordinary Professor (Buitengewoon Hoogleraar), Free
University Brussels (Belgium)
1978-1995 Extraordinary Professor (Buitengewoon Hoogleraar) and
Director (until 1988) of the Laboratory of General Genetics,
State University Gent (Belgium)
1978-2000 Director of the Max-Planck-Institut für Züchtungs-forschung,
Köln (Germany), (Emeritus since 01.08.2000)
1980- Honorary Professor at the University of Cologne (Germany)
10/1994-09/1998 Professeur de biologie moléculaire des plantes, Collège de
France, Paris (France)
10/1998- Professeur honoraire, Collège de France, Paris (France)
1977 - Chairman and Organizer of the “Sessions on Crown gall” of the 3rd
Gordon Research Conference on Plant Cell and Tissue Culture”
- Offer to become a scientific member of the Max-Planck-Gesellschaft”
1978 - Offer to become Full Professor of Biology at Harvard University (Cambridge,
USA)
- Appointment as Guest Professor at the Aarhus University (Institut for
Molekyaer Biologi og Plantenfysiologi), Aarhus (Denmark)
- Nomination as Editorial Board Member of Molecular and General Genetics
(Springer Verlag)
Director of the Max-Planck-Institut für Züchtungsforschung, Köln
(Germany) (emeritus since 01.08.2000)
1979 - Award of the Francquiprize (Belgium)
- Election as Active Member of The New York Academy of Sciences
1980 - Nomination as Editorial Board Member of Journal of Molecular and Applied
Genetics (Raven Press)
- Nomination as Editorial Board Member of Plant Cell Reports (Springer
Verlag) until June 1991
- Co-organizer of EMBL Symposium on “Molecular Biology Looks at Green
Plants”
- Honorary Professor at the University of Köln (FRG)
1981 - Appointment as Guest Professor at the University of California, Riverside
(Department of Biochemistry), Riverside, CA (USA)
- Elected member of the “Deutsche Akademie der Naturforscher Leopoldina”,
Halle (GDR)
- Nomination as Editorial Board Member of The EMBO Journal (IRL Press)
1982 - ONR Lectureship awarded by the “Society for Industrial Microbiology”
- Chairman of the EMBO Fund Committee (until 1984)
1983 - Member of the “Conseil Scientifique de l’ICP” (International Institute of
Cellular and Molecular Pathology), Brussels-Leuven (Belgium)
1984 - Holder of the Belgian Francqui chair at the Katholieke Universiteit Leuven
(K.U.L.), Belgium
- Member of the Scientific Board of the “Gesellschaft Deutscher Naturforscher
und Ärzte”
- Member of “Gesellschaft für Biologische Chemie”, Wuppertal (Germany)
- Chairman of the “Scientific Advisory Board” of the Otto Warburg Center,
Rehovot, Israel (until 2001)
1985 - Award of the “Mendel-Medaille” of the “Deutsche Akademie der
Naturforscher Leopoldina”
- Award of the “Otto Bayer-Preis” of the Otto Bayer-Stiftung
- Award of the “Prix Alexandre de Humboldt” of the Ministère de la
Recherche et de la Technologie and Alexander von Humboldt-Stiftung
Jozef Stefaan Schell 325
- Award of the “Prix Charles Leopold Mayer” of the Academie des Sciences,
Paris, France
- Member of the “Koninklijke Academie voor Wetenschappen, Letteren en
Schone Kunsten van Belgie”, Brussels, Belgium
- Member of the John Innes Council, John Innes Institute, Norwich, G.B.
(until 06/1998)
- Member of the Conseil Scientifique of ORSTOM (Institut Francais de
Recherche Scientifique pour le Développement en Coopération), Paris,
France
- Member of the Peer Advisory Group for Biotechnology Research at the
International Rice Research Institute, Manila, Philippines
- Foreign Member of Biological Research Center, Hungarian Academy of
Sciences, Szeged
- Member of “Arbeitsgruppe Biowissenschaften und Medizin”,
Wissenschaftsrat
- Member of the Editorial Board of “Mechanisms of Development” (MOD),
Elsevier Science Publishers B.V.
- Member of the Editorial Advisory Board of “Critical Reviews in Plant
Sciences”
- Senior Editor of “The Plant Journal” (Blackwell Scientific Publications),
until 10/1998
- Elected Member of the Board of Governors of the Weizmann Institute of
Science, Rehovot, Israel (until 2000)
- Chairman of the EMBO Council (until 1995)
- Chairman AMICA Science Board until 12/1997; since 01/1998 member
1991 - Award of the “Hansen Gold Medal 1991” of the Emil Christian Hansen
Foundation, Denmark
- Member of the International Board of the Shasha Institute for International
Seminars, Jerusalem
- Full member of the Board of Governors of the Hebrew University of
Jerusalem, Israel
- Member of the Scientific Advisory Council of “Alfried Krupp von Bohlen
und Halbach Stiftung”, Essen, Germany
- Chairman of the Founding Committee of the Institute of Plant Biochemistry,
Halle
1992 - Award of the “Feodor Lynen Lecture 1992” medal (Miami Winter
Symposium)
- Member of the Scientific Committee on the Application of Science to
Agriculture, Forestry and Aquaculture (CASAFA)
- Guest Professor at the Collège de France in Paris, from March 18 to April
8, 1992
Jozef Stefaan Schell 327
- Award of the “Sir Hans Krebs Medal” by the Federation of the European
Biochemical Societies, Barcelona (Spain)
1997 - Founding Member of the Euroscience Association, Paris (France)
- “Doctor Philosophiae Honoris Causa”, Tel Aviv University (Israel)
- Nomination as Editorial Board Member of “Molecular Cell” (until
12/2001)
- “Première Grande Médaille d’Or de l’Academie des Sciences”, Paris (France)
1998 - “Japan Prize” for Biotechnology in Agricultural Sciences by the “Science
and Technology Foundation of Japan”
- “Gulden Spoor” in de pijler Wetenschap en bedrijfsevenementen, Comité
2002 Vlaanderen-Europa, Brugge (Belgium)
Member of “Conseil National de la Science”, Ministère de l’Education
Nationale, de la Recherche et de la Technologie, Paris (France), 10/1998-
- Professeur honoraire, Collège de France, Paris (France), 10/1998-
1999 - “Commandeur in de Leopoldsorde”, Brussels (Belgium), 06/1999
“Doctor of Science honoris causa”, University of East Anglia, Norwich
(Great Britain), 07/1999
2000 - Elected Governor Emeritus of the Weizmann Institute of Science, Rehovot,
Israel
16. Schreier, P.H., Reif, H.-J., and Schell, J. Gentechnik. In: “Pflanzenproduktion
im Wandel”, G. Haug, G. Schuhmann, G. Fischbeck (Eds.), VCH Verlags-
gesellschaft, Weinheim, p. 27-39 (1990).
17. Staiger, D., Kaulen, H., and Schell, J. A nuclear factor recognizing a positive
regulatory upstream element of the Antirrhinum majus chalcone synthase
promoter. Plant Physiol. 93, 1347-1353 (1990).
18. Steinbiss, H.-H., Töpfer, R., Pröls, M., and Schell, J. Transient gene expression
in tobacco protoplasts and seed derived embryos of wheat. In: “Proceedings
of the International Congress of Plant Physiology”, New Delhi, India, February
15-20, 1988, S.K. Sinha, P.V. Sane, S.C. Bhargava, P.K. Agrawal (Eds.) Society
for Plant Physiology and Biochemistry, New Delhi, India, Vol. 1, pp. 658-659
(1990).
19. Töpfer, R., Gronenborn, B., Schaefer, S., Schell, J., and Steinbiß, H.-H.
Expression of engineered wheat dwarf virus in seed-derived embryos. Physiol.
Plantarum 79, 158-162 (1990).
20. Töpfer, R., and Schell, J. Ansätze zur Isolation von Genen der de novo
Fettsäurebiosynthese. In: “Pflanzliche Öle im chemisch-technischen Sektor”,
Schriftenreihe des Bundesministers für Ernährung, Landwirtschaft und
Forsten, Reihe A: Angewandte Wissenschaft, Heft 391, Landwirtschaft-sverlag
GmbH., Münster-Hiltrup, pp. 221-225 (1990).
21. Walden, R., Koncz, C., and Schell, J. The use of gene vectors in plant molecular
biology. Meth. Mol. Cell. Biol. 1, 175-194 (1990).
22. Walden, R., and Schell, J. Techniques in plant molecular biology - progress
and problems. Eur. J. Biochem. 192, 563-576 (1990).
23. Wingender, R., Röhrig, H., Höricke, C., and Schell, J. cis-regulatory elements
involved in ultraviolet light regulation and plant defense. The Plant Cell 2,
1019-1026 (1990).
24. Barbier-Brygoo, H., Ephrotikhine, G., Klämbt, D., Maurel, C., Palme, K., Schell,
J. and GUERN, J. Perception of the auxin signal at the plasma membrane of
tobacco mesophyll protoplasts. Plant J. 1, 83-93 (1991).
25. Campos, N., Feldwisch, J., Zettl, R., Boland, W., Schell, J., and Palme, K.
Identification of auxin-binding proteins using an improved assay for
photoaffinity labeling with 5-N3-(7-3H)-indole-3-acetic acid. Technique 3,
69-75 (1991).
26. Davidson, A., Kendiek, J., Peerenbaum, E., Schell, J., and Steinbiss, H.-H.
Genome analysis of Barley Yellow Mosaic Virus RNA1 and RNA2. In: “Barley
Genetics VI” Vol. I, Short Papers, Munck, L., Kirkegaard, K., and Jensen, B.
(Eds.), (Proceedings of the Sixth International Barley Genetics Symposium,
July 22-27, 1991, Helsingborg, Sweden), Munksgaard International Publishers
Ltd., Copenhagen, pp. 608-609 (1991).
Jozef Stefaan Schell 331
27. Davidson, A.D., Pröls, M., Schell, J., and Steinbiss, H.-H. The nucleotide
sequence of RNA 2 of barley yellow mosaic virus. J. Gen. Virol. 72, 989-993
(1991).
28. Estruch, J.J., Chriqui, D., Grossman, K., Schell, J., and Spena, A. The plant
oncogene rolC is responsible for the release of cytokinins from glucoside
conjugates. EMBO J. 10, 2889-2895 (1991).
29. Estruch, J.J., Prinsen, E., van Onckelen, H., Schell, J., and Spena, A. Viviparous
leaves produced by somatic activation of an inactive cytokinin-synthesizing
gene. Science 254, 1364-1367 (1991).
30. Estruch, J.J., Schell, J., and Spena, A. The protein encoded by the rolB plant
oncogene hydrolyses indole glucosides. EMBO J. 10, 3125-3128 (1991).
31. Fraley, R., and Schell, J. Plant biotechnology - Editorial overview. Current
Opinion in Biotechnol. 2, 145-146 (1991).
32. Fritz, C.C., Herget, T., Wolter, F.P., Schell, J., and Schreier, P.H. Reduced steady-
state levels of rbcS mRNA in plants kept in the dark are due to differential
degradation. Proc. Natl. Acad. Sci. USA 88, 4458-4462 (1991).
33. Fritze, K., Staiger, D., Czaja, I., Walden, R., Schell, J., and Wing, D.
Developmental regulation and UV light regulation of the snapdragon chalcone
synthase promoter. The Plant Cell 3, 893-905 (1991).
34. Kammann, M., Matzeit, V., Schmidt, B., Schell, J., Walden, R., and Gronenborn,
B. Geminivirusbased shuttle vectors capable of replication in Escherichia coli
and monocotyledonous plant cells. Gene 104, 247-252 (1991).
35. Kammann, M., Schalck, H.-J., Matzeit, V., Schaefer, S., Schell, J., and
Gronenborn, B. DNA replication of wheat dwarf virus, a geminivirus, requires
two cis-acting signals. Virology 184, 786-790 (1991).
36. Koncz, C., Schmülling, T., Spena, A., and Schell, J. T-DNA gene-functions. In:
“Plant Molecular Biology 2” (Proceedings on Elmau-Meeting, May 13-23,
1990) R.G. Herrmann, and B. Larkins (Eds.) Plenum Press, New York,
pp. 205-209 (1991).
37. Kondorosi, A., Kondorosi, E., John, M., Schmidt, J., and Schell, J. The role
of nodulation genes in bacterium-plant communication. In: “Genetic
Engineering”, Vol. 13, Setlow, J.K. (Ed.), Plenum Press, New York and London,
pp 115-136 (1991).
38. Kondorosi, E., Pierre, M., Cren, M., Haumann, U., Buire, M., Hoffmann, B.,
Schell, J., and Kondorosi, A. Identification of NolR, a negative transacting
factor controlling the nod regulon in Rhizobium meliloti. J. Mol. Biol. 222,
885-896 (1991).
39. Körber, H., Strizhov, N., Staiger, D., Feldwisch, J., Olsson, O., Sandberg, G.,
Palme, K., Schell, J., and Koncz, C. T-DNA gene 5 of Agrobacterium modulates
auxin response by autoregulated synthesis of a growth hormone antagonist
in plants. EMBO J. 10, 3983-3991 (1991).
332 Wolf Prize in Agriculture
40. Leung, J., Fukuda, H., Wing, D., Schell, J., and Masterson, R. Functional
analysis of cis-elements, auxinresponse and early developmental profiles of
the mannopine synthase bidirectional promoter. Mol. Gen. Genet. 230,
463-474 (1991).
41. Matzeit, V., Schaefer, S., Kammann, M., Schalck, H.-J., Schell, J. and
Gronenborn, B. Wheat dwarf virus vectors replicate and express foreign
genes in cells of monocotyledonous plants. The Plant Cell 3, 247-258
(1991).
42. Mayerhoffer, R., Koncz-Kalman, Z., Nawrath, C., Bakkeren, G., Crameri, A.,
Angelis, K., Redei, G.P., Schell, J., Hohn, B., and Koncz, C. T-DNA integration:
a mode of illegitimate recombination in plants. EMBO J. 10, 697-704 (1991).
43. Mukhopadhyay, A., Töpfer, R., Pradhan, A.K., Sodhi, Y.S., Steinbiß, H.-H.,
Schell, J., and Pental, D. Efficient regeneration of Brassica oleracea hypocotyl
protoplasts and high frequency genetic transformation by direct DNA uptake.
Plant Cell Rep. 10, 375-379 (1991).
44. Palme, K., Hesse, T., Moore, I., Campos, N., Feldwisch, J., Garbers, C., Hesse,
F., and Schell, J. Hormonal modulation of plant growth: the role of auxin
perception. Mech. Dev. 33, 97-106 (1991).
45. Palme, K., and Schell, J. Auxin receptors take shape. Current Biology 1,
228-230 (1991).
46. Pechan, P.M., Bartels, D., Brown, D.C.W., and Schell, J. Messenger-RNA and
protein changes associated with induction of Brassica microspore
emryogenesis. Planta 184, 161-165 (1991).
47. Prinsen, E., Chauvaux, N., Schmidt, J., John, M., Wieneke, U., DE Greef, J.,
Schell, J., and van Onckelen, H. Stimulation of indole-3-acetic acid production
in Rhizobium by flavonoids. FEBS Letters 282, 53-55 (1991).
48. Schmidt, J., John, M., Kondorosi, E., Kondorosi, A., and Schell, J. Recent
progress in the elucidation of the function of nodAB and C genes of Rhizobium
meliloti. Israel J. Bot. 40, 165-169 (1991).
49. Schmidt, J., John, M., Wieneke, U., Stacey, G., Röhrig, H., and Schell, J. Studies
on the function of Rhizobium meliloti nodulation genes. In: “Molecular
Genetics of Plant-Microbe Interactions”, Hennecke, H. and Verma, D.P.S.
(Eds.), Kluwer Academic Publishers, Dordrecht, The Netherlands,
pp. 150-155 (1991).
50. Staiger, D., Becker, F., Schell, J., Koncz, C., and Palme, K. Purification of
tobacco nuclear proteins binding to a CACGTG motif of the chalcone synthase
promoter by DNA affinity chromatography. Eur. J. Biochem. 199, 519-527
(1991).
51. Stieger, M., Neuhaus, G., Momma, T., Schell, J., and Kreuzaler, F. Self assembly
of immunoglobulins in the cytoplasm of the alga Acetabularia mediterranea.
Plant Science 73, 181-190 (1991).
Jozef Stefaan Schell 333
52. Walden, R., Hayashi, H., and Schell, J. T-DNA as a gene tag. Plant J. 1,
281-288 (1991).
53. Walden, R., and Schell, J. Tissue culture and the use of transgenic plants to
study plant development. In Vitro Cell. Dev. Biol. 27P, 1-10 (1991).
54. Zettl, R., Campos, N., Feldwisch, J., Schell, J., Boland, W., and Palme, K.
Synthesis and application of 5'-Azido-[3,6-3H2]- naphthylphthalamic acid,
a photo-activatable probe for auxin efflux carrier proteins. Technique 3,
151-158 (1991).
55. Becker, D., Kemper, E., Schell, J., and Masterson, R. New plant binary vectors
with selectable markers located proximal to the left T-DNA border. Plant
Mol. Biol. 20, 1195-1197 (1992).
56. Campos, N., Bako, L., Feldwisch, J., Schell, J., and Palme, K. A protein from
maize labeled with azido-IAA has novel ß-glucosidase activity. Plant J. 2,
675-684 (1992).
57. Feldwisch, J., Zettl, R., Hesse, F., Schell, J., and Palme, K. An auxin-binding
protein is localized to the plasma membrane of maize coleoptile cells:
Identification by photoaffinity labeling and purification of a 23-kDa
polypeptide. Proc. Natl. Acad. Sci. USA 89, 475-479 (1992).
58. Fraley, R., and Schell, J. Plant biotechnology - Editorial overview. Current
Opinion in Biotechnol. 3, 139-140 (1992).
59. Grevelding, C., Becker, D., Kunze, R., von Menges, A., Fantes, V., Schell, J.,
and Masterson, R. High rates of Ac/Ds germinal transposition in Arabidopsis
suitable for gene isolation by insertional mutagenesis. Proc. Natl. Acad. Sci.
USA 89, 6085-6089 (1992).
60. Kemper, E., Grevelding, C., Schell, J., and Masterson, R. Improved method for
the transformation of Arabidopsis thaliana with chimeric dihydrofolate
reductase constructs which confer methotrexate resistance. Plant Cell Rep.
11, 118-121 (1992).
61. Klein, B., Pawlowski, K., Höricke-Grandpierre, C., Schell, J., and Töpfer, R.
Isolation and characterization of a cDNA from Cuphea lanceolata
encoding a ß-ketoacyl-ACP reductase. Mol. Gen. Genet. 233, 122-128
(1992).
62. Koncz, C., Nemeth, K., Redei, G.P., and Schell, J. T-DNA insertional mutagenesis
in Arabidopsis. Plant Mol. Biol. 20, 963-976 (1992).
63. Koncz, C., and Schell, J. Exploitation of Agrobacterium tumefaciens. In:
“Development The Molecular Genetic Approach”, V.E.A. Russo, S. Brody,
D. Cove, and S. Ottolenghi (Eds.) Springer Verlag, Berlin Heidelberg,
pp. 217-224 (1992).
64. Koncz, C., Schell, J., and Redei, G.P. T-DNA transformation and insertion
mutagenesis. In: “Methods in Arabidopsis Research”, C. Koncz, N.-H. Chua,
and J. Schell (Eds.), World Scientific, Singapore, pp. 224-273 (1992).
334 Wolf Prize in Agriculture
65. Logemann, J., Jach, G., Tommerup, H., Mundy, J., and Schell, J. Expression of
a barley ribosome-inactivating protein leads to increased fungal protection
in transgenic tobacco plants. Bio/Technol. 10, 305-308 (1992).
66. Logemann, J., and Schell, J. Das Dilemma der Landwirtschaft. In: “Edition
Zeitthema”, 1/92 Gentechnologie I, Hargitay & List Verlag, Wien, pp. 53-55
(1992).
67. Logemann, J., and Schell, J. Die Landwirtschaft zwischen
Produktivitätssteigerung und zunehmender Umweltbelastung: Kann das
Dilemma durch Gentechnologie gelöst werden? GRUR, Heft 4, VCH
Verlagsgesellschaft mbH., Weinheim, pp. 248-252 (1992).
68. Maas, C., Reichel, C., Schultze, S.C., Matzeit, V., Schell, J., and Steinbiss, H.-H.
Modular structure of monocot expression vectors: A novel stimulatory element
combined with sh 1 intron 1. In: “Technological Impacts and Limiting Factors
of Plant Genetic Transformation” (Biotechnologies’92) Le Conseil Régional
de Picardie (Ed.), pp. 78-88 (1992).
69. Maas, C., Schell, J., and Steinbiss, H.-H. Applications of an optimized monocot
expression vector in studying transient gene expression and stable
transformation of barley. Physiol. Plant. 85, 367-373 (1992).
70. Moore, I., Feldwisch, J., Campos, N., Zettl, R., Brzobohaty, B., Bako, L.,
Schell, J., and Palme, K. Auxin-binding proteins of Zea mays identified by
photoaffinity labelling. Proceedings of “Molecular Biology of Plant Hormones
and their Receptors”, Edinburgh, Sept. 45, 1991. Biochem. Soc. Trans. 20,
70-73 (1992).
71. Nitschke, K., Fleig, U., Schell, J., and Palme, K. Complementation of the cs dis
2-11 cell cycle mutant of Schizosaccharomyces pombe by a protein
phosphatase from Arabidopsis thaliana. EMBO J. 11, 1327-1333 (1992).
72. Palme, K., Dieffenthal, T., Vingron, M., Sander, C., and Schell, J. Molecular
cloning and structural analysis of genes from Zea mays (L.) coding for
members of the ras-related ypt gene family. Proc. Natl. Acad. Sci. USA 89,
787-791 (1992).
73. Palme, K., Feldwisch, J., Peters, W.S., Schell, J., Zettl, R., Campos, N., and
Felle, H. Auxin binding proteins are located in the ER and in the plasma
membrane: Identification by photoaffinity labeling and characterization. In:
“Progress in Plant Growth Regulation”, C.M. Karssen, L.C. Van Loon, and D.
Vreugdenhill (Eds.), Kluwer Academic Publishers, Dordrecht, pp. 73-81
(1992).
74. Palme, K., Hesse, T., Campos, N., Garbers, C., Yanofsky, M.F., and Schell, J.
Molecular analysis of an auxin-binding protein gene located on chromosome
4 of Arabidopsis. The Plant Cell, 4, 193-201 (1992).
75. Peerenbaum, E., Pröls, M., Schell, J., Steinbiss, H.-H., and Davidson, A.D. The
complete nucleotide sequence of RNA 1 of a German isolate of barley yellow
Jozef Stefaan Schell 335
mosaic virus and its comparison with a Japanese isolate. J. Gen. Virol. 73,
1303-1308 (1992).
76. Roberts, D.M., Besl, L., Oh, S.-H., Masterson, R.V., Schell, J., and Stacey, G.
Expression of a calmodulin methylation mutant affects the growth and
development of transgenic tobacco plants. Proc. Natl. Acad. Sci. USA 89,
8394-8398 (1992).
77. Schell, J. Plant biotechnology: A powerful tool to use plant resources and to
improve the environmental impact of agriculture. In: “Natural Resources
and Human Health - Plants of medicinal and nutritional value” Proceedings of
the 1st WHO Symposium on Plants and Health for All: Scientific Advancement,
Kobe, Aug. 26-28, 1991, S. Baba, O. Akerele, and Y. Kawaguchi (Eds.), Elsevier
Science Publishers B.V., Amsterdam, pp. 49-60 (1992).
78. Schell, J. Isolation and characterization of genes involved in phytohormone
activity. In: “Proceedings of the VII International Symposium in Conjunction
with the Awarding of the International Prize for Biology - Cellular Basis of
Growth and Development in Plants, Osaka, Nov. 26-28, 1991”, H. Shibaoka
(Ed.), Osaka University, Osaka, pp. 113-118 (1992).
79. Schell, J. Agrobacterium-Mediated Gene Transfer to Plants: Its Impact
on Plant Breeding, Studies of Gene Expression and Plant Developmental
Biology. In: “Molecular Biology of the Cell” (Final Report of the
Sonderforschungsbereich “Molekularbiologie der Zelle” 1970-1988,
DFG Deutsche Forschungsgemein-schaft), W. Doerfler (Ed.), VCH
Verlagsgesellschaft mbH, Weinheim, pp. 125-143 (1992).
80. Schell, J. Plant biotechnology: A powerful tool to use plant resources and to
improve the environmental impact of agriculture. In: “Basic Life Sciences
and Human Society”, H. Pfeiffer and T. Schmidt-Dörr (Eds.), Proceedings of
the 10th Discoveries Symposium 1991, Bonn. Alexander von Humboldt
Foundation and Honda Foundation. Druckpartner Moser GmbH., Rheinbach,
pp. 141-152 (1992).
81. Schell, J., Koncz, C., Spena, A., Walden, R., John, M., and Schmidt, J.
Agrobacterium and Rhizobium and the genetic control of plant development.
In: “Advances in Gene Technology: Feeding the World in the 21st Century”
W.J. Whelan, F. Ahmad, H. Bialy, S. Black, M.L. King, M.B. Rabin, L.P.
Solomonson, and I.K. Vasil (Eds.), Proceedings of the 1992 Miami Bio/
Technology Winter Symposium. Miami Short Reports Vol. 2, IRL Press at
Oxford University Press, appendix (1992).
82. Schell, J., Koncz, C., Spena, A., Walden, R., and Palme, K. Characterization of
genes affecting plant growth and development. In: “Harnessing Biotechnology
for the 21st Century” M.R. Ladisch, and A. Bose (Eds.), Proceedings of the
Ninth International Biotechnology Symposium and Exposition, Crystal City,
Aug. 16-21, 1992, American Chemical Society, Washington, pp. 496-499
(1992).
336 Wolf Prize in Agriculture
83. Schell, J., and Logemann, J. Pflanzliche Gentechnologie - Perspektiven für die
praktische Anwendung in der Landwirtschaft. Schering Lecture, Heft 6,
Schering Berlin (1992).
84. Schell, J., Schmidt, J., John, M., and Röhrig, H. Potential and limitations of
developing new plant-microbe interactions. In: “Nodulation and Nitrogen
Fixation in Rice - Potential and Prospects”, G.S. Khush and J. Bennett (Eds.),
(Proceedings of the Workshop on Nodulation and Nitrogen Fixation in Rice,
Manila, Febr. 24-27, 1992), IRRI, Manila, pp. 103-105 (1992).
85. Schmülling, T., Schell, J., and Spena, A. Construction of cytokinin
overproducing mutants in higher plants. In: “Physiology and Biochemistry of
Cytokinins in Plants” M. Kaminek, D.W.S. Mok, and E. Zazimalová (Eds.),
Proceedings of the International Symposium on Physiology and Biochemistry
of Cytokinins in Plants, held in Liblice, Czechoslovakia, Sept. 10-14, 1990,
Academic Publishing bv, The Hague, The Netherlands, pp. 87-94 (1992).
86. Spena, A., Estruch, J.J., Aalen, R.D., Prinsen, E., Parets-Soler, A., Nacken, W.,
Sommer, H., Chriqui, D., Grossmann, K., van Onckelen, H., and Schell, J.
Transgenic plants and transgenic plant mosaics for the expression of pathogen
derived genes able to affect phytohormone activity. In: “Progress in Plant
Growth Regulation” C.M. Karssen, L.C. Van Loon, and D. Vreugdenhil (Eds.),
Proceedings of the 14th International Conference on Plant Growth Substances,
Amsterdam, July 21-26, 1991, Kluwer Academic Publishers, Dordrecht,
pp. 724-730 (1992).
87. Spena, A., Estruch, J.J., Hansen, G., Langenkemper, K., Berger, S., and Schell, J.
The Rhizogenes tale: modification of plant growth and physiology by an
enzymatic system of hydrolysis of phytohormone conjugates. In: “Advances
in Molecular Genetics of Plant-Microbe Interactions” Vol. 2, E.W. Nester and
D.P.S. Verma (Eds.), Proceedings of the Sixth International Symposium on
Molecular Plant-Microbe Interactions, July 11-16, 1992, Seattle. Kluwer
Academic Publishers, Dordrecht, pp. 109-124 (1992).
88. Spena, A., Estruch, J.J., and Schell, J. On microbes and plants: new insights in
phytohormonal research. Current Opinion in Biotechnol. 3, 159-163 (1992).
89. Tamas, I.A., Langbridge, W.H.R., Abel, S.D., Crawford, S.W., Randall, J.D.,
Schell, J., and Szalay, A.A. Hormonal control of apical dominance. Studies in
tobacco transformed with bacterial luciferase and Agrobacterium rol genes. In:
“Progress in Plant Growth Regulation” Proceedings of the 14th International
Conference on Plant Growth Substances, Amsterdam, 21-26 July, 1991, C.M.
Karssen, L.C. Van Loon, and D. Vreugdenhil (Eds.), Kluwer Academic
Publishers, Dordrecht, pp. 418-430 (1992).
90. Zettl, R., Feldwisch, J., Boland, W., Schell, J., and Palme, K. Azido-(3,6-3H2)-N-
1-naphthylphthalamic acid, a photoactivatable probe for naphthylphthalamic
acid receptor proteins from higher plants: Identification of a 23-kDa protein
Jozef Stefaan Schell 337
from maize coleoptile plasma membranes. Proc. Natl. Acad. Sci. USA 89,
480-484 (1992).
91. Bachmair, A., Becker, F., and Schell, J. Use of a reporter transgene to generate
Arabidopsis mutants in ubiquitin-dependent protein degradation. Proc. Natl.
Acad. Sci. USA 90, 418-421 (1993).
92. Becker, F., Buschfeld, E., Schell, J., and Bachmair, A. Altered response to
viral infection by tobacco plants perturbed in ubiquitin system. Plant J. 3,
875-881 (1993).
93. Kendiek, J., Davidson, A.D., Schultze, S.C., Schell, J., and Steinbiss, H.-H.
Identification and classification of a resistance breaking strain of barley
yellow mosaic virus. Ann. Appl. Biol. 122, 481-491 (1993).
94. Brzobohaty, B., Moore, I., Kristoffersen, P., Bako, L., Campos, N., Schell, J.,
and Palme, K. Release of active cytokinin by a ß-glucosidase localized to the
maize root meristem. Science 262, 1051-1054 (1993).
95. de Bruijn, F.J., and Schell, J. Regulation of plant genes specifically induced in
developing and mature nitrogen-fixing nodules: Cis-acting elements and trans-
acting factors. In: “Control of Plant Gene Expression”, D.P.S. Verma (Ed.),
CPC Press, Boca Raton, pp. 241-257 (1993).
96. Dehio, C., Grossmann, K., Schell, J., and Schmülling, T. Phenotype and
hormonal status of transgenic tobacco plants overexpressing the rolA gene
of Agrobacterium rhizogenes T-DNA. Plant Mol. Biol. 23, 1199-1210 (1993).
97. Dehio, C., and Schell, J. Stable expression of a single-copy rolA gene in
transgenic Arabidopsis thaliana plants allows an exhaustive mutagenic analysis
of the transgene-associated phenotype. Mol. Gen. Genet. 241, 359-366
(1993).
98. Dehio, C., Schell, J., and Koncz, C. Agrobacterium und Rhizobium -
Bodenbakterien als Pflanzeningenieure. In: “Extremophile” Mikro-organismen
in ausgefallenen Lebensräumen, K. Hausmann, B.P. Kremer (Eds.), VCH
Verlagsgesellschaft, Weinheim, pp. 257-277 (1993).
99. Fraley, R., and Schell, J. Plant biotechnology - Editorial overview. Current
Opinion in Biotechnol. 4, 133-134 (1993).
100. Grevelding, C., Fantes, V., Kemper, E., Schell, J., and Masterson, R. Single-
copy T-DNA insertions in Arabidopsis are the predominant form of integration
in root-derived transgenics, whereas multiple insertions are found in leaf-
discs. Plant Mol. Biol. 23, 847-860 (1993).
101. Grosskopf, E., Cam Ha, D.T., Wingender, R., Röhrig, H., Szecsi, J., Kondorosi,
E., Schell, J., and Kondorosi, A. Enhanced levels of chalcone synthase in
alfalfa nodules induced by a Fix- mutant of Rhizobium meliloti. Molec. Plant-
Microbe Interact. 6, 173-181 (1993).
102. Hesse, T., Garbers, C., Brzobohaty, B., Kreimer, G., Söll, D., Melkonian, M.,
Schell, J., and Palme, K. Two members of the ERabp gene family are expressed
338 Wolf Prize in Agriculture
126. Bako, L., Nuotio, S., Dudits, D., Schell, J., and Koncz, C. Rnapii: A specific
target for the cell cycle kinase complex. In: “Plant Promoters and Transcription
Factors”, (Results and Problems in Cell Differentiation 20), L. Nover (Ed.),
Springer-Verlag, Berlin, Heidelberg, pp. 25-64 (1994).
127. Becker, J., Siegert, H., Logemann, J., and Schell, J. Begleitende
Sicherheitsforschung zur Freisetzung genetisch veränderter Petunien. In:
“Biologische Sicherheit”, Band 3, Ch. Buchholz (Ed.) Projektträger Biologie,
Energie, Ökologie, Forschungszentrum Jülich GmbH., Werka-Druck GmbH.,
Linnich, pp. 563-578 (1994).
128. Campos, N., Schell, J., and Palme, K. In vitro uptake and processing of maize
auxin-binding proteins by ER-derived microsomes. Plant Cell Physiol. 35,
153-161 (1994).
129. Dehio, C., and Schell, J. Identification of plant genetic loci involved in a
posttranscriptional mechanism for meiotically reversible transgene silencing.
Proc. Natl. Acad. Sci. USA 91, 5538-5542 (1994).
130. Hewelt, A., Prinsen, E., Schell, J., van Onckelen, H., and Schmülling, T.
Promoter tagging with a promoterless ipt gene leads to cytokinin induced
phenotypic variability in transgenic tobacco plants: implications of gene
dosage effects. Plant J. 6, 879-891 (1994).
131. Jasper, F., Koncz, C., Schell, J., and Steinbiss, H.-H. Agrobacterium
T-strand production in vitro: Sequence-specific cleavage and 5' protection of
single-stranded DNA templates by purified VirD2 protein. Proc. Natl. Acad.
Sci. USA 91, 694-698 (1994).
132. Koncz, C., Martini, N., Szbados, L., Hrouda, M., Bachmair, A., and Schell, J.
Specialized vectors for gene tagging and expression studies. In: “Plant
Molecular Biology Manual”, S. Gelvin and B. Schilperoort (Eds.), Kluwer
Academic Publishers, Dordrecht, Vol. B2, pp. 1-22 (1994).
133. Koncz, C., Nemeth, K., Redei, G.P., and Schell, J. Homology recognition during
T-DNA integration into plant genome. In: “Homologous Recombination in
Plants”, J. Paszkowski (Ed.), Kluwer Academic Publishers, Dordrecht,
pp. 167-189 (1994).
134. Magrelli, A., Langenkemper, K., Dehio, C., Schell, J., and Spena, A. Splicing of
the rolA transcript of Agrobacterium rhizogenes in Arabidopsis. Science 266,
1986-1988 (1994).
135. Reiss, B., Koncz, C., Moore, I., and Schell, J. A family of binary gene vectors
with low inter-transformant variation. Plant Physiol. (Life Sci. Adv.) 13,
143-149 (1994).
136. Röhrig, H., Schmidt, J., Wieneke, U., Kondorosi, E., Barlier, I., Schell, J., and
John, M. Biosynthesis of lipooligosaccharide nodulation factors: Rhizobium
NodA protein is involved in N-acylation of the chitooligosaccharide backbone.
Proc. Natl. Acad. Sci. USA 91, 3122-3126 (1994).
Jozef Stefaan Schell 341
137. Röhrig, H., Schmidt, J., Wieneke, U., Kondorosi, E., Barlier, I., Schell, J., and
John, M. Role of NodA and NodB proteins in nodulation factor synthesis. In:
Proceedings of “1st European Nitrogen Fixation Conference”, Szeged, 28.8.-
2.9.1994 and “Workshop on Safe Application of Genetically Modified
Microorganisms in the Environment”, Szeged, 2.-3.9.1994, G.B. Kiss, G. Endre
(Eds.), Oficina Press, Szeged, pp. 76-80 (1994).
138. Schell, J. Some Aspects of the EU System of Research Funding. In: “European
Research Structures - Changes and Challenges” Institutional Aspects of
European Research Policy, Ringberg Castle, Tegernsee, October 1993. Max-
Planck-Gesellschaft E1/94, U. Opolka, L. Yakes-Schröder (Eds.), Wendelstein-
Druck, Rosenheim, pp. 42-49 (1994).
139. Schell, J. Closing lecture - ISPMB Congress. Plant Mol. Biol. 26, 1695-1699
(1994).
140. Schell, J. Gentechnik in Deutschland: Ein Verzicht wäre unverantwortlich.
Chemie Heute, Ausgabe 1994/95 (Das Wissenschaftsmagazin des Fonds der
Chemischen Industrie), p. 6-11 (1994).
141. Schulte, W., Schell, J., and Töpfer, R. A gene encoding acetyl-coenzyme A
carboxylase from Brassica napus. Plant Physiol. 106, 793-794 (1994).
142. Sohn, A., Schenk, P., Signoret, P.A., Schmitz, G., Schell, J., and Steinbiss, H.-H.
Sequence analysis of the 3'-terminal half of RNA 1 of wheat spindle streak
mosaic virus. Arch. Virol. 135, 279-292 (1994).
143. Voetz, M., Klein, B., Schell, J., and Töpfer, R. Three different cDNAs encoding
acyl carrier proteins from Cuphea lanceolata. Plant Physiol. 106, 785-786
(1994).
144. Wada, M., Klein, C., Schell, J., and Reiss, B. A functional assay for Taq DNA
polymerase in PCR. BioTechniques 16, 26-30 (1994).
145. Walden, R., Fritze, K., Hayashi, H., Miklashevichs, E., Harling, H., and Schell,
J. Activation tagging: a means of isolating genes implicated as playing a
role in plant growth and development. Plant Mol. Biol. 26, 1521-1528
(1994).
146. Walden, R., Hayashi, H., Fritze, K., and Schell, J. Using T-DNA tagging to
search for genes involved in the mechanism of phytohormone action. In:
“Molecular Botany: Signals and the Environment” (Biochemical Society
Symposium No. 60), D.J. Bowles, P.M. Gilmartin, J.P. Knox, G.G. Lunt (Eds.),
Portland Press, London, pp. 199-206 (1994).
147. Walden, R., and Schell, J. Activation T-DNA tagging - a silver bullet approach
to isolating plant genes. Agro Food Industry Hi-Tech 5 (No. 6), 9-12 (1994).
148. Zettl, R., Schell, J., and Palme, K. Photoaffinity labeling of Arabidopsis thaliana
plasma membrane vesicles by 5-azido-[7- 3 H]-indole-3-acetic acid:
Identification of a glutathione-S-transferase. Proc. Natl. Acad. Sci. USA 91,
689-693 (1994).
342 Wolf Prize in Agriculture
162. Reichel, C., Feltkamp, D., Walden, R., Steinbiss, H.-H., Schell, J., and Rosahl,
S. Inefficient expression of the DNA-binding domain of GAL4 in transgenic
plants. Plant Cell Rep. 14, 773-776 (1995).
163. Schell, J. Plant biotechnology: State of the art in developed countries
and relevant safety considerations. In: “Pan-European conference on the
potential long-term ecological impact of genetically modified organisms”,
Environmental encounters, No. 20, (Proceedings PEC, 24.-26.11.1993,
Strasbourg), Council of Europe (Ed.), Council of Europe Press, Netherlands,
pp. 25-34 (1995).
164. Schell, J. Progress in plant sciences is our best hope to achieve an economically
rewarding, sustainable and environmentally stable agriculture. Plant Tissue
Culture and Biotechnology 1, 10-12 (1995).
165. Schell, J. Statement zur Gentechnologie. In: “Gentechnologie - Ethik und
wissenschaftlicher Fortschritt” (Internationales Forum der Universität zu Köln
gemeinsam mit der Stadt Köln, 23.6.1994, Historisches Rathaus Köln), Band
4, P. Mittelstaedt (Ed.), Bouvier Verlag, Bonn, pp. 31-33 u. 65-66 (1995).
166. Schell, J. Molecular breeding as well as chemical and biological protection
methods have an important role to play to make integrated pest management
a practical and desirable reality. Agro-Food-Industry Hi-Tech Supplement,
Brighton Crop Protection 1995, 4-5 (1995).
167. Schell, J. Crop biotechnology - a necessity for an environmentally friendly
and sustainable agriculture. In: Proceedings, Part I, “Ninth Forum for Applied
Biotechnology” Faculty of Agricultural and Applied Biological Sciences, Gent,
27.-29.9.1995. Med. Fac. Landbouww. Univ. Gent, 60(4a), 1513-1514
(1995).
168. Schell, J., Palme, K., and Walden, R. Molecular approaches to the study of the
mechanism of action of auxins. In: “Plant Hormones - Physiology Biochemistry
and Molecular Biology”, P.J. Davies (Ed.), Kluwer Academic, Dordrecht,
pp. 354-371 (1995).
169. Schultze, M., Staehelin, C., Röhrig, H., John, M., Schmidt, J., Kondorosi, E.,
Schell, J., and Kondorosi, A. In vitro sulfotransferase activity of Rhizobium
meliloti NodH protein: Lipochitooligosaccharide nodulation signals are sulfated
after synthesis of the core structure. Proc. Natl. Acad. Sci. USA 92, 2706-
2709 (1995).
170. Suter-Crazzolara, C., Brzobohaty, B., Gazdova, B., Schell, J., and Reiss, B. T-
DNA integration in a new family of repetitive elements of Nicotiana tabacum.
J. Mol. Evol. 41, 498-504 (1995).
171. Töpfer, R., Hausmann, L., Voetz, M., Walek, J., Schell, J. and Martini, N.
Genes for the improvement of the storage oil content of Brassica napus. In:
“Rapeseed Today and Tomorrow”, Vol. 2 (Proceedings of the 9th International
Rapeseed Congress, 4.-7. July 1995, Cambridge/U.K.), Organising Committee
GCIRC (Eds.), The Dorset Press, Dorchester, pp. 479-481 (1995).
344 Wolf Prize in Agriculture
172. Töpfer, R., Martini, N., and Schell, J. Modification of plant lipid synthesis.
Science 268, 681-686 (1995).
173. Walden, R., Harling, H., Miklashevichs, E., Lubenow, H., and Schell, J.
Activation T-DNA tagging: gene isolation and molecular dissection of complex
biological pathways. In: “Induced Mutations and Molecular Techniques for
Crop Improvement” (Proceedings of an International Symposium on the Use
of Induced Mutations and Molecular Techniques for Crop Improvement,
19-23 June 1995, Vienna), International Atomic Energy Agency, Vienna
(Eds.), IAEA, Austria, pp. 425-431 (1995).
174. Grevelding, C., Suter-Crazzolara, C., von Menges, A., Kemper, E., Masterson,
R., Schell, J., and Reiss, B. Characterisation of a new allele of pale cress and
its role in greening in Arabidopsis thaliana. Mol. Gen. Genet. 251, 532-541
(1996).
175. Keller, M., Sneh, B., Strizhov, N., Prudovsky, E., Regev, A., Koncz, C., Schell, J.,
and Zilberstein, A. Digestion of δ-endotoxin by gut proteases may explain
reduced sensitivity of advanced instar larvae of Spodoptera littoralis to CryIC.
Insect Biochem. Mol. Biol. 26, 365-373 (1996).
176. Kleinmann, J.-W., Kleinow, T., Engelhardt, K., Philipp, C., Wegener, D.,
Schell, J., and Schreier, P. Characterization of two class II chitinase genes
from peanut and expression studies in transgenic tobacco plants. Plant Mol.
Biol. 30, 351-358 (1996).
177. Regev, A., Keller, M., Strizhov, N., Sneh, B., Prudovsky, E., Chet, I.,
Ginzberg, I., Koncz-Kalman, Z., Koncz, C., Schell, J., and Zilberstein, A.
Synergistic activity of a Bacillus thuringiensis δ-endotoxin and a bacterial
endochitinase against Spodoptera littoralis larvae. Appl. Environ. Microbiol.
62, 3581-3586 (1996).
178. Reichel, C., Maas, C., Schultze, S., Schell, J., and Steinbiss, H.-H. Cooperative
binding to nucleic acids by barley yellow mosaic bymovirus coat protein and
characterization of a nucleic acid-binding domain. Virology 77, 587-592
(1996).
179. Reichel, C., Mathur, J., Eckes, P., Langenkemper, K., Koncz, C., Schell, J.,
Reiss, B., and Maas, C. Enhanced green fluorescence by the expression
of an Aequorea victoria green fluorescent protein mutant in mono- and
dicotyledonous plant cells. Proc. Natl. Acad. Sci. USA 93, 5888-5893
(1996).
180. Reinemer, P., Prade, L., Hof, P., Neuefeind, T., Huber, R., Zettl, R., Palme, K.,
Schell, J., Koelln, I., Bartunik, H.D., and Bieseler, B. Three-dimensional
structure of glutathione S-transferase from Arabidopsis thaliana at 2.2 Å
resolution: Structural characterization of herbicide-conjugating plant
glutathione S-transferases and a novel active site architecture. J. Mol. Biol.
255, 289-309 (1996).
Jozef Stefaan Schell 345
181. Reiss, B., Klemm, M., Kosak, H., and Schell, J. RecA protein stimulates
homologous recombination in plants. Proc. Natl. Acad. Sci. USA 93,
3094-3098 (1996).
182. Schell, J. Prof. G. Melchers celebrates his ninetieth (90th) birthday! Mol.
Gen. Genet. 250, 135-136 (1996).
183. Schell, J. Transgenic plants and study of the regeneration of whole plants
from individual cells. In: “Das Ganze und seine Teile” Europäisches Forum
Alpbach 1995, H. Pfusterschmid-Hardtenstein (Ed.), Ibera Verlag, Wien,
pp. 515-516 (1996).
184. Schell, J. Biotechnologies et agriculture: les nouveaux defis. Cahiers de la
Fondation des Treilles, N°7/Année 1995, 167-186 (1996).
185. Strizhov, N., Keller, M., Koncz-Kalman, Z., Regev, A., Sneh, B., Schell, J.,
Koncz, C., and Zilberstein, A. Mapping of the entomocidal fragment of
Spodoptera-specific Bacillus thuringiensis toxin CryIC. Mol. Gen. Genet. 253,
11-19 (1996).
186. Strizhov, N., Keller, M., Mathur, J., Koncz-Kalman, Z., BOSCH, D.,
Prudovsky, E., Schell, J., Sneh, B., Koncz, C., and Zilberstein, A. A synthetic
cryIC gene, encoding a Bacillus thuringiensis δ-endotoxin, confers Spodoptera
resistance in alfalfa and tobacco. Proc. Natl. Acad. Sci. USA 93, 15012-
15017 (1996).
187. Szekeres, M., Nemeth, K., Koncz-Kalman, Z., Mathur, J., Kauschmann, A.,
Altmann, T., Redei, G.P., Nagy, F., Schell, J., and Koncz, C. Brassinosteroids
rescue the deficiency of CYP 90, a cytochrome P450, controlling cell
elongation and de-etiolation in Arabidopsis. Cell 85, 171-182 (1996).
188. Van de Samde, K., Pawlowski, K., Czaja, I., Wieneke, U., Schell, J., Schmidt, J.,
Walden, R., Matvineko, M., Wellink, J., Van Kammen, A., Franssen, H., and
Bisseling, T. Modification of phytohormone response by a peptide encoded
by ENOD40 of legumes and a nonlegume. Science 273, 370-373 (1996).
189. Walden, R., Maas, C., Martini, N., and Schell, J. Transgenic plants. European
Review 4, 393-414 (1996).
190. Gadella, Jr., T.W.J., VerebJr., G., Hardi, A.-E., Röhrig, H., Schmidt, J., John, M.,
Schell, J., and Bisseling, T. Microspectroscopic imaging of nodulation factor-
binding sites on living Vicia sativa roots using a novel bioactive fluorescent
nodulation factor. Biophys. J. 72, 1986-1996 (1997).
191. Jach, G., Pinsdore, E., and Schell, J. Direct isolation of poly A+ RNA from
transgenic tobacco leaves with Oligotex. Qiagen News 1, 20-21 (1997).
192. John, M., Röhrig, H., Schmidt, J., Walden, R., and Schell, J. Cell signalling by
oligosaccharides. Trends Plant Sci. 2, 111-115 (1997).
193. Kendall, H.W., Beachy, R., Eisner, T., Gould, F., Herdt, R., Raven, P.H., Schell,
J.S., and Swaminathan, M.S. (Eds.) “Bioengineering of Crops” Report of the
World Bank Panel on Transgenic Crops. Environmentally and Socially
346 Wolf Prize in Agriculture
family: indication for plastidic localization of at least one isoform. Proc. Natl.
Acad. Sci. USA 94, 3465-3470 (1997).
206. Slabaugh, M.B., Huestis, G.M., Leonard, J., Holloway, J.L., Rosato, C.,
Hongtrakul, V., Martini, N., Toepfer, R., Voetz, M., Schell, J., and Knapp, S.J.
Sequence-based genetic markers for genes and gene families: single-strand
conformational polymorphisms for the fatty acid synthesis genes of Cuphea.
Theor. Appl. Genet. 94, 400-408 (1997).
207. Strizhov, N., Abraham, E., Ökresz, L., Blickling, S., Zilberstein, A., Schell, J.,
Koncz, C., and Szbados, L. Differential expression of two P5CS genes
controlling proline accumulation during salt-stress requires ABA and is
regulated by ABA1, ABI1 and AXR2 in Arabidopsis. Plant J. 12, 557-569
(1997).
208. Walden, R., Reiss, B., Koncz, C., and Schell, J. The impact of Ti-plasmid-
derived gene vectors on the study of the mechanism of action of
phytohormones. Annu. Rev. Phytopathol. 35, 45-66 (1997).
209. First, N.L., Schell, J., and Vasil, I.K. Prospects and Limitations of Agricultural
Biotechnologies: An Update. In: “Agricultural Biotechnology”, A. Altman (Ed.),
Marcel Dekker, New York, pp. 743-748 (1998).
210. Ginzberg, I., Stein, H., Kapulnik, Y., Szbados, L., Strizhov, N., Schell, J., Koncz,
C., and Zilberstein, A. Isolation and characterization of two different cDNAs
of ∆1-pyrroline-5-carboxylate synthase in alfalfa, transcriptionally induced
upon salt stress. Plant Mol. Biol. 38, 755-764 (1998).
211. Mathur, J., Molnar, G., Fujioka, S., Takatsuto, S., Sakurai, A., Yokota, T.,
Adam, G., Voigt, B., Nagy, F., Maas, C., Schell, J., Koncz, C., and Szekeres, M.
Transcription of the Arabidopsis CPD gene, encoding a steroidogenic
cytochrome P450, is negatively controlled by brassinosteroids. Plant J. 14,
593-602 (1998).
212. Mathur, J., Szbados, L., Schaefer, S., Grunenberg, B., Lossow, A., Jonas-Straube,
E., Schell, J., Koncz, C., and Koncz-Kalman, Z. Gene identification with
sequenced T-DNA tags generated by transformation of Arabidopsis cell
suspension. Plant J. 13, 707-716 (1998).
213. Moore, I., Gälweiler, L., Grosskopf, D., Schell, J., and Palme, K. A transcription
activation system for regulated gene expression in transgenic plants. Proc.
Natl. Acad. Sci. USA 95, 376-381 (1998).
214. Nemeth, K., Salchert, K., Putnoky, P., Bhalerao, R., Koncz-Kalman, Z.,
Stankovic-Stangeland, B., Bako, L., Mathur, J., Ökresz, L., Stabel, S.,
Geigenberger, P., STITT, M., Redei, G.P., Schell, J., and Koncz, C. Pleiotropic
control of glucose and hormone responses by PRL1, a nuclear WP protein, in
Arabidopsis. Genes & Development 12, 3059-3073 (1998).
215. Ökresz, L., Mathe, C., Horvath, E., Schell, J., Koncz, C., and Szbados, L.
T-DNA trapping of a cryptic promoter identifies an ortholog of highly
348 Wolf Prize in Agriculture
238. Miklashevichs, E., Röhrig, H., Schell, J. and Schmidt, J. Perception and Signal
Transduction of Rhizobial NOD Factors. Critical Reviews in Plant Sciences,
20(4):373-394 (2001).
239. Jach, G., Binot, E. Frings, S., Luxa, K. and Schell, J. Use of red fluorescent
protein from Discosoma sp. (dsRED) as a reporter for plant gene expression.
The Plant Journal 28(4), 483-491 (2001).
240. Jach G, Binot E, Frings S, Luxa K, Schell J. Use of red fluorescent protein from
Discosoma sp. (dsRED) as a reporter for plant gene expression. Plant J. 28(4):
483-91 (2001).
241. Ferrando A, Koncz-Kálmán Z, Farràs R, Tiburcio A, Schell J, Koncz C. Detection
of in vivo protein interactions between Snf1-related kinase subunits with
intron-tagged epitope-labelling in plants cells. Nucleic Acids Res. 29(17):
3685-93 (2001).
242. Farrás R, Ferrando A, Jásik J, Kleinow T, Okrész L, Tiburcio A, Salchert K, del
Pozo C, Schell J, Koncz C. SKP1-SnRK protein kinase interactions mediate
proteasomal binding of a plant SCF ubiquitin ligase. EMBO J. 20(11): 2742-
56 (2001).
243. Ríos G, Lossow A, Hertel B, Breuer F, Schaefer S, Broich M, Kleinow T, Jásik J,
Winter J, Ferrando A, Farrás R, Panicot M, Henriques R, Mariaux JB,
Oberschall A, Molnár G, Berendzen K, Shukla V, Lafos M, Koncz Z, Rédei GP,
Schell J, Koncz C. Rapid identification of Arabidopsis insertion mutants by
non-radioactive detection of T-DNA tagged genes. Plant J. 32(2):243-53
(2002).
244. Henriques R, Jásik J, Klein M, Martinoia E, Feller U, Schell J, Pais MS, Koncz
C. Knock-out of Arabidopsis metal transporter gene IRT1 results in
iron deficiency accompanied by cell differentiation defects. Plant Mol Biol.
50(4-5):587-97 (2002).
245. Markmann-Mulisch U, Hadi MZ, Koepchen K, Alonso JC, Russo VE,
Schell J, Reiss B. The organization of Physcomitrella patens RAD51 genes
is unique among eukaryotic organisms. Proc Natl Acad Sci USA 99(5):
2959-64 (2002).
246. Rohrig H, Schmidt J, Miklashevichs E, Schell J, John M. Soybean ENOD40
encodes two peptides that bind to sucrose synthase. Proc Natl Acad Sci
USA 99(4):1915-20 (2002).
247. Bakó L, Umeda M, Tiburcio AF, Schell J, Koncz C. The VirD2 pilot protein
of Agrobacterium-transferred DNA interacts with the TATA box-binding
protein and a nuclear protein kinase in plants. Proc Natl Acad Sci USA
100(17):10108-13 (2003).
248. Röhrig, H., Schmidt, J., Miklashevichs, E., Schell, J. and John. M. Soybean
ENOD40 encodes two peptides which bind to sucrose synthase. PNAS
(in press)
Jozef Stefaan Schell 351
Editor’s Choice
Editor’s Choice
bacteria. These studies were followed by ones on tion he and his colleagues made to the development
modifications of phages and restrictions by their and propagation of plant transformation and molec-
hosts. His first paper, coauthored with Marc Van ular genetics. However, the expansion of a technol-
Montagu, on Agrobacterium-related DNA was pub- ogy and its adoption into another area of science
lished in 1972 and focused on Agrobacterium phages. worldwide takes not only skilled experiments and
With Rob Schilperoort, he described in 1973 the de- provocative publications. It takes teaching of a vision
tection of Agrobacterium DNA in sterile crown gall to scientists, research funding agencies, industries
tissue cultures. In 1974, his group showed that a large and governments, and the training of a new genera-
plasmid of Agrobacterium was essential for crown tion of scientists. Jeff did all of this, unceasingly, for
gall induction, and this was followed by a series of twenty years. He had a major impact on students,
papers on the plasmids. The isolation of the Ti plas- established scientists, plant breeders, industries,
mid was described in 1976, and this breakthrough funding agencies, governments, as well as scientific
opened up many new opportunities to study its societies and the general public. During the 1980s
DNA. By 1977, several groups including those of and 1990s, he sustained a punishing schedule of lec-
Schell and Van Montagu in Gent, Mary-Dell Chilton tures around the world. In the 1980s he was sought
in Eugene Nester’s laboratory in Seattle, and Schilp- by nearly every plant molecular biology conference,
eroort’s lab in Leiden were actively testing the idea and he wanted to attend them all to learn, to teach, to
that the Ti plasmid of Agrobacterium might provide inspire, and to open up the practice of plant molec-
a means of inserting genes into plants. In 1978, Jeff ular genetics.
and his colleagues (including Marc Van Montagu) If he gave much to plant science everywhere by his
published 18 papers covering the origins of crown teaching, he gave much to European science by ac-
gall by Ti plasmid transfer and the manipulation of cepting in 1978 the Directorship of the Max Planck
the Ti plasmid by various bacterial genetic tricks, Institute for Plant Breeding Research in Köln, Ger-
including transposon mutagenesis, cointegration, many. This appointment gave Jeff the opportunity to
and transfections to reveal more about the plasmid, establish a very large team of scientists, to develop
the genes it contained, and the requirements for the new approaches to plant science, and to train
crown gall induction. In 1980 the Gent group pub- students and postdocs. The position also gave him
lished 20 papers, many being symposium articles, to enhanced authority to talk to European governmen-
get the Ti–plant cell transformation message out to tal agencies and the European Union (EU). In the
receptive scientists. By the time the seminal papers early years of plant genetic engineering, he also ad-
were published in 1983, showing the expression of vised Monsanto, the company that was to become the
novel chimeric genes introduced into plant cells us- foremost crop genetic engineering multinational. He
ing a Ti plasmid-derived vector, Jeff and his col- also forged a long-term, mutually beneficial, relation-
leagues had published some 150 papers. Then fol- ship between the Max Planck Institute and Bayer, a
lowed many details of the Ti plasmid genes, their role company located close to the Max Planck Institute in
in oncogenesis, and the behavior of these genes when Germany.
transferred into plants. Some 23 papers were pub- Jeff was a true European. The EU started to offer
lished in 1984 describing the development of useful substantial funds for research in the 1980s, and Jeff
vectors and the transfer of several more foreign genes and colleagues at the Max Planck Institute in Köln
into plants. During this time he also studied the Ri saw the value and necessity for this pan-European
plasmid, the equivalent of the Ti plasmid, in Agrobac- resource to spearhead the research that could help
terium rhizogenes and its function after transfer to address the substantial agricultural, economic, and
plants cells. Once the subject was truly well estab- environmental problems of the future. He taught and
lished, the concept of plant genetic engineering be- lobbied EU scientific leaders about the new plant
came a topic of worldwide debate, and the plant science. To improve the funding and management of
scientific community embraced these technologies to EU plant science programs, the Max Planck Institute
literally transform the whole field of plant science. and the John Innes Centre (Norwich, England) –the
During this period, Jeff and many colleagues, includ- two largest centers for plant molecular genetics in
ing students and postdocs, and collaborators in other Europe –formed a legal entity under Jeff’s chairman-
laboratories published a large number of papers (an- ship. Much was changed during the 1990s via this
other 449 until his death in 2003) on Ti plasmid organization and Jeff’s tireless and influential lead-
biology, transformation vectors, plant gene discovery ership. He saw the needs of European and global
especially in Arabidopsis, promoter analyses, hor- societies and was not afraid to articulate the oppor-
mone biology, Rhizobium genetics, and plant genetic tunities, needs, and requirements for change to
engineering. anyone.
The enormous number of papers that carried his While being a Director of a team of well over 100
name, first from Gent and then from his very large scientists, an international leader, and an advocate
group in the Max Planck Institute for Plant Breeding, for European science and plant molecular biology for
Köln, Germany, are a witness to the huge contribu- agriculture and the environment in general, Jeff
1120 Plant Physiol. Vol. 132, 2003
Jozef Stefaan Schell 353
Editor’s Choice
found time to do many of the ordinary things that Hansen Foundation, Denmark (1991), and the
senior scientists do write and review grant proposals Wilhelm-Exner-Medaille of Vienna (1995). Several
and papers and mentor students and postdocs in his universities awarded him an honorary doctorate de-
laboratory. He was a senior editor of The Plant Jour- gree, including the Hebrew University, Israel (1994),
nal from 1990 to 1998. He also held the great distinc- Tel Aviv University, Israel (1997), University of East
tion of being the Chairman of the European Molecu- Anglia, Norwich, United Kingdom (1997), and Uni-
lar Biology Organisation (EMBO) Council from 1990 versity Louis Pasteur, Strasbourg (1992). One of his
to 1995. EMBO is an organization much revered in most treasured appointments was of Professeur
Europe and via his service to EMBO, Jeff gave much Honoraire, College de France, Paris (1998).
to molecular biology as a whole. Of course, he sat on One cannot look at plant science journals today and
many boards and committees around the world to their back issues and not recognize that plant science
give wisdom and guidance to those who sought to has been transformed since the early 1980s following
further science and plant molecular biology in the activities of those few who believed that crown
particular. galls held some valuable biological secrets. Jeff Schell
Plant science is not renowned for having individ- was very much at the helm as he went around the
uals honored by science and society, but Jeff collected world giving such transforming talks, stimulating so
many honors. He was elected to be a Foreign Asso- many experiments, writing so many papers, inform-
ciate of the US National Academy of Sciences (1985), ing politicians and industries, and taking the time
of the Indian National Science Academy (1998), of the here and there to confront “the greens” and skeptics.
Royal Swedish Academy (1989), and of the Hungar- Two titles of his papers in recent years describe his
ian Academy of Sciences (1993). He won many key vision: “Progress in plant science is our best hope to
prizes, including the Mendel-Medaille of the Deut- achieve an economically rewarding, sustainable and
sche Akademie der Naturforscher Leopoldina (1985), environmentally stable agriculture” (1995) and “Crop
the Otto Bayer-Preis of the Otto-Bayer-Stiftung Biotechnology - a necessity for an environmentally
(1985), Prix Alexandre de Humboldt (1985), the Rank friendly and sustainable agriculture” (1995). Let us
Prize for Nutrition (1987), the IBM Europe Science hope that as we go forward, his vision will be
and Technology Prize (1987), the Wolf Prize in Agri- realized.
culture (1990), Prix Charles Leopold Mayer of the
Academie des Sciences, Paris (1990), the Japan Prize
for Biotechnology in Agriculture Sciences by the Sci- LITERATURE CITED
ence and Technology Foundation of Japan (1998),
Premiere Grande Medaille d’Or de l’Academie des Schell J (1995) Progress in plant science is our best hope to achieve an
Sciences, Paris (1997), the Australia Prize of the Aus- economically rewarding, sustainable and environmentally stable agricul-
ture. Plant Tissue Culture and Biotechnology 1: 10–12
tralian Academy of Science (1990), Prix Charles Schell J (1995) Crop Biotechnology - a necessity for an environmentally
Leopold Mayer of the Academy des Sciences, Paris friendly and sustainable agriculture. Proceedings Part 1. Ninth Forum for
(1990), the Hansen Gold Medal of the Emil Christian Applied Biotechnology University of Gent. 60: 1513–1514
Richard Flavell
Chief Scientific Officer
Ceres-inc.
Malibu, California 90265
1932–2007
CURRICULUM VITAE
Education
B.S. National Taiwan University (in Agricultural Chemistry) 1956
M.S. National Taiwan University (in Agricultural Chemistry) 1958
Ph.D. Utah State University (in Plant Biochemistry) 1962
Postdoctoral Work
University of California Davis (with Prof. P.K. Stumpt) 1962-63
New York University Medical School (with Prof. B.N. LaDu) 1963-64
University of California, San Diego (with Prof. AA. Benson) 1964-65
355
356 Wolf Prize in Agriculture
Scientific Societies
American Society for Biochemistry and Molecular Biology, American Society of
Plant Physiologists, American Society for Horticultural Science, Phytochemical Society
of North America, International Plant Growth Substances Association.
Research Activities
Biosynthesis and biochemical action of ethylene in relation to plant senescence;
postharvest biochemistry of fruits and vegetables; interaction of ethylene and other
plant hormones on plant growth; biochemistry of sulfite and sulfur amino acids.
Honors
American Institute of Biological Sciences - Campbell Award, 1969
J.S. Guggenheim Fellow, 1982
International Plant Growth Substances Association Research Award, 1985
Member of the National Academy of Sciences, USA, 1990
Wolf Prize in Agriculture, 1991
University of California - Davis Faculty Research Lecturer, 1991
American Society for Horticultural Science Outstanding Research Award, 1992
Member of Academia Sinica, 1992
Professional Service
Editorial Board of Plant Physiology, 1974-1992
Associate Editor of Journal Plant Growth Regulation, 1981-1993
Editorial Board of Plant and Cell Physiology, 1987-1991
Editorial Board of Plant Physiology and Biochemistry, 1988-1993
Editorial Board of Acta Phytophysiologica Sinica, 1988-1993
Assays for ACC were quickly developed and physiological studies into the
regulation of ethylene biosynthesis accelerated. For example, Shang Fa’s group
demonstrated that under low oxygen conditions, such as root flooding, ACC could
accumulate and be transported in the xylem to the shoot and subsequently converted
to ethylene, serving as a translocatable signal of root stress to the shoots. His group
also discovered that ACC could be conjugated to malonate, resulting in an alternative
pool of ACC in plant tissues. It had been noted that methionine pools in plants are
too low to sustain the observed rates of ethylene synthesis, but Shang Fa and his
students resolved this controversy by demonstrating that the methylthio group
released from SAM during the synthesis of ACC is recycled to replenish methionine
levels. The reactions of this recycling pathway were subsequently christened the
Yang Cycle in plant biochemistry texts. As the tools became available for cloning
and characterizing the genes responsible for the steps in ethylene biosynthesis,
Shang Fa contributed to many studies of the regulation of those genes in fruit
ripening, plant growth, wounding and stress responses. He wrote numerous reviews
and book chapters that defined ethylene biosynthesis and its role in plant biology
for a generation of students and researchers.
In all his work, Shang Fa continually linked his discoveries to practical
applications in postharvest biology and plant growth regulation. He used what he
knew about physiology to learn more about ethylene biosynthesis, and he applied
what he learned about ethylene biosynthesis to contribute to improvements in
postharvest storage conditions. He was known for his clarity of thought and the
ability to identify and design critical experimental tests of hypotheses. Shang Fa
always maintained an open mind and was willing to challenge accepted ideas, even
his own, when they proved untenable in the face of experimental data.
Shang Fa had an uncommon faith in humanity and urged his students to
always expect the best of people. The coupling of an affable nature and a genuine
concern for his students and colleagues enabled Shang Fa to assemble a powerful
and effective research group that shared his vision and strove to match his intensity.
He also developed an extensive and international network of friends and colleagues.
Despite his many honors, he remained humble and always willing to share credit
for the many discoveries coming out of his lab or to acknowledge the priority of
other groups.
Shang Fa figured prominently at many national and international research
conferences over the years and served on the editorial boards of leading journals
and as a member of several learned societies. He won many awards and honors,
including the Campbell Award of the American Institute of Biological Sciences in
1969, a Guggenheim Fellowship in 1982, the International Plant Growth Substances
Association Research Award in 1985, and the Outstanding Researcher Award from
the American Society of Horticultural Science in 1992. Shang Fa was named the
UC Davis Faculty Research Lecturer in 1992. In 1990 and 1992, he was elected to
Shang Fa Yang 359
the National Academy of Sciences, USA and to the Academia Sinica, Taiwan,
respectively. In 1991, he received the prestigious international Wolf Prize in
Agriculture.
After taking early retirement from University of California in 1994, Shang
Fa served as Professor in the Department of Biology at Hong Kong University of
Science and Technology from 1994 to 1997, where he established an active
research group, and as a Distinguished Research Fellow and the Director of the
Institute of Botany at Academia Sinica, Taipei, Taiwan. From 1996 to 1999, he
was Vice President of the Academia Sinica and directed its numerous research
institutes.
Shang Fa is survived by his wife Eleanor and two sons, Albert and Bryant, who
have pursued careers in engineering and chemistry, respectively. While future
plant biologists will know of Shang Fa through the Yang Cycle and his many other
contributions to our field, students and colleagues who were fortunate enough to
know him personally will also remember his humor, his humanity, and his sparkling
intellect.
Kent J. Bradford
Alan B. Bennett
John M. Labavitch
Mikal E. Saltveit
LIST OF PUBLICATIONS
1. Yang, S.F. and H.-K. Ho. 1958. Biochemical studies on post-ripening of banana.
J. Chinese Chem. Soc. 5: 71-85.
2. Yang. S.F. and G.W. Miller. 1963. Biochemical studies on the effect of fluoride
on higher plants. 1. Metabolism of carbohydrates, organic acids and amino
acids. Biochem. J. 88: 505-509.
3. Yang, S.F. and G.W. Miller. 1963. Biochemical studies on the effect of fluoride
on higher plants. 2. The effect of fluoride on sucrose-synthesizing enzymes
from higher plants. Biochem. J. 88: 509-516.
4. Yang, S.F. and G.W. Miller. 1963. Biochemical studies on the effect of fluoride
on higher plants. 3. The effect of fluoride on dark carbon dioxide fixation.
Biochem. J. 88: 517-522.
5. Yang, S.F. and P.K. Stumpf. 1965. Fat metabolism in higher plants. XXI.
Biosynthesis of fatty acids by avocado mesocarp enzyme systems. Biochim.
Biophys. Acta 98: 19-26.
6. Yang, S.F. and P.K. Stumpf. 1965. Fat metabolism in higher plants. XXll.
Enzymic synthesis of 14-hydroxyl-11-eicosenoic acid by particulate
preparations of avocado mesocarp. Biochim. Biophys. Acta 98: 27-35.
360 Wolf Prize in Agriculture
7. Yang, S.F., H.S. Ku, and H.K. Pratt. 1966. Ethylene production from methionine
as mediated by flavin mononucleotide and light. Biochem. Biophys. Res.
Commun. 24: 739-743.
8. Yang, S.F., S. Freer, and A.A. Benson. 1967. Transphosphatidylation by
phospholipase D. J. Biol. Chem. 242: 477-484.
9. Ku, H.S., S.F. Yang, and H.K. Pratt. 1967. Enzymic evolution of ethylene
from methional by a pea seedling extract. Arch. Biochem. Biophys. 118:
756-758.
10. Welkie, G.W., S.F. Yang, and G.W. Miller. 1967. Metabolite changes induced
by cucumber mosaic virus in resistant and susceptible strains of cowpea.
Phytopathology 57: 472-475.
11. Rappaport, L., A. Hsu, R. Thompson, and S.F. Yang. 1967. Fate of 14C-
gibberellin A3 in plant tissues. Annals of the New York Academy of Sciences
144: 211-218.
12. Yang, S.F., H.S. Ku, and H.K. Pratt. 1967. Photochemical production of ethylene
from methionine and its analogues in the presence of flavin mononucleotide.
J. Biol. Chem. 242: 5274-5280.
13. Yang, S.F. 1967. Biosynthesis of ethylene. Ethylene formation from methional
by horseradish peroxidase. Arch. Biochem. Biophys. 122: 481-487.
14. Yang, S.F. 1968. Biosynthesis of ethylene. In P. Wightman, and G. Setterfield
(eds.), Biochemistry and Physiology of Plant Growth Substances. The Runge
Press. Ltd., Ottawa. pp. 1217-1288.
15. Yang, S.F. 1969. Phospholipase D from savoy cabbage. In S.P. Colowick, and
N.O. Kaplan (eds.), Methods in Enzymology. Academic Press. New York.
pp. 208-211.
16. Ku, H.S., S.F. Yang, and H.K. Pratt. 1969. Ethylene formation from alpha-
keto-gamma-methylthiobutryrate by tomato fruit extracts. Phytochemistry
8: 567-573.
17. Baur, A. and S.F. Yang. 1969. Ethylene production from propanal. Plant
Physiol. 44: 189-192.
18. Yang, S.F. 1969. Further studies on ethylene formation from alpha-keto-
gamma-methylthiobutyric acid or beta-methylthiopropionaldehyde by
peroxidase in the presence of sulfite and oxygen. J. Biol. Chem. 244: 4360-
4365.
19. Yang, S.F. 1969. Ethylene evolution from 2-chloroethylphosphonic acid. Plant
Physiol. 44: 1203-1204.
20. Baur, A.H. and S.F. Yang. 1969. Precursors of ethylene. Plant Physiol. 44:
1347-1349.
21. Yang, S.F. and A.H. Baur. 1969. Pathways of ethylene biosynthesis. Qualitas
Plantarum Materiae Vegetabiles 19: 201-220.
22. Yang, S.F. 1969. A simple method for assay of 14C-formic acid. Anal. Biochem.
32: 519-521.
Shang Fa Yang 361
23. Ku, H.S., S.F. Yang, and H.K. Pratt. 1970. Inactivity of apoperoxidase in
indoleacetic acid oxidation and in ethylene formation. Plant Physiol. 45:
358-359.
24. Ku, H.S., S.F. Yang, and H.K. Pratt. 1970. Ethylene production and perioxidase
activity during tomato ripening. Plant Cell Physiol. 11: 241-246.
25. Yang, S.F. 1970. Photosensitized conversion of ethionine to ethylene by flavin
mononucleotide. Photochem. Photobiol. 12: 419-422.
26. Yang, S.F. 1970. Sulfoxide formation from methionine or its sulfide analogs
during aerobic oxidation of sulfite. Biochemistry 9: 5008-5014.
27. Hyodo, H. and S.F. Yang. 1971. Ethylene-enhanced synthesis of phenylalanine
ammonia-lyase in pea seedlings. Plant Physiol. 47: 765-770.
28. Baur, A.H., S.F. Yang, and H.K. Pratt. 1971. Ethylene biosynthesis in fruit
tissues. Plant Physiol. 47: 696-699.
29. Hyodo, H. and S.F. Yang. 1971. Ethylene-enhanced formation of cinnamic
acid 4-hydroxylase in excised pea epicotyl tissue. Arch. Biochem. Biophys.
143: 338-339.
30. Yamaguchi, M., C.W. Chu, and S.F. Yang. 1971. The fate of 14C (2-chloroethyl)
phosphonic acid in summer squash, cucumber, and tomato. J. Amer. Soc.
Hort. Sci. 96: 606-609.
31. Yang, S.F. and A.H. Baur. 1972. Biosynthesis of ethylene in fruit tissues.
In D.J. Carr (ed.), Plant Growth Substances 1970, Springer-Verlag, Heidelberg,
pp. 510-517.
32. Baur, A.H. and S.F. Yang. 1972. Formation of ethionine from homocysteine
and of S-methylmethionine in apple tissue. Phytochemistry 11: 2503-2502.
33. Baur, A.H. and S.F. Yang. 1972. Methionine metabolism in apple tissue in
relation to ethylene biosynthesis. Phytochemistry 11: 3207-3214.
34. Yang, S.F. and M.A. Saleh. 1973. Destruction of indole-3-acetic acid during
the aerobic oxidation of sulfite. Phytochemistry 12: 1463-1466.
35. Lau, O-L. and S.F. Yang. 1973. Mechanism of a synergistic effect of kinetin
on auxin-induced ethylene production: suppression of auxin conjugation.
Plant Physiol. 51: 1011-1014.
36. Horng, A.J. and S.F. Yang. 1973. Peroxidase-catalyzed oxidation of indole-3-
acetaldehyde to 4-hydroxyquinoline in the presence of bisulfite ion:
elimination of pyrrole ring C2 as formic acid. Biochim. Biophys. Acta 321:
456-460.
37. Yang, S.F. 1973. Destruction of tryptophan during the aerobic oxidation of
sulfite ions. Environ. Res. 6: 395-402.
38. Chou, T.W. and S.F. Yang. 1973. The biogenesis of ethylene in Penicillium
digitatum. Arch. Biochem. Biophys. 157: 73-82.
39. Hyodo, H. and S.F. Yang. 1974. The effect of ethylene on the development of
phenylalanine ammonia-lyase in potato tuber disks. Zeitschrift fur
Pflanzenphysiologie 71: 76-79.
362 Wolf Prize in Agriculture
40. Yang, S.F. 1974. The biochemistry of ethylene: biogenesis and metabolism.
In V.C. Runeckles (ed.), Recent Advances in Phytochemistry, Vol. 7. The
Chemistry and Biochemistry of Plant Hormones. Academic Press, New York,
pp. 131-164.
41. Lau, O-L. and S.F. Yang. 1974. Synergistic effect of calcium and kinetin on
ethylene production by the mungbean hypocotyl. Planta 118: 1-6.
42. Mukerji, S.K. and S.F. Yang. 1974. Phosphoenolpyruvate carboxylase from
spinach leaf tissue. Plant Physiol. 53: 829-834.
43. Lau, O-L., S.F. Yang, and K-H. Yung. 1974. Suppression of IAA conjugation
by kinetin in relation to a synergistic effect of kinetin on IAA-induced ethylene
production. In Plant Growth Substances 1973. Hirokawa Publishing Company,
Tokyo, pp. 364-393.
44. Lau, O-L., D.P. Mure, and S.F. Yang. 1974. Effect of 2,4-dinitrophenol on
auxin-induced ethylene production and auxin conjugation by mung bean
tissue. Plant Physiol. 54: 182-185.
45. Yang, S.F. 1974. Ethylene biosynthesis in fruit tissues. In Facteurs et Regulation
de la Maturation des Fruits. Centre National de la Recherche Scientifique,
No. 238, Paris, pp. 171-175.
46. Mure, D.P. and S.F. Yang. 1975. Inhibition of in vivo conversion of methionine
to ethylene by L-canaline and 2,4-dinitrophenol. Plant Physiol. 55: 79-82.
47. Bowen, J.R. and S.F. Yang. 1975. Photosensitized deamination of sulfur-amino
acids by flavin mononucleotide. Photochem. Photobiol. 21: 201-203.
48. Lau, O-L. and S.F. Yang. 1975. Interaction of kinetin and calcium in relation
to their effect on stimulation of ethylene production. Plant Physiol. 55:
738-740.
49. Horng, A.J. and S.F. Yang. 1975. Aerobic oxidation of indole-3-acetic acid
with bisulfite. Phytochemistry 14: 1425-1428.
50. Mure, D.P. and S.F. Yang. 1975. Conversion of 5-methylthioadenosine to
methionine by apple tissue. Phytochemistry 14: 1291-1292.
51. Lau, O-L. and S.F. Yang. 1976. Stimulation of ethylene production in mung
bean hypocotyls by cupric ion, calcium ion, and kinetin. Plant Physiol. 57:
88-92.
52. Lau, O-L. and S.F. Yang. 1976. Inhibition of ethylene production by cobaltous
ion. Plant Physiol. 58: 114-117.
53. Lau, O-L., W.W. John, and S.F. Yang. 1977. Effect of different cytokinins on
ethylene production by mung bean hypocotyls in the presence of indole-3-
acetic acid or calcium ion. Physiol. Plant. 39: 1-3.
54. Peiser, G.D. and S.F. Yang. 1977. Chlorophyll destruction by the bisulfite-
oxygen system. Plant Physiol. 60: 277-281.
55. Adams, D.O. and S.F. Yang. 1977. Methionine metabolism in apple tissue.
Implication of S-adenosylmethionine as an intermediate in the conversion of
methionine to ethylene. Plant Physiol. 60: 893-896.
Shang Fa Yang 363
56. Peiser, G.D. and S.F. Yang. 1978. Chlorophyll destruction in the presence of
bisulfite and linonenic acid hydroperoxide. Phytochemistry 17: 79-84.
57. Lau, O-L., W.W. John, and S.F. Yang. 1978. Inactivity of oxidation products
of indole-3-acetic acid on ethylene production in mung bean hypocotyls.
Plant Physiol. 61: 68-71.
58. Hyodo, H., H. Kuroda, and S.F. Yang. 1978. Induction of phenylalanine
ammonia-lyase and increase in phenolics in lettuce leaves in relation to
the development of russet spotting caused by ethylene. Plant Physiol. 62:
31-35.
59. Yang, S.F. and H.K. Pratt. 1978. The physiology of ethylene in wounded plant
tissue. In G. Kahl (ed.), Biochemistry of Wounded Plant Tissues. De Gruyter
Co., Berlin, pp. 595-622.
60. Yu, Y-B., D.O. Adams, and S.F. Yang. 1979. Regulation of auxin-induced
ethylene production in mung bean hypocotyls. Plant Physiol. 63: 589-590.
61. Peiser, G.D. and S.F. Yang. 1979. Ethylene and ethane production from sulfur
dioxide-injured plants. Plant Physiol. 63: 142-145.
62. Yu, Y-B. and S.F. Yang. 1979. Auxin-induced ethylene production and its
inhibition by aminoethoxy-vinylglycine and cobalt ion. Plant Physiol. 64:
1074-1077.
63. Lizada, M.C.C. and S.F. Yang. 1979. A simple and sensitive assay for
l-aminocyclopropane-l-carboxylic acid. Anal. Biochem. 100: 140-145.
64. Adams, D.O. and S.F. Yang. 1979. Ethylene biosynthesis: Identification of
1-aminocyclopropane-1-carboxylic acid as an intermediate in the conversion
of methionine to ethylene. Proc. Natl. Acad. Sci. USA 76: 170-174.
65. Peiser, G.D. and S.F. Yang. 1979. Sulfite-mediated destruction of beta-carotene.
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66. Cameron, A.C., C.A. L. Fenton, Y. Yu, D.O. Adams, and S.F. Yang. 1979. Increased
production of ethylene by plant tissues treated with l-aminocyclopropane-l-
carboxylic acid. HortScience 14: 178-180.
67. Yu, Y-B., D.O. Adams, and S.F. Yang. 1979. 1-aminocyclopropane-carboxylate
synthase, a key enzyme in ethylene biosynthesis. Arch. Biochem. Biophys.
198: 280-286.
68. Bradford, K.J. and S.F. Yang. 1980. Xylem transport of 1-amino-cyclopropane-
1-carboxylic acid, an ethylene precursor, in waterlogged tomato plants. Plant
Physiol. 65: 322-326.
69. Bradford, K.J. and S.F. Yang. 1980. Stress-induced ethylene production in the
ethylene-requiring tomato mutant diageotropica. Plant Physiol. 65: 327-330.
70. Hoffman, N.E. and S.F. Yang. 1980. Changes of 1-aminocyclopropane-1-
carboxylic acid content in ripening fruits in relation to their ethylene
production rates. J. Amer. Soc. Hort. Sci. 105: 492-495.
71. Yu, Y-B. and S.F. Yang. 1980. Biosynthesis of wound ethylene. Plant Physiol.
66: 281-285.
364 Wolf Prize in Agriculture
72. Yu, Y-B., D.O. Adams, and S.F. Yang. 1980. Inhibition of ethylene production
by 2,4-dinitrophenol and high temperature. Plant Physiol. 66: 286-290.
73. Yang, S.F. and D.O. Adams. 1980. Biosynthesis of ethylene. In P. K. Stumpf
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74. Yang, S.F., D.O. Adams, C. Lizada, Y. Yu, K.J. Bradford, A.C. Cameron, and
N.E. Hoffman. 1980. Mechanism and regulation of ethylene biosynthesis.
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75. Fujino, D.W., M.S. Reid, and S.F. Yang. 1980. Effects of aminooxyacetic
acid on postharvest characteristics of carnation. Acta Horticulturae 113:
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76. Yang, S.F. 1980. Regulation of ethylene biosynthesis. HortScience 15:
238-243.
77. Bufler, G., Y. Mor, M.S. Reid, and S.F. Yang. 1980. Changes in 1-
aminocyclopropane-1-carboxylic-acid content of cut carnation flowers in
relation to their senescence. Planta 150: 439-442.
78. Yu, Y-B, S.F. Yang, J. Corse, J.A. Kuhnle, and S-S. Hua. 1980. Structures of
cytokinins influence synergistic production of ethylene. Phytochemistry 20:
1191-1195.
79. Lizada, M.C.C. and S.F. Yang. 1981. Sulfite-induced lipid peroxidation. Lipids
15: 189-194.
80. Bradford, K.J. and S.F. Yang. 1981. Physiological responses of plants to
waterlogging. HortScience 16: 25-30.
81. Apelbaum, A. and S.F. Yang. 1981. Biosynthesis of stress ethylene induced by
water deficit. Plant Physiol. 68: 594-596.
82. Adams, D.O. and S.F. Yang. 1981. Ethylene the gaseous plant hormone:
mechanism and regulation of biosynthesis. Trends Biochem. Sci. 6:
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83. Yang, S.F. 1981. Biosynthesis of ethylene and its regulation. In J. Friend (ed.),
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84. Hoffman, N.E., S.F. Yang, and T. McKeon. 1982. Identification of 1-
(malonylamino)cyclopropane-1-carboxylic acid as a major conjugate of
1-aminocyclopropane-1-carboxylic acid, an ethylene precursor in higher
plants. Biochem. Biophys. Res. Commun. 104: 765-770.
85. Yung, K.H., S.F. Yang, and F. Schlenk. 1982. Methionine synthesis from
5-methylthioribose in apple tissue. Biochem. Biophys. Res. Commun. 104:
771-777.
86. Hoffman, N.E. and S.F. Yang. 1982. Enhancement of wound-induced ethylene
synthesis by ethylene in preclimacteric cantaloupe. Plant Physiol. 69:
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87. Riov, J. and S.F. Yang. 1982. Autoinhibition of ethylene production in citrus
peel discs: suppression of 1-aminocyc1opropane-1-carboxylic acid synthesis.
Plant Physiol. 69: 687-690.
88. Yang, S.F., N.E. Hoffman, T. McKeon, J. Riov, C.H. Kao, and K.H. Yung. 1982.
Mechanism and regulation of ethylene biosynthesis. In P.P. Wareing (ed.),
Plant Growth Substances, Academic Press, London, pp. 239-248.
89. Peiser, G.D., M.C.C. Lizada, and S.F. Yang. 1982. Sulfite-induced lipid
peroxidation in chloroplasts as determined by ethane production. Plant Physiol.
70: 994-998.
90. Riov, J. and S.F. Yang. 1982. Effect of cycloheximide on ethylene production
in intact and excised citrus fruit tissues. J. Plant Growth Regul. 1: 95-104.
91. Cameron, A.C. and S.F. Yang. 1982. A simple method for the determination
of resistance to gas diffusion in plant organs. Plant Physiol. 70: 21-23.
92. Riov, J. and S.F. Yang. 1982. Effects of exogenous ethylene on ethylene
production in citrus leaf tissue. Plant Physiol. 70: 136-141.
93. Riov, J. and S.F. Yang. 1982. Stimulation of ethylene production in citrus leaf
discs by mannitol. Plant Physiol. 70: 142-146.
94. Hoffman, N.E., S.F. Yang, A. Ichihara, and S. Sakamura. 1982. Stereospecific
conversion of 1-aminocyclopropane-carboxylic acid to ethylene by plant
tissue: conversion of stereoisomers of 1-amino-2-ethylenecyclopropane-
carboxylic acid to 1-butene. Plant Physiol. 70: 195-199.
95. Peiser, G.D., M.C.C. Lizada, and S.F. Yang. 1982. Dark metabolism of carbon
dioxide and lettuce leaf discs. Plant Physiol. 70: 397-400.
96. Kao, C.H. and S.F. Yang. 1982. Light inhibition of the conversion of 1-
aminocyclopropane-1-carboxylic acid to ethylene in leaves is mediated
through carbon dioxide. Planta 155: 261-266.
97. McKeon, T.A., N.E. Hoffman, and S.F. Yang. 1982. The effect of plant hormone
pretreatments on ethylene production and synthesis of 1-aminocyclopropane-
1-carboxylic acid in water-stressed wheat leaves. Planta 155: 437-443.
98. Bradford, K.J., T.C. Hsiao, and S.F. Yang. 1982. Inhibition of ethylene
synthesis in tomato plants subjected to anaerobic root stress. Plant Physiol.
70: 1503-1507.
99. Yang, S.F. 1983. Mechanism and regulation of ethylene biosynthesis.
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100. Hoffman, N.E., Y. Liu, and S.F. Yang. 1983. Changes in 1-(malonylamino)
cyc1opropane-1-carboxylic acid content in wilted wheat leaves in relation to
their ethylene production rates and 1-aminocyclopropane-1-carboxylic acid
content. Planta 157: 518-523.
101. Hoffman, N.E., J-R. Fu, and S.F. Yang. 1983. Identification and metabolism of
1-(malonyl-amino)cyclopropane-1-carboxylic acid in germinating peanut
seeds. Plant Physiol. 71: 197-199.
366 Wolf Prize in Agriculture
102. Liu, Y., N.E. Hoffman, and S.F. Yang. 1983. Relationship between the
malonylation of 1-aminocyclopropane-1-carboxylic acid and D-amino acids
in mung-bean hypocotyls. Planta 158: 437-441.
103. Yang, S.F. 1983. Regulation of ethylene biosynthesis in relation to plant
senescence. Bulletin Plant Growth Regulator 11: 9-11.
104. Aharoni, N. and S.F. Yang. 1983. Auxin-induced ethylene production as
related to auxin metabolism in leaf discs of tobacco and sugar beet. Plant
Physiol. 73: 598-604.
105. Fu, J.R. and S.F. Yang. 1983. Release of heat pretreatment-induced dormancy
in lettuce seeds by ethylene or cytokinin in relation to the production of
ethylene and the synthesis of 1-aminocyclopropane-1-carboxylic acid during
germination. J. Plant Growth Regul. 2: 185-192.
106. McKeon, T.A. and S.F. Yang. 1983. A comparison of the conversion of
1-amino-2-ethylcyclopropane-1-carboxylic acid stereoisomers to 1-butene
by pea epicotyls and by a cell free system. Planta 160: 84-87.
107. Kao, C.H. and S.F. Yang. 1983. Role of ethylene in the senescence of detached
rice leaves. Plant Physiol. 73: 881-885.
108. Sisler, E.C. and S.F. Yang. 1984. Ethylene, the gaseous plant hormone.
Bioscience 34: 234-238.
109. Beyer, E.M., P.W. Morgan, and S.F. Yang. 1984. Ethylene. In M.B. Wilkins
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110. Yang, S.F. and N.E. Hoffman. 1984. Ethylene biosynthesis and its regulation
in higher plants. Ann. Rev. Plant Physiol. 35: 155-189.
111. Peiser, G.D., T. Wang, N.E. Hoffman, S.F. Yang, H. Liu, and C.T. Walsh. 1984.
Formation of cyanide from carbon 1 of 1-aminocyclopropane-1-carboxylic
acid during its conversion to ethylene. Proc. Natl. Acad. Sci. USA 81:
3059-3063.
112. Liu, Y., L. Su, and S.F. Yang. 1984. Metabolism of 1-aminoisobutyric acid in
mungbean hypocotyls in relation to metabolism of 1-aminocyclopropane-1-
carboxylic acid. Planta 161: 439-443.
113. Lau, O.L., Y. Liu, and S.F. Yang. 1984. Influence of storage atmospheres and
procedures on 1-aminocyclopropane-1-carboxylic acid concentration in
relation to flesh ripeness in ‘Golden Delicious’ apple. HortScience 19:
425-426.
114. Su, L., T. McKeon, D. Grierson, M. Cantwell, and S.F. Yang. 1984. Development
of 1-aminocyclo-propane-1-carboxylate synthase and polygalacturonase
activities during the maturation and ripening of tomato fruits in relation to
their ethylene production rates. HortScience 19: 576-578.
115. Yang, S.F. 1984. The formation of ethylene from 1-aminocyclopropane-1-
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130. Sisler, E.C., M.S. Reid, and S.F. Yang. 1986. Effect of antagonists of ethylene
action on binding of ethylene in cut carnations. Plant Growth Regul. 4:
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131. Nieder, M., W-K. Yip, and S.F. Yang. 1986. Interferences and specificity of
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132. Yang, S.F., Y. Liu, and O.L. Lau. 1986. Regulation of ethylene biosynthesis in
ripening apple fruits. Acta Horticulturae 197: 711-710.
133. Jiao, X-Z., S. Philosoph-Hadas, L-Y. Su, and S.F. Yang. 1986. The conversion
of 1-(malonylamino)cyclopropane-1-carboxylic acid to l-aminocyclopropane-
1-carboxylic acid in plant tissues. Plant Physiol. 81: 637-641.
134. Tong, C.B., J.M. Labavitch, and S.F. Yang. 1986. The induction of ethylene
production from pear cell culture by cell wall fragments. Plant Physiol. 81:
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135. Philosoph-Hadas, S., N. Aharoni, and S.F. Yang. 1986. Carbon dioxide enhances
the development of the ethylene forming enzyme in tobacco leaf discs. Plant
Physiol. 82: 925-929.
136. Yang. S.F. 1986. Regulation of plant growth by ethylene and related regulators.
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137. Miyazaki, J.H. and S.F. Yang. 1987. The methionine salvage pathway
in relation to ethylene and polyamine biosynthesis. Physiol. Plant. 69:
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138. Wang, T-T. and S.F. Yang. 1987. The physiological role of lipoxygenase in
ethylene formation from 1-aminocyclopropane-1-carboxylic acid in oat leaves.
Planta 170: 190-196.
139. Miyazaki, J.H. and S.F. Yang. 1987. Metabolism of 5-methylthioribose to
methionine. Plant Physiol. 84: 277-281.
140. Yang. S.F. 1987. The role of ethylene synthesis in fruit ripening. In W.W.
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142. Yang. S.F. 1987. Regulation of biosynthesis and action of ethylene. Acta
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144. Miyazaki, J.H. and S.F. Yang. 1987. Inhibition of the methionine cycle enzymes.
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145. Jiao, X-Z., W-K. Yip, and S.F. Yang. 1987. The effect of light and phytochrome
on 1-aminocyclopropane-1-carboxylic acid metabolism in etiolated wheat
seedling leaves. Plant Physiol. 85: 643-647.
146. Tong, C.B.S. and S.F. Yang. 1987. Chilling-induced ethylene production by
beans and peas. J. Plant Growth Regul. 6: 201-208.
147. Sisler, E.C. and S.F. Yang. 1987. Ethylene. In D.W. Newman and K.G. Wilson
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148. Adams, D.O. and S.F. Yang. 1987. 1-aminocyclopropane-1-carboxylate
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150. Yip, W-K. and S.F. Yang. 1988. Cyanide metabolism in relation to ethylene
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151. Yip, W-K., X-Z. Jiao, and S.F. Yang. 1988. Dependence of in vivo ethylene
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152. Satoh, S. and S.F. Yang. 1988. S-adenosylmethionine-dependent inactivation
and radiolabeling of 1-aminocyclopropane-1-carboxylate synthase isolated
from tomato fruits. Plant Physiol. 88: 109-114.
153. Fujino, D.W., D.W. Burger, S.F. Yang, and K.J. Bradford. 1988. Characterization
of an ethylene overproducing mutant of tomato (Lycopersicon esculentum)
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154. Satoh, S. and S.F. Yang. 1989. Specificity of S-adenosyl-L-methionine in the
inactivation and the labeling of 1-aminocyclopropane-1-carboxylate synthase
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155. Riov, J. and S.F. Yang. 1989. Enhancement of adventitious root formation in
mung bean cuttings by 3,5-dihalo-4-hydroxybenzoic acids and 2,4-
dinitrophenol. Plant Growth Regul. 8: 277-281.
156. Riov, J. and S.F. Yang. 1989. Ethylene and auxin-ethylene interaction in
adventitious root formation in mung bean (Vigna radiata) cuttings. J. Plant
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157. Wien, H.C., A.D. Turner, and S.F. Yang. 1989. Hormonal basis for low light
intensity-induced flower bud abscission of pepper. J. Amer. Soc. Hort. Sci.
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158. Satoh, S. and S.F. Yang. 1989. Inactivation of 1-aminocyclopropane-1-
carboxylate synthase by L-vinylglycine as related to the mechanism-based
370 Wolf Prize in Agriculture
171. Dong, J-G., W.T. Kim, W.K. Yip, G.A. Thompson, L. Li, A.B. Bennett, and
S.F. Yang. 1991. Cloning of a cDNA encoding 1-aminocyclopropane-1-
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172. Yang, S.F., W-K. Yip, J-G. Dong, and S. Satoh. 1991. Characterization of
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biosynthesis: active site peptide sequencing. In T. Fukui, H. Kagamiyama,
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174. Kim, W.T., A. Silverstone, W-K. Yip, J-G. Dong, and S.F. Yang. 1992.
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175. Dong, J-G., D. Olson, A. Silverstone, and S.F. Yang. 1992. Sequence of a
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176. Dong, J-G., J.C. Fernández-Maculet, and S.F. Yang. 1992. Purification and
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177. Fernández-Maculet, J.C. and S.F. Yang. 1992. Extraction and partial
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178. Kim, W.T. and S.F. Yang. 1992. Turnover of 1-aminocyclopropane-1-
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179. Yip, W.-K. and S.F. Yang. 1993. Ethylene metabolism. In P.J. Lea (ed.), Methods
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372 Wolf Prize in Agriculture
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185. Moussatos, V.V., S.F. Yang, B. Ward, and D.G. Gilchrist. 1994. AAL-toxin
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190. Mizutani, P., J.G. Dong, and S.F. Yang. 1995. Effect of pH on CO2-activated
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191. Liu, Y., A.L. Silverstone, Y.M. Wu, and S.F. Yang. 1995. Formation of
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192. Wu, Y.M., A.L. Silverstone, Y. Liu, and S.F. Yang. 1995. Partial purification
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195. Li, N., X.N. Jiang, G.P. Cai, and S.F. Yang. 1996. A novel bifunctional fusion
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196. Shaw, J.P., Y.S. Chou, R.C. Chang, and S.F. Yang. 1996. Characterization of
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197. Kim, W.T., A. Campbell, T. Moriguchi, H.C. Yi, and S.F. Yang. 1997. Auxin
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200. Oetiker. J.H., D.C. Olson, O.Y. Shiu, and S.P. Yang. 1997. Differential induction
of seven 1-aminocyclopropane-1-carboxylate synthase genes by elicitor in
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201. Charng. Y.Y., Y. Liu, J.G. Dong, and S.F. Yang. 1997. On 1-aminocyclopropane-
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374 Wolf Prize in Agriculture
207. Lin, C.T., K.Y. Fu, M.T. Lin, S.F. Yang, and J.P. Shaw. 1998. Cloning and
characterization of a cDNA for 1-aminocyclopropane-1-carboxylate synthase
from papaya fruit (Accession No. Y1l357) (PGR98-046). Plant Physiol. 116:
1193.
208. Chen, H.H., Y.Y. Charng, S.F. Yang, and J.P. Shaw. 1998. Molecular cloning
and sequencing of a broccoli cDNA (Accession No. AF047476) encoding an
ETR-type ethylene receptor. (PGR98-088). Plant Physiol. 117: 717.
209. Chen, H.H., Y.Y. Charng. S.F. Yang, and J.P. Shaw. 1998. Isolation and
characterization of a broccoli cDNA (Accession No. AF047477) encoding
and ERS-type ethylene receptor. (PGR98-123). Plant Physiol. 117: 1126.
210. Yang, S.F. and Y.Y. Charng. 1998. The gaseous plant hormone ethylene: from
biosynthesis and action to biotechnology. In C.H. Chou and K.T. Shao (eds.),
“Plenary Lecture, Frontiers in Biology: The Challenges of Biodiversity,
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pp. 25-34.
211. Yip, W.K. and S.F. Yang. 1998. Ethylene biosynthesis in relation to cyanide
metabolism. Bot. Bull. Acad. Sin. 39: 1-7.
212. Yang, S.F. and J.H. Oetiker. 1998. Molecular biology of ethylene biosynthesis
and its application in horticulture. J. Japan. Soc. Hort. Sci. 67: 1209-1214.
213. Kung. S. D. and S. F. Yang (eds.).1998. Discoveries in Plant Biology. Vol. II.
World Scientific, Singapore, 380 pp.
214. Yang, X.X., H.W. Choi, S.F. Yang, and N. Li. 1999. A UV-light activated
cinnamic acid isomer regulates plant growth and gravitropism via an ethylene
receptor-independent pathway. Aust. J. Plant Physiol. 26: 325-335.
215. Wong WS, W. Ning, P.L. Xu, S.D. Kung, S.F. Yang, and Li N. 1999. Identification
of two chilling-regulated 1-aminocyclopropane-1-carboxylate synthase genes
from citrus (Citrus sinensis Osbeck) fruit. Plant Mol. Biol. 41 (5): 587-600.
216. Zhou H.Q., H.W. Wang, K. Zhu, S.F. Sui, P.L. Xu, S.F. Yang, and Li N. 1999.
The multiple roles of conserved arginine 286 of 1-aminocyclopropane-1-
carboxylate synthase. Coenzyme binding, substrate binding, and beyond. Plant
Physiol. 121: 913-919
217. Ge, L., J.Z. Liu, W.S. Wong, W.L.W. Hsiao, K. Chong, Z.K. Xu, S.F. Yang, S.D.
Kung, and Li, N. 2000. Identification of a novel multiple environmental
factor-responsive 1-aminocyclopropane-1-carboxylate synthase gene,
NT-ACS2, from tobacco. Plant Cell and Environ. 23: 1169-1182.
218. Charng, Y., S.J. Chou, W.T. Jiaang, S.T. Chen, and S.F. Yang. 2001. The catalytic
mechanism of 1-aminocyclopropane-1-carboxylic acid oxidase. Arch.
Biochem. Biophys. 385: 179-185.
219. Wang, N.N., S.F. Yang, and Y.Y. Charng. 2001. Differential expression of 1-
aminocyclopropane-1-carboxylate synthase genes during orchid flower
senescence induced by the protein phosphatase inhibitor okadaic acid. Plant
Physiol. 126: 253-260.
Shang Fa Yang 375
220. Chen, L.F.O., J.Y. Hwang, Y.Y. Charng, C.W. Sun, and S.F. Yang.
2001. Transformation of broccoli (Brassica oleracea var. italica) with
isopentenyltransferase gene via Agrobacterium tumefaciens for post-harvest
yellowing retardation. Molecular Breeding 7: 243-257.
221. Peng, H.P., C.S. Chan, M.C. Shih, and S.F. Yang. 2001. Signaling events in the
hypoxic induction of alcohol dehydrogenase gene in Arabidopsis. Plant Physiol.
126: 742-749.
222. Chung, M.C., S.J. Chou, L.Y. Kuang, Y.Y. Charng, and S.F. Yang. 2002.
Subcellular localization of 1-aminocyclopropane-1-carboxylic acid oxidase
in apple fruit. Plant and Cell Physiol. 43: 549-554.
223. Yang, C.Y., F.H. Chu, Y.T. Wang, Y.T. Chen, S.F. Yang, and J.F. Shaw. 2003.
Novel broccoli 1-aminocyclopropane-1-carboxylate oxidase gene (BoACO3)
associated with the late stage of postharvest floret senescence. J. Agric. Food
Chem. 51: 2569-2575.
224. Chen, Y.T., Y.R. Lee, C.Y. Yang, Y.T. Wang, S.F. Yang, and J.F. Shaw. 2003.
A novel papaya ACC oxidase gene (CpACO2) associated with late stage fruit
ripening and leaf senescence. Plant Science 164: 531-540
225. Grichko, V, M. Serek, C.B. Watkins, and S.F. Yang. 2006. Father of 1-MCP -
Edward C. Sisler. Biotechnol. Advances 24 (4): 355-356.
226. Huang, L.C., U.L. Lai, S.F. Yang, M.J. Chu, C.I. Kuo, M.F. Tsai, C.W. Sun. 2007.
Delayed flower senescence of Petunia hybrida plants transformed with
antisense broccoli ACC synthase and ACC oxidase genes. Postharvest Biol.
and Technol. 46 (1): 47-53
376 Wolf Prize in Agriculture
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Abstract
Shang Fa Yang was born in Taiwan in 1932. After receiving his B.S. and M.S. degrees in Agricultural Chemistry from the National Taiwan
University, he came to the United States in 1958 to pursue a Ph.D. degree at Utah State University. Following three postdoctoral years, he was hired
at the University of California, Davis, in 1966 where he was a biochemist and professor for 28 years. After retiring early from UC Davis, he
subsequently established a research program at the Hong Kong University of Science and Technology and served as Vice President of Academia
Sinica in Taiwan. Yang’s research achievements included discovering the biochemical pathway for the synthesis of ethylene by identifying the key
steps by which S-adenosylmethionine (SAM) is converted into 1-aminocyclopropane-1-carboxylic acid (ACC) and subsequently into ethylene.
Yang further demonstrated how the methylthio and ribose moieties from SAM were recycled back into methionine in order to sustain high rates of
ethylene synthesis, as in ripening fruits. This recycling pathway is now known as the Yang Cycle. Yang also contributed to the isolation,
characterization and cloning of ACC synthase and ACC oxidase, the two enzymes in the ethylene biosynthetic pathway, and to the elucidation of
their structure and reaction mechanisms. He made important contributions to auxin, cytokinin, cyanide and sulfur metabolism in plants as well. His
work formed the basis for subsequent research that has established ethylene as the most thoroughly characterized of the hormonal biosynthesis and
signaling pathways in plants.
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distortions in growing plants, generally associated with leaking (C2H4). At the time that Yang entered the field, Morris
gas mains. However, research primarily at the Boyce Lieberman and others had evidence that amino acids,
Thompson Institute in the 1930s showed that plants produce particularly methionine, might be a precursor for ethylene
ethylene and that it is broadly involved in regulating plant production [4,5]. Various in vitro systems were being used to
growth and development, including seed germination, root and convert methionine and other potential precursors into
shoot growth, responses to environmental stresses, flowering, ethylene, and Yang contributed significantly to these studies,
fruit ripening and senescence or death of plant tissues and using his knowledge of chemistry to explore different
organs [3]. However, little was known about the pathway of reaction mechanisms [6]. He subsequently utilized both in
ethylene biosynthesis in plants, and armed with Pratt’s gas vitro and in vivo approaches to explore potential inter-
chromatograph and his broad knowledge of chemistry and mediates of and mechanisms for the conversion of
biochemistry, Yang set out to explore plant ethylene biology. methionine to ethylene [7,8]. He also studied the mechanism
of formation of ethylene from 2-chloroethylphosphonic acid
2. Ethylene biosynthesis in plant tissues [9]. This compound, under the generic name
of ethephon, enabled the commercial application of ethylene
Ethylene is a simple gaseous compound containing two for agricultural purposes, as it is taken up by plants
carbon and four hydrogen atoms and one double bond and converted into ethylene. It has been widely used as a
Please cite this article in press as: K.J. Bradford, Shang Fa Yang: Pioneer in plant ethylene biochemistry, Plant Sci. (2008),
doi:10.1016/j.plantsci.2008.01.005
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fruit-ripening agent, to loosen fruits for harvest and for An early dilemma in ethylene biosynthesis was that
defoliation of cotton before harvest [3]. methionine pools were generally too low in plant tissues to
Continuing studies on the biogenesis of ethylene showed sustain the observed rates of ethylene synthesis. Some had
that methionine was converted to S-adenosylmethionine (SAM) suggested that following ACC formation, the methylthio group
and that SAM was a precursor of ethylene. When apple tissues from methionine would be attached to an existing homo-
were supplied with 14C-SAM under anaerobic conditions that cysteine molecule to form a new methionine molecule, thus
prevented ethylene formation, a labeled compound accumu- recycling only the methylthio group. However, Yang’s and
lated in tissues [10]. This discovery stimulated active Lieberman’s groups demonstrated that after ACC is released
competition among various groups to identify the unknown from SAM, the methylthio group and the ribose moiety from
intermediate between SAM and ethylene that culminated in the the remaining methylthioadenosine (MTA) are both recycled to
identification by Adams and Yang of 1-aminocyclopropane-1- replenish methionine levels and sustain ethylene biosynthesis
carboxylic acid (ACC) as the final in vivo precursor of ethylene [37–39]. This cycle has been christened the Yang Cycle in plant
(Fig. 2; [11]). Yang’s group quickly developed a sensitive assay biochemistry texts (Fig. 2; [40]).
for ACC via its chemical conversion to ethylene [12], which
facilitated wide-ranging studies by his group on the regulation 4. Other activities and honors
of ACC and ethylene biosynthesis in plant growth, fruit
ripening, and stress responses [13–15]. While most widely known for his work on ethylene
The ethylene biosynthetic pathway requires only two biosynthesis and action, Yang also maintained active research
specific steps, the conversion of SAM to ACC and of ACC programs in other areas of plant growth and metabolism,
to ethylene (Fig. 2), and attention quickly turned to the including on auxin metabolism and action [41–43], on
identification of the enzymes responsible for them. Yang’s cytokinin action [44,45], on the biological effects of sulfite
group and that of Hans Kende at Michigan State University and sulfur dioxide [46,47], and on cyanide generation and
soon reported the properties of 1-aminocyclopropane carbox- metabolism in plants [48].
ylate synthase, the enzyme responsible for the conversion of A theme throughout all of Yang’s work is a linkage to
SAM to ACC [16–18]. The subsequent cloning of ACC practical applications in postharvest biology and plant growth
synthase, in which the Yang, Kende and A. Theologis regulation. He applied his knowledge of ethylene biosynthesis
laboratories were involved, is described in detail by Kende to contribute to improvements in postharvest storage conditions
[19]. Isolation of the ethylene-forming enzyme, or ACC [49–52]. Yang figured prominently at many national and
oxidase, was a more difficult task, as the enzyme appeared to be international research conferences and served on the editorial
membrane-associated and activity was lost upon cellular boards of leading journals and as a member of several learned
disruption and fractionation. The gene coding for the enzyme societies. He won many awards and honors, including a
was eventually cloned by D. Grierson’s group based upon Guggenheim Fellowship in 1982, the International Plant
expression of a ripening-related cDNA and demonstration that Growth Substances Association Research Award in 1985,
its suppression blocked ethylene synthesis [19–21]. Yang and the Outstanding Researcher Award from the American
contributed to the characterization of the multiple alleles in the Society for Horticultural Science in 1992. In 1992, he was
ACC synthase and ACC oxidase families and to structure- named the UC Davis Faculty Research Lecturer, the highest
function studies of the reaction mechanisms of the two enzymes honor given by that institution for excellence in research. He
[22–29]. was elected to the US National Academy of Sciences in 1990
and to the Academia Sinica, Taiwan, in 1992. In 1991, Yang
3. ACC metabolism and the Yang Cycle received the prestigious international Wolf Prize in Agriculture,
which many consider to be the ‘‘Nobel Prize’’ for agricultural
Ethylene synthesis exhibits multiple points of regulation, research.
including transcriptional control of multiple ACC synthase and After a distinguished career as professor and biochemist at
ACC oxidase genes and the activation state and/or stability of the University of California, Davis, Yang took early retirement
the enzymes themselves [30]. For example, light, carbon in 1994 to accept a Distinguished Professorship at the Hong
dioxide, oxygen, and water stress all influence the conversion of Kong University of Science and Technology, where he
ACC to ethylene [31]. The conversion of ACC to ethylene is established an active plant research group in the Department
also stereospecific, as demonstrated by the differential of Biology. In 1995, he was recognized as a Distinguished
conversion of stereoisomers of 1-amino-2-ethylcyclopropane Research Fellow by the Institute of Botany of the Academia
carboxylic acid to 1-butene [32]. This provided an important Sinica in Taipei, Taiwan, and returned to Taiwan in 1996 to
test for the enzymatic conversion of ACC to ethylene versus serve as Vice President of Academia Sinica from 1996 to 1999.
non-specific chemical conversion [33] that was important in the In this position he directed its numerous research institutes,
subsequent isolation and cloning of ACC oxidase [34,35]. In including the establishment of a new Institute of Agricultural
addition to being converted to ethylene, Yang’s group Biotechnology (subsequently renamed the Agricultural Bio-
determined that ACC could be malonylated and that this pool technology Research Center). In both Hong Kong and Taiwan,
of conjugated ACC was largely unavailable for conversion to Yang played important leadership roles in advancing plant
ethylene (Fig. 2; [36]). biology and agricultural biotechnology. Following his tenure at
Please cite this article in press as: K.J. Bradford, Shang Fa Yang: Pioneer in plant ethylene biochemistry, Plant Sci. (2008),
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Fig. 2. The Yang Cycle and formation of ethylene and other products from ACC. See text for details.
Please cite this article in press as: K.J. Bradford, Shang Fa Yang: Pioneer in plant ethylene biochemistry, Plant Sci. (2008),
doi:10.1016/j.plantsci.2008.01.005
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Yang maintained an open mind and was willing to challenge [17] T. Boller, R.C. Herner, H. Kende, Assay for and enzymatic formation of an
ethylene precursor, 1-aminocyclopropane-1-carboxylic acid, Planta 145
accepted ideas, even his own, when they proved untenable in
(1979) 293–303.
the face of experimental data. As a mentor, Yang was [18] H. Kende, Enzymes of ethylene biosynthesis, Plant Physiol. 91 (1989) 1–
demanding and rigorous yet positive, encouraging and 4.
supportive. Many graduate students and postdoctoral associates [19] H. Kende, Ethylene biosynthesis, Annu. Rev. Plant Physiol. Plant Mol.
benefitted greatly from both his mentorship and his subsequent Biol. 44 (1993) 283–307.
support for their careers. Students and colleagues will [20] C.J.S. Smith, A. Slater, D. Grierson, Rapid appearance of an mRNA
correlated with ethylene synthesis encoding a protein of molecular weight
remember and miss his humor, his humanity, and his sparkling 35000, Planta 168 (1986) 94–100.
intellect. This special issue is a fitting tribute to Shang Fa [21] A.J. Hamilton, G.W. Lycett, D. Grierson, Antisense gene that inhibits
Yang’s pioneering contributions to our understanding of the synthesis of the hormone ethylene in transgenic plants, Nature 346 (1990)
role of ethylene in plant biology. 284–287.
[22] J.G. Dong, W.T. Kim, W.K. Yip, G.A. Thompson, L.M. Li, A.B. Bennett,
S.F. Yang, Cloning of a cDNA encoding 1-aminocyclopropane-1-carbox-
Acknowledgements ylate synthase and expression of its messenger RNA in ripening apple
fruit, Planta 185 (1991) 38–45.
The author is grateful to Eleanor, Albert and Bryant Yang for [23] W.T. Kim, S.F. Yang, Structure and expression of cDNAs encoding 1-
aminocyclopropane-1-carboxylate oxidase homologs isolated from
sharing personal information and thanks Yee-Yung Charng,
excised mung bean hypocotyls, Planta 194 (1994) 223–229.
Neil E. Hoffman, John H. Miyazaki, Galen D. Peiser, Mikal E. [24] W.-K. Yip, T. Moore, S.F. Yang, Differential accumulation of transcripts
Saltveit and Wing Kin Yip for contributing additional for four tomato 1-aminocyc1opropane-1-carboxylate synthase homologs
information. under various conditions, Proc. Natl. Acad. Sci. U.S.A. 89 (1992) 2475–
2479.
[25] D.C. Olson, J.H. Oetiker, S.F. Yang, Analysis of LeACS3, a 1-amino-
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[44] L.F.O. Chen, J.Y. Hwang, Y.Y. Charng, C.W. Sun, S.F. Yang, Transforma- Delayed flower senescence of Petunia hybrida plants transformed with
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doi:10.1016/j.plantsci.2008.01.005
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JOHN E. CASIDA
University of California
Berkeley, California, USA
k
Preface
John E. Casida was a major contributor to the Golden Age of insecticide research.
This was an era of glory. DDT, invented by Nobel Laureate Paul Müller, greatly
increased crop yields and brought devastating insect-transmitted human diseases
under control. Many other insecticides quickly followed to eliminate practically all
pest insect problems. It was also a time, as pointed out by Rachel Carson in her
book “Silent Spring”, when broadscale application of toxic and persistent pesticides
damaged natural ecosystems with an impact on life in general. Industries pursued
more effective insecticides and governments regulated their safe use. Fundamental
knowledge was needed from academic research. This was the origin of the fields
of Insect Toxicology and Pesticide Chemistry and Toxicology.
383
384 Wolf Prize in Agriculture
and research positions became available for his father Lester Earl Casida (Professor
of Reproductive Physiology, University of Wisconsin, Madison). He has an older
brother (Lester Earl Casida Jr., Professor of Bacteriology, Pennsylvania State
University) and younger sister (Betty Ruth Damerau, violinist, Denver Symphony
Orchestra). He attended grade and high schools and University in Madison. John
was allowed great freedom to explore, living close to a large arboretum and setting
up a basement science laboratory. He participated in all aspects of the Boy Scouts
program and high school science and photography clubs. At University he was
captain of the fencing team and medalist in saber. Four summers were spent in
forestry nursery and vegetable research stations and in programs for seed
certification and insect rearing and toxicity testing. Graduate school was interrupted
after one year by service in the Korean War as a First Lieutenant in the United
States Air Force specializing in Medical Entomology at Camp Detrick (Frederick,
Maryland).
Academic Career
John graduated from the University of Wisconsin at Madison with a B.S. degree in
Entomology in 1951, an M.S. in Biochemistry in 1952 and a Ph.D. jointly in
Biochemistry, Entomology and Plant Physiology in 1954. He began research in
entomology and on insecticides in high school and as an undergraduate in college
by field evaluation of botanicals and isolation of their insecticidal ingredients. His
Ph.D. thesis was on the metabolic activation of systemic organophosphorus
insecticides. Dr. Casida was appointed Assistant Professor of Entomology (Insect
Toxicology) at the University of Wisconsin at Madison in 1954 and was promoted
to Associate Professor in 1957 and Full Professor in 1961. In 1964 he moved to
John E. Casida 385
Organophosphorus Toxicants
Research in the 1950s and 1960s on organophosphorus and methylcarbamate
insecticides, such as schradan and carbaryl, laid the background for safety
evaluations by synthesis, structure-activity and metabolism studies on perhaps 50
candidate or commercial anticholinergic pesticides. During this period John
discovered the insecticide and anthelmintic butonate, the first specifically-designed
selective proinsecticide based on species differences in activation and detoxifying
enzymes. Studies with tri-o-cresyl phosphate (the casual agent for 40,000 cases of
human peripheral neuropathy) identified a saligenin cyclic phosphate as the highly
potent activated metabolite and established later that the biochemical lesion involved
inhibition of a lysophosphatidylcholine hydrolase designated neuropathy target
esterase. Kynurenine formamidase was shown to be the target for organophosphate
and methylcarbamate insecticides that are extremely potent in producing embryonic
abnormalities (teratogenic effects) in avian embryos. Later studies at Berkeley
established major enzymes involved in organophosphate toxicant detoxification
and multiple effects on the cannabinoid system and lipid biochemistry and signaling.
These are just a few of many sensitive serine hydrolase targets shown to have
toxicological significance.
GABAergic Insecticides
A major advance in insecticide toxicology came from a chance observation of very
simple and highly toxic bicyclophosphates and related bicycloortho-carboxylates
with a totally unexpected mode of action, followed by fundamental research on
neurobiology. Radioligands were prepared (now available commercially as [35S]TBPS,
[3H]TBOB and [3H]EBOB) and receptor binding studies combined with physiological
assays established that the new toxicants are noncompetitive blockers of the
γ-aminobutyric acid (GABA)-gated chloride channel. The binding site of TBPS and
the bicycloorthocarboxylates was discovered to be the same as, or closely coupled
to, that of the polychlorocycloalkane insecticides (such as dieldrin, lindane and
toxaphene) so finally establishing their mode of action after 3 billion pounds
had been used. As further evidence for this mode of action, toxaphene was examined
and shown to be a mixture of >188 isomeric polychlorobornanes which were
fractionated and the principal toxic hepta- and octachlorobornanes identified. The
trace isomers most toxic to insects, fish and mammals are the same as those most
active in vitro at the receptor. A rational approach to totally new classes of
insecticides then led to 1,4-disubstituted-2,6,7-trioxabicyclo[2.2.2]octanes and to
2,5-disubstituted-1,3-dithianes, non-halogenated and non-phosphorus compounds
of unusually simple structure, some of which are more active than many commercial
insecticides. These new heterocyclic insecticides were optimized for high potency,
selective toxicity, and use as photoaffinity ligands and affinity columns. Studies
with [3H]EBOB in Drosophila established that resistance to nine classes of
insecticides is due to a single amino acid change in the chloride channel.
Demonstration that the noncompetitive antagonist site is highly expressed and
ultra sensitive in a human β3 subunit GABAA receptor homopentamer ultimately
allowed a model for the precise binding site of multiple classes of natural and
synthetic toxicants blocking the chloride channel. The arylpyrazole insecticide
fipronil acting at the GABA receptor undergoes a totally unexpected yet facile
photodesulfinylation reaction to become much more toxic and persistent. α-Thujone,
the principal active ingredient of the aperitif absinthe, was shown to act as a
noncompetitive antagonist of the GABA receptor solving a century-old mechanistic
puzzle.
Neonicotinoid Insecticides
Nicotine was used for centuries to control insect pests with marginal effectiveness
and considerable health risk. The discovery of synthetic alternatives, the
neonicotinoids, allowed optimization for potency, safety, and photostability,
ultimately leading to the neonicotinoids, currently the third most important class
of insecticides. [3H]Neonicotinoids prepared as radioligands showed the unique
sensitivity of the nicotinic acetylcholine receptor of insects compared with mammals.
388 Wolf Prize in Agriculture
Herbicide Research
Herbicide research established that: (a) the metabolic activation of important
thiocarbamates involves previously-unrecognized thiocarbamate sulfoxides; (b) the
action of dichloroacetamides as safeners lowering the toxicity of thiocarbamates to
maize but not to weeds involves inducing the synthesis of glutathione and glutathione
S-transferase that rapidly detoxify the metabolically-activated herbicide; (c) major
thiocarbamate herbicides and the fungicide benomyl are potent acetaldehyde
dehydrogenase inhibitors and may therefore sensitize agricultural workers to
ethanol intoxification; (d) the mutagenic and carcinogenic activity of chloroallyl
thiocarbamates and dibromochloropropane are attributable to 2-haloacrolein
metabolites; (e) the stereospecific metabolic conversion of phenoxyaryloxypropionates
to their coenzyme A esters greatly enhances their potency as inhibitors of acetyl-
coenzyme A carboxylase; (f) the inhibition of protein phosphatase 2A in plants by
endothal accounts for its phytotoxicity, as with cantharidin and its effects in mammals;
(g) a 3H-labeled N-aryltetrahydrophthalimide developed as a radioligand effectively
quantitates the protoporphyrinogen IX oxidase site which is very similar in plants
and mammals.
Investigator, first in Madison and then at Berkeley, supported one of the longest
continuous major academic programs on pesticide science in the United States.
Dr. Casida was a member of the United States National Advisory Environmental
Health Sciences Council and currently serves as a consultant in the United States
and abroad on agrochemical research. He is a major contributor to international
congresses and meetings and had joint research programs in Israel and
Hungary and served as advisor to the Ministers of Agriculture and Irrigation
in Egypt.
The lifetime contributions of Professor John Casida have been recognized by
the Jeffrey Lectureship at the University of New South Wales, Australia in 1983,
the Messenger Lectureship at Cornell University in Ithaca in 1985, the Yabuta
Lectureship in Fukuoka and the Botyu-Kagaku Lectureship in Kyoto in 1987, the
Sterling B. Hendricks Memorial Lectureship of the United States Department of
Agriculture in 1992, the Third World Academy of Sciences Lectureship in Sciences,
University of Buenos Aires in Argentina, 1997, and the University of California at
Berkeley Faculty Research Lectureship (the highest distinction the Berkeley campus
can bestow in honoring research accomplishments) in 1998. He received the
Distinguished Service Award for research by the United States Department of
Agriculture in 1988, the J.E. Bussart Award and the title Fellow by the Entomological
Society of America in 1989, the Kôrô-sho Prize of the Pesticide Science Society of
Japan in 1995 and was named Honorary Member of the Society of Toxicology in
1996 and the Pesticide Science Society of Japan in 2005. Professor Casida was
elected as a member of the United States National Academy of Sciences in 1991, as
a foreign member of the Royal Society (United Kingdom) in 1998, as a member of
the European Academy of Sciences in 2004, and received the Wolf Foundation
Prize in Agriculture in 1993.
Education/Training
University of Wisconsin, Madison: B.S. 1951 Entomology, M.S. 1952 Biochemistry,
Ph.D. 1954 Entomology/Biochemistry
Professional Positions
1954-63 Assistant, Associate and Full Professor of Entomology, University of
Wisconsin, Madison
1958-59 Visiting Researcher, University of Stockholm and Several European
Laboratories
1970-71 Visiting Researcher, Cambridge University, University of Stockholm,
and Kyoto University
1986-87 Visiting Researcher, University of Crete (Greece) and Centre National
de Recherche Scientifique (Gif-sur-Yvette, France)
390 Wolf Prize in Agriculture
Award(s)
1958 Haight Fellow
1970 Guggenheim Fellow
1970 International Award for Research in Pesticide Chemistry, American Chemical
Society
1978 Spencer Award for Research in Agricultural & Food Chemistry, American
Chemical Society
1988 Distinguished Service Award for Research, USDA
1989 J.E. Bussart Award and Fellow of the Entomological Society of America
1991 National Academy of Sciences (U.S.A.), Member
1992 Sterling B. Hendricks Memorial Lectureship, Agricultural Research Service,
USDA & American Chemical Society
1993 Wolf Foundation Prize in Agriculture
1994 Founders Award, Society of Environmental Toxicology and Chemistry
1995 Koro-Sho Prize, Pesticide Science Society of Japan
1997 Honorary Member, Society of Toxicology
1997 Honorary Doctor Degree and Third World Academy of Sciences Lectureship,
University of Buenos Aires, Argentina
1998 Faculty Research Lecturer, U.C. Berkeley
1998 Royal Society of the United Kingdom, Fellow (FRS) and Foreign Member
2004 European Academy of Sciences, Member
2005 Honorary Member, Pesticide Science Society of Japan
P5. 1962. Esenther, G.R., T.C. Allen, J.E. Casida and R.D. Shenefelt. Decayed
wood extract as termite attractant. U.S. Pat. 3,070,495. Dec. 25,
1962.
P6. 1975. Tilles, H. and J.E. Casida. Method of making carbonoyl sulfoxide
derivatives. U.S. Pat. 3,896,169. July 22, 1975.
P7. 1975. Tilles, H. and J.E. Casida. Carbamoyl sulfoxide derivatives. U.S. Pat.
3,928,436. Dec. 23, 1975.
P8. 1985. Palmer, C.J. and J.E. Casida. Pesticides comprising 1,4-bis-substituted
2,6,7-trioxabicyclo[2.2.2]octanes. PCT Int. Appl., 34 pp. CA103(23):
191474k
P9. 1986. Dureja, P., J.E. Casida and L.O. Ruzo. Additives for improved pesticide
photostability. U.S. Pat. 4,622,315. Nov. 11, 1986, 6 pp.
P10. 1987. Casida, J.E., C.J. Palmer, J.P. Larkin and I.H. Smith. Trioxabicyclo
[2.2.2]octanes as insecticides and acaricides. Eur. Pat. Appl., 65 pp.
CA106(21): 176403e
P11. 1987. Casida, J.E., C.J. Palmer, J.P. Larkin and I.H. Smith. Preparation of
heterobicycloalkane pesticidal compounds. Eur. Pat. Appl., 52 pp.
CA107(1): 2705k
P12. 1988. Palmer, C.J. and J.E. Casida. 1,4-bis-Substituted-2,6,7-trioxabicyclo
(2,2,2)-octanes having ethynyl substituted phenyl group. U.S. Pat.
4,772,624. Sept. 20, 1988, 8 pp.
P13. 1988. Casida, J.E., C.J. Palmer, J.P. Larkin and I.H. Smith. Preparation of
1-aryl-2,6,7-trioxabicyclo[2.2.2]octanes as pesticides. Eur. Pat. Appl.,
25 pp. CA109(25): 231038d
P14. 1988. Casida, J.E., M. Elliott and D.A. Pulman. Preparation and testing of
4-(tertiary butyl)-1,3-dithianes and oxidized derivatives as insecticides,
acaricides, and anthelmintics. Eur. Pat. Appl., 85 pp. CA110(21):
192834j
P15. 1988. Palmer, C.J. and J.E. Casida. 1,4-Bis-substituted 2,6,7-trioxabicyclo
[2.2.2]octanes having an ethynyl-substituted phenyl group, and their
pesticidal compositions. Eur. Pat. appl., 8 pp. CA110(23): 212833p
P16. 1988. Casida, J.E., M. Elliott and D.A. Pulman. Preparation of dithianes as
pesticides and parasiticides. Eur. Pat. Appl., 44 pp. CA110(25):
231651u
P17. 1989. Palmer, C.J. and J.E. Casida. New class of pesticides comprising 1,4-
bis-substituted-2,6,7-trioxabicyclo(2,2,2)octanes. U.S. Pat. 4,876,274.
Oct. 24, 1989, 12 pp.
P18. 1990. Casida, J.E., C.J. Palmer, J.P. Larkin and I.H. Smith. Pesticidal compounds.
U.S. Pat. 4,942,173. July 17, 1990, 10 pp.
P19. 1990. Casida, J.E., C.J. Palmer, J.P. Larkin and I.H. Smith. Pesticidal compounds.
U.S. Pat. 4,965,257. Oct. 23, 1990, 19 pp.
392 Wolf Prize in Agriculture
Selected Publications
(*Reprints included)
Casida, J.E. and T.C. Allen. 1951. A laboratory method for evaluating the
phytotoxicity or phytostimulation of insecticides. Science 113: 553-555.
1
* Casida, J.E., T.C. Allen and M.A. Stahmann. 1953. Enzymatic and chemical
oxidation of dimethylphosphoramides to biologically active dimethyl-
phosphoramide oxides. Nature 172: 243-245.
Casida, J.E. 1955. Isomeric substituted-vinyl phosphates as systemic insecticides.
Science 122: 587-598.
*Casida, J.E. 1958. Phosphorus-32 pentasulfide: preparation by isotopic exchange
and conversion to thiophosphoryl-32 chloride and phosphorus-32 trichloride.
Acta Chem. Scand. 12: 1691-1692.
1
* Casida, J.E., M. Eto and R.L. Baron. 1961. Biological activity of a tris-o-cresyl
phosphate metabolite. Nature 191: 1396-1397.
Dorough, H.W., N.C. Leeling and J.E. Casida. 1963. Nonhydrolytic pathway in
metabolism of N-methylcarbamate insecticides. Science 140: 170-171.
Roger, J-C., H. Chambers and J.E. Casida. 1964. Nicotinic acid analogs: effects on
response of chick embryos and hens to organophosphate toxicants. Science
144: 539-548.
Casida, J.E., J.L. Engel, E.G. Essac, F.X. Kamienski and S. Kuwatsuka. 1966. Methylene-
C14-dioxyphenyl compounds: metabolism in relation to their synergistic action.
Science 153: 1130-1133.
Fukami, J.-I., I. Yamamoto and J.E.Casida. 1967. Metabolism of rotenone in vitro
by tissue homogenates from mammals and insects. Science 155: 713-716.
Tsukamoto, M. and J.E. Casida. 1967. Metabolism of methylcarbamate insecticides by
the NADPH2-requiring enzyme system from houseflies. Nature 213: 49-51.
Ivie, G.W. and J.E. Casida. 1970. Enhancement of photoalteration of cyclodiene
insecticide chemical residues by rotenone. Science 167: 1620-1622.
Casida, J.E. 1970. Mixed-function oxidase involvement in the biochemistry of
insecticide synergists. J. Agric. Food Chem. 18: 753-772.
1
* Casida, J.E., E.C. Kimmel, M. Elliott and N.F. Janes. 1971. Oxidative metabolism
of pyrethrins in mammals. Nature 230: 326-327.
Maddrell, S.H.P. and J.E. Casida. 1971. Mechanism of insecticide-induced diuresis
in Rhodnius. Nature 213: 55-56.
Abernathy, C.O. and J.E. Casida. 1973. Pyrethroid insecticides: esterase cleavage in
relation to selective toxicity. Science 179: 1235-1236.
Bellet. E.M. and J.E. Casida. 1973. Bicyclic phosphorus esters: high toxicity without
cholinesterase inhibition. Science 182: 1135-1136.
2
* Casida, J.E., R.L. Holmstead, S. Khalifa, J.R. Knox, T. Ohsawa, R.J. Palmer and R.Y.
Wong. 1974. Toxaphene insecticide: a complex biodegradable mixture. Science
183: 520-521.
394 Wolf Prize in Agriculture
*2Lay, M.-M., J.P. Hubbell and J.E. Casida. 1975. Dichloroacetamide antidotes
for thiocarbamate herbicides: mode of action. Science 189: 287-289.
*2Schuphan, I., J.D. Rosen and J.E. Casida. 1979. Novel activation mechanism for
the promuta-genic herbicide diallate. Science 205: 1013-1015.
Wing, K.D., A.H. Glickman and J.E. Casida. 1983. Oxidative bioactivation of
S-alkyl phosphorothiolate pesticides: stereospecificity of profenofos insecticide
activation. Science 219: 63-65.
Lawrence, L.J. and J.E. Casida. 1984. Interactions of lindane, toxaphene and
cyclodienes with brain-specific t-butylbicyclophosphorothionate receptor. Life
Sci. 35: 171-178.
Palmer, C.J. and J.E. Casida. 1985. 1,4-Disubstituted 2,6,7-trioxabicyclo
[2.2.2]octanes: a new class of insecticides. J. Agric. Food Chem. 33: 976-980.
Waterhouse, A.L., I.N. Pessah, A.O. Francini and J.E. Casida. 1987. Structural aspects
of ryanodine action and selectivity. J. Med. Chem. 30: 710-716.
Casida, J.E. 1990. Pesticide mode of action: evidence for and implications of a
finite number of biochemical targets. Pp. 11-22 in Pesticides and Alternatives:
Chemical and Biological Approaches to Pest Control. (Casida, J.E., Ed.), Elsevier
Science Publ., New York.
Deng, Y., C.J. Palmer and J.E. Casida. 1991. Housefly brain γ-aminobutyric acid-
gated chloride channel: target for multiple classes of insecticides. Pestic.
Biochem. Physiol. 41: 60-65.
Elliott, M., D.A. Pulman, J.P. Larkin and J.E. Casida. 1992. Insecticidal 1,3-dithianes.
J. Agric. Food Chem. 40: 147-151.
*Li, Y.-M. and J.E. Casida. 1992. Cantharidin-binding protein: identification as
protein phosphatase 2A. Proc. Natl. Acad. Sci. USA 89: 11867-11870.
Liu. M.-Y. and J.E. Casida. 1993. High affinity binding of [3H]imidacloprid in the
insect acetylcholine receptor. Pestic. Biochem. Physiol. 46: 40-46.
Tomizawa, M., B. Latli and J.E. Casida. 1996. Novel neonicotinoid-agarose affinity
column for Drosophila and Musca nicotinic acetylcholine receptors.
J. Neurochem. 67: 1669-1676.
Hainzl, D. and J.E. Casida. 1996. Fipronil insecticide: novel photochemical
desulfinylation with retention of neurotoxicity. Proc. Natl. Acad. Sci. USA 93:
12764-12767.
Casida, J.E. and G.B. Quistad. 1998. Golden age of insecticide research: past, present
or future? Annu. Rev. Entomol. 43: 1-16.
Fang, N. and J.E. Casida. 1998. Anticancer action of cubé insecticide: correlation for
rotenoid constituents between inhibition of NADH:ubiquinone oxidoreductase
and induced ornithine decarboxylase activities. Proc. Natl. Acad. Sci. USA 95:
3380-3384.
Schuler, F., T. Yano, S. Di Bernardo, T. Yagi, V. Yankovskaya, T.P. Singer and J.E.
Casida. 1999. NADH-quinone oxidoreductase:PSST subunit couples electron
John E. Casida 395
transfer from iron-sulfur cluster N2 toquinone. Proc. Natl. Acad. Sci. USA 96:
4149-4153.
*3Höld, K.M., N.S. Sirisoma, T. Ikeda, T. Narahashi and J.E. Casida. 2000.
α-Thujone (the active component of absinthe): γ-Aminobutyric acid type A
receptor modulation and metabolic detoxification. Proc. Natl. Acad. Sci. USA
97: 3826-3831.
Ratra, G.S., S.G. Kamita and J.E. Casida. 2001. Role of human GABAA receptor β3
subunit in insecticide toxicity. Toxicol. Appl. Pharmacol. 172: 233-240.
Winrow, C.J., M.L. Hemming, D.M. Allen, G.B. Quistad, J.E Casida and C. Barlow.
2003. Loss of neuropathy target esterase in mice links organophosphate
exposure to hyperactivity. Nat. Genet. 33: 477-485.
Quistad, G.B, C. Barlow, C.J. Winrow, S.E. Sparks and J.E Casida. 2003. Evidence
that mouse brain neuropathy target esterase is a lysophospholipase. Proc.
Natl. Acad. Sci. USA 100: 7983-7987.
Nomura, D.K., D. Leung, K. Chiang, G.B. Quistad, B.F. Cravatt and J.E. Casida.
2005. A brain detoxifying enzyme for organophosphorus nerve poisons. Proc.
Natl. Acad. Sci. USA 102: 6195-6200.
* Chen, L., K.A. Durkin, and J.E. Casida. 2006. Structural model for γ-aminobutyric
4
1Initial
publication by Nature Publishing Group.
2Permission granted by AAAS.
3Permission granted, copyright (2000) National Academy of Sciences, U.S.A.
4Permission granted, copyright (2006) National Academy of Sciences, U.S.A.
Source: From Science Vol. 183, No. 124, pp. 520-521 (1974). Reprinted with permission from AAAS.
John E. Casida 407
408 Wolf Prize in Agriculture
Source: From Science Vol. 189, No. 4199, pp. 287-289 (1975). Reprinted with permission from
AAAS.
John E. Casida 409
410 Wolf Prize in Agriculture
John E. Casida 411
Source: From Science Vol. 205, No. 4410, pp. 1013-1015 (1979). Reprinted with permission from
AAAS.
412 Wolf Prize in Agriculture
John E. Casida 413
414 Wolf Prize in Agriculture
The toxic effects of ␣-thujone in mammals are well established DMSO, final concentration 1%) and [3H]EBOB (0.7 nM) in
but the mode of neurotoxic action is poorly understood. It is 1.0 ml of 10 mM sodium phosphate, pH 7.5 buffer containing
porphyrogenic, possibly thereby contributing to the absinthe- 200 mM sodium chloride at 37°C for 70 min (25). Scatchard
induced illness of Vincent van Gogh (5, 16). ␣-Thujone is analyses were performed with no inhibitor and with 5 and 25 M
neurotoxic in rats (17), and ingestion of wormwood oil contain- ␣-thujone by using [3H]EBOB at 0.08–26 nM. The inhibitory
ing ␣-thujone recently resulted in human poisoning (18). The potency also was compared for ethanol and absinthe (based on
hypothesis that ␣-thujone activates the CB1 cannabinoid recep- ethanol content) with that for ethanol containing 5 M ␣-thu-
tor, based on the structural similarity of thujone enol with jone. The incubated mixtures were filtered through GF兾C glass
tetrahydrocannabinol (19), was not supported experimentally fiber filters, then rinsed twice with 5 ml of ice-cold 0.9% sodium
(20). The convulsant action led to multiple speculations on chloride, by using a cell harvester. Specific binding was consid-
mechanisms, one of which was antagonism of the ␥-aminobutyric ered to be the difference between total binding and nonspecific
acid (GABA) receptor system (20), a proposal that was not binding determined in the presence of 5 M ␣-endosulfan {a
explored further. ␣- and -Thujone are reduced in rabbits from potent GABA type A (GABAA) receptor antagonist and specific
the ketones to the corresponding alcohols (thujol and neothujol) inhibitor of [3H]EBOB binding}.
(21) of unknown toxicity but no other metabolites are identified.
The goals of this study are to define the mechanism of Effect on GABA-Induced Whole-Cell Currents. Rat dorsal root gan-
neurotoxicity of ␣-thujone and identify its major metabolites glion neurons were prepared and cultured as described (26).
(Fig. 1) and their role in the poisoning process. Emphasis is Currents were induced by 10-msec pulses of 300 M GABA and
placed on the hypothesis that the convulsant action is caused by recorded by using the whole-cell patch clamp technique. The
modulating the GABA-gated chloride channel. GABA-induced inward current of this preparation was carried
by chloride ions through open chloride channels (27). Each cell
Materials and Methods was tested for the degree of suppression caused by bath appli-
Chemicals. Sources were: ␣-thujone (⬇99% purity) from Fluka; cation of ␣-thujone to determine the concentration for 50%
APPLIED BIOLOGICAL
wormwood oil (3.2% ␣- and 35% -thujone) from Lhasa Karnak inhibition (IC50).
(Berkeley, CA) and absinthe with 0.4 ppm ␣-thujone, 5 ppm
SCIENCES
-thujone, and 50% (vol兾vol) ethanol labeled Herring Absenta GC-MS Identification and Analysis of ␣-Thujone and Metabolites.
(Zaragoza, Spain) with concentrations based on analyses in this Standard analytical methods of GC-MS and derivatization of
laboratory; picrotoxinin, diazepam, and sodium phenobarbital alcohol and ketone functionalities were applied to ␣-thujone and
from Sigma; dieldrin and ␣-endosulfan from Chem Service its metabolites. Analyses used the DB-5 fused silica gel capillary
(West Chester, PA); [3H]ethynylbicycloorthobenzoate ([3H]E- column (30 m, 0.25 mm i.d., 0.25 m film thickness) (J&W
BOB) (38 Ci兾mmol) from NEN. Although not detailed here, Scientific, Folsom, CA). The initial column temperature of 80°C
7-hydroxy- ␣ -thujone, 4-hydroxy- ␣ -thujone, 4-hydroxy-  - was programmed to 200°C at the rate of 5°C兾min, followed by an
thujone, 7,8-dehydro-␣-thujone, and a thujol兾neothujol mixture increase at 20°C兾min to 300°C where it was maintained for 2 min.
were synthesized as standards for comparison with metabolites. The carrier gas and reagent gas were helium and methane,
respectively. Temperatures of the injection port and detector
Toxicity to Mice. Male albino Swiss–Webster mice (22–28 g) were were 250°C and 280°C, respectively. The mass spectrometer was
treated i.p. with the test compound by using propylene glycol (2 operated in the positive chemical ionization mode. One micro-
l兾g body weight) as the carrier vehicle. Prophylactic i.p. treat- liter was injected splitless onto the column. For quantitation, the
ments also were examined for their effect on ␣-thujone toxicity GC-MS was operated in the selected ion monitoring (SIM)
(100 mg兾kg) individually with ethanol (0.5 or 1.0 g兾kg as 20% mode, measuring m兾z 135 for ␣-thujone and m兾z 151 for the
and 40% solutions in saline, 20 min pretreatment), diazepam (1 hydroxythujones, dehydrothujone, and (S)-(⫺)-carvone (inter-
mg兾kg, 15 min pretreatment), or phenobarbital (15 mg兾kg, 15 nal standard). The concentration of each analyte was deter-
min pretreatment). mined from least-squares equations generated from peak-
area ratios of ␣-thujone, 7-hydroxy-␣-thujone, and the internal
Toxicity to Drosophila. Fruit flies (Drosophila melanogaster) were standard. Identification of ␣-thujone and metabolites involved
used in two types of assays: comparing two strains known to be comparison with standards by cochromatography and MS frag-
different in sensitivity to insecticidal chloride channel blockers mentation patterns as parent compounds and two derivatives.
and comparing ␣-thujone and its metabolites for toxicity to the Trimethylsilyl ethers were formed on reaction of alcohols with
susceptible strain. The median lethal concentration (LC50) was N-methyl-N-trimethylsilyltrifluoroacetamide and methyloximes
determined for ␣-thujone and dieldrin with two strains of on coupling ketones with methoxyamine. These derivatization
Drosophila: a dieldrin-resistant RdlMD-RR strain (22, 23) (ob- procedures and MS fragmentation patterns also allowed assign-
tained from the Bloomington Drosophila Stock Center at Indi- ment of some metabolites as hydroxythujones without specifying
ana University, Bloomington) and the Canton-S, wild-type sen- the position of hydroxylation.
sitive (S) strain. The test chamber was a glass tube (12 ⫻ 75 mm)
containing a filter paper strip (Whatman no. 1, 8 ⫻ 65 mm). Five Enzymatic Metabolism. Rabbit or mouse liver cytosol (1 mg
adult flies were placed in the tube, which then was closed with protein) or washed mouse liver microsomes (1 mg protein) and
a single layer of parafilm. A solution of ␣-thujone or dieldrin in NADPH (or other cofactor, 1 mM final concentration) were
propylene glycol (5 l) was injected with a 10-l syringe through incubated with ␣-thujone (30 g, 0.2 M final concentration) in
the parafilm onto the filter paper after which the tube was 100 mM phosphate, pH 7.4 buffer (1 ml) for 1 h at 37°C. For
covered with a second piece of parafilm. Mortality was recorded analysis the internal standard S-carvone (0.05 g) was added in
after 8 h at 25°C as flies that could not move. The experiment ethanol (10 l), and the mixture was saturated with sodium
was repeated four times to prepare dosage mortality curves for chloride and extracted with ethyl acetate (3 ml) for 30 min by
calculation of resistance ratios (LC50 Rdl兾LC50 S). gentle rocking. The organic extract, recovered by centrifugation
at 900 g, was almost completely evaporated (but never to
Effect on [3H]EBOB Binding in Mouse Brain Membranes. Mouse brain dryness) under a stream of nitrogen at room temperature and
membranes were prepared and depleted of GABA as described reconstituted in ethyl acetate (50 l) for GC-MS analysis.
(24). For inhibitor potency assays, the membranes (200 g Recovery values by this procedure for ␣-thujone and the major
protein) were incubated with the test compound (added in metabolite were ⬎60% with no degradation during GC.
Results
␣-Thujone Is a Convulsant. The i.p. LD50 of ␣-thujone in mice is
about 45 mg兾kg, generally with 0% and 100% mortality at 30 and
60 mg兾kg, respectively. Mice at the higher dose undergo a tonic Fig. 3. ␣-Thujone and 7-hydroxy-␣-thujone inhibit [3H]EBOB binding to
convulsion leading to death within 1 min whereas at 30–45 mouse brain membranes. (A) IC50 determination for ␣-thujone and 7-hydroxy-
mg兾kg they exhibit tail-raising within the first 2 min, followed by ␣-thujone (mean ⫾ SEM, n ⫽ 4). (B) Scatchard plots as average of duplicate
flexion of the trunk and clonic activity of the forelimbs, pro- measurements for [3H]EBOB alone (Kd 2.8 nM and Bmax 1,700 fmol兾mg pro-
gressing to generalized and protracted tonic兾clonic convulsions tein) and with ␣-thujone at 5 M (Kd 4.1 and Bmax 1,700) and 25 M (Kd 7.2 and
that ultimately result in death or recovery. Intraperitoneal Bmax 1,700).
administration of diazepam or phenobarbital 15 min before
␣-thujone at 100 mg兾kg results in almost all of the mice surviving ␣-Thujone Modulation of the GABAA Receptor-Chloride Channel. The
this otherwise lethal dose. Ethanol i.p. pretreatment at 1 g兾kg currents induced by 300 M GABA are suppressed with 30 M
(but not at 0.5 g兾kg) also protects against the lethal effects of bath-applied ␣-thujone and there is full reversal on washing with
␣-thujone at 100 mg兾kg. ␣-thujone-free solution (Fig. 4 A and B). The IC50 for ␣-thujone
is 21 M in suppressing the GABA-induced currents (Fig. 4C).
␣-Thujone Cross-Resistance in Drosophila Strain Resistant to Dieldrin.
Flies of the Rdl strain (⬎55-fold resistant to dieldrin; LC50 ⬎275 Absinthe, Ethanol, and Ethanol Containing ␣-Thujone as Inhibitors of
g兾tube for Rdl versus 5 g兾tube for S) are 5-fold resistant to [3H]EBOB Binding. The inhibitory effects on [3H]EBOB binding
␣-thujone (LC50 65 g兾tube for Rdl versus 12 g兾tube for S) were compared for absinthe, ethanol, and ethanol containing
(Fig. 2). This finding establishes moderately high insecticidal ␣-thujone to help understand their independent and combined
activity for ␣-thujone and cross-resistance in the dieldrin- actions on the chloride channel. The IC50 for absinthe (based on
resistant strain. ethanol content) is 263 ⫾ 47 mM and for ethanol is significantly
higher at 370 ⫾ 4 mM (Fig. 5A). There is no significant
␣-Thujone Inhibition of [3H]EBOB Binding. The IC50 of ␣-thujone for interaction between the effects of ethanol and ␣-thujone (Fig.
[3H]EBOB binding in mouse brain membranes is 13 ⫾ 4 M 5B), i.e., ␣-thujone (5 M) inhibition is 20–30% independent of
(Fig. 3A). The binding of ␣-thujone is competitive with that of ethanol concentration up to 300 mM.
[3H]EBOB based on Scatchard analysis (Fig. 3B). For compar-
ison, other IC50 values are 29 ⫾ 8 M for -thujone, 37 ⫾ 8 M Metabolism of ␣-Thujone by Liver Enzymes. Incubation of ␣-thujone
for wormwood oil (calculated as molecular weight of thujone), with rabbit (but not mouse) liver cytosol gives thujol and
and 0.6 ⫾ 0.1 M for picrotoxinin (inhibition curves not shown). neothujol, identified by GC-MS comparison with authentic
APPLIED BIOLOGICAL
SCIENCES
Fig. 5. Absinthe, ethanol, and ethanol containing ␣-thujone inhibit [3H]E-
BOB binding to mouse brain membranes. (A) Comparison of an absinthe
preparation (based on ethanol content) with ethanol (average of duplicate
measurements or mean ⫾ SD, n ⫽ 6). (B) Comparison of ethanol with ethanol
containing 5 M ␣-thujone (average of duplicate measurements).
Fig. 4. Suppression of GABA-induced peak currents by bath application of methylsilyl and methyloxime derivatives, indicating the presence
␣-thujone. Currents were induced by 300 M GABA (10 msec) pulses. The peak
of both an alcohol substituent and a ketone functionality.
amplitude of current decreased with 30 M ␣-thujone and recovered after
washing with ␣-thujone-free solution. (A) Time course of 30 M ␣-thujone- Synthesis of various hydroxythujones and their comparison
induced changes in peak current amplitude. (B) Representative current with the metabolites (directly, and as trimethylsilyl ethers and
records. (C) Concentration-response relationship (mean ⫾ SD, n ⫽ 4 –5). methyloximes) identifies the major product as 7-hydroxy-␣-
thujone and two minor metabolites as the diastereomers of
4-hydroxythujone.
standards per se and by forming trimethylsilyl (but not methyl-
oxime) derivatives. This enzymatic reduction depends on Metabolites in the Brain of ␣-Thujone-Treated Mice. The brain
NADPH but occurs in small yield. Metabolism in mouse liver contains ␣-thujone, dehydro-␣-thujone, and four hydroxythu-
microsomes is a much more facile reaction and gives no thujol or jones (7-hydroxy-␣ major plus 4-hydroxy-␣, 4-hydroxy-, and
neothujol but instead different products. ␣-Thujone is stable on one other) also observed in the liver P450 system (Fig. 6).
incubation with mouse liver microsomes alone but is almost Identifications are based on retention times and MS fragmen-
completely metabolized when NADPH (but not NADP, NADH, tation patterns both direct and as trimethylsilyl and methyloxime
or NAD) also is added. Six NADPH-dependent microsomal derivatives. The brain levels of ␣-thujone and 7-hydroxy-␣-
metabolites are evident by GC-MS, each at higher retention time thujone are dose- and time-dependent after i.p. injection of
than the parent ␣-thujone (Fig. 6). The first-eluting metabolite ␣-thujone (Fig. 7). Importantly, ␣-thujone appears at much
is identical in GC and MS features to synthetic 7,8-dehydro-␣- lower levels and is less persistent than 7-hydroxy-␣-thujone. At
thujone. The next five metabolites each are converted to tri- severely toxic ␣-thujone doses (40–60 mg兾kg) the levels in brain
at 30 min are 0.3–1.0 ppm for ␣-thujone and 1.5–8.4 ppm for
7-hydroxy-␣-thujone (Fig. 7A) with much higher levels (11 and Fig. 7. Brain levels of ␣-thujone and 7-hydroxy-␣-thujone as a function of
29 ppm for ␣-thujone and 7-hydroxy-␣-thujone, respectively) at dose and time in mice treated i.p. with ␣-thujone. Average of determinations
on two mice except for a single determination at 5 min. (A) Dose studies at 30
2.5 min (Fig. 7B) when the poisoning signs are most intense. The
min after treatment. (B) Time studies at 50 mg兾kg.
minor hydroxythujone metabolites are detectable only up to 20
min after the 50 mg兾kg ␣-thujone dose.
toxicity by diazepam, phenobarbital, and ethanol (28, 29). Dro-
Biological Activities of Metabolites. Synthetic standards of the sophila with a single point mutation in the Rdl GABA receptor
metabolites shown in Fig. 1 except the 4-hydroxy-␣-thujone subunit of Ala302 to Ser conferring resistance to dieldrin (22, 23)
diastereomers were compared with ␣-thujone for potency as is also resistant to ␣-thujone, albeit to a lesser degree. ␣-Thujone
toxicants to mice and Drosophila and inhibitors of [3H]EBOB is a competitive inhibitor of [3H]EBOB binding, i.e., of the
binding. The discriminating levels used were 50 mg兾kg i.p. for noncompetitive blocker site of the GABA-gated chloride chan-
mice and 50 g兾tube for the S strain of Drosophila. With mice, nel (25). Most importantly, electrophysiological studies establish
␣-thujone is lethal, whereas 7-hydroxy-␣-thujone, dehydro-␣- that in dorsal root ganglion neurons ␣-thujone is a reversible
thujone, and thujol兾neothujol are not lethal. With Drosophila, modulator of the GABAA receptor.
␣-thujone gives complete mortality, dehydro-␣-thujone gives Absinthe and wormwood oil contain not only ␣-thujone as
70% mortality, and 7-hydroxy-␣-thujone and thujol兾neothujol their purported active ingredient but also many other candidate
give about 30% mortality. In the [3H]EBOB binding assay, toxicants, including -thujone and ethanol in the case of ab-
7-hydroxy-␣-thujone gives an IC50 value of 730 ⫾ 265 M versus sinthe. -Thujone is less toxic than ␣-thujone to mice (10) and
13 ⫾ 4 M for ␣-thujone (Fig. 3A), whereas the value for Drosophila and in addition is 2.3-fold less potent in the [3H]E-
dehydro-␣-thujone is 149 ⫾ 10 M (inhibition curve not shown). BOB assay (this investigation). Ethanol also enhances neuronal
GABAA receptor function (30) and therefore might suppress the
Discussion blocking action of ␣-thujone in absinthe. However, ethanol does
This study establishes that ␣-thujone modulates the GABAA not alter the inhibitory action of ␣-thujone on [3H]EBOB
receptor based on four observations. Comparison with picro- binding. The ␣- and -thujone content of the absinthe sample
toxinin, the classical GABAA receptor antagonist, revealed examined here (0.4 and 5 ppm or 2.6 and 33 M, respectively)
similar poisoning signs and in both cases alleviation of the may be a contributing factor in the somewhat greater potency of
absinthe (based on ethanol content) than of ethanol per se in the cants, ␣-thujone and its 7-hydroxy metabolite. The 7-hydroxy
[3H]EBOB assay. However, the 10 ppm (66 M) upper limit of compound is present in brain at much higher levels than the
the European Commission (6) and particularly the 260 ppm parent ␣-thujone, suggesting possible conversion in situ, but this
(1710 M) thujone content of old absinthe (6) would give a oxidation was not observed on incubation of ␣-thujone with
detectable to major inhibitory effect beyond that of the ethanol brain microsomes and NADPH. ␣-Thujone compared with
content. Current low levels of ␣- and -thujone in absinthe are 7-hydroxy-␣-thujone is 56-fold more potent in the [3H]EBOB
of much less toxicological concern than the ethanol content (6). binding assay and much more toxic to mice and houseflies. It
␣-Thujone as other monoterpenes is easily metabolized. The appears that all of the metabolites studied here are detoxifica-
single report on metabolism identifies thujol and neothujol tion products, i.e., less toxic than ␣-thujone. However, the level
probably as conjugates in the urine of thujone-treated rabbits
in brain of 7-hydroxy-␣-thujone is several-fold greater than that
(21). We find enzymatic reduction (possibly by a cytosolic ketone
of ␣-thujone (e.g., 29 and 11 ppm, respectively, at the time of
reductase) (31) of ␣-thujone to thujol and neothujol in low yield
by rabbit but not mouse liver cytosol with NADPH. The mouse severe poising signs), suggesting that either one or both may
liver microsomal P450 system rapidly converts ␣-thujone to contribute to the toxic manifestations.
7-hydroxy- ␣ -thujone (major), the diastereomers of 4-hy- This study establishes that ␣-thujone acts at the noncompet-
droxythujone (minor), and other hydroxythujones (minor). In- itive blocker site of the GABAA receptor and is rapidly detox-
terestingly, the major sites of P450 hydroxylation at the 4- and ified, thereby providing a reasonable explanation for some of the
7-positions are those involving intermediate tertiary radicals that actions of absinthe other than those caused by ethanol, and
are more stable than secondary and primary radicals. Dehydro- allowing more meaningful evaluation of risks involved in the
␣-thujone also is observed and may arise from dehydration of the continued use of herbal medicines containing ␣-thujone.
7-hydroxy compound as a biological reaction because this pos-
sible conversion is not an artifact during the extraction and Helpful comments were provided by Wilfred Arnold (University of
analysis procedure. The various hydroxythujones probably are
APPLIED BIOLOGICAL
Kansas Medical Center, Kansas City) and Jeffrey Bloomquist (Virginia
not the terminal metabolites because they are expected to Polytechnic Institute, Blacksburg). We thank our former coworker Neil
SCIENCES
undergo conjugation and excretion. However, the presence of Jacobson for initiating our interest in ␣-thujone action and our labora-
hydroxythujones in the brain suggests their potential importance tory colleagues Gary Quistad and Susan Sparks for help and advice. This
in the neurotoxicity. project was supported by National Institute of Environmental Health
Metabolic detoxification is a dominant feature of ␣-thujone Sciences Grant R01 ES08419 to J.E.C. and National Institute of Neu-
neurotoxicity in mice. There are two principal candidate toxi- rological Disorders and Stroke Grant R01 NS14143 to T.N.
1. Vogt, D. D. & Montagne, M. (1982) Int. J. Addict. 17, 1015–1029. Biochem. Pharmacol. 43, 2359–2368.
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Greene, Y. J., Lambrecht, R. W., Srivastava, K. K. & Arnold, W. N. (1992) Jakoby, W. B. (Academic, Orlando, FL), Vol. 1, pp. 281–293.
Several major insecticides, including ␣-endosulfan, lindane, and block chloride flux so the target is referred to as the GABA
fipronil, and the botanical picrotoxinin are noncompetitive antag- receptor NCA-binding site. Vertebrate GABA receptors consist of
onists (NCAs) for the GABA receptor. We showed earlier that ␣, , ␥, , and other subunits in various combinations, for example,
human 3 homopentameric GABAA receptor recognizes all of the ␣12␥2 as a heteropentamer and 1 as a homopentamer (13–15).
important GABAergic insecticides and reproduces the high insec- The molecular localization of the NCA site defined here (Fig. 1B)
ticide sensitivity and structure-activity relationships of the native was first indicated by mutagenesis studies (16) as A2⬘ (17–20), T6⬘
insect receptor. Despite large structural diversity, the NCAs are (21, 22), and L9⬘ (23, 24) in the cytoplasmic half of the transmem-
proposed to fit a single binding site in the chloride channel lumen brane 2 domain of the channel (Fig. 2). Drosophila resistant to
lined by five transmembrane 2 segments. This hypothesis is ex- dieldrin (RDL) have a mutation conferring GABA receptor in-
amined with the 3 homopentamer by mutagenesis, pore structure sensitivity identified as A2⬘S (17). The NCA target of the GABAA
studies, NCA binding, and molecular modeling. The 15 amino acids receptor requires a  subunit, and a 3 homopentamer is sufficient
in the cytoplasmic half of the pore were mutated to cysteine, for binding (9, 26). Importantly, the 3 subunit from human brain,
serine, or other residue for 22 mutants overall. Localization of when expressed in insect Sf9 cells, assembles to form a receptor
A-1ⴕC, A2ⴕC, T6ⴕC, and L9ⴕC (index numbers for the transmembrane sensitive to all of the important GABAergic insecticides (9) and,
2 region) in the channel lumen was established by disulfide surprisingly, reproduces the insecticide sensitivity and structure-
cross-linking. Binding of two NCA radioligands [3H]1-(4-ethynyl- activity relationships of the native insect receptor (27). Studies of
phenyl)-4-n-propyl-2,6,7-trioxabicyclo[2.2.2]octane and [3H] 3,3- the GABA receptor NCA site are therefore simplified by using this
bis-trifluoromethyl-bicyclo[2,2,1]heptane-2,2-dicarbonitrile was dra- highly sensitive 3 homopentamer, an approach verified by showing
matically reduced with 8 of the 15 mutated positions, focusing here that Cys and Ser or Phe mutations in 3 at each of the 2⬘, 6⬘,
attention on A2ⴕ, T6ⴕ, and L9ⴕ as proposed binding sites, consistent and 9⬘ positions greatly reduce or destroy NCA radioligand binding.
with earlier mutagenesis studies. The cytoplasmic half of the 3
This study tested the hypothesis that insecticides and convul-
homopentamer pore was modeled as an ␣-helix. The six NCAs
sants of many chemical types act at the same GABA receptor site
listed above plus t-butylbicyclophosphorothionate fit the 2ⴕ to 9ⴕ
in the same way to initiate insecticidal action and mammalian
pore region forming hydrogen bonds with the T6ⴕ hydroxyl and
toxicity. The goal was to characterize the GABA receptor–NCA
hydrophobic interactions with A2ⴕ, T6ⴕ, and L9ⴕ alkyl substituents,
interaction by using the human GABAA receptor recombinant
thereby blocking the channel. Thus, widely diverse NCA structures
3 homopentamer as a model. The first step was to prepare Cys
fit the same GABA receptor  subunit site with important impli-
and other mutations to scan the cytoplasmic half of M2 and the
cations for insecticide cross-resistance and selective toxicity be-
flanking region (⫺4⬘ to 10⬘), overall 22 mutants involving 15
tween insects and mammals.
positions. The mutants were used to identify Cys residues
3 homopentamer 兩 transmembrane 2 兩 insecticide 兩 disulfide trapping 兩
undergoing disulfide cross-linking as a guide to channel pore
receptor model
structure (28). Next, [3H]EBOB and [3H]BIDN were used to
identify positions where mutation altered binding (6, 8). Finally,
PHARMACOLOGY
modeling of the NCA-binding domain (29, 30) was applied to the
P est insect control in the past 60 years was achieved, in part,
by application of ⬎3 billion (3 ⫻ 109) pounds of polychlo-
rocycloalkane insecticides, including cyclodienes (e.g., ␣-en-
3 homopentamer to determine whether the wide diversity of
NCAs could fit the same site.
dosulfan and dieldrin), lindane and its isomers, and others, which Results
are now highly restricted or banned except for endosulfan and Mutagenesis and Protein Expression. The transfection efficiency of
some uses of lindane (1–3). One of the replacement compounds each recombinant baculovirus was examined by PCR analysis.
is the phenylpyrazole fipronil. All of these insecticides and the The nonrecombinant virus would give one 839-bp band of its
botanical picrotoxinin (PTX) have widely diverse chemical
polyhedrin region and the recombinant virus incorporating the
structures but appear to act at the same nerve target. It is
1,425-bp 3 cDNA would appear at 2.3 kb. Each extracted
therefore important to understand how these compounds work
recombinant virus gave only one 2.3-kb band (Fig. 3A), indicat-
in mammals and insects, or how they do not work when resistant
ing a recombination efficiency for the target gene of nearly 100%
insect strains appear.
for all mutants and the WT. Further, all PCR products from
The GABA-gated chloride channel is the target for the insecti-
cides and toxicants referred to above based on radioligand binding
and electrophysiology studies (3–10). Important radioligands in Conflict of interest statement: No conflicts declared.
these developments are [ 3 H]dihydroPTX (4, 11), [ 35 S]t- Abbreviations: BIDN, 3,3-bis-trifluoromethyl-bicyclo[2,2,1]heptane-2,2-dicarbonitrile;
butylbicyclophosphorothionate (TBPS) (5, 12), [3H]1-(4-ethynyl- Cu:phen, copper:phenanthroline; EBOB, 1-(4-ethynylphenyl)-4-n-propyl-2,6,7-trioxabicyclo-
phenyl)-4-n-propyl-2,6,7-trioxabicyclo[2.2.2]octane (EBOB) (6), [2.2.2]octane; M2, transmembrane 2; NCA, noncompetitive antagonist; PTX, picrotoxinin;
and [ 3 H]3,3-bis-trif luoromethyl-bicyclo[2,2,1]heptane-2,2- RDL, resistant to dieldrin; TBPS, t-butylbicyclophosphorothionate.
dicarbonitrile (BIDN) (8) (Fig. 1A). All of these compounds act in ‡To whom correspondence should be addressed: E-mail: ectl@nature.berkeley.edu.
mammals and insects as noncompetitive antagonists (NCAs) to © 2006 by The National Academy of Sciences of the USA
Fig. 1. Structure-activity relationships of seven GABA receptor noncompetitive antagonists. (A) Structures of three important insecticides (lindane, fipronil,
and ␣-endosulfan) and four radioligands (asterisk designates labeling position). The high potencies of each compound with the 3 homopentamer are indicated
by the 2.7 nM Kd for [3H]EBOB on direct binding and 0.47–59 nM IC50 values for the other compounds in displacing [3H]EBOB binding (9). (B) Models of four
antagonists positioned as in A showing their proposed 3 homopentamer M2 binding sites in the channel lumen. A, L, and T refer to the side chains of the
interacting 2⬘, 6⬘, and 9⬘ residues, respectively.
virus extraction were sequenced and confirmed as the right cases of a molecular mass increase from 55 kDa to ⬇130 kDa for
mutations. the monomer and dimer, respectively, as detected by SDS兾
The expression levels of the WT and mutant 3 subunits were PAGE and immunoblotting (Fig. 4). Cys substituents at ⫺1⬘, 2⬘
determined by Western blotting. The monoclonal anti--chain (weak), 6⬘, and 9⬘ formed disulfide-linked dimers in the presence
antibody recognized a very specific band at ⬇55 kDa with similar but not in the absence of Cu:phen. Only trace amounts of the ⫺1⬘
intensity for membrane extracts of the WT and each mutant (Fig. and 9⬘ monomers are left with Cu:phen indicating more exten-
3B). Equal protein transfer levels were determined by Ponceau sive reaction possibly due to higher flexibility at these positions.
S staining. As exceptions, two mutants (L3⬘C and L3⬘F) were not Dimers were not detected under the same conditions for Cys
expressed. substituents at 0⬘, 1⬘, 4⬘, 5⬘, 7⬘, 8⬘, and 10⬘, although in some cases
there were apparent losses in receptor levels on oxidation.
Disulfide Cross-Linking Profiles. Oxidation of the Cys mutants with Disulfides at ⫺1⬘, 2⬘, and 9⬘ were completely reversed with DTT,
copper sulfate:1,10-phenanthroline (Cu:phen) resulted in four but the one at 6⬘ was only partially reversed.
the cell surface because, on the Western blot, they all have a
protein of similar size and presumably maturely glycosylated.
Most importantly, on an overall basis, the results are essentially
the same with [3H]EBOB and [3H]BIDN.
Two methanethiosulfonate (MTS) sulfhydryl-modification re-
agents provided further information on the NCA site by com-
paring their effect on [3H]EBOB binding for the Cys active
mutants 1⬘, 7⬘, and 10⬘ compared with the WT. With both
sulfhydryl reagents, there was a site-dependent effect on [3H]E-
BOB binding with little inhibition for the T7⬘C mutant, moder-
ate for T10⬘C, and almost complete for V1⬘C.
Structural Model for NCA Binding. Fig. 6 Upper Left shows a model
of the channel lumen from the 2⬘ to 9⬘ positions with five 3
␣-helices and lindane docked into the putative binding site,
which it clearly fills to block the pore. Similar models of the six
other NCAs also show filling of the pore space.
Attention was focused on A2⬘, T6⬘, and L9⬘, because these
residues are in the channel lumen (based on disulfide trapping)
and mutations (Cys versus Ser or Phe in each case) at these sites
greatly reduce or abolish binding. The interacting sites are shown
in Figs. 1B and 6. Docking of EBOB positions the A2⬘ methyls
interacting with the normal-propyl and two O-methylenes, two
T6⬘ hydroxyls interacting with the oxygens (H---O distance
⬇3.1Å), T6⬘ methyls binding to the phenyl moiety, and, at a
slightly longer range, a L9⬘ methyl also interacting with the
ethynyl substituent (evident in Fig. 1B but not Fig. 6). TBPS has
Fig. 4. Disulfide cross-linking profiles. Samples are control without Cu:phen
or DTT (–兾–), oxidized with Cu:phen but not treated with DTT (⫹兾–), or
numerous favorable A2⬘ interactions with the tertiary-butyl
oxidized with Cu:phen then reduced with 10 mM DTT (⫹兾⫹). Reactions were moiety, and the T6⬘ methyls and hydroxyls interact with the
terminated with 10 mM N-ethylmaleimide before SDS兾PAGE-Western blot- sulfur and cage oxygens. PTX has A2⬘ methyl interactions with
ting analysis. the isopropenyl methyl and methylene and three T6⬘ hydroxyl
hydrogen bonding interactions to three PTX oxygens. BIDN has
multiple contact points with A2⬘ methyls and T6⬘ methyls and
conditions and amounts of receptors, the mutants were then hydroxyls. A cyano nitrogen and a fluorine each form hydrogen
compared to the WT for both [3H]EBOB and [3H]BIDN bind- bonds to a T6⬘ hydroxyl. Lindane bridges A2⬘ methyls and T6⬘
ing. The binding activities of A-4⬘S, T7⬘C and T10⬘C were similar hydroxyls and methyls, each interacting with multiple chlorines.
to the WT, whereas A-2⬘S and V1⬘C gave reduced binding (Fig. ␣-Endosulfan and fipronil have multiple interaction sites and
5). All of the rest gave little or no specific binding. It was indeed types, with A2⬘ methyls and T6⬘ methyls and hydroxyls for both
surprising to find that the low binding for mutants involves the compounds reinforced by L9⬘ side chains for fipronil. More
whole segment from A2⬘ to I5⬘, in addition to the expected T6⬘ complete depictions of the 3 homopentamer model and the
to L9⬘, with the two exceptions of L3⬘ not expressed and T7⬘ docked ligands are given in supporting information, which is
normal. More generally, mutations in the lowest region of the published on the PNAS web site.
channel have no (⫺4⬘ or ⫺3⬘) or little (⫺2⬘) influence on
activity, whereas those in the region of ⫺1⬘ to 10⬘, except 1⬘, 7⬘, Discussion
and 10⬘, drastically reduce [3H]EBOB and [3H]BIDN binding. Mutagenesis and Expression. The cytoplasmic half of the M2
This reduction is not due to interference from oxidation of the region contains 11 amino acids (0⬘ to 10⬘), and this number is
extended to 15 (⫺4⬘ to 10⬘) with the flanking region of interest.
PHARMACOLOGY
Cys moiety because (i) DTT did not restore the activity of the
eight low-binding mutants (data not shown), and (ii) the findings Site-specific mutagenesis introduced Cys at 12 sites (A-1⬘C to
are essentially the same with Ser (0⬘, 2⬘, and 9⬘), Leu (5⬘), Phe T10⬘C), Ser at five sites (A-4⬘S, A-2⬘S, R0⬘S, A2⬘S, and L9⬘S),
(6⬘), and Ala (4⬘ and 8⬘) as well as for the corresponding Cys and Phe at two sites (L3⬘F and T6⬘F). In addition, three
mutants. It is assumed that the mutants, which do not bind mutations were introduced with little change in polarity, i.e.,
G4⬘A, I5⬘L and V8⬘A. The 3⬘-position was an exception because
NCAs, form functional channels that are correctly assembled on
L3⬘C and L3⬘F did not show detectable expression by Western
blotting either in the 3 homopentamer studied here or the ␣13
heteropentamer (data not shown).
Chen et al. PNAS 兩 March 28, 2006 兩 vol. 103 兩 no. 13 兩 5187
John E. Casida 423
Fig. 6. Proposed interactions of seven noncompetitive antagonists at the same GABAA receptor 3 homopentamer binding site. (Upper Left) lindane (space
fill, red and blue for partial negative and positive charges of chlorine and carbon, respectively) binds to the GABAA receptor (five 3 ␣-helices shown in green)
to block the channel pore shown as the 2⬘ to 9⬘ positions viewed from the top into the pore. Remaining panels: seven ligands (see Fig. 1 A) docked at their
optimized positions with the perspective chosen for ease of viewing. A, L, and T refer to the side chains of the interacting 2⬘, 6⬘, and 9⬘ residues, respectively.
van der Waals contacts are illustrated in green (see text for discussion of hydrogen bonding). The space filling aspects of all of the ligands are most readily evident
in supporting information.
mobility兾flexibility. For T6⬘, similar findings are obtained with addition, with V2⬘C at the ␣1 subunit of the ␣11␥2 receptor,
the ␣1T6⬘C1T6⬘C receptor but only in the presence of GABA PTX protects against sulfhydryl derivatization (18), and a sulf-
(28), suggesting that the 3 homopentamer of the present hydryl-reactive fipronil analog [-C(O)CH2Br replaces -S(O)CF3]
investigation assumes the spontaneous open state (32). Homol- serves as an irreversible blocker (19). The involvement of 3⬘,
ogy of the GABA receptor 3 homopentamer with the nicotinic directly or by influencing the neighboring A2⬘, is shown by L3⬘F
acetylcholine receptor (33) indicates the narrowest gating region at 3 of the ␣13 receptor almost abolishing TBPS and PTX
of the pore is between 9⬘ and 14⬘, suggesting the positioning of binding (20). The structurally critical apolar pocket in the 3
L9⬘C in the pore (24, 31). In the 1 subunit of the ␣11␥2 homopentamer appears to involve A2⬘, L3⬘, G4⬘, and I5⬘, i.e., a
receptor, A2⬘C, T6⬘C, T7⬘C (slow reaction rate), V8⬘C, L9⬘C, tightly packed and completely hydrophobic region that may play
and T10⬘C are all accessible to a sulfhydryl-modification reagent a role in stabilizing the helical structure (31, 35). Although G4⬘
depending on the state of the channel (31). Methanethiosulfon- is on the backside of the helix, the side chains introduced with
ate reagents in the present 3 homopentamer study show that the G4⬘C and G4⬘A mutants appear to perturb the tightly packed
V1⬘C is transiently available in the channel lumen in contrast to 2⬘-5⬘ region of the channel lumen to disturb NCA binding. The
T7⬘C and T10⬘C, which are not readily accessible. In addition, T6⬘C and T6⬘F mutations in the 3 homopentamer abolish NCA
reaction with the cationic methanethiosulfonate reagent sug- sensitivity, and introducing T6⬘F in 2 (or ␣1 or ␥2) of ␣12␥2
gests that the anion-selective filter may be below V1⬘C. Molec- greatly reduces PTX sensitivity (21). Mutagenesis of the 6⬘
ular modeling of the 3 homopentamer as an ␣-helix (Fig. 6) position of 1 and 2 receptors from rats showed this site to be
places ⫺1⬘, 2⬘, 6⬘, and 9⬘, but not 0⬘, 1⬘, 3⬘, 4⬘, 5⬘, 7⬘, 8⬘, or 10⬘, important in PTX sensitivity (22). T7⬘C and V8⬘C fall outside the
in the channel pore (see supporting information), consistent with pore and, therefore, are not expected to be important binding
the other approaches. sites, yet the 8⬘ mutants block binding, perhaps, by changing the
shape of the pore. For L9⬘, where a mutation can potentially
Sites for NCA Interactions. The interacting residues are considered perturb the gating kinetics (24), the L9⬘C and L9⬘S mutations for
to be A2⬘ (or more generally the A2⬘-I5⬘ hydrophobic pocket) 3 abolish NCA binding here and L9⬘S reduces PTX sensitivity
and T6⬘ (the highly conserved and most important structural in each subunit of ␣12␥2 (24). Finally, with 1, several mutations
determinant) with a supplemental role for L9⬘. A biophysical at L9⬘ also reduce PTX sensitivity (23). Lying outside the pore,
calculation model focused on PTX interactions with A2⬘ and T6⬘ T10⬘C does not affect [3H]EBOB or [3H]BIDN binding.
of the 1 receptor (29). The present study uses site-specific
mutations in the 3 homopentamer to determine the importance Widely Diverse NCA Structures Fit the Same Site. The RDL A2⬘S
of 10 other amino acid residues in NCA binding, i.e., the whole mutation confers cross-resistance of insects to all classes of com-
cytoplasmic half of the M2 region. A-4⬘, S-3⬘, and A-2⬘ are mercial NCA insecticides (10, 17), and this cross-resistance also
apparently outside of the binding site. 3 homopentamer mu- applies to the highly potent model compounds EBOB and BIDN.
tants A-1⬘C, R0⬘C, and R0⬘S block binding, perhaps because of The effect of all mutations is essentially the same with [3H]EBOB
proximity to A2⬘. Sulfhydryl modification at V1⬘C impedes and [3H]BIDN, indicating that they both have the same binding site.
[3H]EBOB binding (this study), possibly by overlapping the More generally, an extremely wide diversity of chemical types, each
sensitive A2⬘ position. Further, for 2⬘, the low sensitivity of the with configurational specificity, appears to act the same way as
Drosophila RDL homomeric receptor to [3H]EBOB with A2⬘S GABA receptor NCAs (3, 9, 36, 37). Figs. 1B and 6 illustrate how
(or A2⬘G) (34) suggests this site for binding with confirmation they, in fact, may all fit the same site by showing the proposed
here from A2⬘C and A2⬘S mutants in the 3 homopentamer. In interactions of seven NCAs with the 3 homopentameric receptor.
Favorable hydrophobic interactions are observed for the A2⬘ Materials and Methods
methyls with the alkyl substituents of EBOB, TBPS, PTX, and Site-Directed Mutagenensis. cDNA encoding the human GABAA
BIDN, the trifluoromethylsulfinyl and pyrazole cyano of fipronil, receptor 3 subunit inserted in the pVL1392 baculovirus transfer
the hexachlorocyclopentenyl moiety of endosulfan, and the hexa- vector was described in ref. 9. Point mutations were introduced
chlorocyclohexane isomer lindane. The T6⬘ methyls interact with with the QuikChange Site-Directed Mutagenesis kit (Strat-
the ethynylphenyl and trifluoromethylphenyl substituents of EBOB agene). Mutagenic oligonucleotides were prepared by Operon
and fipronil, respectively, the trifluoromethyls and cyanos of BIDN, (Huntsville, AL). All mutations were confirmed by double-
and the exocyclic oxygen of ␣-endosulfan. The T6⬘ hydroxyl sub- strand DNA sequencing (DNA Sequencing Facility, University
stituent hydrogen bonds (H-X distance ⬍3.5 Å) to multiple elec- of California, Berkeley).
tronegative sites, i.e., the trioxabicyclooctane oxygens of EBOB and
TBPS; the exocyclic oxygen of endosulfan; the epoxy, hydroxyl, and Cell Culture and Protein Expression. Insect Sf9 cells (serum-free
lactone exocyclic oxygens of PTX; the pyrazole, amino, and cyano adapted, derived from ovaries of Spodoptera frugiperda) were
nitrogens of fipronil; and a cyano nitrogen and fluorine of BIDN. maintained by described methods in refs. 9 and 41. Recombinant
In lindane, four chlorines are ⬍3.5 Å from T6⬘ hydroxyl hydrogens. baculoviruses were constructed by using a Bacfectin-mediated
On calculating the relative energies of the bound ligands by using transfection kit (BD Biosciences Clontech). The Invitrogen
MAESTRO兾MACROMODEL (Schrödinger LLC, Portland, OR), the protocol was used for PCR analysis of recombinant virus. All
most potent ␥ isomer lindane binds in a more stable configuration PCR products were recycled with GelQuick Gel Extraction Kit
than the less active ␣, , and ␦ isomer(s) by ⬎30 kJ兾mol and the (Qiagen, Valencia, CA) and then were sequenced as described
more active ␣-endosulfan versus the less potent -endosulfan by 15 above. Log phase Sf9 cells were infected with recombinant
kJ兾mol. The L9⬘ side chains associate with the phenyl group of baculovirus at a multiplicity of infection of 5–8. Cells were
EBOB and fipronil, the ethynyl of EBOB and the aryl trifluorom- harvested at 65 h after infection. They were pelleted at 1,500 ⫻
ethyl and chloro substituents of fipronil, enhancing the potency of g for 5 min and washed once with PBS (155 mM NaCl兾3.0 mM
these long or extended molecules. The more compact NCAs, NaH2PO4兾1.0 mM K2HPO4, pH 7.4). Cell pellets were stored at
including TBPS, lindane, and BIDN, require only A2⬘ and T6⬘ for ⫺80°C until ready to use.
fit lengthwise or lying across the pore, and this positioning probably
also applies to ␣-endosulfan and PTX. These docking proposals are Membrane Preparation. The pelleted cells were resuspended in
consistent with current structure-activity relationships and may PBS and homogenized in a glass tube with a motor-driven Teflon
help in further ligand optimization. pestle (9, 41). Cellular debris was removed by centrifugation at
The NCAs are chloride channel blockers, i.e., their potency in 500 ⫻ g for 10 min at 4°C. The supernatant was centrifuged at
binding to the NCA site is proportional to their effectiveness in 100,000 ⫻ g for 40 min at 4°C, and the resulting pellet was
inhibiting chloride flux (38, 39). In the proposed binding site resuspended in PBS and stored at ⫺80°C. Protein concentration
model, the NCAs fill up and actually block the pore, although was determined with the detergent-compatible Lowry assay
they also may act allosterically by changing the channel confor- (Bio-Rad).
mation. The internuclear distance across the channel pore is on
Western Blotting. Membrane preparations were mixed with Lae-
the order of 8.5 Å, which is the same as or only slightly longer
mmli sample buffer (1.5% SDS兾5% glycerol兾65 mM Tris䡠HCl,
than the distance across multiple types of NCAs (6–8 Å).
pH 6.8, with or without 10 mM DTT). After boiling at 100°C for
5 min, samples were analyzed by SDS兾PAGE (10% acrylamide)
NCA Potency and Selectivity Conferred by Subunit Specificity. The 3
by using a Mini-PROTEAN II apparatus (Bio-Rad). Proteins
homopentamer has higher NCA sensitivity than other vertebrate
were transferred onto poly(vinylidene difluoride) membranes
GABA receptors and any replacement subunits of those tested
for 2 h at 100 V and 4°C by using the Transblot apparatus
reduce ligand affinity (9). The 3 homopentamer can form a
(Bio-Rad). The membranes were blocked in Tris-buffered saline
spontaneously opening ion channel (32), potentially facilitating (Bio-Rad) containing 2% nonfat dry milk with 0.5% Tween 20
ligand binding. GABA and other agonist modulators affect NCA for 1 h at room temperature and incubated with the mouse
binding with native and ␣1 subunit-containing receptors but not anti-GABAA receptor, -chain monoclonal antibody (Chemicon
with the 3 homopentamer (9, 12, 40). As with related ligand- International, Temecula, CA), at a dilution of 1:1,000, also for
PHARMACOLOGY
gated ion channels the NCA potency profile varies with subunit 1 h at room temperature. After three 5-min washings in TBS with
composition. Selectivity is conferred by these additional subunits 0.5% Tween 20, the blots were incubated with anti-mouse
as evident by comparing native receptors with ␣12␥2 hetero- horseradish peroxidase-linked secondary antibodies (Santa Cruz
pentameric and 3 homopentameric recombinant receptors (9, Biotechnology) at a dilution of 1:2,000 for 1 h at room temper-
27). NCAs with excellent fit for the 3 homopentamer model ature. After extensive washing, immunoreactivity was detected
may show less favorable docking in the heteropentameric native by chemiluminescence kit (PerkinElmer). Finally, the trans-
receptors associated with subunit variation at the 2⬘ position. ferred protein was visualized by incubation in Ponceau S solution
(Bio-Rad).
Concluding Remarks. The human GABAA receptor recombinant
3 homopentamer retains the NCA site in its most sensitive Disulfide Cross-Linking and Sulfhydryl Modification. For disulfide
form, equal to the insect site. Both the 3 homopentamer pore cross-linking, the membrane preparation (100 g of protein) in
and principal radioligand [3H]EBOB are symmetrical, thereby PBS (100 l) was oxidized with Cu:phen (100 M:400 M) (28,
greatly facilitating receptor modeling and ligand positioning. 42) for 5 min at 25°C. The reaction was terminated by adding 10
Ligands of widely diverse structures approach similar potency mM N-ethylmaleimide and 1 mM EDTA (final concentrations).
when optimized. The effect of mutations is the same for [3H]E- After 3 min, the membranes were recovered by centrifugation
BOB and [3H]BIDN binding and possibly for the other NCAs as (20,000 ⫻ g for 15 min at 4°C), resuspended in PBS, mixed with
well. A model for the GABAA receptor M2 region applied to the sample buffer with or without 10 mM DTT, and subjected to
3 homopentamer brings these observations together to propose SDS兾PAGE (10% acrylamide) and Western blot analysis. For
structural aspects of the NCA site. Further test of this proposal sulfhydryl modification, two methanethiosulfonate reagents
requires direct rather than indirect structural analysis of the were used, 2-(trimethylammonium)ethyl methanethiosulfonate
homopentameric and heteropentameric GABA receptors. bromide and sodium (2-sulfonatoethyl)methanethiosulfonate,
Chen et al. PNAS 兩 March 28, 2006 兩 vol. 103 兩 no. 13 兩 5189
John E. Casida 425
under described conditions (18, 43) with analysis for their effect original ␣1 and ␥2 backbone atom positions as a guide, to make
on [3H]EBOB binding. the homopentameric model. The cytoplasmic side of the M2
region and adjacent residues are considered, i.e., A-4⬘ to T10⬘
[3H]EBOB and [3H]BIDN Binding. Assay mixtures contained 1 nM (Fig. 2), with particular attention to A2⬘ to L9⬘.
[3H]EBOB (48 Ci兾mmol; 1 Ci ⫽ 37 GBq) (PerkinElmer) (6, 9) All modeling was done with MAESTRO 6.5 (Schrödinger
or 2.5 nM [3H]BIDN (50 Ci兾mmol) (8) and the recombinant LLC). Macromodel atom types were used to assign partial
expressed receptor (100 g protein) in PBS (500 l final volume) charges (44). van der Waals contacts were defined as C ⫽
(6). After incubation for 90 min at 25°C, the samples were (distance between atomic centers)兾(radius 1st atom ⫹ radius
filtered through GF兾B filters (presoaked in 0.2% polyethylenei- 2nd atom) where good, bad, and ugly contacts are defined as C ⫽
mine for 3 h) and rinsed three times with ice-cold saline (0.9% 1.3, 0.89, and 0.75 Å, respectively. The antagonists were manu-
NaCl). Nonspecific binding was determined in the presence of 1 ally docked into the putative binding site to maximize good
M ␣-endosulfan for [3H]EBOB or 5 M unlabeled BIDN for contacts, and then the ligand geometry and location were
[3H]BIDN by using 5 l dimethyl sulfoxide to add the displacing allowed to optimize relative to the 3 homopentamer, which was
agent immediately before incubation. Each experiment was itself constrained. In this optimization, all settings were left at
repeated three or more times with duplicate samples. The
the default values except a water model (generalized Born兾
binding activity of mutants was expressed as percent (mean ⫾
surface area) was used instead of a gas phase model. In each case,
SD) of that for the WT. Supplemental binding studies were made
sufficient optimization steps were performed as necessary to
by using DTT preincubation (10 mM for 5 min at 25°C) in
deoxygenated PBS continuously bubbling with argon to rule out ensure that the convergence criteria were met.
any spontaneous disulfide formation.
We thank our department colleagues Jung-Chi Liao, Motohiro Tomi-
zawa, Gary Quistad, Daniel Nomura, and Shannon Liang for helpful
Modeling Receptor–Ligand Interactions. Modeling started from the
advice and Myles Akabas, Gerald Brooks, Richard Olsen, and David
␣12␥2 GABAA receptor based on the homologous nicotinic Weiss for important suggestions. This work was supported by the William
acetylcholine receptor and acetylcholine binding protein (30). Mureice Hoskins Chair in Chemical and Molecular Entomology (to
This ␣-helical structure was reconstructed here as the 3 ho- J.E.C.) and National Science Foundation Grant CHE-0233882 (to
mopentamer, i.e., the two 2 subunits were directly replaced by K.A.D.). Molecular modeling was performed on workstations purchased
3, because they have the same M2 sequence, then the two ␣1 with National Science Foundation support matched with an equipment
subunits and one ␥2 subunit were replaced with 3, by using the donation from Dell.
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Contributed by John E. Casida, April 10, 2007 (sent for review March 19, 2007)
cloprid; EPI, epibatidine; IMI, imidacloprid; nAChR, nicotinic acetylcholine receptor; THIA,
analysis of the Torpedo nAChR (3) and x-ray crystallography of thiacloprid.
ACh binding proteins (AChBPs), soluble homologues of the ¶To whom correspondence should be addressed. E-mail: ectl@nature.berkeley.edu.
nAChR extracellular drug-binding domain (4–7). Nicotine and This article contains supporting information online at www.pnas.org/cgi/content/full/
its more potent analog epibatidine (EPI) (Fig. 2) (8) played a 0703309104/DC1.
major role in structural characterization of the agonist-binding © 2007 by The National Academy of Sciences of the USA
Table 3. Tryptic peptides labeled by nitrene identified by FTICR and Met-116 in all cases as the labeled sites (SI Figs. 8 and 9
Molecular mass, Da* showing the CID data for the 1-nitrene-labeled peptides as an
m/z, example). It was observed that some modified fragments may
Nitrene observed (z) Measured Theoretical undergo a gas-phase rearrangement reaction and lose the label
Gln-184 to Lys-203 with Tyr-195 modified
(SI Fig. 8), a process that could hamper the site assignment, but,
1 906.0771 (3) 2,715.2078 2,715.2091 fortunately, even peptides bearing the less stable labels yielded
2 678.5490 (4) 2,710.1647 2,710.1646 sufficient information to identify the modification site.
3 668.5632 (4) 2,670.2215 2,670.2239
4 672.3003 (4) 2,685.1699 2,685.1694 Structural Models for Neonicotinoid and Nicotinoid Binding Interac-
Thr-80 to Arg-122 with Met-116 modified tions. Structural models for interactions of the neonicotinoids
1 1,224.3602 (4) 4,893.4096 4,893.4063 and nicotinoids with the agonist binding domain were estab-
2 1,223.0987 (4) 4,888.3636 4,888.3621 lished by combining (i) the photoaffinity labeling results for
3 970.6924 (4) 4,848.4229 4,848.4213 precisely positioning the chloropyridinyl moiety with (ii) the
4 973.6812 (5) 4,863.3669 4,863.3668 crystal structure of Aplysia AChBP defining the localization and
geometry of all relevant nearby residues (Fig. 4). IMI and THIA
*⌬M are 0.0 to 0.8 ppm. are calculated to dock in the same orientation into the binding
pocket with energies of ⫺7.42 and ⫺7.17 kcal/mol, respectively.
The nearby amino acids are: Tyr-93 (loop A); Trp-147 (loop B);
liquid chromatography/MS/MS. Two-step searches were per- Tyr-188, Ser-189, Cys-190, and Tyr-195 (loop C); Tyr-55 (loop
formed with the CID data, the first with very strict search D); and Ile-106, Met-116, and Ile-118 (loop E). The chlorine
parameters establishing the identity of the protein and the purity atom of IMI and THIA interacts with the loop E segment and
of the preparation. The second search to identify modification(s) particularly makes van der Waals contacts to the backbone
was performed allowing an addition of up to 300 Da on any carbonyl oxygen atoms of Ile-106 (4.2 and 4.3 Å, respectively)
amino acid in the sequence. The labeled peptides were identified and Met-116 (4.8 and 4.3 Å, respectively). The pyridine nitrogen
by the predicted mass shifts corresponding to the different of IMI or THIA is expected to undergo hydrogen-bonding with
labeling molecules (Table 3). MS/MS (CID) data provided the the backbone carbonyl oxygen atoms of Ile-118 (4.3 and 4.2 Å,
final proof, both confirming the identity of the labeled peptides respectively) and Trp-147 (3.7 and 3.2 Å, respectively) via water
and pinpointing the modification site. Assignments were based bridge(s). Tyr-195 and Met-116 are located at spatially suitable
on the presence of the properly shifted fragment ions in the CID positions to be modified by the photoactivated nitrene of the
spectra of the labeled peptides. This evidence identified Tyr-195 probes (distance between nitrene and hydroxyl oxygen of Tyr-
CHEMISTRY
NEUROSCIENCE
Fig. 4. Structural models for neonicotinoid (Upper, IMI and THIA) and nicotinoid (Lower, DNIMI and DCTHIA) binding site interactions based on photoaffinity
labeling and the crystal structure of EPI–AChBP complex (Protein Data Bank ID code 2BYQ) (7). View angle is from an apical and radial location. Residues in yellow
are from the (⫹)-face and in blue are from the (⫺)-face. Loop C (shown as a portion Tyr-188 to Cys-191) flexibility (7) may yield multiple similar conformations
induced by neonicotinoid binding. Tyr-195 is not displayed for the IMI and THIA models because it would visually obscure their nitro and cyano substituents,
respectively, but the positioning is evident from the lower two images (see also SI Fig. 10).
Tomizawa et al. PNAS 兩 May 22, 2007 兩 vol. 104 兩 no. 21 兩 9077
John E. Casida 429
neonicotinoid-bound complex, presumably stabilizing the neo- scintillation beads (Amersham Biosciences), monoclonal anti-
nicotinoid interaction with loop C in the closed-conformation FLAG M2 antibody from mouse (Sigma), and either [3H]ACE
(SI Fig. 12). Thus, the conformational flexibility of the loop C or [3H]EPI were combined in a 100 mM sodium phosphate
segment induced by ligand binding (7) appears to play an buffer (pH 7.0) with or without the test compound in a final
important role for interaction with the electronegative pharma- volume of 100 l. After equilibration for 2 h at room temper-
cophore of the neonicotinoid. In sharp contrast to the neonic- ature the reaction mixtures were measured by liquid scintillation
otinoid, an iminium cation of the nicotinoid DNIMI or counting.
DCTHIA, as with an ammonium moiety of nicotine or EPI,
critically contacts the carbonyl oxygen of the loop B Trp-147 via Photoaffinity Labeling. AChBP (38 or 76 pmol sites) was incubated
hydrogen-bonding and secondarily makes van der Waals contact with 1.2-fold molar excess [3H]5 for 60 min in the dark at 25°C, and
with the -electron of the Trp indole side chain (cation– the reaction mixture (final volume 50 or 100 l of PBS) was then
interaction) (5, 7, 9, 21). Amazingly, these two very different irradiated with 300-nm lamps for 20 s (5-azidopyridinyl moiety of
interactions of neonicotinoids and nicotinoids occur in the same the probe completely photoreacted in this condition). Underivat-
binding pocket, accounting for the similarity of their structure– ized radioligand was removed by ultrafiltration with the YM-30
activity relationships (22, 23). Centricon unit, and the radiolabeled AChBP was recovered to
proceed with SDS/PAGE separation. The radiolabeled protein
Concluding Remarks. Neonicotinoids are selectively toxic to in- band was visualized by fluorography and quantified by scintillation
sects and nicotinoids to mammals attributable in part to the counting. For identification of the modified amino acid(s) involving
relative ease of penetration into the nervous system but mostly MS analysis, unlabeled photoreactive probes (1–4) were used
because of differences in target site interactions (1). New instead of the radiolabeled form 5. AChBP (200 pmol sites) was
approaches are needed in defining the unique aspects of neo- incubated and photoreacted with 1.5-fold molar excess of unlabeled
nicotinoid and nicotinoid binding with the nAChR. The Aplysia photoprobes using the above condition. The derivatized AChBP
AChBP provides an ideal model because it interacts with was loaded onto the YM-30 Centricon unit for quick purification,
neonicotinoids and nicotinoids equally well and the positioning and the reaction buffer PBS was exchanged to 25 mM ammonium
of nicotinoid (EPI) binding is unequivocally established (7). bicarbonate buffer (pH 7.8).
Photoaffinity labeling provided the crucial information that the
chloropyridinyl substituent of the neonicotinoid and nicotinoid MS Analysis. Molecular weights of intact proteins were measured
fits exactly the same site in the same way, focusing attention on on an MDS Sciex QSTAR hybrid quadrupole time-of-flight mass
the orientation of the rest of the molecule. Nicotinoid analogs of spectrometer (Applied Biosystems, Foster City, CA). Samples in
the insecticides dock identically to EPI (7). The neonicotinoids, 10 mM ammonium bicarbonate were separated by a 150 ⫻ 0.1
on the other hand, assume a unique inverted pharmacophore mm Onyx monolithic column (Phenomenex, Torrance, CA)
flowing at 500 nl/min using acetonitrile/water 1:1 with 0.1%
position for the nitro or cyano plus guanidine or amidine
formic acid as the mobile phase. Charge state distributions were
coplanar system compared with the cationic equivalent of their
converted to a normalized zero charge spectrum by using
desnitro and descyano analogs (Fig. 5). These types of neonic-
Analyst 1.1 software. The mass accuracy was ⬇1–2 Da for
otinoid and nicotinoid interactions with AChBP subsites may
proteins in this mass range.
serve as models for their unique positioning with nAChRs of
Each intact protein (⬇1 g in 10 l of ammonium bicarbonate
insects and mammals, leading to selective agonist action and buffer) mixed with an equal volume of acetonitrile was reduced
toxicity. The AChBP model so important in understanding with DTT, alkylated with iodoacetamide, and digested with
nicotinic drug action also helps solve the problem of the elusive trypsin at pH 7.8 for 4 h at 37°C. After removal of the
neonicotinoid binding site. acetonitrile, digested samples were analyzed by nanoflow liquid
Materials and Methods chromatography/MS/MS on the QSTAR instrument using a
self-packed 150 ⫻ 0.1 mm capillary column (Ultro 120, 5-m
Chemicals. The neonicotinoids and related compounds including particle size packing from Peeke Scientific, Redwood City, CA).
photoaffinity probes 1-4, [3H]ACE, and [3H]5 used here were The flow rate was 300–330 nl/min, and the gradient was 5%
available from previous studies in the Berkeley laboratory (12, acetonitrile to 50% acetonitrile in 60 min (all solvents contained
17, 24, 25). Structures are given in SI Fig. 6. [3H]EPI was 0.1% formic acid). Data were acquired in the Information-
purchased from Amersham Biosciences (Piscataway, NJ). (⫾)- Dependent Acquisition mode by using 1.0-s mass acquisitions
EPI and (⫺)-nicotine were from Tocris (Ellisville, MO) and followed by 3.0-s CID analyses of the most abundant multiply
Sigma (St. Louis, MO), respectively. charged ion in the MS survey. The database search was per-
formed with the v4.23.4 in-house version of ProteinProspector.
AChBP and Radioligand Binding. Aplysia AChBP (flanked with an Accurate masses of the labeled peptides were obtained on a
N-terminal FLAG epitope) was expressed from chemically LTQ FT hybrid linear ion trap FTICR mass spectrometer
synthesized cDNA as a soluble exported protein from stably (Thermo Electron, Waltham, MA) with chromatography as
transfected HEK293S cells lacking the N-acetylglucosaminyl- above. The Data-Dependent Acquisition method consisted of
transferase I gene and selected for geneticin resistance (7, 26). one full mass survey scan in the FTICR at 25,000 resolution. The
CHEMISTRY
Culture media containing AChBP were collected at 24- to 36-h three most intense target ions were isolated for accurate mass
intervals and stored at 4°C with 0.02% sodium azide. AChBP was measurement in the FTICR in SIM mode, using a 10-Da mass
purified on immobilized anti-FLAG antibody and dialyzed window at 50,000 resolution. The accuracy of the mass mea-
against 50 mM Tris buffer (pH 7.4) containing 150 mM sodium surements is typically within 2 ppm under such conditions. These
chloride and 0.02% sodium azide. The dialysate was concen- three ions were then fragmented in the linear ion trap by using
trated by ultrafiltration using the YM-50 Centricon unit (Milli- CID. Target values for the three different scan modes were
pore, Bedford, MA), and the Tris buffer dissolving the protein 3,000,000, 50,000, and 30,000, respectively.
was finally exchanged by 50 mM sodium phosphate buffer
NEUROSCIENCE
containing 50 mM sodium chloride (pH 7.5) (PBS) using the Calculation. Structural models for the binding sites of IMI, THIA,
YM-50 Centricon to give 2–3 mg/ml. Potencies of test chemicals DNIMI, and DCTHIA were established based on the present
(Ki values) against the Aplysia AChBP were evaluated by an photoaffinity labeling and crystal structure of the EPI–AChBP
adaptation of a scintillation proximity assay (11) with [3H]ACE complex (Protein Data Bank ID code 2BYQ) (7). EPI was re-
and [3H]EPI. In brief, AChBP, polyvinyltoluene anti-mouse SPA moved, and then the four neonicotinoids and nicotinoids were
Tomizawa et al. PNAS 兩 May 22, 2007 兩 vol. 104 兩 no. 21 兩 9079
John E. Casida 431
individually redocked. Docking calculations were carried out by Yamamoto for important suggestions. J.E.C. was supported by the
using the GALS algorithm in AutoDock 3 (27, 28). The force field William Muriece Hoskins Chair in Chemical and Molecular Entomol-
in AutoDock is derived from Amber and models electrostatic and ogy. M.T. and J.E.C. were supported by National Institute of Environ-
van der Waals effects. Docked molecules were constrained to a mental Health Sciences Grant R01 ES08424. T.T.T. and P.T. were
40-point grid in the region of the binding pocket. Visualizations supported by National Institutes of Health Grants R37-GM18360 and
were done with Maestro 7.5 (Schrödinger, Portland, OR). UO1-NS05846. D.M., K.F.M., and A.L.B. were supported by National
Center for Research Resources Grants RR015084, RR001614, and
We thank Jack Presley and Kazutoshi Fujioka for analytical assistance RR019934. K.A.D. was supported by National Science Foundation
and Dennis Dougherty, Maurice Goeldner, Shinzo Kagabu, and Izuru Grant CHE-0233882.
1. Tomizawa M, Casida JE (2003) Annu Rev Entomol 48:339–364. 16. Shimomura M, Yokota M, Ihara M, Akamatsu M, Sattelle DB, Matsuda K
2. Tomizawa M, Casida JE (2005) Annu Rev Pharmacol Toxicol 45:247–268. (2006) Mol Pharmacol 70:1255–1263.
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(2004) Neuron 41:907–914. 20. Shimomura M, Yokota M, Matsuda K, Sattelle DB, Komai K (2004) Neurosci
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9. Zhong W, Gallivan JP, Zhang Y, Li L, Lester HA, Dougherty DA (1998) Proc
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24. Latli B, Than C, Morimoto H, Williams PG, Casida JE (1996) J Labeled Comp
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J Neurochem 99:1273–1281. AJ (1998) J Comp Chem 19:1639–1662.
CURRICULUM VITAE
Date and Place of Birth: March 11, l929, Hickman, Arkansas
EDUCATION:
1946 - Graduate, Armorel High School, Armorel, Arkansas
1950 - B.S. (Agric.), University of Arkansas, Fayetteville, Arkansas
1954 - M.S. (Agron.), University of Arkansas, Fayetteville, Arkansas
1956 - Ph.D. (Ento.), Kansas State University, Manhattan, Kansas
1963 - USPHS Senior Postdoctoral Fellow, Harvard University
EMPLOYMENT BACKGROUND:
1995- - Chancellor Emeritus and Distinguished Professor Emeritus, Texas A&M,
College Station, Texas
1991-95 - Regents Professor of Entomology, Texas A&M, College Station, Texas
1991-93 - Regents Professor of Entomology, Texas A&M University, College Station,
Texas; and Executive Director, George Bush Presidential Library Center
1986-90 - Chancellor, The Texas A&M University System, College Station, Texas
433
434 Wolf Prize in Agriculture
1983-86 - Deputy Chancellor, The Texas A&M University System, College Station,
Texas
1980-83 - Deputy Chancellor for Agriculture, The Texas A&M University System,
College Station, Texas
1978-80 - Vice President for Agriculture and Renewable Resources, Texas A&M
University, College Station, Texas
1979-95 - Distinguished Professor of Entomology, Department of Entomology, Texas
A&M University, College Station, Texas
1967-78 - Head, Department of Entomology, Texas A&M University, College Station,
Texas
1963-67 - Professor, Department of Entomology, Texas A&M University, College
Station, Texas
1958-63 - Associate Professor, Department of Entomology, Texas A&M University,
College Station, Texas
1956-58 - Assistant Professor, Department of Entomology, University of Missouri,
Columbia, Missouri
1950 - Instructor (Veterans Program), Vocational Agriculture, Blytheville High
School, Blytheville, Arkansas
MILITARY EXPERIENCE:
1950-53 - Enlisted Man, U.S. Army Medical Corps
1953-79 - Senior Scientist, U.S. Public Health Services, Commissioned Reserve Corps
HONORS:
1965 - Faculty Distinguished Achievement Award for Research, Texas A&M
University
1967 - J. Everett Bussart Award for Outstanding Research in Economic
Entomology, Entomological Society of America
1967 - Award for Outstanding Service, Plains Cotton Growers, Inc., Lubbock,
Texas
1977 - Outstanding Entomologist Award, Central Texas Chapter, American
Registry of Professional Entomologists
1978 - Man of the Year in Service to Texas Agriculture, Awarded by Progressive
Farmer Magazine
1979 - Elected to National Academy of Sciences
1979 - Adventurer in Science Award, International Plant Protection Congress,
Washington, D.C.
1979 - Award for Distinguished Service to Professional Entomology, American
Registry of Professional Entomologists
Perry L. Adkisson 435
SIGNIFICANT GRANTS:
National Science Foundation:
Photoperiodic Control of Insect Diapause, 1961-1966
Cotton Incorporated:
Systems Approach to Cotton Insect Control, 1970-1973
Development of Juvenile Hormone as a Third Generation Insecticide, 1970-
1973
U.S. Department of Agriculture:
Seasonal Biology of the Boll Weevil in the Texas High Plains, 1965-1968
Potential Attractants for Use in Controlling the Tobacco Budworm, 1966-1968
Improvement of Methods for the Total Population Suppression of the Boll
Weevil, 1971-1974
Rockefeller Foundation:
Development of Insect-Resistant Cotton Varieties to Minimize Insecticide
Treatments for Control of Bollworms (Heliothis spp.), 1971-1976
International Biological Program/National Science Foundation-Environmental
Protection Agency Project:
The Principles, Strategies and Tactics of Pest Population Regulation and
Control in Major Crop Ecosystems: Director, Cotton Project; Member, Project
Executive Committee, 1970-1978
Environmental Protection Agency/USDA Project:
Development of Comprehensive, Unified, Economically and Environmentally
Sound Systems of Integrated Pest Management for Major Crops, 1979-1985
MEMBERSHIPS:
National Academy of Sciences
American Academy of Arts and Sciences
American Association for the Advancement of Science
Entomological Society of America (President, 1974)
International Organization for Biological Control
Southwestern Entomological Society
Sigma Xi
American Registry of Professional Entomologists (President, 1977)
Phi Kappa Phi
Gamma Sigma Delta
Perry L. Adkisson 437
PROFESSIONAL ACTIVITIES:
Member, Plains Cotton Growers, Inc., Technical Advisory Committee on Boll Weevil
Control, 1968-72
Member, UN/FAO Panel of Experts on Integrated Control, 1968-82
Chairman, Committee on Memorial Lectures, Entomological Society of America
(ESA), 1969-71
Chairman, Scientific Advisory Committee to Governor of Texas on Uses of
Agricultural Chemicals, 1970-72
Chairman, Texas Pesticide Advisory Committee, 1970-78
Consultant, US/EPA Office of Water Programs, 1971-72
Southern Agricultural Experiment Station Directors’ Representative to the Southern
Regional Pest Management Working Group, 1971-72
Consultant on Pesticide Use, US/EPA Hazardous Advisory Committee, 1971-72
Member, Governing Board, Entomological Society of America (ESA), 1971-75
Adviser on Pesticides to Texas Legislature, 1972
Member, Texas Structural Pest Control Board, 1972-78
Member, NAS Committee on Biologies of Pest Species (Chairman, 1975-79)
1973-79
President, Entomological Society of America, (ESA), 1974
Member, Executive Committee, N AS Environmental Sciences Board Study Group
on Problems of Pest Control, and Member, Cotton Study Team of this Project,
1974-76
Member, Editorial Board, Annual Review of Entomology, 1974-78
Chairman, Governing Council, American Registry of Professional Entomologists,
1975-76
Vice-Chairman, NAS Committee on Acquisition and Use of Scientific and Technical
Information in Regulatory Decision-Making, 1976-77
Member, NAS Committee on World Food and Nutrition, 1976-77
President, American Registry of Professional Entomologists, 1977
Member, NRC Committee for the International Union of Biological Sciences,
1979-1985
Member, NAS Class Ill Membership Committee, 1980-83
Member, NAS ad hoc Committee on Relationships between Universities and the
U.S. Government 198 1-83
greatly reducing the use of chemical insecticides. The decreased use of insecticides
has lessened the exposure of farm workers to these toxic chemicals and has
decreased pesticide pollution of the environment.
Among Adkisson’s most important contributions was the development of one
of the first semi-synthetic diets for the laboratory rearing of phytophagous insects.
He and E. S. Vanderzant developed the Vanderzant-Adkisson Wheat Germ Diet
that is widely used for rearing plant-feeding caterpillars in the laboratory under
controlled conditions. This diet has greatly facilitated basic studies in insect
physiology, developmental biology, toxicology, ecology and genetics. Research in
these areas previous to the diet had been very difficult to do when the insects had
to be reared on plants or plant parts.
In research using insects reared on artificial diets, Adkisson and his students
made many important contributions, elucidating the role of photoperiod,
temperature and diet in controlling growth and development of the most important
insect pests of cotton in the United States. Especially important was their work
demonstrating the role of photoperiod and temperature in controlling the onset
and termination of diapause in certain of these pests. Among the most important
of these studies was research conducted with Prof. C. M. Williams (Harvard
University) which showed that the photoperiod has its primary action on certain
receptors in the insect brain by regulating the flow of hormones that control insect
growth and development. Adkisson later showed that the onset of diapause in field
populations of pest insects whose diapause is photoperiodically controlled can be
predicted with considerable precision.
The fundamental knowledge gained in these studies was utilized by Adkisson
and co-workers to develop improved 1PM systems for cotton. These were designed
to make more effective use of cultural and phytosanitation measures for suppressing
pests. When these practices were combined with the limited use of insecticides,
maximum suppression of the primary pests could be obtained while preserving
the natural enemies of key secondary pests.
These programs were implemented on millions of acres of cotton and proved
effective in increasing yields. They also saved farmers millions of dollars by greatly
reducing the amounts of insecticides used on the crop. The reduced use of
insecticides prevented the release of hundreds of tons of these chemicals that
otherwise would have entered the environment.
Based on the early success of 1PM, Adkisson, Carl Huffaker (co-recipient of
the 1994-95 Wolf Prize in Agriculture) and several colleagues organized, gained
funding for, and managed the first large national 1PM research program in the
United States. This project involved more than 250 scientists from seventeen
universities who focused their efforts on developing new and improved 1PM systems
for managing the arthropod pests of six major crops. This project, known as the
Huffaker project, was successfully completed in the late 1970’s. It was succeeded
Perry L. Adkisson 439
LIST OF PUBLICATIONS
1. Adkisson, Perry L. 1954. The influence of hybridity and boll load upon the
incidence of verticillium wilt of cotton. Masters Thesis, University of Arkansas.
Fayetteville, AK.
2. Adkisson, Perry L. 1956. The relative susceptibility of the various development
stages and instars of the rice weevil, Sitophilus oryza (I.) to certain fumigants.
Ph.D. Dissertation, Kansas State University. Manhattan, KS.
3. Adkisson, Perry L. 1957. The relative susceptibility of the life history stages
of the rice weevil to certain fumigants. J. Econ. Entomol. 50(6):761-764.
4. Adkisson, Perry L. 1957. Influence of irrigation and fertilizer on populations of
three species of minds attacking cotton. FAO Plant Protect. Bull. 6(3):33-36.
5. Adkisson, Perry L. 1957. Cotton insect research in Southeast Missouri in
1956. 1956 Progress Report, SEMO Exp. Fields, Mo. Agr. Ext. Sta. Ann. Rpt.
1936:29-36.
6. Adkisson, P. L., S. Kyd and G. W. Thomas. 1957. Cotton insect controls for
Missouri. Mo. Agr. Exp. Sta. Ext. Cire. 680.
7. Adkisson, Perry L. 1958. Cotton insect research, 1957. Mo. Agr. Exp. Sta.
Bull. 700:4-11.
8. Adkisson, Perry L. 1958. An inexpensive unit for converting a vacuum cleaner
to an insect aspirator. J. Kans. Entomol. Soc. 31(4):284-285.
9. Adkisson, Perry L. 1958. Field tests of materials for control of the cotton
leafworm. J. Econ. Entomol. 51(2):259.
440 Wolf Prize in Agriculture
10. Adkisson, Perry L. 1958. Seed treatment of cotton with systemic insecticides
alone and in combination with a fungicidal treatment. J. Econ. Entomol.
5l(5):697-700.
11. Adkisson, Perry L. 1958. The influence of fertilizer applications on populations
of Heliothis zea (Boddie), and certain insect predators. J. Econ. Entomol.
51(6):757-759.
12. Adkisson, P. L., S. Kyd and G. W. Thomas. 1958. 1958 Cotton insect control:
Recommendations for Missouri. Mo. Agr. Exp. Sta. Ext. Circ. 685.
13. Adkisson, P. L., L. H. Wilkes and S. P. Johnson. 1958. Chemical, cultural and
mechanical control of the pink bollworm. Tex. Agr. Exp. Sta. Bull. 920. 16 p.
14. Wilkes, L. H., P. L. Adkisson and B. J. Cochran. 1959. Effect of spray nozzle
types on cotton insect control. Tex. Agr. Exp. Sta. PR 2078.
15. Adkisson, P. L., L. H. Wilkes and B. J. Cochran. 1959. Relative efficiencies
of certain spray nozzles for cotton insect control. J. Econ. Entomol.
52(5):985-991.
16. Wilkes, L. H., P. L. Adkisson and B. J. Cochran. 1959. Stalk shredder tests for
pink bollworm control, 1958. Tex. Agr. Exp. Sta. PR 2095.
17. Adkisson, Perry L. 1959. The effect of various humidity levels on hatchability
of pink bollworm eggs. J. Kans. Entomol. Soc. 32(4): 189-190.
18. Adkisson, P. L., L. H. Wilkes and B. J. Cochran. 1960. Stalk shredding and
plowing as measures for controlling the pink bollworm, Pectinophora
gossypiella. J. Econ. Entomol. 53(3):436-439.
19. Adkjsson, P. L., E. S. Vanderzant, D. L. Bull and W. E. Allison. 1960.
A wheat germ medium for rearing the pink bollworm. J. Econ. Entomol.
53(5):759-762.
20. Adkisson, P. L., D. L. Bull and W. B. Allison. 1960. A comparison of certain
artificial diets for laboratory cultures of the pink bollworm. J. Econ. Entomol.
53(5):791-793.
21. Bull, D. L. and P. L. Adkisson. 1960. Certain factors influencing diapause in the
pink bollworm, Pectinophora gossypiella. J. Econ. Entomol. 53(5):793-798.
22. Adkisson, Perry L. 1960. The effect of pink bollworm infestations on cotton
produced under high moisture conditions. Tex. Agr. Exp. Sta. PR 2156.
23. Adkisson, Perry L. 1960. The influence of fertilization and irrigation practices
on field populations of certain insects of the USA. Proc. XI Intl. Congress
Entomol. (Vienna). Band III. (Symposium): 141.
24. Adkisson, P. L. and J. C. Gaines. 1960. Pink bollworm control as related to
the total cotton insect control program of Central Texas. Tex. Agr. Exp. Sta.
Misc. Publ. 444. 7 p.
25. Reed, D. K. and P. L. Adkisson. 1961. Short-day cotton stocks as possible
source of host plant resistance to the pink bollworm. J. Econ. Entomol.
54(3):484-486.
Perry L. Adkisson 441
26. Cochran, B. J., L. H. Wilkes and P. L. Adkisson. 1961. Primary tillage practices
for pink bollworm control. Tex. Agr. Exp. Sta. PR 2194.
27. Graham, H. M., O. T. Robertson, P. L. Adkisson and L. H. Wilkes. 1961.
Further tests of the effectiveness of cotton stalk shredders for controlling the
pink bollworm. J. Econ. Entomol. 54(5):1057-1058.
28. Wilkes, L. H., P. L. Adkisson and B. J. Cochran. 1961. Use of an air-carrier
sprayer for cotton insect control. Tex. Agr. Exp. Sta. PR 2205.
29. Adkisson, Perry L. 1961. Fecundity and longevity of adult female pink
bollworm reared from natural and synthetic diets. J. Econ. Entomol.
54(6):1224-1227.
30. Adkisson, Perry L. 1961. Effect of larval diet on the seasonal occurrence of
diapause in the pink bollworm. J. Econ. Entomol. 54(6):7107-7l 12.
31. Bull, D. L. and P. L. Adkisson. 1962. Fat content of the larval diet as a factor
influencing diapause and growth-rate of the pink bollworm. Ann. Entomol.
Soc. Amer. 53(3):499-502.
32. Adkisson, P. L., R. L. Hanna and C. F. Bailey. 1962. Cotton yield and quality
losses resulting from various size populations of bollworms. Tex. Agr. Exp.
Sta. PR 2235.
33. Wellso, S. G. and P. L. Adkisson. 1962. The morphology of the reproductive
system of the female pink bollworm moth, Pectinophora gossypiella (Saund.).
S. Kansas Entomol. Soc. 35(2):233-235.
34. Adkisson, P. L. and S. G. Wellso. 1962. Effect of DDT poisoning on
the longevity and fecundity of the pink bollworm. J. Econ. Entomol.
55(6):842-845.
35. Adkisson, Perry L. 1962. Timing defoliants and desiccants to reduce
populations of the pink bollworm in diapause. J. Econ. Entomol. 55(6):
949-951.
36. Wilkes, L. H., B. J. Cochran and P. L. Adkisson. 1962. Further studies of the
effectiveness of certain spray nozzle types and sizes and arrangements for
controlling boll weevils and bollworms. Tex. Agr. Exp. Sta. PR 2264.
37. Adkisson, P. L., S. Hacskaylo, R. L. Hanna and S. K. Walker. 1962. Flowering
rate, boll maturity and yield of cotton treated with various insecticides. Tex.
Agr. Exp. Sta. PR 2245.
38. Graham, H. M., L. C. Fife, 0. T. Robertson and P. L. Adkisson. 1962. Pink
bollworm population dynamics. In A Summary of Recent Research Basic to
the Cultural Control of the Pink Bollworm. Tex. Agr. Exp. Sta. Misc. Publ.
579. pp. 5-10.
39. Lukefahr, M. S., L. C. Pife and P. L. Adkisson. 1962. Pink bollworm
diapause studies. In A Summary of Recent Research Basic to the Cultural
Control of the Pink Bollworm. Tex. Agr. Exp. Sta. Misc. Publ. 579.
pp. 11-13.
442 Wolf Prize in Agriculture
40. Adkisson, P. L., 0. T. Robertson and L. C. Fife. 1962. Planting date as a factor
involved in pink bollworm control. In A Summary of Recent Research Basic
to the Cultural Control of the Pink Bollworm. Tex. Agr. Exp. Sta. Misc. Publ.
579. pp. 16-20.
41. Adkisson, P. L., L. H. Wilkes, B. J. Cochran and R. L. Hanna. 1962. Spray
nozzle arrangements, types and rates of application for cotton insect control.
Tex. Agr. Exp. Sta. Misc. PubI. 595. 12 p.
42. Adkisson, P. L., C. F. Bailey and G. A, Niles. 1962. Cotton stocks screened for
resistance to the pink bollworm, 1960-61. Tex. Agr. Exp. Sta. Misc. Publ.
606.
43. Adkisson, P. L., R. A. Bell and S. G. Wellso. 1963. Environmental factors
controlling the induction of diapause in the pink bollworm, Pectinophora
gossypiella (Saunders). J. Insect Physiol. 9:299-310.
44. Bull, D. L. and P. L. Adkisson. 1963. Absorption and metabolism of C 14-
labeled DDT by DDT-susceptible and DDT-resistant pink bollworm adults.
J. Econ. Entomol. 56(5):641-643.
45. Adkisson, P. L., J. R. Brazzel and J. C. Gaines. 1963. Yield and quality losses
resulting from pink bollworm damage to cotton. Tex. Agr. Exp. Sta. Misc.
Publ. 632.
46. Adkisson, Perry L. 1963. Timee measurement in the photoperiodic induction
of diapause in the pink bollworm. Tex. Agr. Exp. Sta. PR 2274. 4 p.
47. Adkisson, Perry L. 1963. Insect control, a continuing challenge in the
Southwest. Cotton Gin and Oil Mill Press. April 13. pp. 9, 10, 34.
48. Adkisson, Perry L. 1963. Time measurement in the photoperiodic control
of diapause in an insect. Proc. XVI Intl. Congress of Zoology (Washington,
D.C.) 2:51.
49. Henry, P. and P. L. Adkisson. 1963. Relative numbers of bollworms and
tobacco bollworms in cotton of Central Texas in 1963. Tex. Agr. Exp. Sta. PR
2289.
50. Adkisson, Perry L. 1963. Relative susceptibility of various geographical
races of the pink bollworm to certain insecticides. Tex. Agr. Exp. Sta. PR
2300.
51. Weliso, S. G. and P. L. Adkisson. 1964. Photoperiod and moisture as factors
involved in the termination of diapause in the pink bollworm, Pectinophora
gossypielki. Ann. Entomol. Soc. Amer. 57(2):l70-173.
52. Adkisson, P. L., C. F. Bailey and R. L. Hanna. 1964. Effect of the bollworm,
Heliothis zea, on yield and quality of cotton. J. Econ. Entomol. 57(4):
448-450.
53. Adkisson, P. L., R. L. Hanna and C. F. Bailey. 1964. Estimates of the number
of Heliothis larvae per acre in cotton and their relation to the fruiting cycle
and yield of the host. J. Econ. Entomol. 57(5):657-663.
Perry L. Adkisson 443
82. Adkisson, Perry L. 1966. Internal clocks and insect diapause. Science,
154:234-241.
83. Nemec, S. J. and P. L. Adkisson. 1966. Comparative effectiveness of low
volume concentrate and water emulsion sprays of certain insecticides for
cotton insect control. Tex. Agr. Exp. Sta. Misc. Publ. 813. 6 p.
84. Wellso, S. G. and P. L. Adkisson. 1966. A long-day short-day effect in the
photoperiodic control of the pupal diapause of the bollworm, Heliothis zea
(Boddie). J. Insect Physiol. 12:1455-1465.
85. Adkisson, Perry L. 1966. Diapause boll weevil control on the High Plains of
Texas. Agr. Chem. 21(5):56, 57, 60, 62.
86. Adkjsson, P. L., D. R. Rummel, W. L. Sterling and W. L. Owen, Jr. 1966.
Diapause boll weevil control: A comparison of two methods. Tei. Agr. Exp.
Sta. Bull. 1054. 11 p.
87. Ankersmit, G. W. and P. L. Adkisson. 1967. Photoperiodjc responses of certain
geographical strains of Pectinophora gossypiella (Lepidoptera). J. Insect
Physiol. 13:553-564.
88. Adkisson, P. L. and S. J. Nemec. 1967. Effectiveness of certain
organophosphorous insecticides against chlorinated hydrocarbon resistant
bollworm and tobacco budworm larvae. J. Econ. Entomol. 60:268-270.
89. Adkisson, Perry L. 1967. Development of resistance by the tobacco
budworm to mixtures of toxaphene or Strobane plus DDT. J. Econ. Entomol.
60(3):788-79l.
90. Adkisson, P. L. and S. J. Nemec. 1967. Insecticides for controlling the bollworm,
tobacco budworm and boll weevil. Tex. Agr. Exp. Sta. Misc. PubI. 837. 7 p.
91. Nemec, S. J. and P. L. Adkisson. 1967. Effects of adding molasses to certain
insecticidal sprays for bollworm control. Tex. Agr. Exp. Sta. PR 2436. 4 p.
92. Hanna, R. L., J. K. Walker, Jr., P. L. Adkisson and S. J. Nemec 1967. Results of
field and laboratory studies on the control of cotton insects, 1966. Tex. Agr.
Exp. Sta. Dept. Entomol. Tech. Rpt. No. 6. 55 p.
93. Sterling, W. L., P. L. Adkisson, and D. R. Rummel, 1967. Effects of the diapause
boll weevil control program on the size and rate of increase of boll weevil
populations in the High Plains of Texas. Tex. Agr. Exp. Sta. Dept. Entomol.
Tech. Rpt. No. 7. 11 p.
94. Rummel, D. R., W. L. Sterling and P. L. Adkisson. 1967. Evaluation of the
1966 diapause boll weevil control program on the High Plains of Texas. Tex.
Agr. Exp. Sta. Dept. Entomol. Tech. Rpt. No. 8. 11 p.
95. Adkisson, Perry L. 1968. Development of resistance by the tobacco budworm
to endrin and carbaryl. J. Econ. Entomol. 61(l):37-40.
96. Adkisson, Perry L. 1968. Problems and progress in controlling diapausing
boll weevils. Proc. Beltwide Cotton Prod. and Mech. Conf. (Hot Springs, Ark.).
pp. 18-20.
446 Wolf Prize in Agriculture
technique. Proc. Intl. Atomic Energy Agency Conf. (Bogota, Colombia - May
1970). pp. 5-16.
112. Rummel, D. R. and P. L. Adkisson. 1971. A two-phased control program
designed for maximum suppression of the boll weevil in the High and Rolling
Plains of Texas. J. Econ. Entomol. 64(4):919-922.
113. Adkisson, Perry L. 1971. Weak-links in the population dynamics and diapause
of Heliothis zea (Boddie) which might be exploited by the sterile-insect release
technique. Proc. Intl. Atomic Energy Agency Symp. on Use of Sterility Principle
for Insect Eradication and Control. (Athens, Greece - Sept. 14-17, 1970).
IAEA-SM-138/21: pp. 355-364.
114. Adkisson, P. L. and S. H. Roach. 1971. A mechanism for seasonal
discrimination in the photoperiodic induction of pupal diapause in the
bollworm Heliothis zea (Boddie). Biochronometry (Ed. by Menaker, M.).
National Academy of Science. U.S. Natl. Acad. of Sciences. Washington, D.C.
pp. 272-281.
115. Sterling, W. U. and P. L. Adkisson. 1971. Seasonal biology of the boll weevil
in the High and Rolling Plains of Texas as compared with the previous
biological studies of this insect. Tex. Agr. Exp. Sta. MP 993. 12 p.
116. Roach, S. H. and P. L. Adkisson. 1971. Termination of pupal diapause of the
bollworm. J. Econ. Entomol. 64(5):1057-1060.
117. Adkisson, Perry L. 1971. Combining chemical and non-chemical techniques
in a cotton insect control program. Proc. Western Cotton Prod. Conf. (El
Paso, Texas - March 3-4, 1971). p. 20-22.
118. Adkisson, Perry L. 1971. Objective uses of insecticides in agriculture.
Proc. Symp. Agric. Chem. Harmony or Discord for Food, People and the
Environment. Univ. of Calif. (Sacramento, CA, Feb. 16-17, 1971). pp. 43-51.
119. Bottrell, D. G., D. R. Rummel and P. L. Adkisson. 1972. Spread of the boll
weevil into the High Plains of Texas. Environ. Entomol. l(2):136-l40.
120. Meisch, M. V., S. J. Nemec and P. L. Adkisson. 1972. Effects of temperature
and photoperiod on the toxicity of azinphosmethyl and malathion to the boll
weevil. J. Econ. Entomol. 65(4):1021-l023.
121. Graham, H. M., P. D. Lingren, C. G. Lincoln and P. L. Adkisson. 1972. The
economic threshold of infestations for Heliothis spp. on cotton. In Distribution,
Abundance and Control Heliothis spp. in Cotton and Other Host Plants,
1964-1971. South. Coop. Serv. Bull. 169:7-15
122. Adkisson, P. L., S. H. Roach and J. R. Phillips. 1972. Diapause. In Distribution,
Abundance and Control of Heliothis spp. in Cotton and Other Host Plants,
1964-1971. South. Coop. Serv. Bull. 169:33-41.
123. Adkisson, Perry L. 1972. The integrated control of the insect pests of cotton.
Proc. Tall Timbers Conf. on Ecological Animal Control by Habitat Management.
Tall Timbers Res. Sta. (Tallahassee, FL) 4:175-188.
448 Wolf Prize in Agriculture
124. Adkisson, Perry L. 1972. Use of cultural practices in insect pest management.
In Implementing Practical Pest Management Strategies. Proc. Natl. Ext. Pest
Management Workshop. Purdue Univ. (Lafayette, IN - March 14-16, 1972).
pp. 37-50.
125. Adkisson, P. L. and R. Haney. 1972. Cut cotton insecticide costs with integrated
control. Tex. Agr. Exp. Sta. Progress. 18(4):4-7.
126. Adkisson, Perry L. 1973. The principles, strategies and tactics of pest control
in cotton. In Insects: Studies in Population Management. Ed., P. W. Gier. Ecol.
Soc. Austr. Mem. 1:274-282.
127. Cole, C. L., P. L. Adkisson and R. E. Fye. 1973. Seasonal abundance of Heliothis
larvae on cotton in Presidio, Texas, area. J. Econ. Entomol. 66(2):524-526.
128. Adkisson, Perry L. 1973. Educational, extension and legislative requirements
for integrated control. Proc. FAO Conf. Ecol. in Relation to Plant Pest Control.
(Rome, Italy - Dec. 11-14, 1972). PP. 299-304.
129. Nemec, S. J. and P. L. Adkisson. 1973. Organophosphate resistance levels in
tobacco budworm and bollworm populations in Texas. In Investigations of
Chemicals for Control of Cotton Insects in Texas. Tex. Agr. Exp. Sta. Dept.
Entomol. Tech. Rpt. 73-2. pp. 18-25.
130. Adkisson, P. L. and R. F. Smith. 1973. Pest management in perspective. Proc.
Natl. Ext. Pest Management Workshop. (Baton Rouge, LA - March 12-13,
1973).
131. Sterling, W. L. and P. L. Adkisson. 1974. Seasonal incidence of diapause and
reproduction in boll weevils inhabiting the High and Rolling Plains of Texas.
Tex. Agr. Exp. Sta., Misc. Publ. 1145. 9 p.
132. Adkisson, Perry L. 1974. Opportunities for professional entomologists in
private practice. Bull. Entomol. Soc. Amer. 20:277-278.
133. Adkisson, Perry L. 1974. Southern pine beetle research in the Texas
Agricultural Experiment Station. Proc. Southern Pine Beetle Symp., Tex. Agr.
Exp. Sta. and A.S.F.S. Southern Forest Exp. Sta. (March 1974). p. 6.
134. Smith, R. F., C. B. Huffaker, P. L. Adkisson and L. D. Newsom. 1974. Progress
achieved in the implementation of integrated control projects in the USA and
tropical countries. EPPO Bull. 43(3):221-239.
135. Adkisson, Perry L. 1975. Entomology, the important profession. Bull. Entomol.
Soc. Amer. 21(7):3-4.
136. Rummel, D. R., D. G. Bottrell, P. L. Adkisson and R. C. McIntyre. 1975. An
appraisal of a 10-year effort to prevent the westward spread of the boll
weevil. Bull. Entomol. Soc. Amer. 21:6-11.
137. Reynolds, H. T., P. L. Adkisson and R. F. Smith. 1975. Cotton Insect Pest
Management. j11 Introduction to Insect Pest Management. W. H. Luckmann
and R. L. Metcalf, Eds. John Wiley and Sons, NY. pp. 379-443.
Perry L. Adkisson 449
150. Eden, W. G., P. L. Adkisson (and 14 others). 1977. Analytical studies for the
U.S. Environmental Protection Agency. Vol. VII. Pesticide Decision Making.
Natl. Acad. Sci. (Washington, D.C.). 100 p.
151. Smith, R. F., J. H. Perkins, P. L. Adkisson, B. E. Day, J. B. Siddall and H. D.
Thurston. 1977. Pest control. In Vol. I. Supporting Papers: World Food and
Nutrition Study. Natl. Acad. Sci. (Washington, D.C.). pp. 76-138.
152. Meola, R. W. and P. L. Adkisson. 1978. Endocrine aspects of diapause
regulation in Heliothis zea. Folin Mexicana 39-40:56-57.
153. Sterling, W. L. and P. L. Adkisson. 1978. Population dynamics of the boll
weevil inhabiting the High and Rolling Plains of Texas. Environ. Entomol.
7(3):439-444.
154. Adkisson, Perry L. 1978. Systems approach to integrated pest management.
Proc. UC/AID-ICA Conf. on Pest Management and the Environment (Bogota,
Colombia - Feb. 19-24, 1978).
155. Adkisson, Perry L. 1978. Selective use of insecticides in integrated pest
management. Proc. UC/AID-ICA Conf. on Pest Management and the
Environment (Bogota, Colombia - Feb. 19-24, 1978).
156. Adkisson, P. L., R. B. Frisbie and R. D. Lacewell. 1978. The need for cost/
benefit analyses in the development of integrated pest management programs.
Proc. Generale de Central Integrado de Plagas y Enfermedade con Enfasis en
Maiz and Saya. Natl. Agr. Univ. de Peru. (Lima, Peru - April 17-May 26,
1978). Vol. I.
157. Adkisson, P. L. and R. F. Smith. 1978. Measurement of crop losses caused by
insects. Proc. Generale de Central Integrado de Plagas y Enfermedade con
Enfasis en Maiz and Saya. Natl. Agr. Univ. de Peru (Lima, Peru - April 17-
May 26, 1978). Vol. I.
158. Adkisson, Perry L. 1978. Development of the systems approach to integrated
pest management. Proc. Generale de Central Jntegrado de Plagas y
Enfermedade con Enfasis en Maiz and Saya. Nati. Agr. Univ. de Peru (Lima,
Peru - April 17-May 26, 1978). Vol. II.
159. Dean, H. A. and P. L. Adkisson. 1978. Imported parasites destroy rhodesgrass
scale. Tex. Agr. Prog. 24(2):25-26.
160. Smith, R. F. and P. L. Adkisson. 1979. Expanded horizons of integrated pest
control in crop production. Proc. Opening Session and Plenary Session
Symposium IX Intl. Congress Plant Prot, (Washington, D.C. - Aug. 5-11,
1979). pp. 29-30.
161. Adkisson, P. L., R. B. Frisbie, J. K. Walker and W. L. Sterling. 1980. Integrated
control of the insect pest of cotton. Proc. Entretiens Ecologiques de Dijon.
Dijon, France.
162. Adkisson, Perry L. 1980. An ecological approach to the integrated
control of the insect pests of sorghum. Proc. Intl. Workshop on Sorghum
Perry L. Adkisson 451
Shoot-fly. Intl. Cent. Ins. Physiol. and Ecol. (Nairobi, Kenya, May 5-8, 1980).
pp. 12-16.
163. Adkisson, Perry L. 1980. Organizational constraints affecting research and
development in integrated pest management. Proc. Intl. Org. Biol. Control on
Conference on Future Trends of Integrated Pest Management. (Bellagio, Italy
- May 30-June 4, 1980).
164. Adkisson, Perry L. 1980. Desarrollo e implementacion de programs de control
de plagas. Proc. CICP/OAS/AID Curso de Control Integrado de Plagas.
(Tapachula, Mexico, Sept. 2-14, 1980). 20 p.
165. Adkisson, Perry L. 1980. The essential role of pest control in human health
and food production. Proc. Seminar and Workshop on Pest arid Pesticide
Management in the Caribbean. (Bridgetown, Barbados - Nov. 3-7, 1980).
8 p.
166. Adkisson, P. L. and V. A. Dyck. 1980. Resistant varieties in pest management
systems. In Breeding Plants Resistant to Insects. F. Maxwell and P. Jennings,
Eds. John Wiley and Sons, NY. pp. 233-251.
167. Phillips, J. R., A. P. Gutierrez and P. L. Adkisson. 1980. General
accomplishments toward better insect control in cotton. In New Technology
of Pest Control. Chap. 5. John Wiley and Sons, NY. pp. 123-153.
168. Adkisson, Perry L. 1981. The systems approach to livestock production. Proc.
Symp. Systems Approach to Animal Health and Production. (Univ. of Kentucky,
Lexington, KY - March 31-April 2, 1981). pp. 18-30.
169. Adkisson, P. L., R. E. Frisbie, J. U. Thomas and G. M. McWhorter. 1981,
Organization and implementation of an integrated pest management system.
Southwestern Entomol. 6(4):279-287.
170. Cole, C. L. and P. L. Adkisson. 1981. Life history and fecundity of the boll
weevil reared in constant and variable temperature regimens. Southwestern
Entomol. 6(4):298-302.
171. Adkisson, Perry L. 1981. The approaching crisis in sustaining high-yielding
agricultural systems. Plant Disease. 65:940-942.
172. Adkisson, P. L., G. A. Niles, J. K. Walker, L. S. Bird and H. B. Scott. 1982.
Controlling cotton’s insect pests: A new system. Science, 2 16:19-22.
173. Adkisson, Perry L. 1982. Science and the individual. Texas Journal of Science.
34(2):101-103.
174. Reynolds, H. T., P. L. Adkisson, R. F. Smith and R. E. Frisbie. 1982. Cotton
insect pest management. In Introduction to Insect Pest Management (2nd
Edition). R. L. Metcalf and W. H. Luckmann, Eds. John Wiley and Sons, NY.
pp. 375-441.
175. Cole, C. L. and P. L. Adkisson. 1982. Effects of constant and variable
temperature regimens on the survival and rate of increase of the boll weevil.
Southwestern Entomol. 7(1):50-55.
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189. Adkisson, Perry L. 1 990. Integrated Pest Management: Past and Future. Proc.
Entomol. Soc. of Amer. Cent. Natl. Symp. Annual Mtg. San Antonio, TX
December 12-13. pp 3-4.
190. Adkjsson, Perry L. 1990. Warning: Eating May be Harmful to Your Health.
William Henry Hatch Memorial Lecture, National Association of State
Universities and Land Grant Colleges. Kansas City, Missouri. Nov. 12.
9 pp.
191. Adkisson, Perry L. 1991. Integrated Pest Management: Past and Future. In
Entomology Serving Society: Emerging Technologies and Challenges - S. B.
Vinson and R. L. Metcalf, Eds. Entomological Society of America, Lanham,
MD. pp. 320-322.
454 Wolf Prize in Agriculture
The above article is reprinted with permission from the Entomological Society of America, MD
20706-4876, USA.
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Source: From Science Vol. 216, No. 4541, pp. 19-22 (1982). Reprinted with permission from AAAS.
474 Wolf Prize in Agriculture
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Carl B. Huffaker
University of California
Berkeley, California, USA
k
1914–1995
Professor Carl B. Huffaker performed basic research on crop pests, and subsequently
broadened their work to develop biologically-based systems for pest control. Both
were major contributors to the development and implementation of integrated
pest management, or IPM, systems for crop production. Huffaker successfully
demonstrated the efficacy of biological control agents for the control of weeds and
insects.
Huffaker was one of the leaders in large-scale programs that demonstrated the
effectiveness of IPM systems for crop protection, and that led to the establishment
of IPM as an environmentally beneficial and economically favorable system for
crop production that is today used throughout the world.
CURRICULUM VITAE
General
Born September 30, 1914, Monticello, Kentucky (USA)
Attended public schools in Monticello, Kentucky, 1920-1933
479
480 Wolf Prize in Agriculture
Positions
1940-41 Teaching Assistant in Zoology, Ohio State University
1941 U.S. Department of Agriculture, Bureau of Entomology (summer)
1941-43 Assistant Entomologist, University of Delaware
1943-45 Associate Entomologist, then Entomologist, Health and Sanitation
Division, Institute of Inter-American Affairs (Colombia, Haiti and
Dominican Republic)
1946-1984 Assistant Entomologist, then Associate Entomologist, then Entomologist
and Professor of Entomology, University of California, Berkeley
1970-1983 Director, International Center for Integrated and Biological Control,
University of California, Berkeley and Riverside
1985- Emeritus Professor, Division of Biological Control, University of
California, Berkeley
Research Interests
Population ecology, integrated control, natural balance and regulating mechanisms,
biological control, roles of predators, parasites and environmental conditions in
population dynamics. Use of experimental models and analysis of performance
and functions of components. Integrated pest management.
General
Dr. Huffaker’s services to education and governmental agencies have included, in
addition to those listed above, consultations and briefs to the National Science
Foundation, the President’s Office of Science and Technology, the Agricultural
Research Policy Advisory Committee, and various state and national investigative
committees. He has acted as consultant to and prepared background papers for
the United Nations Environmental Programme, the United Nations Development
Program, Food and Agricultural Organization of the United Nations, and the
Consortium for International Crop Protection. As a member of various committees
within the College of Agriculture, University of California, Berkeley, he has helped
to upgrade existing courses and to help establish new courses, especially in the
field of pest management.
He has served as reviewer for NSF, NIH and EPA grant proposals and has
reviewed both scientific and popular articles and books on ecology and pest
management for U.S. and foreign publishers. He was a member of the Editorial
Advisory Committee for Agro-Ecosystems (Elsevier Publ. Co.). Author of over 200
scientific publications, Dr. Huffaker is editor of and contributor to the book
“Biological Control” (Plenum Press, N.Y., 1971), co-editor with P.S. Messenger and
contributor to “Theory and Practice of Biological Control” (Academic Press, 1976),
editor of and contributor to “New Technology of Pest Control” (Wiley, N.Y., 1980),
and co-editor with R L Rabb and contributor to Ecological Entomology (Wiley,
N.Y., 1984).
been one of the leading advocates of integrating biological control, along with
other selective alternatives to the wholesale use of toxic pesticides, in a holistic
system now known as 1PM, and has been instrumental in implementing this
approach in California in his capacity as Director of the Statewide International
Center for Biological and Integrated Control (1970-83). His unrelenting efforts,
and the increasing realization that unilateral chemical control will not solve pest
problems, eventually led to the endorsement of 1PM by government agencies in
the USA and elsewhere. The turning point was the initiation of a huge nation-wide
project, known as the “Huffaker Project”, which he directed during 1972-78.
Engaging some 300 scientists from 18 universities and funded jointly by the NSF,
EPA and USDA, it was aimed at promoting the development of integrated pest
management programs in major agro-ecosystems including soybeans, cotton, alfalfa,
pome and stone fruits, citrus and coniferous forests. Summarized in his book,
“New Technology of Pest Control” (1980), this unprecedented project had a
tremendous impact on the philosophy and practice of plant protection world-
wide, and the resulting reduction in pesticide use has already affected environmental
quality in many parts of the world.
Professor Huffaker continues to be highly active in both theoretical and practical
research, and his seminal publications will continue to influence the world’s
agricultural and scientific communities for generations to come.
1971. Editor. Biological Control. Plenum Press, New. York, 511 pp.
1976. Editor (with P.S. Messenger). Theory and Practice of Biological Control.
Academic Press, New York, 788 pp.
1980. Editor. New Technology of Pest Control. Wiley, New York, 500 pp.
1983. (with D. Rosen). An overview of desired attributes of effective biological
control agents, with particular emphasis on mites, pp. 2-11. In: M.A. Hoy,
G.L. Cunningham and L. Knutson, ed., Biological Control of Pests by Mites.
Univ. Calif. Div. Agric. Publ. 3304.
1984. Editor (with R.L. Rabb). Ecological Entomology. Wiley, New York, 844 pp.
1990. (with A.P. Gutierrez). Natural enemies and prey population regulation,
pp. 183-195, and: Evaluation of efficiency of natural enemies in biological
control, pp. 473-495, In: O. Rosen, ed., Armored Scale Insects: Their Biology,
Natural Enemies and Control. Elsevier, Amsterdam.
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Morris Schnitzer
Centre for Land and Biological Resources Research
Agriculture Canada, Ottawa, Ontario, Canada
k
CURRICULUM VITAE
Born February 4, 1922 in Bochum, Germany
Immigrated to Canada in May, 1947
DEGREE RECEIVED
B. Sc. Agr. (Soil Chemistry), 1951 - McGill University, Montreal, Canada.
M. Sc. Soil Chemistry, 1952 - McGill University, Montreal, Canada.
Ph.D. Soil Chemistry, 1955 - McGill University, Montreal, Canada.
Postdoctorate studies in organic chemistry, 1961-62, Imperial College of Science
and Technology, London, England, with Prof. D. H. R. Barton, Nobel Laureate.
487
488 Wolf Prize in Agriculture
PROFESSIONAL PUBLICATIONS
a) Books Written
Schnitzer, M. and S.U. Khan. 1972. Humic Substances in the Environment.
Marcel Dekker, New York. 327 Pages.
b) Books edited
- Schnitzer, M. and S.U. Khan. 1978. Soil Organic Matter. Elsevier, Amsterdam,
319 pages.
- Huang, P.M. and M. Schnitzer. 1984. Interaction of Soil Minerals with Natural
Organics and Microbes. Soil Science Society of America, Madison, Wisconsin,
606 pages.
c) Scientific papers
355 refereed scientific papers.
EDITORIAL WORK
Served on the editorial boards of the Canadian Journal of Soil Science, Soil Science,
Geoderma, and Plant and Soil.
SCIENTIFIC PUBLICATIONS
1. Schnitzer M. and W.A. Delong. A note on the podzolization process. Sci. Agr.
32: 680-681, 1952.
2. Schnitzer M. and W.A. Delong. Note on the reaction of 2,2’-dipyridyl
with iron in the presence of organic matter. Cand. J. Agr. Sci. 34: 324-325,
1954.
490 Wolf Prize in Agriculture
19. Schnitzer, M. and J.R. Wright. Nitric acid oxidation of the organic matter of
a podzol. Soil Sci. Soc. Proc. 24: 273-276, 1960.
20. Schnitzer, M. and J.R. Wright. Studies on the oxidation of the organic matter
of the AO and Bh horizons of a podzol. Proc. 7th Intl. Congr. of Soil Sci. II:
112-119, 1960.
21. Wright, J.R. and M. Schnitzer. Functional groups in the organic matter of the
AO and Bh horizons of a podzol. Proc. 7th Intl. Congr. of Soil Sci. II: 120-127,
1960.
22. Wright, J.R. and M. Schnitzer. An estimate of the aromaticity of the organic
matter of a Podzol soil. Nature 190: 703-704, 1961.
23. Schnitzer, M. and I. Hoffman. Thermogravimetry of the organic matter of a
Podzol soil. Chem. & Ind. 1397-1398, 1961.
24. Schnitzer, M. and J.G. Desjardins. Cryoscopic molecular weight apparatus.
Chemist-Analyst 50: 117, 1961.
25. Turner, R.C. and M. Schnitzer. Thermogravimetry of the organic matter of a
Podzol. Soil Sci. 93: 225-232, 1962.
26. Schnitzer, M., J.R. Wright and I. Hoffman. High temperature thermo-
gravimetry of chloride and sulphates. Anal. Chim. Acta. 26: 371-377,
1962.
27. Schnitzer, M. and J.G. Desjardins. Molecular and equivalent weights of the
organic matter of a Podzol. Soil. Soc. Am. Proc. 26: 362-365, 1962.
28. Barton, D.H.R. and M. Schnitzer. A new experimental approach to the humic
acid problem. Nature 198: 417-418, 1963.
29. Wright, J.R. and M. Schnitzer. Metallo-organic interactions associated with
podolization. Soil Sci. Soc. Am. Proc. 27: 171-176, 1963.
30. Schnitzer, M. and S.I.M. Skinner. Organo-metallic reactions in soils. 1: Soil
Sci. 96: 86-93, 1963.
31. Schnitzer, M. and S.I.M. Skinner. Organo-metallic reactions in soils. 2. Soil
Sci. 96: 181-186, 1963.
32. Hoffman, I. and M. Schnitzer. Thermogravimetry: A. valuable analytical tool,
Chemistry in Canada 16: 30-32, 1964.
33. Schnitzer, M., R.C. Turner and I. Hoffman. A thermogravimetric study of
organic matter of representative Canadian podzol soils. Can. J. Soil Sci. 44:
7-13, 1964.
34. Schnitzer, M. and U.C. Gupta. Some chemical characteristics of the organic
matter extracted from the O and B2 horizon of the grey wooded soil. Soil Sci.
Soc. Am. Proc. 28: 374-377, 1964.
35. Schnitzer, M. and I. Hoffman. Pyrolysis of soil organic matter. Soil Sci. Soc.
Am. Proc. 28: 520-525, 1964.
36. Schnitzer, M. and S.I.M. Skinner. Organometallic reactions in soil. 3: Soil Sci.
98: 197-203, 1964.
492 Wolf Prize in Agriculture
153. Senesi, N., S.M. Griffith and M. Schnitzer. Binding of Fe+3 by humic materials.
Geochim. Cosmochim. Acta. 41: 969-976. 1977.
154. Schnitzer, M. recent findings on the characterization of humic substances
extracted from soils from widely different climatic zones. Soil Organic Matter
Studies. II International Atomic Energy Agency, Vienna, pp. 117-132, 1977.
155. Neyroud, J.A. and M. Schnitzer. Sur la structure des acides humiques at
fulviques du sol. Soil Organic Matter Studies II. International Atomic Energy
Agency, Vienna, pp. 157-169, 1977.
156. Senesi, N., Y. Chen and M. Schnitzer. Aggregation-dispersion phenomena in
humic substances. Soil Organic Matter Studies II. International Atomic Energy
Agency, Vienna, pp. 143-155, 1977.
157. Senesi, N., Y. Chen and M. Schnitzer. The role of free radicals in the oxidation
and reduction of fulvic acid. Soil Biol. Biochem. 397-403, 1977.
158. Gamble, D.S., M. Schnitzer and D.S. Skinner. Mn(II)-fulvic acid complexing
equilibrium measurements by electron spin resonance spectrometry. Can. J.
Soil Sci. 57: 47-53, 1977.
159. Griffith, S.M. and M. Schnitzer. Organic compounds formed by the hydrogen
peroxide oxidation of soils. Can. J. Soil Sci. 57: 223-231, 1977.
160. Sowden, F.J., Y. Chen and M. Schnitzer. The nitrogen distribution in soils
formed under widely differing climatic conditions. Geochim. et Cosmochim.
Acta. 41: 1524-1526, 1977.
161. Chen, Y., F.J. Sowden and M. Schnitzer. Nitrogen in Mediterranean soils.
Agrochimica. 21: 7-14, 1977.
162. Kodama, H. and M. Schnitzer. Effect of fulvic acid on the crystallization of
Fe(III) oxides. Geoderma 19: 279-291, 1977.
163. Chen, Y., N. Senesi and M. Schnitzer. The chemical degradation of humic
and fulvic acids extracted from Mediterranean soils. J. Soil Sci. 29: 350-359,
1978.
164. Chen, Y. and M. Schnitzer. The surface tension of soil humic substances. Soil
Sci. 125: 7-15, 1978.
165. Chen, Y., N. Senesi and M. Schnitzer. Chemical and physical characteristics
of humic anf fulvic acids extracted from the soils of Mediterranean region.
Geoderma 20: 87-104, 1978.
166. Whitby, L.M. and M. Schnitzer. Humic and fulvic acids in sediments and
soils of agricultural watersheds. Can. J. Soil Sci. 58: 161-168, 1978.
167. Schnitzer, M. Reactions of humic substances with the minerals in the
soil environment. Chapter 51 in Environmental Biogeochemistry and
Geomicrobiology. Vol II: 639-647, 1978.
168. Senesi, M. and M. Schnitzer. Free radicals in humic substances. Chapter 36
in Environmental Biogeochemistry and geomicrobiology. Vol II: 467-481,
1978.
500 Wolf Prize in Agriculture
169. Chen, Y., S.U. Khan and M. Schnitzer. UV irradiation of dilute fulvic acid
solutions. Soil Sci. Soc. Am. J. 42: 292-296, 1978.
170. Mathur, S.P. and M. Schnitzer. A chemical and spectroscopic characterization
of some synthetic analogues of humic acids. Soil Sci. Soc. Am. J. 42:
591-596, 1978.
171. Schnitzer, M. Some observations on the chemistry of humic substances.
Agrochimica XXII: 216-226, 1978.
172. Schnitzer, M. The chemistry of humic substances. Intern. Congress of soil
Sci., Edmonton, Alberta, Canada, I: 51-52, 1978.
173. Schnitzer, M. and M. Levesque. A novel approach to assessing the degree of
humification of peats. Soil Sci. 127: 140-145, 1979.
174. Iverson, K.C. and M. Schnitzer. The biodegradability of the “unknown” soil
nitrogen. Can. J. Soil Sci. 59: 277-286, 1979.
175. Schnitzer, M. Effect of low pH on the chemical structure and reactions of
humic substances. Pages 202-222 in Effects of acid precipitation on terrestrial
ecosystems. Edt. by T.C. Hutchinson and M. Havas. Plenum Press, New York,
1979.
176. Schnitzer, M. the chemistry and reactions of humic substances. Pages 807-
819 in Ecology and Coal Resource Development (M.K. Wali ed.) Pergamon
Press, New York. Vol. II, 1979.
177. Cortez, J. and M. Schnitzer. Nucleic acid bases in soils and their association
with organic and inorganic soil constituents. Can. J. Soil Sci. 59: 277-286,
1979.
178. Cortez, J. and M. Schnitzer. Purines and pyrimidines in soils and humic
substances. Soil Sci. Soc. Am. J. 43: 958-961, 1979.
179. Ghosh, K. and M. Schnitzer. Macromolecular structure of humic substances.
Soil Sci. 129: 266-276, 1980.
180. Ghosh, K. and M. Schnitzer. Some recent advances in the chemistry and
reactions of humic substances. Indian J. Chem. LVI: 1090-1093, 1979.
181. Ghosh, K. and M. Schnitzer. UV and visible absorption spectroscopic
investigations in relation to macromolecular characteristics of humic
substances. J. Soil Sci. 30: 735-745, 1979.
182. Kerndorff, H. and M. Schnitzer. Humic and fulvic acids as indicators
of soil and water pollution. J. Water, Air and Soil Pollution 12: 319-329,
1979.
183. Biederbeck, V.O., C.A. Campbell, K. E. Bowren, M. Schnitzer and R.N. McIver.
Effect of burning cereal straw on soil properties and grain yields in
Saskatchewan. Soil Sci. Soc. Am. J. 44: 103-111, 1980.
184. Schnitzer, M., H. Kodama and K.C. Iverson. Interaction of “protein-like”
materials in fulvic acid with montmorillonite, Zeitschr. Pflanzenernahr.
Bodenk. 143: 334-343, 1980.
Morris Schnitzer 501
231. Biederbeck, V.O., C.A. Campbell and M. Schnitzer. Effects of wheat rotation
and fertilization on microorganisms and biochemical properties of a brown
loam in Saskatchewan. Transactions XIIIth International Congress of Soil
Science (Hamburg) II: 552-553, 1986.
232. MacCarthy, P., R.L. Malcom, M.H.B. Hayes, R.S. Swift, M. Schnitzer and W.L.
Campbell. Establishment of a collection of standard humic substances.
Transactions XIIIth International Congress of Soil Science (Hamburg) II:
378-379, 1986.
233. Campbell, C.A., M. Schnitzer, J.W.B. Stewart, V.O. Biederbeck and F. Selles.
Effects of manure and P fertilizers on properties of a black charnozem in
southern Saskatchewan. Can. J. Soil Sci. 66: 601-614, 1986.
234. Arshad, M.A. and M. Schnitzer. The chemistry of a termite fungus comb.
Plant and Soil 98: 247-256, 1987.
235. Arshad, M.A., M. Schnitzer and C. Preston. Characteristics of a termite fungus
comb studied by chemical and spectroscopic methods. Soil Biol. Biochem.
19: 227-230, 1987.
236. Schnitzer, M. and C.M. Preston. The supercritical gas extraction of a soil
with solvents of increasing polarities. Soil Sci. Soc. Am. J. 51: 639-646,
1987.
237. Catroux, G. and M. Schnitzer. Chemical, spectroscopic and biological
characteristics of the organic matter in particle size fractions separated from
an Aquoll. Soil Sci. Soc. Am. J. 51: 1200-1207, 1987.
238. Preston, C.M. and M. Schnitzer. 13C NMR of humic substances: pH and
solvent effects. J. Soil Sci. 38: 667-678, 1987.
239. Arshad, M.A. and M. Schnitzer. Characteristics of the organic matter
in a slightly and in a severely crusted soil. Z Pflanzenernachr. Bodenk. 150:
412-416, 1987.
240. Byler, D.M., W.V. Gerasimowicz, H. Susi and M. Schnitzer. FTIR spectra of
soil constituents: Fulvic acid and fulvic acid complex with ferric ions. Applied
Spectroscopy 41: 1428-1430, 1987.
241. Baldock, J.A., M. Schnitzer and B.D. Kay. Influence of selected cropping
treatments and aggregate sizes on soil carbohydrates. Can. J. Soil Sci. 67:
489-499, 1987.
242. Schnitzer, M, J.A. Ripmeester and H. Kodama. Characterization of the organic
matter associated with a soil clay. Soil Sci. 145: 448-454, 1988.
243. Schnitzer, M. Selected methods for the characterization of soil humic
substances. Humic substances in Soil and Crop Sciences; selected Readings.
Special publication of the Soil Sci. Soc. Am. Madison, Wisconsin, 65-89,
1990.
244. Campbell, C.A., V.O. Biederbeck, M. Schnitzer, F. Selles and R.P. Zentner.
Effect of 6 years of zero tillage and N fertilizer management on changes in
Morris Schnitzer 505
258. Dinel, H., M. Schnitzer and G. Mehuys. Soil lipids: origin, nature, content,
decomposition and effect on soil physical properties. Soil Biochem. 6:
397-429, 1990.
259. Schulten, H.-R. and M. Schnitzer. Aliphatics in soil organic matter associated
with clays. Soil Sci. Soc. Am. J. 54: 98-105, 1990.
260. Schnitzer, M. Soil organic matter and soil quality. Agriculture Canada, 1991.
261. De Kempe, C.R. and M. Schnitzer. Low-temperature ashing of humic and
fulvic acids. Soil Sci. Soc. Am. J. 54: 399-403, 1990.
262. Schnitzer, M., C. Tarnocia, P. Schuppli and H.-R. Schulten. Nature of the
organic matter in tertiary Palescols in the Canadian Arctic. Soil Sci. 149:
257-267, 1990.
263. Schnitzer, M. C. Tarnocia, P. Schuppli, R. Hempfling, R. Muller and H.-R.
Schulten. Paleoenvironmental organic indicators in Eocene paleosols from
Arctic Canada. Fresenius Z. Anal. Chem. 337: 1882-1884, 1990.
264. Schnitzer, M., H.-R. Schulten, P. Schuppli and D.A. Angers. Extraction of
organic matter from soils with water at high pressure and temperatures Soil
Sci. Soc. Am. J. 55: 102-108, 1991.
265. Schnitzer, M. Soil organic matter — the next 75 years. Soil Sci. 149:
257-267, 1991.
266. Mathur S.P., M. Schnitzer and P. Schuppli. The distribution of nitrogen in
peat-based composts of manure slurries and fisheries wastes. Biol. Agr. Hort.
7: 153-163, 1990.
267. Campbell, C.A., M. Schnitzer, G.P. Lafond, R.V. Zentner and J.E. Knipel. Effects
of crop rotations and management practices on soil- and amino- N in a Thin
Black Chernozem. Soil Sci. Soc. Am. J. 55: 739-745, 1991.
268. Arshad, M.A., M. Schnitzer, D.A. Angers and J.A. Ripmeester. Effects of till vs
no-till on the quality of soil organic matter. Soil Biol. Biochem. 22: 595-599,
1990.
269. Schnitzer, M., H. Kodama and J.A. Ripmeester. Determination of aromaticity
of humic substances by X-ray diffraction analysis. Soil Sci. Soc. Am. J. 55:
745-750, 1991.
270. Schnitzer, M. Significance of soil organic matter in soil formation, transport
processes in soils and in the formation of soil structures. Barichte uber
Landwirtschaft. 206: 63-81, 1992.
271. Schulten, H.-R, B. Plage and M. Schnitzer. A chemical structure for humic
substances. Naturwissenschaften 78: 311-312, 1991.
272. Schulten, H.-R and M. Schnitzer. A contribution to solving the puzzle of the
chemical structure of humic substances: pyrolysis soft-ionization mass
spectrometry. Sci. Tot. Environ. 117/118: 27-39, 1992.
273. Schulten, H.-R and M. Schnitzer. Structural studies of Bainsville humic acids
by Curie-point pyrolysis gas chromatography/mass spectrometry. Soil Sci.
153: 205-224, 1992.
Morris Schnitzer 507
287. Preston, C.M., R. Hempfling, H.-R. Schulten, M. Schnitzer, J.A. Torfymow and
D.E. Axelson. Characterization of organic matter in the soil profile in a
young Douglas-fir plantation. Plant and Soil 158: 69-82, 1994.
288. Schnitzer, M. Organic-inorganic interactions in soil and their effects in soil
quality. Chapter 1 in Environmental Impact of Soil Component Interactions,
edt. by P.M. Huang, J. Berthelin, J.-M. Bollag, W.B. McGill and A.L. Page.
pp. 3-19, Lewis Publishers Boca Raton, 1995.
289. Schnitzer, M. A chemical structure for humic acid. Chemical, 13C NMR,
colloid-chemical and electron microscopic evidence in Humic Substances in
the Global Environment and Implications on Human Health, edt. by N. Senesi
and T.M. Miano. pp. 57-59, Elsevier, 1994.
290. Schnitzer, M., H. Kodama and H.-R. Schulten. Mineral effects on the pyrolysis-
field ionization mass spectrometry of fulvic acid. Soil Sci. Soc. Am. J. 58:
1100-1107, 1994.
291. Monreal, C.M., M. Schnitzer, H.-R. Schulten, C.A. Campbell and D.W.
Anderson. Soil organic structures in macro and microaggregates of a cultivated
brown chernozem. Soil Biol. Biochem. 27: 845-853, 1995.
292. Righi, D., H. Dinel, H.-R. Schulten and M. Schnitzer. Characterization of clay
organic matter complexes resistant to oxidation by peroxides. Europ. J. Soil
Sci. 46: 423-429, 1995.
293. Sorge, C., M. Schnitzer, P. Leinweber, and H.-R. Schulten. Molecular chemical
characterization of organic matter in whole soil and particle-size fractions
of a spodosol by pyrolysis-field ionization mass spectrometry. Soil Sci. 158:
189-203, 1994.
294. Schnitzer, M. Research on prodzolization at MacDonald College from 147 to
1959. Can. J. Soil Sci. 75: 155-159, 1995.
295. Schulten, H.-R., C.M. Monreal and M. Schnitzer. Effect of long-term cultivation
on the chemical structure of soil organic matter. Naturwissenschaften 82:
42-44, 1995.
296. Schulten, H.-R., C. Sorge and M. Schnitzer. Structural studies on soil nitrogen
by Currie-point pyrolysis-gas chromatography/mass spectrometry with
nitrogen selective detection. Biol. Fert. Soil 20: 174-184, 1995.
297. Liang, B.C., E.G. Gregorich and M. Schnitzer and R.P. Voroney. Carbon
mineralization in soils of different textures as affected by water soluble
organic carbon extracted from composted dairy manure. Biol. Fert. Soil 21:
10-16, 1996.
298. Van Bochove, E., D. Conillard, M. Schnitzer and H.-R. Schulten. Pyrolysis-
field ionization mass spectrometry of the four phases of cow manure
composting. Soil Sci. Soc. Am. J. 60: 1781-1786, 1996.
299. Schnitzer, M. and H.-R. Schulten. Analysis of organic matter in soil
extracts and whole soils by pyrolysis mass spectrometry. Adv. Agronomy. 55:
167-217, 1995.
Morris Schnitzer 509
300. Gregorich, E.G., M. Schnitzer, C.M. Monreal and H.-R. Schulten. Management
effects and transformation of plant residues into soil organic matter: plant
tissues, isolated soil fractions and whole soils. Soil. Sci. 161: 693, 1996.
301. Liang, B.C., E.G. Gregorich and M. Schnitzer. Mineral nitrogen accumulations
in soils as affected by water soluble organic carbon extracted from composted
dairy manure. Commun. Soil Sci. Plant Anal. 26: 2711-2723, 1995.
302. Dinel, H., M. Schnitzer and S.U. Dumontet. Compost maturity: chemical
characteristics of extractable lipids. Compost Sci. Util. 4: 16-25, 1996.
303. Schulten, H.-R. and M. Schnitzer. Three-dimensional models for humic acids
and soil organic matter. Naturwissenschaften 82: 487-498, 1995.
304. Liang, B.C., E.G. Gregorich, M. Schnitzer and H.-R. Schulten. Chemical
and spectroscopic characterization of water extracts of stockpiled and
composted dairy manures and their adsorption by soils. Soil Sci. Sco. Am. J.
60: 1758-1763, 1996.
305. Schnitzer, M. and H.-R. Schulten. New ideas on the chemical make-up of soil
organic matter in Future Prospects of Soil Chemistry SSSA Special publication
No: 155, pp 153-177, Madison, Wisconsin 1998.
306. Dinel, H., M. Schnitzer and S. Dumontet. Compost maturity: extractable
lipids as indicators of organic matter stability. Compost Sci. Util. 4: 6-12, 1996.
307. Monreal, C.M., H. Dinel, M. Schnitzer, D.S. Gamble and V.O. Biederbeck.
Impact of carbon sequestration on indicators of soil quality as influenced by
management in sustainable agriculture in Soil Process and the Carbon Cycle
(ed. By R. Lal, J. Kimble and B.A. Stewart). Adv. in Soil. Sci. 435-458, 1997.
308. Celi, L. M. Schnitzer and M. Negre. Determination of carboxyl groups in
humic acids by wet chemical, Fourier-transform infrared spectroscopic, and
13C NMR spectroscopic methods: A comparative study. Soil Sci. 162-189:
190-197, 1997.
309. Schulten H.-R. and M. Schnitzer. Chemical model structures for soil organic
matter and soils. Soil Sci. 162: 115-130, 1997.
310. Celi, L., M. Gennari, S.U. Khan and M. Schnitzer. Extractable and non-
extractable (bound) residues of scifluorfen in an organic soil. Arg. Foof
Chem. 45: 36773680, 1997.
311. Celi, L., M. Gennari, S.U. Khan and M. Schnitzer. Mechanism of acifluorfen
interaction with humic acid. Soil Sci. Soc. Am. J. 61: 1659-1665, 1997.
312. Schulten, H.-R., C. Sorge-Lewin and M. Schnitzer. The structure of “unknown”
soil nitrogen investigated by analytical pyrolysis. Biol. Fert. Soils 24:
249-254, 1997.
313. Schulten H.-R. and M. Schnitzer. The chemistry of soil nitrogen: a review.
Biol. Fert. Soils 26: 1-15, 1998.
314. Schulten H.-R., P. Leinweber and M. Schnitzer. Analytical pyrolysis and
computer modeling of humic and soil particles, in Environmental Particles,
Vol. 4, IUPAC, 281-324, 1998.
510 Wolf Prize in Agriculture
315. Liang, B.C., A.F. MacKenzie, M. Schnitzer, C.M. Monreal, P.R. Voroney.
Management induced chemical change in labile organic matter under
continuous corn in eastern Canadian soils. Biol. Fert. Soils 26: 88-94, 1998.
316. Pare, T., H. Dinel, M. Schnitzer and S. Dumontet. Transformation of carbon
and nitrogen during composting of animal manure and shredded paper. Bio.
Fert. Soils 26: 173-178, 1998.
317. Dinel, H., C.M. Monreal and M. Schnitzer. Extractable lipids and organic
matter status as influenced by tillage in two catenas. Geoderma 86:
279-293, 1998.
318. Dinel, H. M. Schnitzer and H.-R. Schulten. Chemical and spectroscopic
characteristics of colloidal fractions separated from liquid pig manures. Soil
Sci. 163: 665-673, 1998.
319. Arshad, C., M. Schnitzer and H.-R. Schulten. Effect of high magnetic field on
the chemical composition of soil organic matter. World Congr. Soil Sci.,
France, Symp. 7, No. 1128: 1-4, 1998.
320. Liang, B.C., E.G. Gregorich, A.F. MacKenzie, M. Schnitzer, P.R. Voroney C.M.
Monreal and R.P. Beyaert. Reaction and turn-over of corn residue carbon in
eastern Canadian soils. Soil Sci. Soc. Am. J. 62: 1361-1366, 1998.
321. Pare, T., H. Dinel and M. Schnitzer. Extractability of trace metals during
co-composting of biosolids and municipal solid wastes. Biol. Fert. Soils 29:
31-37, 1999.
322. Dinel, H., M. Schnitzer, T. Pare, L. Lemee, A. Ambles, E. Topp and N. Pelzer.
Effects of direct land application of calcitic lime, and lime- and cement kiln
dust-sensitized biosolids on the chemical and spectroscopic characterization
of soil lipids. Soil Sci. 164: 322-330, 1999.
323. Dinel, H., T. Pare, M. Schnitzer and N. Pelzer. Direct land application of
cement kiln dust and lime-sensitized biosolids: extractability of trace metals
and organic matter quality. Geoderma 96: 307-320, 2000.
324. Schnitzer, M. A lifetime perspective on the chemistry of soil organic matter.
Adv. Agron. 68: 1-58, 2000.
325. Dinel, H., T. Pare and M. Schnitzer. Carbon and nitrogen mineralization in
soil amended with non-tabletized and tebletized poultry manure. Can. J. Soil
Sci. 80: 271-276, 2000.
326. Schnitzer, M. The in-situ analysis of organic matter in solids. Can. J. Soil. Sci.
81: 249-254, 2001.
327. Schnitzer, M., H. Dinel, H.-R. Schulten, T. Pare and S. Lafond. Humification
of duck farm wastes in Humic Substances, Versatile Components of Plants,
Soils and Waters (edt. by E.A. Ghabbour and G. Davies), pp 21-35, Royal
Society of Chemistry, Cambridge, UK, 2000.
328. Dumontet, S., H. Dinel, M. Schnitzer, T. Pare and A. Scopa. Composting
organic residues: trace metals and microbial pathogens. Can. J. Soil. Sci. 81:
357-367, 2001.
Morris Schnitzer 511
329. Dinel, H., M. Schnitzer, T. Pare, L. Lemee, A. Ambles and S. Lafond. Changes in
lipids and sterols during composting. J. Environ. Sci. Health B 36: 651-665,
2001.
330. Tudoret, M.-J., M. Schnitzer and H. Dinel. Some aspects of organic and
analytical chemistry of lignite-like materials: A literature review. Agriculture
and Agri-Food Canada, 2000.
331. Saharinen, M., H. Dinel and M. Schnitzer. Effects of humic substances on
plants and soil properties. Agriculture and Agri-Food Canada, 2000.
332. Lafond, S., T. Pare, H. Dinel, M. Schnitzer, J. Chambers and A. Jaonich.
Composting duck excreta-enriched wood shavings: C and N transformations
and bacterial pathogen reductions. J. Environ. Sci. Health B 35: 651-665,
2000.
333. Chen, Y., J. Katan, A. Gamliel, T. Aviad and M. Schnitzer. Involvement of
soluble organic matter in increased plant growth in solarized soils. Biol. Fert.
Soils 32: 28-34, 2000.
334. Schnitzer, M. the chemistry of nitrogen in soils (in E. Keinan and I. Schechter
eds.) chemistry for the 21st century. pp 117-128, Wiley-VCH, Weinheim,
2001.
335. Ozdoba, D.M., J. C. Blyth, K.F. Eugler, H. Dinel and M. Schnitzer. Leonadite
and humified organic matter in Humic Substances: Structures, Models and
Functions (edt. by E.A. Ghabour and G. Davies), pp 309-313, Royal Society
of Chemistry, Cambridge, UK, 2001.
336. Schnitzer, M., H. Dinel, T. Pare, H.-R. Schulten and D. Ozdoba. Some chemical
and spectroscopic characteristics of six organic ores, in Humic Substances:
Structures, Models and Functions (edt. by E.A. Ghabour and G. Davies),
pp 315-328, Royal Society of Chemistry, Cambridge, UK, 2001.
337. Dinel, H., M. Schnitzer, H.-R. Schulten, D. Ozdoba and T. Marche. Interaction
by principal component analysis of pyrolysis-field ionization mass spectra of
lignite ores, Humic Substances: Structures, Models and Functions (edt. by
E.A. Ghabour and G. Davies), pp 315-328, Royal Society of Chemistry,
Cambridge, UK, 2001.
338. Pare, T., M. Saharinen, M.J. Tucloret, H. Dinel, M. Schnitzer and D. Ozdoba.
Response of alfalfa to calcium lignite fertilizer, in Humic Substances:
Structures, Models and Functions (edt. by E.A. Ghabour and G. Davies),
pp 315-328, Royal Society of Chemistry, Cambridge, UK, 2001.
339. Dinel, H., M. Schnitzer, T. Pare, L. Lemee, A. Ambles, and S. Lafond. Changes
in lipids and sterols during composting. J. Environ. Sci. Health B 36(5):
651-665, 2001.
340. McArrthur, D.F.E., H.-R. Schulten, M. Schnitzer, L.M. Kozak and P.M. Huang.
Impact of long term cultivation on the status of organic matter in selected
Canadian Prerie Soils. Proceedings of the 17th World Congress of Soil Science,
Bangkok, 2002.
512 Wolf Prize in Agriculture
353. Schnitzer, M., C.M. Monreal and G. Jandl. The conversion of chicken manure
to bio-oil by fast pyrolysis III. Analysis of chicken manure, bio-oils and char
by Py-FIMS and Py-FDMS. J. Environ. Sci. Health B, 43, 81-95, 2008.
354. Munoz, C., C.M. Monreal, M. Schnitzer and E. Zagal. Influence of Acacia
Caven (mol) coverage on carbon distribution in soil organic carbon fractions
in a Mediterranean climate region. Geoderma, 144: 352-360, 2008.
355. Das, D.D., M. Schnitzer, C.M. Monreal and P. Mayer. Acid-base separation of
bio-oil derived by fast pyrolysis of chicken manure. Bioresource Technology
(In press) 2008.
for humins and whole soils in the solid state. Valuable information on the chemical
structure of humic substances can be obtained by combining 13C NMR with
chemical methods. In this manner, effects on the chemical structure of humic
substances of different extractants, methylations, hydrolysis, oxidation and reduction
can be evaluated.
Electron spin resonance measurement of humic substances showed that these
materials are rich in free radicals (unpaired electrons). These free radicals participate
in inorganic-organic and organic-organic interactions. From the electron spin
resonance spectra it appears that the prominent free radicals in humic substances
are semiquinones or substituted semiquinones. We found two types of free radicals
in humic substances: (a) permanent ones with long lifetimes; and (b) transient free
radicals with relatively short lives (several hours). Transient free radicals in humic
materials can be generated by chemical reduction, irradiation or increase in pH.
Permanent free radicals appear to stabilize the complex chemical substances of
humic materials. Electron spin resonance spectroscopy also provides important
information on the co-ordination and symmetry of paramagnetic metal ions
complexed by humic substances.
During the 1990’s, I collaborated with H.-R. Schulten of the Fresenius Technical
University in Wiesbaden, Germany, on developing methods for the mass
spectrometric analysis of humic substances. We found that pyrolysis-field ionization
mass spectrometry (Py-FIMS) and Curie point-gas chromatography-mass
spectrometry (Cp-GC/MS) were suitable for this purpose. The components of
humic substances, which could be identified by these methods, were: carbohydrates,
phenols, lignin monomers, lignin dimers, n-fatty acids, unsaturated fatty acids,
n-alkanes, alkenes, n-alkyl monoesters, n-alkyl diesters, n-alkyl benzenes,
naphthalenes, phenanthrenes and N- and S-containing compounds. The methods
could be applied to humic acids, fulvic acids, humins, and whole soils. It was no
longer necessary to extract the humic substances and de-ashing them prior to
chemical analyses.
One important application of Py-FIMS and Cp-GC/MS was in structural studies
on humic substances. By both methods we found that benzene, substituted benzenes,
and C1 to C13 n-alkyl benzenes were the major products. From these data we
concluded that n-alkylbenzenes were the main structural features in humic
substances and that the aromatic rings were linked covalently by aliphatic chains,
which were broken during pyrolysis. We then developed an averaged two
dimensional model humic acid structure based on n-alkylbenzenes with oxygen
containing functional groups (CO2H, OH, C=O) on the outside. From the two
dimensional structure a three-dimensional structure was derived with the aid of
computational chemistry. Throughout our structural work, we saw to it that our
model humic acid structure was in harmony with chemical, colloid-chemical,
electron microscopic, 13C NMR, X-ray and mass spectrometric data, which my
516 Wolf Prize in Agriculture
co-workers and I had obtained over a period of many years as well as with
exhaustive consultations of the voluminous literature. One of the main features of
our model humic acid structure is that it is perforated by voids of various
dimensions, which can trap or retain both inorganic and organic molecules such
as clays, pesticides, herbicides, carbohydrates and peptides. Another significant
point is that we do not consider carbohydrates and proteinaceous material
components of the humic acid structure. We believe that the latter are either
retained by the voids or adsorbed on the humic acid surfaces. The reason for this is
that these compounds are rapidly removed by acid hydrolysis, leaving the humic
acid structure intact. Our model structure contains relatively large numbers of
CO2H and OH groups so that there are many opportunities for interacting with
metal ions, metal oxides and minerals. There are also many possibilities for the
formation of both inter and intra-molecular H-bonds. So far we have developed
model structures for humic acids, soil organic matter and a whole soil. As new
information on these materials becomes available, it may be necessary to modify
and refine the model structures. We hope that our research on model soil organic
matter structures will be beneficial to the development of a sustainable agriculture
and to the protection of the environment.
To obtain information on the “unknown” N in soils and humic substances, my
co-workers and I used Py-GC/MS, with the gas chromatograph being equipped
with a N-selective detector. This means that only N-containing compounds
were transferred to the mass spectrometer for identification. Employing this
approach, we identified over 100 N-containing compounds, many of which were
N-heterocyclics. The data included pyrroles, pyrrolidines, imidazoles, pyridines,
pyrazines, indoles, quinolines, benzothiazoles, and pyrimidines. In another invention
with scientists at the University of Saskatchewan we studied the catalysis of the
Maillard reaction by δ-MnO2 and found that N-heterocyclics could be formed
abiotically in this manner.
After the year 2000, I worked only half time and joined a group of scientists at
Agriculture and Agri-Food Canada here in Ottawa, who worked on composting of
all kinds of organic residues. My job was to elucidate the chemical reactions,
which were controlling the decomposition of the organics. One of the interesting
conclusions of this research was that the biological and chemical reactions during
composting focused mainly on the degradation of the aliphatic structures while
the aromatic structures were preserved. In recent years I have worked on the
chemistry of bio-oils produced by the fast pyrolysis of chicken manure. The bio-
oils produced in this manner contained at least 500 organic compounds. So far we
have identified by 13C NMR and by mass spectrometric methods about 350 of
these compounds.
My lifetime research was concerned with the chemistry and reactions of soil
humic substances and with many N-containing compounds in agricultural soils. I
Morris Schnitzer 517
spent close to 50 years on this research. During that time I had many co-workers.
These were 30 postdoctoral fellows, from 15 different countries, 10 technicians,
and 20 scientists from Canada and outside Canada. Each person who worked with
me became a co-author of the paper reporting the research, which he or she did
with me (see list of publications). I am most grateful to each of them for their
contributions. I learned as much from them as they learned from me.
Finally, let us remember that the soil is our most important resource. It is the
presence of organic matter, by acting as a habitat for the microbes, fungi, small
animals, etc., which distinguishes the soil from a mass of rock particles and allows
it to become a living system. Our ability to produce sufficient food for our expanding
population and, at the same time, protect the environment, demands a
comprehensive understanding of the physical, chemical, and biological properties
of the soil and soil organic matter. I hope that the research, which my colleagues
and I have done, will contribute to attaining these objectives.
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Frank J. Stevenson
University of Illinois, Urbana, Illinois, USA
k
CURRICULUM VITAE
519
520 Wolf Prize in Agriculture
and zinc) and toxic heavy metals (i.e., lead and cadmium) with soil organic
substances. For the latter, a continuous distribution model was proposed for the
calculations of stability constants of metal-humate complexes.
Regarding the chemical properties of humic substances, infrared studies on
humic and fulvic acids and their methylated/acetylated derivatives showed that
the acidity of the different reactive groups overlapped; accordingly, methods for
the determining of functional groups based on acidity are not specific and values
obtained there from cannot be used as an absolute measure for functional group
content.
In cooperation with others, research was carried out on the mineralization-
immobilization turnover of biologically and chemically fixed nitrogen in soil, using
the stable isotope nitrogen-15 as a tracer. This research was of importance in that
as much as one-third of the nitrogen applied to temperate zone soils as fertilizer is
retained in organic forms after the growing season and that an equal amount is
lost from the soil-plant system in as yet unexplained ways. Findings of the research
have been of use in modeling fertilizer nitrogen transformations in soil, thereby
leading to improvements in the management of fertilizer nitrogen for increasing
crop yields while at the same time protecting the environment by reducing
movement of nitrates into water supplies.
BOOKS EDITED:
Eliott, L. F. and F. J. Stevenson (Co-Editor). 1976. Soils for Management of Organic
Wastes and Waste Waters. Amer. Soc. Agron., Madison, WI, 650 pp.
F. J. Stevenson (Editor). 1982. Nitrogen in Agricultural Soils. Amer. Soc. Agron.,
pp. 940.
BOOK CHAPTERS:
F. J. Stevenson. 1964. Soil Nitrogen. In: Fertilizer Nitrogen: Its Chemistry and
Technology. Amer. Chem. Soc. Monog. 161, Reinhold, pp. 18-39.
522 Wolf Prize in Agriculture
F. J. Stevenson. 1965. Origin and distribution of nitrogen in soil. In: Soil Nitrogen.
Amer. Soc. Agron., Madison, WI, pp. 1-42.
F. E. Broadbent and F. J. Stevenson.1966. Organic matter interactions. In: Agricultural
Anhydrous Ammonia: Amer. Soc. Agron., Madison, WI, pp. 169-187.
F. J. Stevenson. 1967. Organic acids. In: Soil Bioch. Marcel Dekker, NY,
pp. 119-146.
F. J. Stevenson. 1969. Nitrogen origin and distribution. In: McGraw Hill Yearbook
Science and Technology for 1969, pp. 313-315.
F. J. Stevenson and J. H. A. Butler. 1969. Chemistry of humic acids and related
pigments. In: Organic Geochemistry: Methods and Results. Springer-Verlag,
NY, pp. 534-557.
F. J. Stevenson and G. H. Wagner, 1970, Chemistry of nitrogen in soils.
In: Agricultural Practices and Water Quality. Iowa State Univ. Press, Ames, IA,
pp. 125- 141.
F. J. Stevenson and M. S. Ardakani. 1972. Organic matter interactions involving
micronutrients in soil. In: Micronutrients in Agriculture. Amer. Soc. Agron.,
pp. 79-114.
F. J. Stevenson and A. Fitch. 1981. Reactions with soil organic matter, In: Copper in
Soils and Plants. Academic Press, NY, pp. 69-95.
F. J. Stevenson. 1982. Origin and distribution of nitrogen in soil. In: Nitrogen in
Agricultural Soils. Amer. Soc. Agron., Madison, WI, 1-42. Also: Organic forms,.
pp. 67-122.
F. J. Stevenson. 1983. Trace metal-organic matter interactions in geologic
environments. In: The Significance of Trace Elements in Solving Petrogenetic
Problems and Controversies. Theophrastus Publications, Athens, pp. 671-691.
A. Fitch and F. J. Stevenson. 1983. Stability constants of metal-organic matter
complexes. Ibid., pp. 645-669.
F. J. Stevenson. 1985. Geochemistry of soil humic substances. In: Organic
Geochemistry: Vol. 1, Wiley, pp. 13-52.
F. J. Stevenson. 1985. Nitrogen transformations in soil: A perspective. In: Nitrogen
and the Environment. Nuclear Institute for Agriculture, Faisaisbad, Pakistan,
pp. 7-26.
F. J. Stevenson and K. A. Kelley. 1985. Stabilization, chemical characteristics,.and
availability of immobilized nitrogen in soil. Ibid, pp. 239-259.
Y. Chen and F. J. Stevenson. 1986. Soil organic matter interactions with trace
elements. In: The Role of Organic Matter in Modern Agr. Martinus Nijhoff,
The Netherlands, pp. 73-116.
F. J. Stevenson and A. Fitch. 1986. Chemistry of complexation of metal ions by soil
solution organics. In: Interaction of Soil Minerals with Natural Organics and
Microbes. Amer. Soc. Agron., pp. 29-58.
F. J. Stevenson. 1989. Reductive degradation of humic substances. In: Organic
Geochemistry: Vol. 2, Wiley, pp. 122-142.
Frank J. Stevenson 523
F. J. Stevenson and X-T. He. 1989. Nitrogen in humic substances as related to soil
fertility. In: Advances in Humic Substances Research Related to Soil and Crop
Sciences. Soil Sci. Soc. Amer. Special Pubi., pp. 91-110.
F. J. Stevenson and G. F. Vance. 1989. Naturally occurring aluminum-organic
complexes. In: The Environmental Chemistry of Aluminum. CRC Press, Boca
Raton, FL, pp. 117-146.
F. J. Stevenson and E. T. Elliott. 1990. Methodologies for assessing organic matter
quality. In: Dynamics of Organic Matter in Tropical Soils. Univ. Hawaii Press,
173-199
F. J. Stevenson. 1991. Organic matter-micronutrient reactions in soils. In:
Micronutrients in Agriculture. 2nd Ed., Amer. Soc. Agron., pp. 145-186.
G. F. Vance, F. J. Sikora, and F. J. Stevenson. 1995. Aluminum-organic matter
complexes. In: The Environmental Chemistry of Aluminum. CRC Press, Boca
Raton, FL.
K. A. Kelley and F. J. Stevenson, 1996. Organic forms of Soil N. In: Humic Substances
in Agricultural Ecosystems. Elsevier, Chapter 10.
RESEARCH ARTICLES:
Over 100 articles in a wide variety of research journals. These papers are cited in
reviews and book chapters, as noted above.
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Neal L. First
University of Wisconsin
Madison, Wisconsin, USA
k
Professor Neal L. First has made pioneering contributions to animal genetics by the
development of systems of bovine embryo cloning, gene transfer, and
in-vitro production of livestock embryos. His research has resulted in major advances
in the application of biotechnology to reproduction in farm animals, eliminating
the need for brood cows in beef cattle breeding.
Nuclear transfer of genetic material is being developed to clone farm animal
embryos. His ground-breaking research in this field has led to significant discoveries
in reproductive biology. Prof. First’s laboratory pioneered the application of this
technique to the genetic selection of livestock. He and his colleagues defined the
conditions leading to optimal in-vitro fertilization and high frequency embryo
development. They developed the defined growth media that are widely used to
culture bovine embryos. The procedures developed in Prof. First’s laboratory have
led to major discoveries throughout the world concerning embryo development in
domestic animals.
The new knowledge provided by Prof. First’s research is having a major impact
on the production of genetically advanced strains of livestock throughout the
world. Advances include improved reproduction, growth, lactation, and disease
resistance.
525
526 Wolf Prize in Agriculture
CURRICULUM VITAE
Date and Place of Birth: October 9, 1930, Michigan
EDUCATION:
1948-1952 - B.S. Michigan State College, Animal Husbandry
1954-1957 - M.S. Michigan State University, Animal Science
1957-1959 - Ph.D. Michigan State University, Animal Science and Physiology of
Reproduction (Ph.D. Thesis Title: Fertility of Frozen Ram Semen)
PROFESSIONAL EXPERIENCE:
1956-1960 - Instructor, Michigan State University-East Lansing.
1960-1964 - Assistant Professor, Department of Meat and Animal Science,
University of Wisconsin-Madison.
1964-1968 - Associate Professor, Department of Meat and Animal Science,
University of Wisconsin-Madison.
1968-present - Member, Endocrinology-Reproductive Physiology Program,
University of Wisconsin.
1968-present - Professor, Department of Meat and Animal Science, University of
Wisconsin-Madison.
1987-1990 - Joint Appointment in Dept. of Obstetrics and Gynecology, Medical
School.
1990 - Director USDA-CSRS-ARS National Animal Genome Mapping
Program.
NATIONAL COMMITTEES:
Acting Director, National Program in Mapping the Genome of Domestic Animals,
1992
National Academy of Science, Institute of Laboratory Animal Research,
1991-1993
NAS Institute of Medicine Committee on Fetal Research and Application,
1992-1993
NAS Class Membership Committee, 1992-1993
National Advisory Board on Ethics in Reproduction, 1994
in vitro fertilize in vitro matured oocytes and the first to define many of the
conditions for efficient IVF such as temperature and timing.
III. In embryo culture his laboratory was the first to characterize the in vitro block
to bovine embryo development, to describe the timing of bovine embryonic cell
cycles and to describe the time of initiation of embryonic transcription, and
the mechanisms initiating bovine embryonic transcription. His students were
the first to show that bovine embryos could be cultured past the period of
blocked development by co- culture with oviduct epithelial cells or their
conditioned media. His laboratory then developed a defined medium CR1aa
that is widely used to culture bovine embryos.
Together these discoveries led to an efficient system for producing embryos in vitro
that is being used to eliminate the brood cow in beef cattle breeding, to propagate
valuable animals and to make possible more efficient nuclear and gene transfer.
An explosion of information concerning early development and differentiation in
domestic animals has resulted in laboratories around the world, stemming directly
from the systems developed by Dr. First’s laboratory. Dr. First’s laboratory was also
original in developing cloning of cattle embryos and pigs. It was the first to produce
offspring from cultured bovine embryonic stem cells. These two technologies when
combined with in vitro production of embryos and homologous DNA recombination
provide the basis for effective gene transfer or gene deletion in cattle. The enormous
impact on research and commercial animal production will long be felt due to the
work of Dr. First and his laboratory for developing systems necessary to the in
vitro production of bovine embryos.
SELECTED PUBLICATIONS
Schoff, P.K. and N.L. First. 1995. Manipulation of bovine sperm metabolism and
motility using anoxia and phosphodiesterase inhibitors. Cell Motility and the
Cytoskeleton 31:140-146.
Jones, J.M. and N.L. First. 1995. Expression of cell cycle control protein cdc25 in
cleavage stage bovine embryos. Zygote. 3: May, 1995.
Hageman, L.J., Hillery, F.L., Leibfried-Rutledge, M.L. and First, N.L. 1994. Activation
of murme oocytes with Ca2-i- ionophore and cycloheximide. J. Exper. Zool.
271:57-61.
Susko-Parrish, J., Leibfried-Rutledge, M.L., Northey, D., Schutzkus, V. and First, N.L.
1994. Inhibition of protein kinases after an induced calcium transient causes
transition of bovine oocytes to embryonic cycles without meiotic completion.
Developmental Biology. 166: 729-739.
Rosenkrans, C.F. and First, N.L. 1994. Effect of free amino acids and vitamins on
cleavage and developmental rate of bovine zygotes in vitro. J. Anim. Sci. 72:
434-437.
Neal L. First 529
First, N.L., Sims, M.M., Park, S.P., and Kent-First, M.J. 1994. Systems for production
of calves from cultured bovine embryonic cells. Reprod. Fertil. and Dev. 6:
553-562.
Sims, M.M. and First, N.L. 1994. Production of calves by transfer of nuclei from
cultured inner cell mass cells. Proc. Natl. Acad. Sci. USA 90: 6143-6147.
Navara, C.S., First, N.L. and G. Schatten. 1994. Microtubule organization in the
cow during fertilization, polyspermy, paerthenogenesis and nuclear transfer:
The role of the sperm aster. Dev. Biol. 162:29-40.
Sims, M.M., and First, N.L. 1994. Production of calves by transfer of nuclei from
cultured inner cell mass cells. Proc. Natl. Acad. Sci. USA 90:6143-6147.
First, N.L., and Leibfried-Rutledge M.L. 1993. Nuclear Transfer in Mammals. lit “In
vitro fertilization and Embryo Transfer in Primates” (D.P. Wolf, Stouffer R.L.,
and Brenner, R.M., Eds.), Springer-Verlag New York, Inc. 3 17-330.
Fulka Jr., J., Leibfried-Rutledge M.L, and First, N.L. 1993. Control of germinal
vesicle breakdown in bovine x murine hybrid oocytes. Reprod. Nutr. Dev.
33:411-417.
Kim, T., M.L. Leibfried-Rutledge and First, N.L. 1993. Gene transfer in bovine
oocytes using replication-defective retroviral vectors packaged with Gibbon
ape leukemia virus envelopes. Molec. Reprod. Dev. 35: 105-113.
Kim, T., M.L. Leibfried-Rutledge and First, N.L. 1993. Gene transfer in bovine
oocytes using replication-defective retroviral vectors packaged with Gibbon
ape leukemia virus envelopes. Molec. Reprod. Dev. 35: 105-113.
Prather, R.S. and First, N.L. 1993. Cell-to-cell coupling in early-stage bovine embryo:
a preliminary report. Theriogenology 39:561-567.
Takahashi, Y. and First, N.L. 1993. In vitro fertilization of bovine oocytes in the
presence of theophylline. Anim. Reprod. Sci. 34:1-8.
Takahashi, Y. and First, N.L. 1993. In vitro culture of bovine one-cell embryos
fertilized in vitro using synthetic oviduct fluid medium supplemented with
fetal calf serum. Anim. Reprod. Sci. 3 1:33-47.
Rosenkrans, Jr., C.F., G.Q. Zeng, G.T. McNamara, P.K. Schoff and First, N.L. 1993.
Development of bovine embryos in vitro as affected by energy substrates.
Biol. Reprod. 49:459-462.
Rosenkrans, Jr., CF., and First, N.L. 1993. Effect of free amino acids and vitamins
on cleavage and developmental rate of bovine zygotes in vitro. J. Anim. Sci.
72:434-437.
Yom, H.C., R.D. Bremel and First, N.L. 1993. Mouse mammary tumor virus promoter
directs high-level expression of bovine S1 casein in the milk of transgenic
heterozygous and homozygous mice. Anim. Biotech. 4(1):89-107.
Hagemann, L.J. and First, N.L. 1992. Embryonic cytoplasmic extracts rescue murine
androgenones to the blastocyst stage. Development 114(4): 997-1001.
530 Wolf Prize in Agriculture
Patents
Co-culture of embryos with other cell types
CRlaa embryos cultural system
Nuclear transfer process
Patents Pending
A. Bovine embryonic stem cell derivation
B. Production of bovine parthenotes
Ilan Chet
Faculty of Agriculture
The Hebrew University of Jerusalem
Rehovot, Israel
k
CURRICULUM VITAE
PERSONAL
Date and place of birth: April 12, 1939, Haifa, Israel.
Nationality: Israeli.
Marital status: Married, with five children.
531
532 Wolf Prize in Agriculture
UNIVERSITY DEGREES
1962 - B.Sc. with honors. The Hebrew University of Jerusalem, Faculty of
Agriculture, Rehovot.
1964 - M.Sc. with honors. The Hebrew University of Jerusalem.
1968 - Ph.D. The Hebrew University of Jerusalem.
Thesis: The structure and behaviour of the fungus Sclerotium rolfsii.
1988 Invited speaker to the VII International Congress on the Global Impact
of Microbiology (GIAM), Hong Kong
1988 Invited speaker to the International Congress of Plant Pathology,
Kyoto, Japan
1988 Invited speaker to the Annual Meeting of the British Society of Plant
Pathology, Reading, England
1989 Invited speaker to the UCLA Symposium on Molecular and Ecological
Aspects of Biological Control, Frisco, Colorado
1989-1993 Chairman, The Special Projects Committee of the International
Society for Plant Pathology (ISPP)
1989-1994 Member of the Scientific Advisory Committee of the Institute for
Cereal Crops Improvement. Tel Aviv University
1989 Invited speaker to the Biotechnology Workshop, Rutgers University,
New Jersey, USA
1989 Invited speaker to the Beltsville Symposium on the Rhizosphere and
Plant Growth, Beltsville, USA
1990 Invited speaker to the Fourth International Mycological Congress,
Regensburg, F.R.G.
1990 Invited speaker to the German Botanical Society Meeting, Regensburg,
F.R.G.
1990 Invited speaker to the Workshop on Biological Control of Postharvest
Diseases, Sheperdstown, West Virginia, USA
1990- Member, International Editorial Board, Crop Protection
1991 Invited speaker to the XII International Plant Protection Congress,
Rio de Janeiro, Brazil
1991 Invited speaker to the International Symposium on Application of
Biotechnology to tree culture protection and utilization. Columbus,
Ohio, USA
1991- Advisory Editor, Wiley Series in Ecological and Applied Microbiology
1992 Invited overview speaker to the International Seminar on “Impact
of Biotechnology on Agriculture and Food in Developing Countries”,
Madras, India
1992 Teaching Course, University of Torino, Italy
1992 Invited speaker, The Center of Bioengineering, Academy of Sciences
of Russia
1992-1994 Member, The Committee for Reduction of Pesticides, Ministry of
Agriculture
1993-1999 Member, The Scientific Committee of “Med-Campus” of the
EEC
1993 Invited speaker, International Conference on Agricultural and
Environmental Biotechnology, Torino, Italy
Ilan Chet 537
GRANTS
Since 1973, Professor Chet has received research grants from the following:
Agencies and foundations:
Israel Ministry of Agriculture
Deutsche Forschungsgemeinschaft (DFG)
National Council for Research and Development (NCRD)
Federal German Ministry for Science and Technology (BMFT)
Ministry of Niedersachsen for Science, Germany
Binational Agricultural Research and Development Fund (BARD)
The Wolfson Foundation, England
Agency for International Development (AID)
Kay Foundation for Biotechnology
The Szold Foundation
GIFRID Foundation, Germany
German-Israeli Agricultural Research and Development (GIARA)
German-Israeli Foundation (GIF)
Volkswagen Foundation (VW)
Dutch-Israel Agricultural Foundation (DIAF)
Charles Wolfson Charitable Trust
The Chais Family Charitable Fund
European Union (E.U)
Belgian Trilateral Governmental Fund
Federal German Ministry for Education and Science (BMBF)
Horowitz Association
Industry:
Teva Pharmaceutical Industry
Galil Advanced Technologies
Bio Technology General Ltd. (BTG)
Gadot Petrochemical Industries
Biotechnology Application (BA)
Makhteshim
Migal, Israel
FRM Agricultural Sciences Partnership
Ilan Chet 541
Yissum, Israel
Ecogen, USA
Ieves Rocher, France
Israel Biotechnology Research (IBR)
Haifa Chemicals
Mycontrol
Biotechnological Integrated Technologies BIT, (Italy)
Algatech
PATENTS
1. Henis, Y. and I. Chet. 1971. Method for drug detection. Israel Patent 38,233.
2. Mitchell, R. and I. Chet. 1977. Magnetic separation method. U.S. Patent
4,00-197.
3. Sivan, A., I. Chet and Y. Elad. 1985. Fungicidal compositions and method for
using them. Israel Patent 68124.
4. Sivan, A. and I. Chet. 1985. Antifungal compositions containing Trichoderma
active against Fusarium. Israel Patent 69368.
5. Chet, I., A. Sivan, and Y. Elad. 1985. Novel isolates of Trichoderma fungicidal
compositions containing said isolates and use thereof. European Patent 133878.
6. Shoseyov, O., B.A. Bravdo, R. Ikan and I. Chet. 1987. Production and utilization
of a specific endo-beta-glucosidase for food, wine and perfume industries.
Israel Patent No. 82980-2, 1987, U.S. Patent Pending, 1988.
7. Chet, I. and Sivan, A. 1988. Antifungal compositions containing Trichoderma
active against Fusarium. US Patent 4,784,021.
8. Oppenheim, A., I. Chet and R. Shapira. 1988. A system for excretion of
foreign protein, enzymes or polypeptides from E. coli into the culture media,
especially for the production and purification of the enzyme chitinase. Israel
Patent Pending 85408/3.
9. Eyal, Z., I. Chet and M. Fleishman. 1988. Method of controlling foliar diseases
caused by fungal pathogens with fungicidal bacteria and novel pure cultures
of fungicidal bacteria. Israel Patent Pending 87322.
10. Spiegel, I., I. Chet and E. Cohn. 1988. Method and composition for combating
soil nematodes comprising chitinolytic bacteria and novel pure cultures of
certain such bacteria. Israel Patent 87388.
11. Spiegel, I., I. Chet, Cohen, E. and Galper, S. 1989. Nematocidal microorganisms,
their isolation and their use as bionematocides. European Patent 89114460.2.
12. Eyal, Z., I. Chet, M. Fleishman and E. Levy. 1989. Method of controlling foliar
diseases caused by fungal pathogens with fungicidal bacteria and novel pure
cultures of fungicidal bacteria. European Patent No. 89114108.7.
13. Chet, I., A. Sivan and Y. Elad. 1990. A novel isolate of Trichoderma fungicidal
compositions containing said isolate and use thereof. U.S. Patent 4,915,944.
542 Wolf Prize in Agriculture
LIST OF PUBLICATIONS
I. Books
1. Chet, I. 1967. The structure and behaviour of the fungus Sclerotium rolfsii.
Ph.D. Thesis. Supervised by Prof. Y. Henis.
2. Huttermann, A. and I. Chet (eds.). 1986. Soil-Plant Interaction. Gottingen
Bodenkundische Berichte, FRG, pp. 269.
3. Chet, I. (ed.). 1987. Innovative Approaches to Plant Disease Control. John
Wiley & Sons, N.Y. pp. 372.
4. Chet, I. (ed.) 1993. Biotechnology in Plant Disease Control. John Wiley &
Sons, N.Y. pp. 373.
22. Chet, I.S. Fogel and R. Mitchell. 1971. Chemical detection of microbial prey
by bacterial predators. J. Bacteriol. 106:863-867.
23. Huttermann, A. and I. Chet. 1971. Activity of some enzymes in Physarum
polycephalum III. During spherulation (differentiation) induced by mannitol.
Arch. Microbiol. 78:180-192.
24. Mitchell, R., S. Fogel and I. Chet. 1972. Bacterial chemoreception: an
important ecological phenomenon inhibited by hydrocarbons. Water Research
6:1137-1140.
25. Okon, Y., I. Chet and Y. Henis. 1972. Lactose-induced synchronous sclerotium
formation in Sclerotium rolfsii and its inhibition by ethanol. J. Gen. Microbiol.
71:465-470.
26. Assa, Y., B. Gestetner, I. Chet and Y. Henis. 1972. Fungistatic activity
of lucerne saponins and digitonin as related to sterols. Life Sciences
11:637-647.
27. Chet, I., N. Retig and Y. Henis. 1972. Changes in total soluble proteins and in
some enzymes during morphogenesis of Sclerotium rolfsii. J. Gen. Microbiol.
72:451-456.
28. Chet, I. and Y. Henis. 1972. The response of two types of Sclerotium rolfsii
to factors affecting sclerotium formation. J. Gen. Microbiol. 73:483-486.
29. Okon, Y., I. Chet and Y. Henis. 1973. Effect of lactose, ethanol and
cycloheximide on the translocation pattern of radioactive compounds and on
sclerotial formation in Sclerotium rolfsii. J. Gen. Microbiol. 74:251-258.
30. Chet, I., N. Retig and Y. Henis. 1973. Changes in ribonucleases during
differentiation (spherulation) of Physarum polycephalum. Biochim. Biophys.
Acta 294:343-347.
31. Kislev, N. and I. Chet. 1973. Scanning electron microscopy of sporulating
cultures of the myxomycete Physarum polycephalum. Tissue and Cell
5:349-357.
32. Chet, I., Y. Henis and R. Mitchell. 1973. The effect of biogenic amines and
cannabinoids on bacterial chemotaxis. J. Bacteriol. 115:1215-1218.
33. Tirosh, R., A. Oplatka and I. Chet. 1973. Motility in a “cell sap” of the slime
mold Physarum polycephalum. FEBS Letters 34:40-42.
34. Chet, I., Y. Zilberstein and Y. Henis. 1973. Chemotaxis of Pseudomonas
lachrymans to plant extracts and to water droplets collected from the
leaf surfaces of resistant and susceptible plants. Physiol. Plant Pathol.
3:473-479.
35. Henis, Y., Y. Okon and I. Chet. 1973. The relationship between early hyphal
branching and formation of sclerotia in Sclerotium rolfsii. J. Gen. Microbiol.
79:147-150.
36. Chet, I. and N. Kislev. 1973. Scanning electron microscopy of spherules of
Physarum polycephalum. Tissue and Cell 5:545-551.
546 Wolf Prize in Agriculture
37. Okon, Y., I. Chet, N. Kislev and Y. Henis. 1974. Effect of lactose on soluble
glucan production and on the ultrastructure of Sclerotium rolfsii Sacc. grown
in submerged culture. J. Gen. Microbiol. 81:145-149.
38. Kislev, N. and I. Chet. 1974. Scanning electron microscopy of freeze-fractured
sclerotia of Physarum polycephalum. Tissue and Cell 6:209-214.
39. Retig, N. and I. Chet. 1974. Catechol-induced resistance of tomato plants to
Fusarium wilt. Physiol. Plant pathol. 4:469-475.
40. Assa, Y., I. Chet, B. Gestetner, R. Govrin, Y. Birk and A. Bondi. 1975. The
effect of alfalfa saponins on growth and lysis of Physarum polycephalum.
Arch. Microbiol. 103:77-81.
41. Mitchell, R. and I. Chet. 1975. Bacterial attack of corals in polluted seawater.
Microbiol. Ecol. 2:227-233.
42. Okon, Y., I. Chet and Y. Henis. 1975. The effect of glucose and lactose on
β-D-galactosidase activity and formation of sclerotia in Sclerotium rolfsii.
Can. J. Microbiol. 21:1123-1126.
43. Lisker, N., J. Katan, I. Chet and Y. Henis. 1975. Release of cell-bound
polygalacturonase and cellulase from mycelium of Rhizoctonia solani. Can.
J. Microbiol. 21:521-526.
44. Chet, I. 1975. Ultrastructural basis of sclerotial survival in soil. Microbiol.
Ecol. 2:194-200.
45. Spiegel, S., Y. Henis, I. Chet and G. Messer. 1975. The effect of heat shock on
differentiation of germinating conidia of Trichoderma viride. Can. J. Bot.
53:2274-2281.
46. Chet, I., P. Asketh and R. Mitchell. 1975. Repulsion of bacteria from marine
surfaces. Appl. Microbiol. 30:1043-1045.
47. Chet, I. and R. Mitchell. 1976. Petroleum hydrocarbons inhibit decomposition
of organic matter in seawater. Nature 261:308-309.
48. Kritzman, G., Y. Okon, I. Chet and Y. Henis. 1976. L-threonine metabolism
in Sclerotium rolfsii and it relationship to morphpgenesis. Isr. J. Med. Sci.
12:696-697.
49. Kritzman, G., Y. Okon, I. Chet and Y. Henis. 1976. Metabolism of
L-threonine and its relationship to sclerotium formation in Sclerotium rolfsii.
J. Gen. Microbiol. 95:78-86.
50. Chet, I. and R. Mitchell. 1976. An enrichment technique for isolation of
marine chemotactic bacteria. Microbiol. Ecol. 3:75-78.
51. Chet, I. and R. Mitchell. 1976. The relationship between chemical structure
of attractants and chemotaxis by a marine bacterium. Can. J. Microbiol.
22:1206-1208.
52. Segal, L. A., I. Chet and Y. Henis. 1977. A simple quantitative assay for
bacterial motility. J. Gen. Microbiol. 98:329-337.
53. Chet, I. and A. Huttermann. 1977. Germination inhibitor in the slime mold
Physarum polycephalum. FEMS Microbiol. Letts. 1:149-152.
Ilan Chet 547
54. Chet, I., D. Timar and Y. Henis. 1977. Physiological and ultrastructural changes
occurring during germination of sclerotia of Sclerotium rolfsii. Can. J. Bot.
55:1137-1142.
55. Kritzman, G., I. Chet and Y. Henis. 1977. Effect of carbon dioxide on
growth and carbohydrate metabolism in Sclerotium rolfsii. J. Gen. Microbiol.
100:167-175.
56. Kritzman, G., I. Chet and Y. Henis. 1977. The relationship between rhythmic
hyphal growth and circadian formation of sclerotia in Sclerotium rolfsii.
Sacc. Can. J. Microbiol. 23:959-963.
57. Chet, I. and A. Huttermann. 1977. Melanin biosynthesis during differentiation
of Physarum polycephalum. Biochem. Biophys. Acta 499:148-155.
58. Chet, I., A. Naveh and Y. Henis. 1977. Chemotaxis of Physarum polycephalum
towards carbohydrates, amino acids and nucleotides. J. Gen. Microbiol.
102:145-148.
59. Chet, I., A. Naveh and Y. Henis. 1977. Chemotaxis and movement of Physarum
polycephalum and its response to some neurotransmitters and psychomimetic
compounds. J. Mechanochem. and Cell Motility 4:177-187.
60. Chet, I., D. Havkin and J. Katan. 1977. The role of catechol in inhibition of
Fusarium wilt. Phytopathol. Z. 91:60-66.
61. Kritzman, G., I. Chet and Y. Henis. 1977. The role of oxalic acid in the
pathogenic behaviour of Sclerotium rolfsii Sacc. Exper. Mycol. 1:280-285.
62. Huttermann, A., Y. Henis and I. Chet. 1978. Okologische Modellversuche in
der Petrischale. Biologie Uns. Z. 8:57-60.
63. Kritzman, G., I. Chet and Y. Henis. 1978. Localization of β-(1,3)-glucanase
in mycelium of Sclerotium rolfsii. J. Bacteriol. 134:470-475.
64. Kritzman, G., I. Chet, Y. Henis and A. Huttermann. 1978. The use of the
brightener “calcofluor white M2R new” in the study of fungal growth. Isr. J.
Bot. 27:138-146.
65. Sjoblad, R.D., I. Chet and R. Mitchell. 1978. Chemoreception in the green
alga Danaliella tertiolecta. Curr. Microbiol. 1:305-308.
66. Hadar, Y., I. Chet and Y. Henis. 1979. Biological control of Rhizoctonia
solani damping-off with wheat bran culture of Trichoderma harzianum.
Phytopathology 69:64-68.
67. Netzer, D., G. Kritzman and I. Chet. 1979. β-(1-3) glucanase and quantity of
the fungus in relation to Fusarium wilt in resistant and susceptible near
isogenic lines of musk melon. Physiol. Plant Pathol. 14:47-55.
68. Grinstein, A., Y. Elad, J. Katan and I. Chet. 1979. Control of Sclerotium
rolfsii by means of a herbicide and Trichoderma harzianum. Plant Dis. Reprt.
63:823-826.
69. Ruben, D.M., Z.R. Frank and I. Chet. 1979. Factors affecting behaviour
on developmental synchrony of germinating oospores of Pythium
aphanidermatum. Phytopathology 70:54-59.
548 Wolf Prize in Agriculture
70. Bitton, G., D.A. Chuckran, I. Chet and R. Mitchell. 1979. Resistance of
bacterial chemotaxis to blockage in petroleum waters. Marine Pollution Bull.
10:48-49.
71. Sjoblad, R.D., I. Chet and R. Mitchell. 1979. A quantitative assay for algal
chemotaxis. Appl. Environ. Microbiol. 36:847-850.
72. Kritzman, G., I. Chet and Y. Henis. 1979. Isolation of extracellular
polysaccharides from Sclerotium rolfsii. Can. J. Bot. 57:1855-1859.
73. Huttermann, A., M. Gebauer and I. Chet. 1979. Studies on isoenzyme pattern
during differentiation (spherulation) of Physarum polycephalum. Arch.
Microbiol. 120:113-123.
74. Zweck, S., A. Huttermann and I. Chet. 1978. A convenient method for
preparing inocula of homogenized mycelium. Exper. Mycol. 2:377-378.
75. Chet, I. and A. Huttermann. 1980. Chemical composition of hyphal walls of
Fomes annosus. Eur. J. Forest Pathol. 10:65-70.
76. Elad, Y., I. Chet and J. Katan. 1980. Trichoderma harzianum: A biocontrol
agent effective against Sclerotium rolfsii and Rhizoctonia solani. Phytopathology
70:119-121.
77. Chet, I. and R. Baker. 1980. Induction of suppressiveness to Rhizoctonia
solani in soil. Phytopathology 70:994-998.
78. Stzejnberg, A., Z. Madar and I. Chet. 1980. Induction and quantification of
sclerotia in Rosellinia necatrix. Phytopathology 70:525-527.
79. Elad, Y., J. Katan and I. Chet. 1980. Physical, biological and chemical
control integrated for soilborne diseases in potatoes. Phytopathology
70:418-422.
80. Okon, Y., L. Cakmakci, I. Nur and I. Chet. 1980. Aerotaxis and chemotaxis
of Azospirillum brasilense. Microbial. Ecol. 6:277-220.
81. Paster, N. and I. Chet. 1980. Effect of environmental factors on growth and
sclerotium formation in Aspergillus ochraceus. Can. J. Bot. 58:1844-1850.
82. Kritzman, G., D. Gilan and I. Chet. 1980. Germination inhibitor in Botrytis
allii spores. Phytoparasitica 8:73-76.
83. Kritzman, G. and I. Chet. 1980. The role of phenols in pathogenicity of
Botrytis allii. Phytoparasitica 8:27-37.
84. Harman, G.E., I. Chet and R. Baker. 1980. Trichoderma hamatum effects on
seed and seedling disease induced in radish and pea by Pythium spp. or
Rhizoctonia solani. Phytopathology 70:1167-1172.
85. Hadar, Y., Y. Henis and I. Chet. 1981. The potential for the formation of
sclerotia in submerged mycelium of Sclerotium rolfsii. J. Gen. Microbiol.
122:137-141.
86. Chet, I. and R. Baker. 1981. Isolation and biocontrol potential of Trichoderma
hamatum from soil naturally suppressive to Rhizoctonia solani. Phytopathology
71:286-290.
Ilan Chet 549
87. Chet, I., G.E. Harman and R. Baker. 1981. Trichoderma hamatum: its
hyphal interactions with Rhizoctonia solani and Pythium spp. Microbiol.
Ecol. 7:29-38.
88. Elad, Y., I. Chet and Y. Henis. 1981. A selective medium for improving
quantitative isolation of Trichoderma spp. from soil. Phytoparasitica 9:59-67.
89. Elad, Y., Y. Hadar, E. Hadar, I. Chet and Y. Henis. 1981. Biological control of
Rhizoctonia solani by Trichoderma harzianum in carnation. Plant Disease
65:675-677.
90. Kritzman, G., I. Chet and D. Gilan. 1981. Spore germination and penetration
of Botrytis allii into Allium cepa host. Botanical Gazette 142:151-155.
91. Harman, G.E., I. Chet and R. Baker. 1981. Factors affecting efficacy of
Trichoderma hamatum applied to seeds as a biocontrol agent. Phytopathology
71:569-572.
92. Ruben, D.M., Z.R. Frank, I. Chet and D. Sklan. 1981. Changes in chemical
composition of oospores of Pythium aphanidermatum during biomodal
germination. Can. J. Microbiol. 27:536-543.
93. Elad, Y., I. Chet and Y. Henis. 1981. Biological control of Rhizoctonia solani
in strawberry fields by Trichoderma harzianum. Plant and Soil 60:245-254.
94. Huttermann, A., W.E. Kuhl and I. Chet. 1981. The effect of L-threonine on
oxalic acid synthesis by Fomes annosus. Eur. J. Forest Pathol. 10:339-344.
95. Haars, A., I. Chet and A. Huttermann. 1981. Effect of phenolic compounds
and tannin on growth and laccase activity of Fomes annosus. Eur. J. Forest
Pathol. 11:67-76.
96. Gerson, U. and I. Chet. 1981. Are allochthonous and autochthonous soil
microorganisms r- and k-selected? Rev. Ecol. Biol. Sol. 18:285-289.
97. Platt, M.W., I. Chet and Y. Henis. 1981. Lignocellulose degradation during
growth of the mushroom Pleurotus sp. Eur. J. Appl. Microbiol. and Biotech.
13:194-195.
98. Platt, M., Y. Bashan, I. Chet and Y. Henis. 1981. Two media for the rapid
growth of Pleurotus sp. Mushroom Newsletter for Tropics 1:2-5.
99. Elad, Y., Hadar, Y., Hadar, E., I. Chet and Y. Henis, 1981. Biological control
of Rhizoctonia solani by Trichoderma harzianum in carnation. Plant Dis.
65:675-677.
100. Paster, N. and I. Chet. 1982. Influence of controlled atmospheres on the
formation and ultrastructure of Aspergillus ochraceus sclerotia. Trans. Brit.
Mycol. Soc. 78:315-322.
101. Elad, Y., I. Hadar, I. Chet and Y. Henis. 1982. Prevention with Trichoderma
harzianum Rifai aggr. of reinfestation of Trichoderma harzianum and
Rhizoctonia solani Kuhn of soil fumigated with methyl bromide and
improvement of disease control in tomatoes and peanuts. Crop Protection
1:199-211.
550 Wolf Prize in Agriculture
102. Hadar, E., Y. Elad, Y. Hadar and I. Chet. 1982. Build-up and decline
of Rhizoctonia solani inoculum under field conditions. Plant and Soil
65:303-307.
103. Chet, I. and A. Huttermann. 1982. De novo synthesis of polyphenol oxidase
(laccase) during formation of sclerotia in Sclerotium rolfsii. FEMS Microbiol.
Letts. 14:211-215.
104. Elad, Y., I. Chet and Y. Henis. 1982. Degradation of plant pathogenic fungi
by Trichoderma harzianum. Can. J. Microbiol. 28:719-725.
105. Hadar, Y., Y. Elad, I. Chet and Y. Henis. 1982. Induction of macroscopic
strand formation in Sclerotium rolfsii by Trichoderma harzianum. Isr. J. Bot.
30:156-164.
106. Elad, Y., A. Kalfon and I. Chet. 1982. Control of Rhizoctonia solani in cotton
by seed coating with Trichoderma spores. Plant and Soil 66:279-281.
107. Elad, Y., I. Chet, P. Boyle and Y. Henis. 1983. Parasitism of Trichoderma
spp.on Rhizoctonia solani and Sclerotium rolfsii — SEM studies and
fluorescence microscopy. Phytopathology 73:85-88.
108. Chet, I., Y. Elad, A. Kalfon, Y. Hadar and J. Katan. 1982. Integrated
control of soilborne and bulb-borne pathogens in iris. Phytoparasitica
10:229-236.
109. Elad, Y. and I. Chet. 1983. Improved selective media for isolation of
Trichoderma or Fusarium spp. Phytoparasitica 11:55-58.
110. Chet, I. and Y. Elad. 1982. Prevention of plant infection by biological means.
La Selection des Plantes, Bordeaux, France, INRA Publ. 195-204.
111. Hadar, Y., M. Pines, I. Chet and Y. Henis. 1983. The regulation of sclerotium
initiation in Sclerotium rolfsii by glucose and cyclic AMP. Can. J. Microbiol.
28:21-26.
112. Elad, Y., Z. Sadovski and I. Chet. 1983. Detection of mycoparasitism by
infra-red photomicrography. Microbial Ecol. 9:185-187.
113. Zilberstein, Y., I. Chet and Y. Henis. 1983. Effect of atmosphere on germination
of microsclerotia of Verticillium dahliae Kleb. Isr. J. Bot. 32:33-36.
114. Lisker, N., N. Paster and I. Chet. 1983. Ochratoxin production by Aspergillus
ochraceus as affected by methionine and structurally related compounds.
Can. J. Microbiol. 29:536-540.
115. Paster, N., N. Lisker and I. Chet. 1983. Ochratoxin A production by Aspergillus
ochraceus Wilhelm grown under controlled atmospheres. Appl. Environ.
Microbiol. 45:1136-1139.
116. Stzejnberg, A., H. Azaizia and I. Chet. 1983. The possible role of phenolic
compounds in resistance of horticultural crops to Dematophora necatrix
Hartig. Phytopathology Z. 107:318-326.
117. Kapulnik, E., A.K. Matoo, E. Chalutz and I. Chet. 1983. The relationship
between the production of ethylene by certain fungi, their enzyme
Ilan Chet 551
133. Sivan, A., Y. Elad and I. Chet. 1984. Biological control of Pythium
aphanidermatum by a new isolate of Trichoderma harzianum. Phytopathology
74:498-501.
134. Shapira, R., D. Sklan, Y. Henis and I. Chet. 1984. Changes in fatty acids during
morphogenesis in Sclerotium rolfsii. J. Gen. Microbiol. 130:1183-1191.
135. Platt, M.W., Y. Hadar and I. Chet. 1984. Fungal activities involved in
lignocellulose degradation by Pleurotus. Appl. Microbiol. and Biotechnol.
20:150-154.
136. Baker, R., Y. Elad and I. Chet. 1984. The controlled experiment in the
scientific method with special emphasis on biological control. Phytopathology
74:1019-1021.
137. Strashnov, Y., Y. Elad, A. Sivan, Y. Rudich and I. Chet. 1985. Control of
Rhizoctonia solani fruit rot of tomatoes by Trichoderma harzianum Rifai.
Crop Protection 4:359-364.
138. Strashnov, Y., Y. Elad, A. Sivan and I. Chet. 1985. Integrated control of
Rhizoctonia solani Kuhn on bean and carrot seedlings by methyl bromide
and Trichoderma harzianum Rifai Aggr. Plant Pathol. 34:146-151.
139. Barak, R., Y. Elad, D. Mirelman and I. Chet. 1985. Lectins: a possible
basis for specific recognition in Trichoderma-Sclerotium rolfsii interaction.
Phytopathology 75:458-462.
140. Sadeh, R., G. Kritzman and I. Chet. 1985. The ultrastructure of resting and
germinating sclerotia of Botrytis alli. Bot. Gaz. 146:302-307.
141. Platt, M.W., Y. Hadar and I. Chet. 1985. The decolorization of the polymeric
dye poly blue (Poly vinalamine sulfonate anthroquinone) by lignin degrading
fungi. Appl. Microbiol. and Biotechnol. 21:394-396.
142. Spiegel, Y. and I. Chet. 1985. Chitin synthetase inhibitors and their potential
to control the root-knot nematode, Meloidogyne javanica. Nematologica 31:
480-482.
143. Zamir, D. and I. Chet. 1985. Application of enzyme electrophoresis
for identification of isolates in Trichoderma harzianum. Can. J. Microbiol.
31:578-580.
144. Chet, I., J. Trojanowsky and A. Huttermann. 1985. Decolorization of the
dyepoly-B-411 and its correlation with lignin degradation by fungi. Microbiol.
Letters 29:37-43.
145. Barak, R., A. Maoz and I. Chet. 1985. Antigenic differences among several
Trichoderma isolates. Can. J. Microbiol. 31:810-816.
146. Paster, N., N. Lisker and I. Chet. 1985. Accumulation of ochratoxin A in
sclerotia of Aspergillus ochraceus. Can. J. Bot. 63:661-662.
147. Lisker, N., N. Paster and I. Chet. 1985. Effects of cysteine and structurally
related compounds on ochratoxin production by Aspergillus ochraceus. Can.
J. Microbiol. 31:973-977.
Ilan Chet 553
163. Elad, Y., I. Chet and R. Baker. 1987. Increased growth response of plants
induced by rhizobacteria antagonistic to soilborne pathogenic fungi. Plant
and Soil 98:325-330.
164. Sivan, A. and I. Chet. 1987. Biological control of Fusarium crown rot of
tomato by Trichoderma harzianum under field conditions. Plant Disease
71:587-592.
165. Spiegel, Y., I. Chet and E. Cohn. 1987. Use of chitin for controlling plant
parasitic nematodes. II. Mode of action. Plant and Soil 98:337-345.
166. Elad, Y. and I. Chet. 1987. The role of competition for nutrients in biocontrol
of Pythium damping off by bacteria. Phytopathology 77:190-195.
167. Sztejnberg, A., S. Freeman, I. Chet and J. Katan. 1987. Control of Rosellinia
necratrix in soil and apple orchard by solarization and Trichoderma
harzianum. Plant Disease 71:365-369.
168. Peleg, Y., S. Kader, J. S. Rokem, I. Chet and I. Goldberg. 1987. Phaseolin
production in cell suspensions and whole plants of Phaseolus vulgaris. Plant
Cell, Tissue and Organ Culture 9:207-215.
169. Ordentlich, A., Y. Elad and I. Chet. 1987. Rhizosphere colonization by
Serratia marcescens for the control of Sclerotium rolfsii. Soil Biol. Biochem.
19:747-751.
170. Ordentlich, A., Y. Elad and I. Chet. 1988. The role of chitinase of Serratia
marcescens in biocontrol of Sclerotium rolfsii. Phytopathology 78:84-88.
171. Shoseyov, O., B. A. Bravdo, R. Ikan and I. Chet. 1988. Endo-β-glucosidase
from Aspergillus niger grown on a monoterpene glycoside-containing
medium. Phytochemistry 7:1973-1976.
172. Sivan, A. and I. Chet. 1988. Degradation of fungal cell walls by lytic enzymes
of Trichoderma harzianum. J. Gen. Microbiol. 135:675-682.
173. Spiegel, Y., I. Chet, E. Cohn, S. Galper and E. Sharon 1988. Use of chitin for
controlling plant parasitic nematodes. III. Influence of temperature on
nematicidal effect, mineralization and microbial population buildup. Plant
and Soil 109:251-256.
174. Levy, E., Z. Eyal and I. Chet. 1988. Suppression of Septoria tritici blotch
and leaf rust on wheat seedling leaves by Pseudomonas. Plant Pathology
37:551-557.
175. Sivan, A. and I. Chet. 1989. The possible role of competition between
Trichoderma harzianum and Fusarium oxysporum on rhizosphere
colonization. Phytopathology 79:198-203.
176. Dackman. C., I. Chet and B. Nordbring-Hertz. 1989. Fungal parasitism of
the cyst nematode Heterodera schachtii: infection and enzymatic activity.
FEMS Ecology Letters 62:201-208.
177. Sivan, A. and I. Chet. 1989. Cell wall composition of Fusarium oxysporum.
Soil Biol. Biochem. 21:869-876.
Ilan Chet 555
178. Shapira, R., A. Altman, Y. Henis and I. Chet. 1989. Polyamines and ornithine
decarboxylase activity during growth and differentiation in Sclerotium rolfsii.
J. Gen. Microbiol. 135:1361-1367.
179. Kless, H., Y. Sitrit, I. Chet and A.B. Oppenheim. 1989. Cloning of gene
coding for chitobiase of Serratia marcescens. Molec. & Gen. Genetics
217:471-473.
180. Levy, E., Z. Eyal, S. Carmeli, Y. Kashman and I. Chet. 1989. Suppression of
Septoria tritici and Puccinia recondita of wheat by an antibiotic-producing
fluorescent pseudomonad. Plant Pathol. 38:564-570.
181. Spiegel, Y., E. Cohn and I. Chet. 1989. Use of chitin for controlling Heterodera
aveenae and Tylenchulus semipenetrans. J. Nematol. 21:419-422.
182. Shapira, R., A. Ordentlich, I. Chet and A.B. Oppenheim. 1989. Control of
plant diseases by chitinase expressed from cloned DNA in Escherichia coli.
Phytopathology 79:1246-1249.
183. Galper, S., E. Cohn, Y. Spiegel and I. Chet. 1989. Nematicidal effect of collagen-
amended soil and the influence of protease and collagenase. Review de
Nematologie 13:67-71.
184. Persson, Y., I. Chet and B. Nordbring-Hertz. 1990. The effect of polyoxin D
on morphogenesis of the nematode-trapping fungus Arthrobotrys oligospora.
Mycol. Res. 94:196-200.
185. Pe’er, S. and I Chet. 1990. Trichoderma protoplast fusion: a tool for improving
biocontrol agents. Can. J. Microbiol, 36:6-9.
186. Patterson, N.A., I. Chet and Y. Kapulnik. 1990. Effect of mycorrhizal
inoculation on nodule initiation activity and contribution to legume
productivity. Symbiosis 8:9-20.
187. Barak, R. and I. Chet. 1990. Lectins of Sclerotium rolfsii: Its purification and
possible function in fungal interaction. J. Appl. Bacteriol. 69:101-112.
188. Ordentlich, A., A. Nachmias and I. Chet, 1990. Integrated control of
Verticillium dahliae in potato by Trichoderma harzianum and captan. Crop
Protection 9:363-366.
189. Roby, D., K. Broglie, R. Cressman, P. Biddle, I. Chet and R. Broglie. 1990.
Activation of a bean chitinase promoter in transgenic tobacco plants by
phytopathogenic fungi. Plant Cell 2:999-1007.
190. Chet, I., A. Ordentlich, R. Shapira and A.B. Oppenheim. 1990. Mechanisms
of biocontrol of soil-borne plant pathogens with rhizobacteria. Plant and
Soil 129:85-92.
191. Inbar , J. and I. Chet. 1991. Detection of chitinolytic activity in the rhizosphere
using image analysis. Soil Biol. Biochem. 23:239-242.
192. Galper, S., Y. Spiegel, I. Chet and E. Cohn. 1991. A collagenolytic fungus,
Cunninghamella elegans, for biological control of plant-parasitic nematodes.
J. Nematol. 23:269-274.
556 Wolf Prize in Agriculture
193. Spiegel, Y., E. Cohn, S. Galper, E. Sharon, and I. Chet. 1991. Evaluation of a
newly isolated bacterium, Pseudomonas chitinolytica sp. nov. for controlling
the root-knot nematode Meloidogyne javanica. Biocontrol Science and Technol.
1:115-125.
194. Inbar, J. and I. Chet. 1991. Evidence that chitinase produced by Aeromonas
caviae is involved in the biological control of soil plant pathogens. Soil Biology
and Biochemistry, 23:973-978.
195. Broglie, K., I. Chet, M. Holliday, R. Cressman, P. Biddle, S. Knowlton,
C.J. Mauvals, and R. Broglie. 1991. Transgenic plants with enhanced resistance
to the fungal pathogen Rhizoctonia solani. Science 254:1194-1197.
196. Pe’er, S., Z. Barak, O. Yarden and I. Chet. 1991. Stability of Trichoderma
harzianum amdS transformants in soil and rhizosphere. Soil Biol. Biochem.
23:1043-1046.
197. Ordentlich, A., Q. Migheli and I. Chet. 1991. Biological control activity
of three Trichoderma isolates against Fusarium wilts of cotton and
muskmelon and lack of correlation with their lytic enzymes. J. Phytopathology
133:177-186.
198. Ordentlich, A., Z. Weisman, H.E. Gottlieb, M. Cojocaru and I. Chet. 1992.
Inhibitory furanone produced by the biocontrol agent Trichoderma
harzianum. Phytochemistry 31:485-486.
199. Inbar, J. and I. Chet. 1992. Biomimics of fungal cell-cell recognition by the
use of lectin-coated nylon fibers. J. Bacteriol. 174:1055-1059.
200. Jach, G., S. Logemann, G. Wolf, A. Oppenheim, I. Chet, J. Schell and
J. Logemann. 1992. Expression of a bacterial chitinase leads to improved
resistance of transgenic tobacco plants against fungal infection. Biopractice
1:33-36.
201. Kleifeld, O. and I. Chet. 1992. Trichoderma — Plant interaction and its effect
on increased growth response. Plant and Soil 144:267-272.
202. Levy, E., Z. Eyal, I Chet and A. Hochman. 1992. Resistance mechanisms of
Septoria tritici to antifungal products of Pseudomonas. Physiol. Mol. Plant
Pathol. 40:163-171.
203. Paster, N., A. Pushinsky, M. Menasherov and I. Chet. 1992. Inhibitory effect
of Aspergillus ochraceus and Aspergillus flavus, and on Aflatoxin formation.
J. Sci. Food Agric. 58:589-591.
204. Amir, R., D. Levanon, Y. Hadar and I. Chet. 1992. Formation of sclerotia by
Morchella esculenta: relationship between media composition and turgor
potential in the mycelium. Mycol. Res. 96:943-948.
205. Benhamou, N., K. Broglie, R. Broglie and I. Chet. 1992. Antifungal effect of
bean endochitinase on Rhizoctonia solani: ultrastructural changes and
cytochemical aspects of chitin breakdown. Can. J. Microbiol. 39:318-328.
Ilan Chet 557
206. Oppenheim, A.B. and I. Chet. 1992. Biological control of soilborne plant
pathogenic fungi by cloned chitinase. Trends Biotechnol. 10:392-394.
207. Amir, R., D. Levanon Y. Hadar and I. Chet. 1993. Morphology and physiology
of Morchella esculenta during sclerotial formation. Mycological Res.
97:683-689.
208. Elad, Y., G. Zimand, Y. Zaqs, S. Zuriel and I. Chet. 1993. Use of Trichodrma
harzianum in combination or alternation with fungicides to control cucumber
grey mould (Botrytis cinerea) under commercial greenhouse conditions. Plant
Pathol. 42:324-332.
209. Ocka, Y., I. Chet and Y. Speigel. 1993. Control of the root-knot
nematode Meloidogyne javanica by Bacillus cereus. Biocontrol Sci. Technol.
3:115-126.
210. Sivan, A. and I. Chet. 1993. Integrated control of fusarium crown and root
rot of tomato with Trichoderma harzianum in combination with methyl
bromide or soil solarization. Crop Protection 12:380-386.
211. Fridlender, M., J. Inbar and I. Chet. 1993. Biological control of soilborne
plant pathogens by a β-1,3-glucanase producing Pseudomonas cepacia. Soil
Biol. Biochem. 25: 1211-1221.
212. Sitrit, Y., Z. Barak, Y. Kapulnik, A. Oppenheim and I. Chet. 1993. Expression
of Serratia marcescens chitinase gene in Rhizobium meliloti during symbiosis
on alfalfa roots. Mol. Plant Microbe Inter. 6:293-298.
213. Haran, S., H. Schickler, S. Pe’er, S. Logemann, A. Oppenheim and I. Chet.
1993. Increased constitutive chitinase activity in transformed Trichoderma
harzianum. Biol. Control 3:101-108.
214. Zimand, G., L. Valinsky, S. Manulis, Y. Elad and I. Chet. 1993. DNA
fingerprinting of a Trichoderma isolate by RAPD procedure. IOBD Bull.
16:173-176.
215. Benhamou, N. and I. Chet. 1993. Hypal interactions between Trichoderma
harzianum and Rhizoctonia solani: ultrastructure and gold cytochemistry of
the mycoparasitic process. Phytopathology 83:1062-1071.
216. Benhamou, N., K. Broglie, I. Chet and R. Broglie. 1993. Cytology of infection
of 35S-bean chitinase transgenic canola plants by Rhizoctonia solani:
cytochemical aspects of chitin breakdown in vivo. The Plant Journal
4:295-305.
217. Inbar, J. and I. Chet. 1994. A newly isolated lectin from the plant pathogenic
fungus Sclerotium rolfsii: purification, characterization and role in
mycoparasitism. Microbiology 140:651-657.
218. Zimand, G., L. Valinsky, Y. Elad, I. Chet and S. Manulis. 1994. Use of the
RAPD procedure for the identification of Trichoderma strains. Mycol. Res.
98: 531-534.
558 Wolf Prize in Agriculture
219. Amir, R., D. Levanon, Y. Hadar and I. Chet. 1994. The role of source sink
relationships in translocation during sclerotia formation by Morchella
esculenta Mycological Res. 98:1409-1414.
220. Inbar, J., Abramsky, M., Cohen, D. and I. Chet. 1994. Plant growth
enhancement and disease control by Trichoderma harzianum grown under
commercial conditions. Eur. J. Plant Pathol. 100:337-346.
221. Koby, S., H. Schickler, I. Chet and A.B. Oppenheim. 1994. The chitinase
encoding Tn-7-based ChiA gene endows Pseudomonas fluorescens with the
capacity to control plant pathogens in soil. Gene 147:81-83.
222. Perrakis, A., I. Tews, Z. Dauter, A.B. Oppenheim, I. Chet, K.S. Wilson and
C.E. Vorgias. 1994. Crystal structure of a bacterial chitinase at 2.3 Å
resolution. Structure 2:1169-1180.
223. Zimand, G., Y. Elad, G. Kritzman, and I. Chet. 1994. Control of Botyris
cinerea by Trichoderma harzianum (T39): Does the biocontrol agent produce
inhibitory substances? Phytoparasitica 22:150-151.
224. Haran, S., H. Schickler, A. Oppenheim and I. Chet. 1995. New components
of the chitinolytic system of Trichoderma harzianum. Mycol. Research.
99:441-446.
225. Amir, R., D. Levanon, Y. Hadar and I. Chet. 1995. Factors affecting
translocation and sclerotia formation by Morchella esculenta. Exp. Mycol.
19: 61-70.
226. Amir, R., E. Steudle, D. Levanon, Y. Hadar and I. Chet. 1995. Turgor changes
in Morchella esculenta during translocation and sclerotia formation. Exp.
Mycol. 19: 129-136.
227. Sitrit, Y., Vorgias, C.E., I. Chet and A.B. Oppenheim. 1995. Cloning and
primary structure of the chiA gene from Aeromonas caviae. J. Bacteriol.
177:4187-4189.
228. Salt, D.E., M. Blaylock, N.P.B.A. Kumar, V. Dushenkov, B.D. Ensley, I. Chet,
and I. Raskin. 1995. Phytoremediation: A novel strategy for the removal
of toxic metals from the environment using plants. Nature Biotechnology
13:468-474.
229. Chernin, L., Z. Ismailov, S. Haran and I. Chet. 1995. Chitinolytic Enterobacter
agglomerans antagonistic to fungal plant pathogens. Appl Environ. Microbiol.
61:1720-1726.
230. Inbar, J. and I. Chet. 1995. The role of recognition in the induction of
specific chitinases during mycoparasitism by Trichoderma harzianum.
Microbiology 141:2823-2829.
231. Pleban, S., F. Ingel and I. Chet. 1995. Control of Rhizoctonia solani and
Sclerotium rolfsii in the greenhouse using endophytic Bacillus spp. Eur. J.
Plant Pathol. 101:665-672.
Ilan Chet 559
232. Spiegel, Y., I. Chet, I. Mor, O. Kleifeld, E. Sharon and M. Bar-Eyal. 1995.
Trichoderma Reparation as biological control of the root-knot nematode
Meloidogyne javanics: Evaluation of the nematocidal activity and preliminary
studies of the mechanism. ARO publication, series E, 1995m no. 1210
(appeared in Hebrew in Hassadeh 75:67-72).
233. Benhamou, N. and Ilan Chet. 1996. Parasitism of sclerotia of Sclerotium
rolfsii by Trichoderma harzianum: ultrastructural and cytochemical aspects
of the interaction. Phytopathology 86:405-416.
234. Chernin, L., A. Brandis, Z. Ismailov and I. Chet. 1996. Pyrrolnitrin production
by an Enterobacter agglomerans strain with a broad spectrum of antagonistic
activity towards fungal and bacterial phytopathogens. Current Microbiol.
32:208-212.
235. Inbar, J., A. Menendez, and I. Chet. 1996. Hyphal interactions between
Trichoderma harzianum and Sclerotinia sclerotiorum and its role in biological
control. Soil Biol. & Biochem. 28:757-763.
236. Zimand, G., Y. Elad and I. Chet. 1996. Effect of Trichoderma harzianum on
Botrytis cinerea pathogenicity. Phytopathology 86:1255-1262.
237. Rousseau, A., N. Benhamou, I. Chet and Y. Piché, 1996. Mycoparasitism of
the extramatrical phase of Glomus intraradices by Trichoderma harzianum.
Phytopathology 86, 434-443.
238. Haran, S., H. Schickler, A. Oppenheim and I. Chet. 1996. Differential
expression of Trichoderma harzianum chitinases during mycoparasitism.
Phytopathology 86:980-985.
239. Regev, A., M. Keller, N. Strizhov, B. Sneh, E. Prudovsky, I. Chet, I. Ginzberg,
Z. Koncz-Kalman, C. Koncz, J. Schell, and A. Zilberstein. 1996. Synergistic
activity of a Bacillus thuringiensis δ-endotoxin and a bacterial endochitinase
against Spodoptera littoralis larvae. App. Environ. Microbiol. 62:3581-3586.
240. Chernin, L.S., L. de la Fuente, V. Sobolev, S. Haran, C.E. Vorgias, A.B.
Oppenheim and I. Chet. 1997. Molecular cloning, structural analysis, and
expression in Escherichia coli of a chitinase gene from Enterobacter
agglomerans. Appl. Environ. Microbiol. 63:834-839.
241. Maas, C., C.G. Simpson, P. Eckes, H. Schickler, J.W.S. Brown, B. Reiss,
K. Salchert, I. Chet, J. Schell and C. Reichel. 1997. Expression of intron
modified NPT II genes in monocotyledonous and dicotyledonous plant cells.
Mol. Breeding 3:15-28.
242. Benhamou, N. and I. Chet. 1997. Cellular and molecular mechanisms involved
in the interaction between Trichoderma harzianum and Pythium ultimum.
Appl. Environ. Microbiol. 63:2095-2099.
243. Flores, A., I. Chet and A. Herrera-Estrella. 1997. Improved biocontrol activity
of Trichoderma harzianum by overexpression of the proteinase-encoding
gene prb1. Current Genetics 31:30-37.
560 Wolf Prize in Agriculture
244. Pleban, S., L. Chernin and I. Chet. 1997. Chitinolytic activity of endophytic
strain of Bacillus cereus. Lett. Appl. Microbiol. 25:284-288.
245. Sela, S., H. Schickler, I. Chet and Y. Spiegel. 1997. Purification and
characterization of a Bacillus cereus collagenolytic/proteolytic enzyme
and its effect on Meloidogyne javanica cuticular proteins. Eur. J. Plant Path.
104:59-67.
246. Gomez, I, I. Chet and A. Herrera-Estrella. 1997. Genetic diversity and
vegetative compatibility among Triochoderma harzianum isolates. Mol. Gen.
Genetics 256:127-135.
247. Schickler, H. and I. Chet. 1997. Heterologous chitinase gene expression to
improve plant defense against phytopathogenic fungi. J. Industrial Microbiol.
& biotechnol. 19:196-201.
248. Oka, Y., I. Chet and Y. Spiegel. 1997. An immunoreactive protein to
wheat-germ agglutinin antibody is induced in oat roots following invasion
of the cereal cyst nematode Heterodera avenae, and by Jasmonate. Mol.
Plant-Microbe Interactions 10:961-969.
249. Oka, Y., I. Chet, More, M. and Y. Spiegel. 1997. A fungal parasite of
Meloidogyne javanica eggs: Evaluation of its use to control the root-knot
nematode. Biocontrol Science and Technol. 7:489-497.
250. Oka, Y., I. Chet and Y. Spiegel. 1997. Are pathogenesis-related poteins
induced by Meloidogyne javanica or Heterodera avenae invasion? J. Nematol.
29:501-508.
251. Oka, Y., I. Chet and Y. Spiegel. 1997. Accumulation of lectins in cereal roots
invaded by the cereal cyst nematode Heterodera venae. Physiol. Mol. Olant
Pathol. 51:333-345.
252. Zoubenko, O., F. Uckun, Y. Hur, I. Chet and N. Tumer. 1997. Plant resistance
to fungal infection induced by nontoxic pokeweed antiviral protein mutants.
Nature Biotechnol. 15(10):992-996.
253. Oka, Y., I. Chet, Mor, M. and Y. Spiegel. 1997. A fungal parasite of
Meloidogyne javanica eggs: Evaluation of its use to control the root-knot
nematode. Biocontrol Science and Technol. 7:489-497.
254. Danai, O., Olenik, I., Hadar, Y., I. Chet and D. Levanon. 1998. The role of
light in the morphogenesis of Pleurotus ostreatus. Int. J. Mushroom Sci.
2:31-37.
255. Oka, Y., I. Chet and Y. Spiegel. 1997. Accumulation of lectins in cereal roots
invaded by the cereal cyst nematode Heterodera avenae. Physiol. Mol. Plant
Pathol. 51:333-345.
256. Dickman, M.B. and I. Chet. 1998. Biodegradation of oxalic acid: A potential
new approach to biological control. Soil Biol. Biochem. 30:1195-1197.
257. Schickler, H., B-C. Danin-Gehali, S. Haran and I. Chet. 1998. Electro-phoretic
characterization of chitinases as a tool for the identification of Trichoderma
harzianum strains. Mycol. Res. 102:373-377.
Ilan Chet 561
258. Sela, S., H. Schickler, I. Chet and Y. Spiegel. 1998. Putrification and
characterization of Bacillus cereus collagenolytic/proteolytic enzymes and
its effect on Meloidogyne juvanica cuticular proteins. Eur. J. Plant Pathol.
104: 59-67.
259. Schickler, H., Haran, S., A. Oppenheim and I. Chet. 1998. Induction of the
Trichoderma harzianum chitinolytic system is triggered by the chitin
monomer, N-acetylglucosamine. Mycol. Res. 102:1224-1226.
260. Chernin, L.S., M.K. Winson, J.M. Thompson, S. Haran, B.W. Bycroft,
I. Chet, P. Williams and G.S.A.B. Stewart. 1998. Chitinolytic activity in
Chromobacterium violaceum: Substrate analysis and regulation by quorum
sensing. J. Bacteriol. 180:4435-4441.
261. Cortes, C., A. Gutierrez, V. Olmedo, J. Inbar, I. Chet and A. Herrera-Estrella.
1998. The expression of genes involved in parasitism by Trichoderma
harzianum is triggered by a diffusable factor. Mol. Gen. Gen. 260:218-225.
262. Yedidia I., N. Benhamou, and I. Chet. 1999. Induction of defense responses
in cucumber plants (Cucumis sativus L.) by the biocontrol agent Trichoderma
harzianum. Appl. Environ. Microbiol. 65:1061-1070.
263. Carsolio C., N. Benhamou, Haran, S., C. Cortés, A. Gutiérrez, I, Chet and
A. Herrera-Estrella. 1999. Role of the Trichoderma harzianum endochitinase
gene, ech42, in mycoparasitism. Appl. Environ. Microbiol. 65:929-935.
264. Salt, D.E., N. Benhamou, M. Leszczyniecka, I. Raskin and I. Chet. 1999.
Rhizobacterial enhanced Cadmium A accumulation in plants. Int. J.
Phytoremed. 1:67-79.
265. Cohen-Kupiec, R., K.E. Broglie, D. Friesem, R.M. Broglie and I. Chet. 1999.
Molecular characterization of a novel β-1,3 exo-glucanase related to
mycoparasitism of Trichoderma harzianum. Gene 226:147-154.
266. Omero, C., J. Inbar, V. Rocha-Ramirez, A. Herrera-Estrella, I. Chet and
B.A. Horwitz. 1999. G protein activators and cAMP promote mycoparasitic
behaviour in Trichoderma harzianum. Mycol. Res. 103:1637-1642.
267. Shaul, O., S. Galili, H. Volpin, I. Ginzberg, Y. Elad, I. Chet and Y. Kapulnik.
1999. Mycorrhiza-induced changes in disease severity and PR protein
expression in tobacco leaves. Mol. Plant Microbe Interaction 12:1000-1007.
268. Ramot, O., R. Cohen-Kupiec and I. Chet. 1999. Regulation of β-1,3-glucanase
by carbon starvation in the mycoparasite Trichoderma harzianum. Mycol.
Res. 104:415-420.
269. Oka, Y., H. Koltai, M. Bar-Eyal, M. Mor, E. Sharon, I. Chet and Y. Spiegel.
2000. New strategies for the control of plant-parasitic nematodes. Pest Manag.
Sci. 56:983-988.
270. Yedidia I., N. Benhamou, Y. Kapulnik and I. Chet. 2000. Induction and
accumulation of PR proteins activity during early stages of roots colonization
by the mycoparasite Trichoderma harzianum strain T-203. Plant Physiol.
Biochem. 38:863-873.
562 Wolf Prize in Agriculture
296. Meziane, H., S. Gavriel, Z. Ismailov, I. Chet, L. Chernin and M. Höfte. 2006.
Control of green and blue mould on orange fruit by Serratia plymuthica
strains IC14 and IC1270 and putative modes of action. Postharvest Biology
and Technology 39:125-133.
297. Viterbo A. and I. Chet. 2006. TasHyd1, a new hydrophobin gene from the
biocontrol agent Trichoderma apenellum, is involved in plant root
colonization. Molecular plant pathology (in press).
298. Shoresh M., A. Gal-On, D. Leibman, and I. Chet. 2006. Charecterization of a
MAPK gene from cucumber required for Trichoderma-conferred plant
resistance. Plant Physiol. (in press).
299. Sharon, E, I. Chet, A. Viterbo, M. Bar-Eyal, H. Nagan, G.J. Samuels and
Y. Spiegel. 2007. Parasitism of Trichoderma on Meloidogynejavanica and
role of the gelatinous matrix. Eur. Plant Pathol. (in press).
300. Viterbo, A., E. Wiest, Y. Brotman, I. Chet and C. Kenerley. 2007. The 18mer
peptaibols from Trichoderma virens Elicit plant defense responses. Molecular
Plant Pathol. 8:737-746.
301. Makovitzki, A., A. Viterbo, Y. Brotman, I. Chet and Y. Shai. 2007. Inhibition
of fungal and bacterialplant pathogens in vitro and in vivo with ultrashot
cationic lipopeptide. Appl. Environ. Microbiol. 73:6629-6636.
321. Chet, I., E. Cohen and I. Elster. 1985. The role of chitinase and chitin
synthetase inhibitors in controlling plant pathogenic fungi, pp. 237-240.
In: R.A.A. Muzzarelli, C. Jeuniaux and G.W. Gooday (eds.), “Chitin in Nature
and Technology, Plenum Publishing Corp., N.Y.
322. Chet, I. and Y. Elad. 1985. Biological and integrated control of soilborne
plant pathogens: mechanisms and application. Proceedings of Gottingen
Symposium.
323. Nordbring-Hertz, B. and I. Chet. 1986. Fungal lectins and agglutinins,
pp. 393-408. In: D. Mirelman (ed.), “Microbial Lectins and Agglutinins”.
John Wiley & Sons, N.Y.
324. Sivan, A. and I. Chet. 1986. Trichoderma harzianum: an effective biocontrol
agent of Fusarium spp. In: V. Jansen, A. Kjoller and L.H. Sorenson (eds.).
“Microbial Communities in Soil”. Elsevier Applied Science Publishers, London,
New York.
325. Chet, I. 1986. Saprophyte-parasite interaction of Trichoderma in the
rhizosphere, pp. 489-492. In: F. Megusar and M. Gantar (eds.) Proc. IV
ISME, “Perspectives in Microbial Ecology”.
326. Shoseyov, O., B.A. Bravdo, R. Ikan and I. Chet. 1987. Monoterpene glycoside
hydrolysis by a beta-glucosidase from immobilized Aspergillus niger,
pp. 63-71. In: A. Scienza and G. Versini (eds.), Proc. Int. Symp. The Aromatic
Substances in Grapes and Wines.
327. Chet, I. (ed).1987. Trichoderma-application, mode of action and potential
use as a biocontrol agent of soilborne plant pathogenic fungi, pp. 137-160.
In: I. Chet (ed.), “Innovative Approaches to Plant Disease Control”, John
Wiley & Sons, NY.
328. Chet, I. 1990. Biological control of soilborne pathogens with fungal antagonists
in combination with soil treatments. pp. 15-25. In: D. Hornby, R.J. Cook,
Y. Henis, W.H. Ko, A.D. Rovira, B. Schippers and P.R. Scott (eds.), “Biological
Control of Soilborne Pathogens”. CAB Publishing House, N.Y.
329. Chet, I. 1989. Mycoparasitism — recognition, physiology and ecology,
pp. 723-733. In: R. Baker and P. Dunn (eds.), “New Directions in Biological
Control”, Vol. 112, Alan R. Liss Inc., New York.
330. Levy, E., Z. Eyal, S. Carmely, Y. Kashman and I. Chet. 1989 Suppression of
Septoria tritici blotch and leaf rust on wheat seedling leaves by antibiotic
producing Pseudomonas. In: Septoria diseases of cereals. Proceedings of the
3rd International Septoria Workshop. Zurich-Reckenhold, Switzerland.
331. Sivan, A. and I. Chet. 1992. Microbial control of plant diseases, pp. 335-
354. In: R. Mitchell (ed.), “New Concepts in Environmental Microbiology”,
Wiley-Liss, New York.
332. Oppenheim, A.B., Hirsch, Y., Koby, S. and I. Chet. 1992. Protein secretion in
biotechnology, pp. 69-76. In: M.D. White, S. Revveny and A. Sahafferman
Ilan Chet 567
344. Chet, I., J. Inbar, S. Haran, S. Pleban and H. Schickler. 1995. Antagonistic
microorganisms in biological control. Proceedings of the “Year of Louis
Pasteur” Symposium, Microbes, Enviornment, Biotechnology, Tahiti, May
1995. pp.
345. Chet, I. and Inbar, J. 1994. Biological control of fungal pathogens. Applied
Biochemistry and Biotechnology. 47:37-43.
346. Shaul, O., Y. Elad, S. Gallili, H. Volpin, Y. Kapulnik and I. Chet. 1995.
Peroxidase and pathogenicity-related proteins in plant tissues infected by
Botrytis cinerea. Aspects of Appl. Biol., Physiological Responses of Plants to
Pathogens, 42:1-8.
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Ilan Chet 571
A D A V I T E R B O 1 * , A R I C W I E S T 2 † , YA R I V B R OT M A N 1 , I L A N C H E T 1 A N D C H A R L E S K E N E R L E Y 2
1
Department of Plant Science, The Weizmann Institute of Science, 76100, Rehovot, Israel
2
Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX 77843, USA
INTRODUCTION
S U M M A RY
Peptaibols are a class of linear, short-chain-length (≤ 20 residues)
Peptaibols, the products of non-ribosomal peptide synthetases peptides of fungal origin. Typical features are the presence of
(NRPS), are linear peptide antibiotics produced by Trichoderma α-amino isobutyric acid moieties, acetylation of the N-terminus
and other fungal genera. Trichoderma virens strain Gv29-8, a of the peptide chain and reduction of the C-terminus to an amino
well-known biocontrol agent and inducer of plant defence alcohol (Grigoriev et al., 2003). In a previous study we have
responses, produces three lengths of peptaibols, 11, 14 and 18 shown that tex1, a non-ribosomal peptide synthetase (NRPS), is
residues long, with several isoforms of each. Disruption of the responsible for peptaibol production in Trichoderma virens
NRPS gene, tex1, encoded by a 62.8-kb uninterrupted open (Wiest et al., 2002). NRPSs are large proteins capable of enzy-
reading frame, results in the loss of production of all forms of matic assembly of small peptides, characterized by an ordered
18-residue peptaibols. Tex1 is expressed during all Trichoderma modular arrangement. Each module consists of approximately
developmental stages (germinating conidia, sporulating and 1100-amino-acid residues (Marahiel et al., 1997) and catalyses
non-sporulating mycelia) examined on solid media. Expression the incorporation of a single amino acid residue. Modular order
analysis by reverse transcriptase PCR shows that in Gv29-8 parallels the order of the residues in the product (Marahiel et al.,
wild-type the abundance of tex1 transcript is greater during co- 1997). A single module of an NRPS, at minimum, consists of three
cultivation with cucumber seedling roots than when grown alone. domains: adenylation (A-domain), thiolation (T-Domain) and
Cucumber plants co-cultivated with T. virens strains disrupted in condensation (C-domain). The coding sequence of tex1 repre-
tex1 show a significantly reduced systemic resistance response sents the largest known open reading frame (ORF), 62.8kb, and
against the leaf pathogen Pseudomonas syringae pv. lachrymans, contains modules for the incorporation of 18 amino acids as well
and reduced ability to produce phenolic compounds with inhib- as motifs for the acetylation of the N-terminus and the reduction
itory activity to the bacteria as compared with plants grown in of the C-terminus. Even though a peptaibol database (http://
the presence of wild-type. Two synthetic 18-amino-acid peptaibol public-1.cryst.bbk.ac.uk/peptaibol/home.shtml) provides docu-
isoforms (TvBI and TvBII) from Gv29-8 when applied to cucumber mentation of over 300 subfamilies of these unique compounds,
seedlings through the transpiration stream can alone induce their role in biological systems has only been partially demon-
systemic protection to the leaf pathogenic bacteria, induce strated for a few individual compounds (Chug and Wallace, 2001;
antimicrobial compounds in cucumber cotyledons and up-regulate Rebuffat et al., 2000; Schirmbock et al., 1994). The in vitro
hydroxyperoxide lyase (hpl ), phenylalanine ammonia lyase (pal1) antibacterial activity of several peptaibols has been shown
and peroxidase ( prx) gene expression. These data strongly (Szekeres et al., 2005), suggesting a role for these compounds as
suggest that the 18mer peptaibols are critical in the chemical competitive inhibitors of other microbes in soil or the rhizosphere.
communication between Trichoderma and plants as triggers of The antibiotic functions of peptaibols arise from their membrane
non-cultivar-specific defence responses. insertion and pore-forming abilities (Duclohier and Wroblewski,
2001). There are a few reports indicating that peptaibols may also
represent a novel class of plant elicitors. Exogenous application
of the 20-residue peptaibol alamethicin, produced by T. viride,
has been shown to induce defence responses in Phaseolus
*Correspondence: Tel.: +972 8 9344466; Fax: +972 8 9344112; lunatus (lima bean) (Engelberth et al., 2000) and Arabidopsis
E-mail: ada.viterbo@weizmann.ac.il.
†Present address: Fungal Genetics Stock Center-School of Biological Sciences, SBS 404,
thaliana (Chen et al., 2003). In lima bean these responses include
University of Missouri, Kansas City, MO 64110, USA. biosynthesis of volatile compounds and salicylate biosynthesis.
A. thaliana showed increased production of methyl salicylate. 11- and 14-residue isoforms were still produced, while all forms
Furthermore, Chrysospermin, a 19-residue peptaibol from of the 18-residue peptaibol were absent. These results, similar to
Apiocrea chrosospermin, afforded Nicotiana tabacum protection the findings of Wei et al. (2005), suggest that the NRPS TEX1 is
from tobacco mosaic virus infection (Kim et al., 2000). only responsible for the production of 18-residue isomers.
As a ubiquitous soil inhabitant and rhizosphere competent To explore the significance of peptaibol production in T. virens,
fungus, T. virens has been used successfully as a biological we examined the expression patterns of tex1 in an axenic plant
control agent for the management of plant pathogens (Fravel, system along with several developmental and nutrient conditions.
2005; Lumsden et al., 1996). Several mechanisms of biocontrol We also took advantage of the new tex1 knockouts and synthetic
have been proposed for T. virens , including competition, 18-residue peptaibols to test whether these peptides alone are
mycoparasitism and the induction of plant defence responses due necessary or sufficient to induce plant defence responses in vivo.
to colonization of plant root intercellular spaces (Baek et al., Our data strongly suggest that in addition to their antibacterial activity,
1999; Harman et al., 2004; Howell, 2003; Pozo et al., 2004). The peptaibols also have a role as signalling molecules during the devel-
induction of both local and systemic resistance in cotton has been opment of symbiotic interactions between Trichoderma and plants.
demonstrated by the application of the proteinaceous elicitor SM1
(Djonovic et al., 2006). Considering the large number of metabolites
R E S U LT S
secreted and the intimate contact with the root epidermis by
T. virens, as well as the expanding list of diverse elicitors produced
tex1 expression analysis by RT-PCR
by fungi, peptaibols may be part of a signalling cascade resulting
in greater colonization by T. virens or plant resistance induction. RT-PCR was used to investigate expression of tex1. Northern
Several types of peptaibols are produced by T. virens strain analysis was not attempted because the probability of isolating
GV29-8, including 11-, 14- and 18-amino-acid residue isoforms. intact tex1 mRNA was low as RNA is prone to shearing during
Our initial tex1 disruptant strains (Gv234 and Gv223) were the extraction procedures. Using this indirect technique, tex1
result of multiple integration events, and we have reported that transcript was detected differentially. Intensity of RT-PCR
they produced no peptaibol isoforms (Wiest et al., 2002). In order products was used as a preliminary indicator of tex1 expression
to generate single homologous integration disruptants, the following amplification with primer pairs for module 1 (AW1 and
fungal transformation was repeated. In the present study, AW2). tex1 was expressed at all developmental stages for at
analysis of peptaibols from the new disruptants revealed that the least one of the conditions on solid media (Fig. 1A). Transcript
Fig. 1 (A) Tex1 expression during developmental stages of Gv29-8. Conditions used: germinating spores (gs) for 12 h on solid VMS (lane1) or PDA (lane 2); 2-day-
old non-sporulating mycelia (nsm) on solid VMS (lane 3) or PDA (lane 4); 5-day-old sporulating mycelia (sm) from solid VMS (lane 5) or PDA (lane 6); mycelia of Gv29-
8 indirectly confrontating a culture of R. solani (lane 7); 5-day-old VMS liquid culture of Gv29-8 (lane 8); genomic DNA (lane 9). (B) Tex1 expression in liquid media differing
in carbon and ammonium source. Conditions used: VM (Vogel’s minimal with no carbon), VMR (VM + 0.5% R. solani cell walls), VMG (VM with 1.5% glucose), VMG-N (no
ammonium), VMR-N (0.5% R. solani cell walls without ammonium). (C) Tex1 expression under cucumber co-cultivation (+) and without (–) at 24 h and 48 h. Mycelia
were pooled for RNA extraction from three different hydroponic culture boxes each containing 25 seedlings. Graph illustrates quantification of mRNA fold induction. The values
obtained for each band were normalized to β-tubulin. The results are averages of three independent experiments and the error bars indicate standard deviations.
MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746 © 2007 BLACKWELL PUBLISHING LTD
Ilan Chet 573
level was higher among conidia germinating on the complex by Southern analysis using NcoI- and XhoI-digested genomic
medium PDA than the minimal medium VMS. However, in both DNA. A homologous double crossover event was identified by a
types of mycelia (sporulating and non-sporulating), tex1 was 3.0-kb Nco I fragment and a 3.8-kb Xho I fragment. In the
expressed at higher levels in VMS than in PDA. There appears to wild-type strain or transformants containing ectopic copies of
be transient induction in VM liquid culture at 12 h and consistent the disruption vector, bands of 12.5 kb (NcoI) and 8.5 kb (XhoI)
expression in VMG at 12, 24 and 48 h (Fig. 1B). In the absence of were detected. Three transformants, GvL1, GvC3 and GvC12, con-
a nitrogen source, the levels appear faint, indicating a partial tained single-copy homologous integrations of the disruption
dependence on nitrogen. Simulated mycoparasitism towards vector (Fig. 3). Several ectopic integrants were also detected
Rhizoctonia solani did not induce tex1 expression either on solid (data not shown) but were not analysed further. The three
or in liquid cultures. tex1 transcript was detected at a higher level transformants had growth rates, colony morphology and sporu-
in PGM with cucumber than PGM alone at both 24 and 48 h lation rates comparable with the wild-type (data not shown).
(Fig. 1C). Two of the homologous single-integrant disruptants were
further tested by RT-PCR. RNA isolated from mycelia of Gv29-8,
GvL1 and GvC12 was used as template for cDNA synthesis.
tex1 Disruption and confirmation
Transcripts of tex1 were detected by amplification of cDNA
We previously reported the detection of three lengths of peptaibol fragments using primer pairs located upstream (P1: AW1 and
isoforms in Gv29-8, but not in two other tex1 disruptants AW2 in module 1) and downstream (P2: RT1 and RT2 in module
strains, Gv223 and Gv234 (Wiest et al., 2002). These two original 14; P3: AW3 and AW4 in module 18) of the disruption event
disruption strains were generated using hygromycin resistance as (Fig. 4A). Strains GvL1 and GvC12 yielded PCR products upstream
a selectable marker. These strains contained multiple copies of of the disruption, but not downstream, indicating that tex1 was
the disruption vector and had reduced growth compared with not fully transcribed in these strains (Fig. 4B). In Gv29-8,
Gv29-8. New disruptant strains were generated using comple- transcripts of tex1 were detected using all primer pairs. No prod-
mentation of arginine auxotrophy as a selectable marker. The ucts were amplified in the negative control reactions. To ensure
colossal size of tex1 precluded elimination of the entire gene cDNA preparations were free of genomic DNA contamina-
from the genome; therefore, arg2 was inserted between modules tion, amplification of actin, including genomic introns, was used
10 and 11 to interrupt the reading frame. This region was chosen as a control. All cDNA preparations yielded a single intronless
as it was the furthest upstream module characterized at the time. 576-bp actin PCR product while the genomic DNA control
tex1 disruptants were generated by replacing a 3-kb NdeI yielded a 940-bp PCR product.
fragment of module 10 with the arg2 gene using disruption
vector pSKO2 (Fig. 2). Of 85 stable transformants, 23 were screened
© 2007 BLACKWELL PUBLISHING LTD MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746
574 Wolf Prize in Agriculture
Fig. 5 MALDI-TOF of lyophilized fungal tissue harvested from 9-day-old stationary cultures grown in VMS. Strains GvL1 and GvC12 are disrupted in tex1.
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Ilan Chet 575
Fig. 7 Plant elicitation responses by synthetic 18mer peptaibols TVBI and TVBII. (A) Psl multiplication in cotyledons of cucumber seedlings treated with 9.6 nmol of
TvBI or TvBII (and mock) 48 h prior to bacterial challenge. The results are the average of three independent experiments. (B) Bioassay comparing the antimicrobial
activity of the aglycone fraction obtained by acid hydrolysis of crude phenolics extract of cucumber cotyledons from seedlings treated for 48 h with 9.6 nmol of TvBI
(and mock), 48 h post challenge with Psl. (C) RT-PCR analysis of hpl, pal1 and prx expression in cotyledons 24 h and 48 h after application of TvBII (9.6 nmol) applied
through the transpiration stream. Three seedlings were pooled for each extraction, and three independent extractions were performed per time point. (D) RT-PCR
analysis of hpl, pal1 and prx expression in cotyledons 24 h and 48 h after injection with TvBI (lanes 1 and 3) and TVBII (lanes 2 and 4) (3 nmol). As a control cotyledons
were infiltrated with 0.1% methanol. The gel is representative of one experiment out of five independent injections.
Synthetic TvBI and TvBII peptides when applied through the presented growth-inhibitory activity towards the Psl bacteria at
transpiration stream (9.6 nmol) for 48 h prior to the bacteria a much higher extent than the extract obtained from plants
challenge could mimic the protective effect of wild-type Gv29-8 challenged with the bacteria without prior incubation with the
(Fig. 7A). The phenol aglyconic extract from such plants also two 18mer peptaibols (Fig. 7B). When synthetic TvBI or TvBII
© 2007 BLACKWELL PUBLISHING LTD MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746
576 Wolf Prize in Agriculture
peptides (5, 10, 20 nmol) were incubated overnight with cultures gene induction, suggesting that the products of the NRPS may
of Psl or P. syringae DC3000, no growth inhibition (OD600) could not be directly involved in this specific interaction. All three tex1
be detected (data not shown). disruptants were capable of contacting and coiling around the
hyphae of R. solani, indicating that the peptaibol 18mer isoforms
are not required for the initial stages of mycoparasitism (data not
Induction of defence-related genes expression by
shown). Both synthetic 18mer peptaibols had weak antifungal
synthetic 18mer peptaibols TvBI and TvBII
activity at 200 µg/mL when tested against R. solani on PDA
Induction of hpl, pal1 and prx genes by synthetic TvBI and TvBII plates (data not shown).
was followed in cucumber cotyledons that were directly injected Engelberth et al. (2000) showed that emission of ethylene,
with a peptide solution (3 nmol) or that were harvested from jasmonic acid and volatile compounds related to the octadecanoid
cucumber seedlings after the peptide solution (9.6 nmol) was signalling pathway are inducible responses to treatment of lima
applied through the transpiration stream. In both experimental bean plants with alamecithin, an ion channel-forming peptaibol
systems a clear up-regulation of these genes could be detected from T. viride. Interestingly, these compounds and pathways are
by RT-PCR analysis even after 24 h (Fig. 7C–D). Expression of β- the same as those involved in the Trichoderma plant-induced
glucanase and chitinase1 genes was not affected (data not resistance (Shoresh et al., 2005; Yedidia et al., 2003). These
shown). No phytotoxic symptoms were detected in infiltrated observations led us to verify the direct involvement of peptaibols
cucumber leaves or treated seedlings at these concentrations. in the elicitation of Trichoderma–plant-induced resistance.
tex1 mutants still retain the 11- and 14-amino-acids peptaibols,
but these strains do not provide full systemic protection as the
DISCUSSION
wild-type strain, indicating that the 18-amino-acid peptaibols
Peptaibols are the largest known products produced by peptide have a dominant effect in initiating the very first steps of the
synthetases and over 300 have currently been identified (http:// signalling cascade of the plant defence response, most
www.cryst.bbk.ac.uk/peptaibol/home.shtml) (Chugh and Wallace, probably due to their membrane channel-forming properties
2001). Because of the similar structure of most peptaibols and (Engelberth et al., 2000). Changes in membrane potential are
the evolutionary relatedness of the producing fungi, it is likely the initial responses in many signalling pathways (Ehrhardt
that all peptaibols are produced by NRPS enzymes encoded by et al., 1992).
genes similar to tex1. Our study shows that disruptants of tex1 Yedidia et al. (2003) demonstrated that the inhibition of Psl
still produce 11- and 14-residue peptaibols. This suggests that proliferation in cucumber leaves by application of T. asperellum
there is more than one NRPS responsible for peptaibol production to the roots correlated with the accumulation of antimicrobial
in T. virens. In a recent study (Wei et al., 2005) the tex1 homologue aglyconic phenolic compounds. Concomitantly, expression of
was examined in a commercially relevant T. virens strain, G20. As pal1 and hpl, genes related to the phytoalexin synthesis pathway,
our MALDI-TOF data of the strains disrupted in tex1 indicated, was increased in roots and cotyledons when plants were treated
this gene is only responsible for the production of an 18mer with T. asperellum. Exogenous application of synthetic peptides
peptaibol, and not 11mer and 14mer peptaibols. Two additional of TvBI and TvBII confirmed that these isoforms are effective
NRPS adenylate domains were identified in T. virens, and sequence inducers of defence responses in cucumber against Psl. The defence
comparisons suggest that these might be part of a separate response involves the production of antimicrobial compounds
peptaibol synthesis NRPS gene. It is likely that there are two originating through the signalling cascade of pal1 and hpl path-
additional NRPS genes, one producing the 11-residue isoforms ways (Fig. 7). Jabs et al. (1997) have already suggested that the
and one producing the 14-residue isoforms. The predicted sizes of induction of phytoalexin biosynthesis in plants might be directly
these genes are approximately 38 and 49 kb, respectively. Indeed, linked to ion fluxes and subsequent intracellular signalling. The
the genomic sequence of Trichoderma reesei reveals two large protective effect of the 18-residue peptaibols against the leaf
NRPSs of 14 and 18 modules of approximately 51 and 69.3kb, pathogen is mediated by the induction of the plant disease
respectively. (http://genome.jgi-psf.org/Trire2/Trire2.home.html). defence system. At comparable and even higher concentrations
Tex1 expression is generally detectable in all the fungal of the peptides, no inhibitory effect on growth of the pathogen
developmental stages we examined, being down-regulated by could be seen after 24 h of direct incubation of bacteria with the
low nitrogen levels in liquid cultures and up-regulated during peptides (data not shown). As Psl are Gram-negative bacteria,
plant–root interaction. In a previous study (Schirmbck et al., the lipopolysaccharides of their outer membrane form a strong
1994) it was shown that Botrytis cinerea cell walls trigger the diffusion barrier against hydrophobic molecules as peptaibols.
production of both cell wall hydrolytic enzymes and peptaibols in This effect has been shown for alamethicin (Duclohier and
T. harzianum. In the present study, simulated mycoparasitism of Wroblewski, 2001) and trichokonins from T. koningii (Song et al.,
T. virens with R. solani cell walls or mycelium did not show tex1 2006). As the tex1 transformants are capable of producing the 11
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Ilan Chet 577
and 14mer peptaibols, the interesting quandary is the inability of CMK40B7 flanking the selectable arg2 gene. This plasmid was
these two peptaibols to induce plant defence responses. There is transformed as described (Thomas and Kenerley, 1989) into the
evidence for major differences in the relative membrane activities arginine auxotrophic strain Tv10.4 with selection for arginine
of peptaibols depending on the length of the peptide chain and prototrophy. DNA was extracted as described (Xu et al., 1996)
amino acid composition (Grigoriev et al., 2003). Efficiency of from 23 putative transformants, digested with either NcoI or
membrane pore formation by peptaibols containing > 17 amino XhoI, and screened by Southern blot analysis to identify strains
acids was much greater than for smaller peptaibols. Peptaibols containing the disruption. A 832-bp probe was amplified from
of 11 residues appear to lack the ability to span membranes, pPSK2 using primers PS9348 (5′-TCAACCATCGGCCACCCTGTC-3′)
greatly decreasing their membrane activity (Shenkarev et al., and PS3-11 (5′-GTGACAAGATGGATGGC-3′). Based on tex1 sequence
2004). data, a positive disruption event was expected to yield a 3.0-kb
Therefore, it has been suggested that the capacity of channel band (vs. a 12.5-kb band in the wild-type) for the NcoI digest and
formation decreases in parallel with the decreasing number of 3.8-kb band (vs. a 8.5-kb band in the wild-type) for the XhoI
the constituting amino acids (Berg et al., 2003). digest.
In addition, it is possible that the genes coding for the 11- and
14-amino-acid peptaibols are less affected by the interaction
Growth and sporulation assays
with the plant roots and more involved as component of
mycoparasitism. Transformants and the WT strains were assayed for radial growth
In the communication zone between plant and Trichoderma and colony morphology. Agar plugs from actively expanding
there are many molecules that can act as PAMPs/elicitors that colonies were placed in the centre of VMS, PDA or WA (water
can affect different signalling pathways, in turn affecting agar) plates. Production of aerial hyphae and colony colour and
different classes of pathogens. In T. virens we have already morphology were visually inspected after 7 days and hyphal
shown that the small hydrophobin-like protein SM1 can induce extension recorded at 24 and 48 h at 27 °C. Conidial production
expression of defence responses in cotton and maize (Djonovic among the various strains was assayed by inoculating PDA with
et al., 2006). Here we present genetic and chemical evidence for conidial suspensions (1 × 106 mL–1). After 10 days of incubation,
the first time that also 18mer peptaibols in T. virens are potent three agar plugs (5 mm diameter) were removed from each of
elicitors of defence responses in cucumber against the leaf five repetitions and placed in 10 mL of sterile water containing
pathogen P. syringae pv. lachrymans and can be assumed as 0.1% triton. Conidial concentrations were determined by
major players in the ‘antigenic’ potential of Trichoderma virens to haemacytometer. Surface area of growth for each day was
trigger non-cultivar-specific defence responses. determined by photographing fungal growth and importing
the images into ImageJ software (http://rsb.info.nih.gov/) for
calculations. The treatments contained four repetitions and
E X P E R I M E N TA L P R O C E D U R E S
each experiment was repeated at least twice. Data analysis
were performed by analysis of variance and Fisher’s PLSD test
Strains, culture conditions and fermentation
(P < 0.05).
The wild-type strain (WT) of Trichoderma virens , Gv29-8, was
isolated from a cotton field in College Station, TX. Strain Tv10.4,
Detection of peptaibols by MALDI-TOF
an arginine auxotroph, was created by a point mutation in the
small subunit of a carbamoyl phosphate synthetase ( arg2) after Mycelia harvested from 9-day-old stationary cultures grown in
treatment with 4-nitroquinoline-1-oxide (NQO) (Baek and liquid VMS were filtered, flash frozen in liquid nitrogen and
Kenerley, 1998). Vogel’s minimal medium (Vogel, 1956) was lyophilized overnight. Dried cells were dissolved in water/
used with different carbon sources: VMS (1.5% sucrose), VMR ethanol/acetonitrile (1 : 1 : 1). All samples were analysed in a
(0.5% Rhizoctonia solani cell wall), VMG (glucose 1.5%). saturated alpha-CHCA matrix solubilized in 30% acetonitrile,
Ammonium was omitted from treatments labelled VM-N, VMG-N 0.07% TFA. A mixture of 1 µL matrix and 1 µL sample was prepared
and VMR-N. Stock cultures were stored at –80 °C as conidial directly on the plate. The samples were analysed using a MALDI
suspensions in 30% glycerol. VOYAGER ELITE time-of-flight mass spectrometer with a nitrogen
laser giving a 337-nm output. The ions were accelerated with a
voltage of 20 kV. Measurements were performed in the delayed
tex1 Disruption vectors and Southern analysis
extraction mode, allowing the determination of monoisotopic
The tex1 disruptants were generated by replacing a 3-kb NdeI mass values. The mass spectrometer was used in the positive ion
fragment coding for module 10 with the arg2 gene. The disruption detection and reflector mode (Texas A&M University, College
vector pPSK2 contained 1.65- and 2.4-kb fragments from cosmid Station, TX, and Anagnostec Luckenwalde, Germany).
© 2007 BLACKWELL PUBLISHING LTD MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746
578 Wolf Prize in Agriculture
MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746 © 2007 BLACKWELL PUBLISHING LTD
Ilan Chet 579
© 2007 BLACKWELL PUBLISHING LTD MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746
580 Wolf Prize in Agriculture
Howell, C.R. (2003) Mechanisms employed by Trichoderma species in the dynamics refinement from transhydrogen bond J couplings. Biophys. J.
biological control of plant diseases. Plant Dis. 87, 4–10. 86, 3687–3699.
Jabs, T., Tschope, M., Colling, C., Hahlbrock, K. and Scheel, D. (1997) Shoresh, M., Yedidia, I. and Chet, I. (2005) Involvement of jasmonic
Elicitor-stimulated ion fluxes and O2- from the oxidative burst are essential acid/ethylene signaling pathway in the systemic resistance induced in
components in triggering defense gene activation and phytoalexin cucumber by Trichoderma asperellum T203. Phytopathology, 95, 76–84.
synthesis in parsley. Proc. Natl Acad. Sci. 94, 4800 –4805. Song, X.Y., Shen, Q.T.X.T., Chen, X.L., Sun, C.Y. and Zhang, Y.Z. (2006)
Jones, J.D.G., Dunsmuir, P. and Bedbrook, J. (1985) High level of Broad-spectrum antimicrobial activity and high stability of trichokonins
expression of introduced chimeric genes in regenerated transformed from Trichoderma koningii SMF2 against plant pathogens. FEMS Microbiol.
plants. The EMBO. J. 4, 2411–2418. Lett. 260, 119–125.
Kim, Y.H., Yeo, W.H., Kim, Y.S., Chae, S.Y. and Kim, K.S. (2000) Antiviral Szekeres, A., Leitgeb, B., Kredics, L., Antal, Z., Hatvani, L.,
activity of antibiotic peptaibols, chrysospemins B and D, produced by Manczinger, L. and Vagvolgyi, C. (2005) Peptaibols and related
Apiocrea sp. 14T against TMV infection. J. Microbiol. Biotechnol. 10, peptaibiotics of Trichoderma. A review. Acta Microbiol. Immunol. Hung.
522–528. 52, 137–168.
Lumsden, R.D., Walter, J.F. and Baker, C.P. (1996) Development of Thomas, M.D. and Kenerley, C.M. (1989) Transformation of the mycoparasite
Gliocladium virens for damping-off disease control. Can. J. Plant Pathol. Gliocladium. Curr. Genet. 15, 415–420.
18, 463–468;. Viterbo, A., Harel, M., Horwitz, B.A., Chet, I. and Mukherjee, P.K.
Marahiel, M.A., Stachelhaus, T. and Mootz, H.D. (1997) Modular (2005) Trichoderma MAP-kinase signaling is involved in induction of
peptide synthetases involved in nonribosomal peptide synthesis. plant systemic resistance. Appl. Environ. Microbiol. 71, 6241–6246.
Chem. Rev. 97, 2651–2674. Vogel, H.J. (1956) A convenient growth medium for Neurospora (Medium
Pozo, M.J., Baek, J.M., Garcia, J.M. and Kenerley, C.M. (2004) Functional N). Microbiol. Genet. Bull. 13, 42–43.
analysis of tvsp1, a serine protease-encoding gene in the biocontrol Wei, X., Yang, F. and Straney, D.C. (2005) Multiple non-ribosomal
agent Trichoderma virens. Fungal Genet. Biol. 41, 336–348. peptide synthetase genes determine peptaibol synthesis in Trichoderma
Rebuffat, S., Goulard, C., Hlimi, S. and Bodo, B. (2000) Two unprece- virens. Can. J. Microbiol. 51, 423–429.
dented natural Aib-peptides with the (Xaa-Yaa-Aib-Pro) motif and an Wiest, A., Grzegorski, D., Xu, B.W., Goulard, C., Rebuffat, S., Ebbole,
unusual C-terminus: structures, membrane-modifying and antibacterial D.J., Bodo, B. and Kenerley, C. (2002) Identification of peptaibols from
properties of pseudokonins KL III and KL VI from the fungus Trichoderma Trichoderma virens and cloning of a peptaibol synthetase. J. Biol. Chem.
pseudokoningii. J. Pept. Sci. 10, 519–533. 277, 20862–20868.
Schirmböck, M., Lorito, M., Wang, Y.L., Hayes, C.K., Arisan-Atac, I., Xu, B.W., Wild, J.R. and Kenerley, C.M. (1996) Enhanced expression of a
Scala, F., Harman, G.E. and Kubicek, C.P. (1994) Parallel formation bacterial gene for pesticide degradation in a common soil fungus. J.
and synergism of hydrolytic enzymes and peptaibol antibiotics, Ferment. Bioeng. 81, 473–481.
molecular mechanisms involved in the antagonistic action of Trichode- Yedidia, I., Shoresh, M., Kerem, Z., Benhamou, N., Kapulnik, Y. and
rma harzianum against phytopathogenic fungi. Appl. Environ. Microbiol. Chet, I. (2003) Concomitant induction of systemic resistance to
60, 4364–4370. Pseudomonas syringae pv. lachrymans in cucumber by Trichoderma
Shenkarev, Z.O., Balashova, T.A., Yakimenko, Z.A., Ovchinnikova, T.V. asperellum (T-203) and accumulation of phytoalexins. Appl. Environ.
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MOLECULAR PLANT PATHOLOGY (2007) 8(6), 737–746 © 2007 BLACKWELL PUBLISHING LTD
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Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 129
onistic activity of necrotrophic mycoparasites is rolfsii were tested to determine if they could stim-
attributed to the production of antibiotics, toxins, ulate germination of conidia of S. sclerotivorum,
or hydrolytic enzymes in proportions that cause none of the chemicals alone or a combination of
the death and destruction of their host. Biotrophic all chemicals induced germination (Fravel et al.
mycoparasites, on the other hand, tend to have 2002). The hyphae penetrate the intercellular
a more restricted host range and produce special- matrix of the conidia, which is composed mainly of
ized structures to adsorb nutrients from their host β-glucans (Ayers et al. 1981). The production and
(Manocha 1990). activity of haustoria by the mycoparasite stimulate
The parasitic relationships between fungi and the host sclerotia to increase their glucanase, and
their significance in biological control are the sub- probably other enzyme activities, resulting in the
jects of this chapter. Both types of mycoparasitism degradation of glucan into available glucose (Bul-
are described and discussed in the scope of their lock et al. 1986). The mycoparasite establishes itself
contribution to biological control. We will con- in the sclerotia, where its mycelium grows out into
centrate on the morphological, biochemical and the surrounding soil to infect additional sclerotia
molecular aspects of mycoparasitism in relation to and to produce new macroconidia (Adams et al.
biological control. The ecological aspects of this 1984). The interaction between S. sclerotivorum
phenomenon are discussed in Jeffries (1997). and Sclerotinia minor depends on both the host
and parasite density (Adams 1986). The infection
process is favored by soil pH, water potential and
II. Mycoparasites as Biocontrol Agents temperature (20–22 ◦ C) (Adams and Ayers 1980).
Under field conditions, a single application of
Many comprehensive reviews on mycoparasitism an S. sclerotivorum preparation at a concentra-
in biological control in general have been pub- tion of 102 or 103 macroconidia g–1 soil caused
lished over recent years (Chet 1990; Deacon 1991; a 75–95% reduction in the number of sclerotia of
Elad and Chet 1995; Benitez et al. 2004). Due to S. minor per plot. Control of lettuce drop caused
their nature, only a few examples of biotrophic my- by S. minor in these plots varied from 40–83% in
coparasites as biocontrol agents exist (Ayers and four consecutive lettuce crops (Adams and Ayers
Adams 1981; Sztejnberg et al. 1989; Adams 1990). 1982). These results were significant but not eco-
Necrotrophic mycoparasites, being more common, nomically important (Adams 1990). Adams (1990)
saprophytic in nature, and less specialized in their concluded that “one of the biggest obstacles to prac-
mode of action, are easier to study. As a result, the tical biological control is the large quantity of the
majority of the mycoparasites used as biocontrol agent necessary to achieve biological control when
agents in greenhouse or field trials to date have applied directly to soil in the field.” He therefore
been necrotrophs. In this chapter, we will empha- suggested two alternatives: (1) to add sclerotia of
size those examples in which the research that has S. minor or a nonpathogenic Sclerotinia that is also
been carried out is both applied and basic in nature. a host of S. sclerotivorum that are already infected
by the mycoparasite; (2) to apply a low dosage of the
mycoparasite preparation to a diseased crop, and
A. Biotrophic Mycoparasites then immediately incorporate the treated crop into
1. Sporidesmium sclerotivorum the soil. This latter procedure ensures that a high
percentage of the mycoparasites will be present in
Sporidesmium sclerotivorum is a dermatiacesus the soil in close contact with the sclerotia of the
hyphomycete that was isolated from field soil by pathogen. Although the author assumes that these
Uecker et al. (1980). In nature, the fungus has been alternatives are easier and more practical, neither
found to be an obligate parasite on sclerotia of has been explored to any significant extent (Adams
Sclerotinia sclerotiorum, S. minor, S. trifoliorum, 1990). Field studies were later conducted (Del Rio
Sclerotium cepivorum, and Botrytis cinerea (Ayers et al. 2002) to evaluate the effectiveness of S. sclero-
and Adams 1981). In response to chemicals tivorum to control Sclerotinia stem rot of soybean.
released by the host’s sclerotia, macroconidia of Experimental plots were infested with S. sclerotivo-
S. sclerotivorum in the soil germinate and the rum macroconidia at a rate of 0, 2, or 20 spores per
germ tubes infect the sclerotia. When volatile cm2 . Two years later, the disease was completely
compounds secreted by sclerotia of Sclerotinia suppressed in all plots. S. sclerotivorum was re-
minor, Sclerotinia sclerotiorum, and Sclerotium trieved from all infested plots at all locations 2 years
584 Wolf Prize in Agriculture
after infestation with sclerotia of S. sclerotiorum as by A. quisqualis was studied by scanning electron
bait. This paper constitutes the first report describ- microscopy. Within 24 h after inoculation, the
ing the biocontrol of a disease on field crops that hyperparasite had germinated, and the germ tubes
may be employed economically. had developed appressorium-like structures at
the point of contact with the powdery mildew
host. Both conidia and hyphae were parasitized
2. Ampelomyces quisqualis by penetration. Within 5 days of inoculation,
the hyperparasite had developed pycnidia with
Ampelomyces quisqualis, a hyperparasite on conidia on the powdery mildew hyphae and
Erysiphales, has been reported as a biocontrol conidiophores (Sunhdeim and Krekling 1982).
agent of powdery mildews (Sztejnberg et al. 1989). Hashioka and Nakai (1980) used both transmission
An isolate of A. quisqualis obtained from an Oid- and scanning electron microscopy to study the
ium sp. infecting Catha edulis in Israel proved to be hyphal extension and pycnidial development of the
infective to several powdery mildew fungi belong- mycoparasite A. quisqualis Ces. inside the hyphae,
ing to the genera Oidium, Erysiphe, Sphaerotheca, and conidiophores of several species of powdery
Podosphaera, Uncinula, and Leveillula. In field mildew fungi belonging to Microsphaera, Erysiphe,
trials, A. quisqualis parasitized the powdery and Sphaerotheca. The mycoparasite cells grew
mildews of cucumber, carrot, and mango, and normally inside the host cells, despite gradual
reduced the disease. A. quisqualis was tolerant to degeneration of these latter cells. The invading
many fungicides used to control powdery mildews hyphal cells of the mycoparasite migrated into the
and/or other plant diseases. Treating powdery neighboring host cells by constricting themselves
mildew of cucumber (cv. Hazera 205) with spores of through the host cell’s septal pore. The mycopar-
A. quisqualis alone significantly decreased disease asite extended hyphae inside the conidiophores
severity and increased cucumber yield by approxi- of the hosts, and formed pycnidia consisting of
mately 50%. Combining the fungicide pyrazophos a unicellular outer layer and interior cells that
with the mycoparasite resulted in a larger increase later differentiated into conidiogenous structures
in cucumber yield (Sztejnberg et al. 1989). (Hashioka and Nakai 1980). Recent studies based
Treating powdery mildew-infected zucchini both on morphological and life cycle parameters
leaves with A. quisqualis increased the rates of and ribosomal DNA internal transcribed spacer
photosynthesis from 3.8 µmol CO2 m−2 s−1 in region 1 sequence analysis have shown that isolates
untreated plants to 10.2, compared to 12.8 in un- previously attributed to the genus Ampelomyces
infected healthy plants (Sztejnberg and Abo-Foul were actually isolates of Phoma spp. Phoma
1990). Electron micrographs of leaf sections of dis- glomerata can colonize and suppress development
eased cucumber plants revealed marked deteriora- of powdery mildew on oak, and may have utility as
tion on the morphological organization of chloro- a mycoparasitic agent (Sullivan and White 2000).
plast membranes. Chloroplasts of A. quisqualis-
treated plants seemed undamaged, like those of un-
treated plants (Abo-Foul et al. 1996). Fluorescence B. Necrotrophic Mycoparasites
measurements (e.g., low-temperature fluorescence 1. Pythium nunn
emission spectra, and room-temperature fluores-
cence transients) indicated a disease-correlated Pythium nunn is a mycoparasite isolated from soil
increase in levels of uncoupled chlorophyll (Abo- suppressive to a plant parasitic Pythium sp. When
Foul et al. 1996). A simple, inexpensive medium this mycoparasite was introduced into soil con-
based on potato dextrose broth (PDB) was devel- ducive to Pythium sp., the competitive saprophytic
oped for mass production of infective spores of ability of this isolate was suppressed. An inverse re-
A. quisqualis in fermentation for biological control lationship was found between propagule densities
(Sztejnberg et al. 1990), which was later developed of the plant pathogen and of the antagonist P. nunn
in the commercial biofungicide AQ10. (Lifshitz et al. 1984b).
The interaction between the hyperparasite The modes of hyphal interaction between the
A. quisqualis and its host fungi was studied by mycoparasite P. nunn and several soil fungi were
Hashioka and Nakai (1980) and Sundheim and studied by both phase-contrast and scanning elec-
Krekling (1982). The infection process of the tron microscopy (Lifshitz et al. 1984a). In the zone
cucumber powdery mildew Sphaerotheca fuliginea of interaction, P. nunn massively coiled around
Ilan Chet 585
Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 131
and subsequently lysed hyphae of P. ultimum T. flavus resulted in granulation of the cytoplasm
and P. vexans without penetration. In contrast, and collapse of the cell walls (McLaren et al. 1986).
P. nunn penetrated and eventually parasitized Direct invasion of R. solani hyphae via the pro-
hyphae of R. solani, P. aphanidermatum, Phy- duction of penetration pegs by T. f lavus was ob-
tophthora parasitica, and P. cinnamomi, forming served by Boosalis (1956). These pegs developed
appressorium-like structures. However, P. nunn from either a mycelium coiling around the host hy-
was not mycoparasitic against F. oxysporum f. phae or from a hypha in direct contact with the
sp. cucumerinum or Trichoderma koningnii, and host. Fahima and Henis (1990) applied T. flavus
was destroyed by T. harzianum and T. viride. The as an ascospore suspension to soil naturally in-
authors concluded that P. nunn is a necrotrophic fested with Verticillium dahliae, the causal agent
mycoparasite with a limited host range and of Verticillium wilt in eggplant. Twelve weeks after
differential modes of action among susceptible transplanting, 77% disease reduction was achieved,
organisms (Lifshitz et al. 1984a). compared with the untreated control.
Lysis and penetration of the host cell wall at Scanning electron micrographs showed heavy
the site of interaction with the mycoparasite were fungal colonization and typical T. flavus conidia
demonstrated by Elad et al. (1985). Calcofluor on the surface of the microsclerotia buried in the
White M2R binds to the edges of polysaccharide treated soil, but not in control soils. Transmission
oligomers (Kritzman et al. 1978). Using this electron micrographs of microsclerotia incubated
reagent, the appearance of fluorescence indicated with T. flavus on agar revealed parasitism involving
localized lysis of the host cell wall by P. nunn. invasion of some host cells by means of small pen-
The cell walls of Oomycota are composed of etration pegs; the host cell walls were lysed mainly
β-glucan, cellulose, and less than 1.5% chitin. at their site of contact with the parasite hyphal tips.
Basidiomycota and Ascomycota contain mainly Further colonization of the microsclerotial cells oc-
β-glucan and chitin but no cellulose. P. nunn curred simultaneously with the degradation of the
produced large amounts of β-1-3-glucanase and invaded host cell contents, rather than the cell walls
chitinase in liquid cultures containing cell walls of (Fahima et al. 1992). It was suggested that myco-
pathogenic fungi belonging to the class Basidiomy- parasitism of V. dahliae microsclerotia by T. flavus
cota. This mycoparasite produced cellulase but no hyphae may be involved in the biological control of
chitinase when grown on culture containing cell Verticillium wilt disease. Fravel and Keinath (1991),
walls of two pathogens belonging to the Oomycota however, claimed that T. flavus is known to produce
(Elad et al. 1985). These extracellular hydrolytic compounds that mediate antibiosis, which is there-
enzymes were detected in P. nunn when grown fore suspected of being involved in the control of
in dual culture with six host fungi but not with Verticillium wilt of eggplant and potato. Similarly,
ten nonhost fungi, indicating specificity in the McLaren et al. (1986) observed that hyphal cells of
antagonistic activity of P. nunn (Baker 1987). S. sclerotiorum eventually collapse as a result of in-
fection by T. flavus, but host cell walls remain intact.
2. Talaromyces flavus They suggested that cell wall-degrading enzymes
may not play a major role in the control of S. sclero-
Talaromyces flavus (the perfect stage of Penicillium tiorum by T. flavus, and that antibiotics produced
dangeardii; synonym: P. vermiculatum) is a my- by the parasite may be involved in the deteriora-
coparasite of several soil-borne plant pathogenic tion of the host’s hyphae (McLaren et al. 1986). In
fungi including R. solani (Boosalis 1956), S. scle- a recent work (Duo-Chuan et al. 2005), two chiti-
rotiorum (McLaren et al. 1986) and Verticillium nases (CHIT41 and CHIT32) were isolated from
spp. (Fahima and Henis 1990). Laboratory investi- T. flavus and were shown to be able to decompose
gations using light and electron microscopy indi- chitin in the cell walls of V. dahliae, S. sclerotiorum
cate that T. flavus is a destructive hyperparasite of and R. solani, thus indicating that these enzymes
S. sclerotiorum. In dual culture, hyphae of T. flavus may play an important role in the mycoparasitic
grew toward, and coiled around the host hyphal behavior of T. flavus.
cells. The coiling effect intensified as the hyphae
of T. flavus branched repeatedly on the host sur- 3. Corniothyrium minitans
face. Tips of the hyphal branches often invaded the
host by direct penetration of the cell wall without Corniothyrium minitans has been found to be
formation of appressoria. Infection of host cells by a natural mycoparasite of sclerotia of the plant
586 Wolf Prize in Agriculture
Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 133
Phillips (1986) studied aspects of the biology contact with Botrytis allili to cause severe internal
of G. virens and its parasitism of sclerotia of disorganization of host cells, coagulation of
S. sclerotiorum in soil. G. virens parasitized cytoplasm, vacuolation, and loss of contents from
and decayed sclerotia of S. sclerotiorum, S. mi- organelles. Cultures of B. allili parasitized by G. ro-
nor, Botrytis cinerea, Sclerotium rolfsii, and seum contained considerable β-(1-3)-glucanase
Macrophomina phaseolina on laboratory media, and chitinase, and the cytoplasm coagulated
and caused a reduction in the survival of sclerotia without physical contact. G. virens isolate G1-21
of S. sclerotiorum in soil. However, parasitism of was grown on various solid and liquid media:
the mycelium was not detected. wheat bran and peanut hull meal (PHM), as well
A strain of G. virens isolated from the para- as spent glucose tartrate broth (GTB), Czapek-Dox
sitized hyphae of R. solani by Howell (1982) signif- broth (CDB), and potato dextrose broth (PDB)
icantly suppressed damping-off in cotton seedlings (Lewis et al. 1991). Aqueous extracts of these media
by this pathogen and by Pythium ultimum. Treat- caused leakage of carbohydrates and electrolytes
ment with G. virens more than doubled the num- from hyphae of the soil-borne plant pathogen
ber of surviving cotton seedlings grown in soil in- R. solani, and its mycelial weight was reduced. Size
fested with either pathogen. G. virens parasitized fractionation experiments indicated that it was
R. solani by coiling around, and penetrating the hy- a combination of factors associated with G. virens,
phae. P. ultimum was not parasitized by G. virens, rather than a single one, which induced this
but was strongly inhibited by antibiosis. Treatment phenomenon. Gliotoxin was detected in culture
of soil infested with propagules of R. solani or P. ul- filtrates from G. virens grown on bran and PHM
timum with G. virens resulted in a 63% reduction media. Gliotoxin preparations induced leakage
in the number of viable R. solani sclerotia after of carbohydrates and electrolytes from R. solani,
3 weeks of incubation, whereas oospores of P. ul- and caused a concomitant reduction in mycelial
timum were unaffected. Strains of G. virens were weight, which suggests the action of a leakage
separated into two distinct groups, P and Q, on the factor (Lewis et al. 1991). The authors speculated
basis of secondary metabolite production in vitro that hydrolytic enzymes such as β-1-3-glucanase,
(Howell et al. 1993). β-1-4-glucanase, chitinase, and protease, shown
Gliovirin was very inhibitory to P. ultimum, to be produced by isolates of G. virens (Roberts
but exhibited no activity against R. solani, and and Lumsden 1990), have the potential to act on
strains that produced it (P group) were more ef- R. solani cell walls and membranes. The role of
fective seed-treatment biocontrol agents of disease extracellular chitinase in the biocontrol activity
incited by P. ultimum. Conversely, gliotoxin was of Trichoderma virens was later examined using
more active against R. solani than against P. ulti- genetically manipulated strains of this fungus. The
mum, and strains that produced it (Q group) were T. virens strains in which the chitinase gene (cht42)
more effective seed treatments for controlling dis- was disrupted (KO) or constitutively overexpressed
ease incited by R. solani. Based on these results, (COE) were constructed through genetic trans-
the authors suggested that it may be necessary to formation. Biocontrol activity of the KO and COE
treat seeds with a combination of strains in order strains were significantly decreased and enhanced,
to broaden the disease control spectrum. respectively against cotton seedling disease incited
Howell (1987) isolated mutants of G. virens, by Rhizoctonia solani when compared with the
obtained by irradiation with ultraviolet light, that wild-type strain (Baek et al. 1999).
showed no mycoparasitic activity. The selected mu- More than 60 years ago, Weindling (1932) was
tants retained the same antibiotic complement as the first to demonstrate the mycoparasitic nature
the parent strains. Peat moss-Czapek’s broth cul- of fungi from the genus Trichoderma. He suggested
tures of parent and mutant strains were similarly their potential use as biocontrol agents of plant
effective as biocontrol agents of cotton seedling dis- pathogenic fungi. However, the first report on a bio-
ease induced by R. solani, and as antagonists of logical control experiment using Trichoderma spp.
R. solani sclerotia in soil. In the light of these results, under natural field conditions came 40 years later,
Howell (1987) concluded that mycoparasitism is by Wells et al. (1972) who used T. harzianum grown
not a major mechanism in the biological control of on an autoclaved mixture of ryegrass seeds and soil
R. solani-incited seedling disease by G. virens. to control Sclerotium rolfsii Sace. Since then, more
In addition, Pachenari and Dix (1980) con- Trichoderma isolates have been obtained from nat-
cluded that G. virens need not make intimate ural habitats, and used in biocontrol trials against
588 Wolf Prize in Agriculture
several soil-borne plant pathogenic fungi under T. harzianum was applied as a seed coating or
both greenhouse and field conditions (Chet 1990; as a wheat branpeat (1:1, v/v) preparation intro-
Harman and Lumsden 1990; Harman 2006). duced into the tomato rooting mixture. Tricho-
A seed treatment was developed by Harman derma-treated transplants were better protected
et al. (1980) to reduce the amount of Trichoderma against Fusarium crown rot than untreated con-
added to the soil to control soil-borne plant trols when planted in MB-fumigated or nonfumi-
pathogenic fungi. T. hamatum conidia applied gated infested fields. The total yield of tomatoes in
in the laboratory, to seeds of pea and radish as the T. harzianum-treated plots increased as much
a Methocel slurry, provided protection to seeds as 26.2% over the controls. Integrated control of
and seedlings from Pythium spp. and R. solani, Verticillium dahliae in potato by T. harzianum and
respectively, almost as effectively as fungicide the fungicide Captan was reported by Ordentlich
seed treatment. Establishment of the mycoparasite et al. (1990).
and long-term action were demonstrated, as the
propagules of T. hamatum increased approxi-
mately 100-fold in soils planted with treated seeds. C. Induced Systemic Resistance
Population densities of R. solani and Pythium spp. by Trichoderma spp.
were lower in soils containing T. hamatum than in
soils lacking this antagonist. Replanting these soils Some Trichoderma rhizosphere-competent strains
once, or even twice with untreated seeds yielded colonize entire root surfaces with morphological
lower disease incidence than in soils originally features reminiscent of those seen during myco-
planted with untreated seeds. Addition of chitin parasitism (Yedidia et al. 1999). Penetration of the
or R. solani cell walls to the coating of seeds root tissue is usually limited to the first or second
previously treated with a conidial suspension layers of cells, and occurs only in the intercellu-
increased both the ability of T. hamatum to protect lar spaces. Trichoderma strains capable of estab-
the seeds against Pythium spp. or R. solani, and lishing such interaction induce metabolic changes
the population density of Trichoderma in the soil. in plants that increase resistance to a wide range
T. hamatum with chitin, but without R. solani of plant-pathogenic microorganisms and viruses
cell walls, effectively reduced damping-off caused (Harman et al. 2004; Fig. 8.1). This response seems
by Pythium spp., compared to seed treatment to be broadly effective for many plants, which in-
containing only T. hamatum (Harman et al. dicates that there is little or no plant specificity.
1980). Sivan et al. (1984) applied a peat-bran At least three classes of substances that elicit
mixture (1:1 v/v) preparation of T. harzianum plant defense responses have been identified.
(isolate 315) to either soil or rooting mixture, These elicitors include proteins, peptides, and
and efficiently controlled damping-off induced low-molecular-weight compounds (Harman
by Pythium aphanidermatum in pea, cucumber, et al. 2004; Viterbo et al. 2004). The systemic
tomato, pepper, and gypsophila. Several isolates response in plants occurs through the jasmonic
of T. harzianum and T. hamatum were found to
antagonize and control Macrophomina phaseolina
in beans and melon (Elad et al. 1986). Isolates of
T. harzianum and T. hamatum antagonized and
controlled Rosellinia necatrix in almond seedlings
(Freeman et al. 1986). Sztejnberg et al. (1987) com-
bined sublethal soil heating with an application of
T. harzianum to yield better control of R. necatrix
than that achieved by either treatment alone.
Sivan and Chet (1986) isolated a new Tricho-
derma harzianum isolate (T-35) from the rhizo-
sphere of cotton plants grown in fields infested
with Fusarium. In a further study, the isolate was
tested in biological control trials over two succes- Fig. 8.1. Induced resistance toward the leaf pathogen
Cochliobolus heterostrophus in maize. Seedling roots were
sive growing seasons against Fusarium crown rot infected with germinated Trichoderma spores (105 ml−1 )
of tomato in fields naturally infested with F. oxys- 48 h prior to pathogen leaf infection (800 spores). The
porum f. sp. radici lycopersici (Sivan et al. 1987). symptoms were recorded 72 h after infection
Ilan Chet 589
Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 135
acid/ethylene signaling pathway in a way similar vidually or as a group, are responsible for the sup-
to the rhizobacteria-induced systemic resistance pression effect. Mycoparasitism, the focus of this
(van Loon et al. 1998; Shoresh et al. 2005). Several chapter, is a major mechanism of specific suppres-
studies have shown that root colonization by sion. For example, Chet and Baker (1981) reported
Trichoderma strains results in massive changes in on a soil which was suppressive to R. solani of car-
plant gene expression patterns and metabolome. nation near Bogota, Colombia. This soil contained
Changes in plant metabolism lead to the accumu- high levels of organic matter (35%), was highly
lation of antimicrobial compounds. In cucumber, acidic (pH 5.1), and its main microbiological com-
root colonization by T. asperellum strain T-203 ponent was the antagonistic fungus T. hamatum at
causes an increase in phenolic glucoside levels in a population density of 8 ×105 propagules g−1 . The
leaves, which are strongly inhibitory to a range level of this mycoparasite in mineral-conducive soil
of bacteria and fungi (Yedidia et al. 2003). The was four orders of magnitude lower.
protection afforded by the biocontrol agent is In another, related study, Henis et al. (1978)
associated with the accumulation of mRNA of showed the effect of successive plantings on
two defense genes: the phenylpropanoid pathway the development of suppression. A soil sown
gene phenylalanine ammonia lyase (PAL) and with radish every week became suppressive to
the lipoxygenase pathway gene hydroxyperoxide R. solani by the fourth sowing, and was even more
lyase (HPL) (Yedidia et al. 2003). Increased levels suppressive by the fifth and subsequent sowings.
of other defense-related plant enzymes, such as The population of T. harzianum, antagonistic to
peroxidases, chitinases, and β-1,3-glucanases, have R. solani, increased with successive sowings of
been recorded in Trichoderma-treated cucumber radish (Liu and Baker 1980), possibly in response
seedlings upon pathogen challenge (Shoresh et al. to increases in the amount of R. solani in the
2005). This potentiation in the gene expression soil resulting from its parasitism of the radish
enables Trichoderma-treated plants to be more seedlings. The addition of T. harzianum spores to
resistant to subsequent pathogen infection. The a conducive soil at the same density as that found
MAPK signal transduction pathways, both of the in the suppressive soil caused the conducive soil to
plant and of Trichoderma, are important for the become suppressive. Trichoderma spp. were also
induction of systemic resistance (Viterbo et al. reported to be responsible for the suppression of
2005; Shoresh et al. 2006). the take-all disease caused by Gaeumannomyces
graminis. Low pH conditions were found to
be favorable to Trichoderma, and to enhance
D. Mycoparasitism suppression (Simon and Sivasithamparam 1990).
in Suppressive Environments A practical approach to utilizing suppression
(Soils and Composts) in agriculture is the use of suppressive composts,
mainly in container media. Composting is the
Suppression of soil-borne plant pathogens occurs breakdown of organic waste material by a suc-
in environments such as field soils, and soils cession of mixed populations of microorganisms
amended with organic matter or compost as the in a thermophilic aerobic environment. The final
organic component in media for container-grown product is compost or humus, which is the stabi-
plants. A review on this topic for field soils was lized organic matter populated by microorganisms
recently published by Stone et al. (2004). capable of suppressing soil-borne plant pathogens.
Pathogen suppressiveness has been defined Disease-suppressive effects of composts have
by Cook and Baker (1983) as “soils in which the been investigated intensively over the past two
pathogen does not establish or persist, establishes decades, and were recently reviewed by Noble
but causes little or no damage, or establishes and and Coventry (2005) and Zinati (2005). Compost
causes disease for a while but thereafter the disease of a wide variety of waste materials (hardwood
is less important, although the pathogen may or pine bark, municipal sludge, grape marc, or
persist in the soil.” cattle manure) is an economically and ecologically
Baker and Cook (1974) divided suppression sound alternative to pesticides.
mechanisms into two broad categories defined as The mechanisms of suppression in composts
general and specific. General suppression is a re- do not differ substantially from those described
sult of total microbial activity. In contrast, specific for soils, and can be either general or specific.
suppression applies when bacteria or fungi, indi- Physiological profiling, and the use of DNA-based
590 Wolf Prize in Agriculture
techniques such as denaturing gradient gel elec- 1981). Attachment of a biotrophic mycoparasite
trophoresis (DGGE) may lead to an improved to its host surface is considered to be an essential
understanding of the changes in microbial com- prerequisite step for further penetration of the
munities associated with disease control resulting host by the parasite (Manocha and Chen 1990).
from compost amendment of soil, sand, or peat. P. virginiana attaches to the surface of both
Nelson et al. (1983) identified specific strains of the compatible Choanephora cucurbitarum and
four Trichoderma spp. and isolates of Gliocladium Mortierella pusilla, and the incompatible Phas-
virens as the most effective fungal hyperparasites colomyces articulosus hosts, but not to the surface
of R. solani present in bark compost. A few of the of the nonhost Mortierella candelabrum (Manocha
230 other fungal species also showed activity, but 1985; Manocha et al. 1986). Comparative research
most were ineffective. Kwok et al. (1987) described was performed by Manocha and his coworkers in
synergistic interactions between T. hamatum and an attempt to unravel the molecular basis for speci-
Flavobacterium balustinum. Several other bacte- ficity and recognition in this system. Cytological
rial strains, including Enterobacter, Pseudomonas, and biochemical investigations were carried out
and Xanthomonas spp., also interacted with the to study the structure and chemical composition
Trichoderma isolate in suppression of Rhizoctonia of cell walls of host and nonhost species (Manocha
damping-off (Kwok et al. 1987). Composted grape 1981, 1987). The germ tubes of the biotrophic
marc was effective in suppressing disease caused mycoparasite P. virginiana were found to attach
by S. rolfsii in beans and chickpeas (Gorodecki and to the cell-wall surface of the host, but not to that
Hadar 1990). Hadar and Gorodecki (1991) placed of the nonhost (Manocha 1985; Manocha et al.
sclerotia of S. rolfsii on composted grape mare to 1986). This attachment could be specifically in-
isolate hyperparasites of this pathogen. Viability hibited by chitobiose and chitotriose. The authors
of sclerotia decreased from 100% to less than 10% therefore suggested a possible involvement of
within 40 h. It remained close to 100% for sclerotia carbohydrate-binding proteins in the specificity
placed on a conducive peat mix. Penicillium spp. of this interaction. A comparison of protein and
and Fusarium spp. were observed by scanning glycoprotein profiles of cell-wall extracts revealed
electron microscopy to colonize the sclerotia. Tri- marked differences between host and nonhost
choderma populations in the grape mare compost species. Two high-molecular-weight glycoproteins
were at very low levels (102 cfu g–1 dry weight). were observed only in the extract of host cell walls,
The hyperparasites present in this compost are being absent in that of the nonhost (Manocha 1985;
therefore quite different from those isolated Manocha et al. 1986). Further isolation and charac-
from tree bark compost, where Trichoderma and terization of the host cell surface proteins revealed
Gliocladium isolates predominate. that attachment and appressorium formation by
In conclusion, suppression of soil-borne plant the parasite germ tubes could be inhibited by treat-
pathogens in field soil or container media is ing host cell-wall fragments with 0.1 M NaOH or
brought about by antagonistic microorganisms. pronase E. Furthermore, the two purified glycopro-
Such systems could be a source for mycoparasities teins were able to agglutinate both nongerminated
to be used in biocontrol, or to be incorporated and germinated spores of the mycoparasite.
into integrated disease control programs. The Arabinose, glucose, and N-acetylglucosamine
inoculation of composts with biological control could totally inhibit this agglutination. These
agents may improve the efficacy and reliability of glycoproteins were suggested to be two subunits
disease control obtained. of a carbohydrate-binding agglutinin present
on the host cell surface, and to be involved in
agglutination and attachment of the mycoparasite
germ tubes (Manocha and Chen 1991).
III. Hyphal Interactions Using fluorescein isothiocyanate-labeled
in Mycoparasitism lectin-binding techniques, Manocha et al. (1990)
were able to show differences in the distribution
A. Biotrophs pattern of glycosyl residues at the level of the cell
wall between fungi that are hosts and those that
Piptocephalis virginiana is a haustorial biotrophic are nonhosts of the mycoparasite P. virginiana,
mycoparasite that parasitizes fungi belonging and at the protoplast level between compatible and
to the order Mucorales exclusively (Manocha incompatible hosts.
Ilan Chet 591
Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 137
The cell walls of the compatible hosts (C. cucur- are not a major factor in recognition at this
bitarum and M. pusilla) and the incompatible host level (Manocha et al. 1990). Immunofluorescence
(P. articulosus), as well as that of the mycoparasite microscopy was used to detect, in the mycoparasite
itself, contain glucose and N-acetylglucosamine. P. virginiana, the presence of a complementary
In the nonhost (M. candelabrum), however, other glycoprotein that binds specifically to the host
sugars such as fucose, N-acetylgalactosamine, cell surface glycoproteins. This technique revealed
and galactose could also be detected. These latter surface localization of the protein on the germ
sugars could be detected on both the host and tubes of P. virginiana. Fluorescence was also
the parasite surface after mild treatment with observed at the surface of the germinated spores
proteinase or when grown in liquid medium. and hyphae of the host M. pusilla, after treatment
The researchers speculated that the failure of with complementary protein from P. virginiana,
the mycoparasite to attach to the host cells and with primary antibody prepared against the
after proteinase treatment or in liquid culture complementary protein (Manocha et al. 1997).
may be due to the appearance of galactose and
galactosamine at the host cell surface. The idea
that N-acetylglucosamine and glucose may be B. Necrotrophs
involved in the attachment of P. virginiana to its
host cell surface was supported by the observation As early as 1932, Weindling reported the coiling of
that pretreatment of the mycoparasite germ tubes Trichoderma spp. hyphae around hyphae of other
with N-acetylguclosamine or glucose inhibited fungi. These strains were later shown to actually
their attachment to the host cells. In addition, the be a species of Gliocladium (Webster and Lomas
germ tubes attached to agarose beads coated with 1964). Dennis and Webster (1971a, b, c) published
glucose or with N-acetylglucosamine, but not with an extensive report on the antagonistic properties
N-acetylgalactosamine (Manocha et al. 1990). of species groups of Trichoderma. The hyphal inter-
The protoplast surfaces of compatible hosts action between Trichoderma and plant pathogenic
contained all of the above-listed sugars, and fungi was first comprehensively studied in their
these protoplasts could attach to the germ tube work. Since then, numerous studies on the hy-
of the mycoparasite. Only lectins specific for phal interaction and coiling phenomenon of Tri-
N-acetylglucosamine and glucose were bound choderma around its host hyphae have been carried
at the protoplast surface of the incompatible out with the use of light and electron microscopy
host; these protoplasts did not attach to the (Chet et al. 1981; Elad et al. 1983a; Baker 1987; Inbar
mycoparasite germ tubes. Indications were found and Chet 1992; Omero et al. 1999; Rocha-Ramirez
for different factors being responsible for at- et al. 2002; Fig. 8.2).
tachment and for appressorium formation, as The destructive mode of parasitism in Tricho-
pretreatment of the mycoparasite with glucose derma appears to be a process consisting of sev-
and N-acetylglucosamine inhibited its attachment eral consecutive events initiated by attraction and
to the host cell surface, but had no obvious effect directed growth of Trichoderma toward its host,
on appressorium formation. On the other hand, probably by chemotropism. Positive chemotropism
appressorium formation was inhibited by heat was found in Trichoderma (Chet et al. 1981), as
treatment of host cell-wall fragments that still it could detect its host from a distance and be-
permitted attachment (Manocha et al. 1990). gin to branch in an atypical way. These branches
The authors therefore suggested a model for grew toward the pathogenic host fungi. Similar be-
the recognition between P. virginiana and its havior was also found in Pythium nunn (Lifshitz
host fungi that operates at two levels at least: et al. 1984a), P. oligandrum (Lewis et al. 1989),
the cell wall, and the protoplast surface. At the and in Gliocladium spp. (Huang 1978). This event
cell-wall level, the attachment probably involves is presumably a response of the antagonist to the
carbohydrate-binding agglutinins that recognize chemical gradient of an attractant coming from the
specific sugar residues on the host but not on the host. However, no specific stimuli other than amino
nonhost cell wall. After the initial recognition and acids and simple sugars have thus far been detected
attachment, at the protoplast level, the parasite (R. Barak and I. Chet, unpublished data). Hence,
distinguishes compatible from incompatible hosts. the specificity of the phenomenon is not clear. Ap-
The mechanism of this distinction is not clear. parently, it is not an essential step for mycopara-
Yet, it seems that protoplast membrane sugars sitism, although it may hold some advantage for
592 Wolf Prize in Agriculture
Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 139
amounts of chitin (<1.5%). Therefore, to penetrate ing techniques have been applied to gain a better
the host cell wall, mycoparasites should have and more basic understanding of the system,
a system of hydrolytic enzymes that can degrade as well as to develop superior and improved
these components. Enzymatic degradation of strains of biocontrol agents with enhanced activity
fungal cell walls occurs mainly via the excretion (Mendoza et al. 2003; Brunner et al. 2005). The
of the extracellular enzymes β-1-3-glucanase chitinase gene chiA, encoding one of the chitinases
and chitinase. Indeed, high β-1-3-glucanase and from Serratia marcescens, a well-known biocontrol
chitinase activities were detected in dual cultures agent, was isolated and cloned into E. coli (Shapira
when T. harzianum parasitized S. rolfsii, contrast- et al. 1989). E. coli transformed by the chiA
ing with the low levels found with either fungus gene, under the oLpL operator and promoter of
alone. Cycloheximide prevented antagonism, and bacteriophage, expressed and excreted the corre-
enzymatic activity was diminished (Elad et al. sponding protein into the growth medium. Almost
1983b). Using gold cytochemistry, T. harzianum pure S. marcescens chitinase from E. coli or whole
hyphae were shown to coil around and penetrate viable cells were used in greenhouse experiments
cells of R. solani, causing extensive damage such as against S. rolfsii in beans, and R. solani in cotton.
cell-wall alteration, plasma membrane retraction, Using the chitinase preparation in the irrigation
and cytoplasm aggregation (Benhamou and Chet water effectively reduced the number of diseased
1993). plants. Whole viable cells of transformed E. coli
The involvement and importance of lytic en- were also effective in inhibiting S. rolfsii, but to
zymes, mainly chitinase, in the biological control a lesser degree. The genetically engineered E. coli,
of plant pathogens by both fungi and rhizobacte- a nonsoil bacterium, served here as a model system
rial agents (Ordentlich et al. 1988; Inbar and Chet to demonstrate the role of chitinase in controlling
1991; Sahai and Manocha 1993; Viterbo et al. 2002), a chitin-containing plant pathogen.
as well as their involvement in the defense of plants It is suggested that the introduction of such
against pathogenic infection (Boller 1985; Broglie engineered genes into soil bacteria will increase
et al. 1991) is well documented. control efficiency by combining high expression
of a gene coding for a lytic enzyme with rhi-
zosphere competence. Southern blot analysis of
the chiA gene cloned from S. marcescens showed
IV. Molecular Aspects
homology to one of the Trichoderma chitinase
and Genetic Engineering genes. Based on this, the chiA gene was used as
in Mycoparasitism a probe to isolate a chitinase gene from a cDNA
library prepared from T. harzianum (T-35) grown
Trichoderma is one the most frequently used bio- on chitin (Chet et al. 1993). The chitinase gene
control agents in agriculture. The role of lytic en- from T. harzianum (T-35) was cloned in a Blue-
zymes in its mycoparasitic activity has recently script plasmid under the lac promoter. When
been largely reviewed (Viterbo et al. 2002; Ben- the transformed E. coli was plated on LB+0.2%
itez et al. 2004). The sensing of the host in the chitin plates and induced by 1 mM IPTG, the
Trichoderma mycoparasitic interaction and gene bacteria showed chitinolytic activity. In green-
activation has been the subject of extensive studies house experiments, irrigation of bean seedlings
in the last few years (Inbar and Chet 1995; Zeilinger with 107 cfu g−1 soil day−1 of E. coli XLIBlue,
et al. 1999; Brunner et al. 2003). The pattern of in- transformed with the Trichoderma chitinase gene
duction of different cell wall-degrading enzymes induced by 1 mM IPTG, resulted in significant
differs from one Trichoderma strain to another. It biocontrol activity. Suppression of the disease
is believed that Trichoderma secrete exochitinases caused by S. rolfsii was obvious. The treated plants
constitutively at low levels. When chitinases de- exhibited a better growth rate than untreated con-
grade fungal cell walls, they release oligomers that trols. After 18 days, the growth rate of the plants
induce other chitinases, and attack begins. irrigated with the transformed bacteria was similar
In the last decade, the significance of several to that of uninfected plants (Chet et al. 1993).
newly isolated lytic enzymes has been demon- In an attempt to increase its effectiveness,
strated by overexpression and deletion of the T. harzianum protoplasts were cotransformed
respective genes (Pozo et al. 2004; Hoell et al. using two plasmids: pSL3chiAII, containing a bac-
2005). Molecular approaches and genetic engineer- terial chitinase gene from S. marcescens under
Ilan Chet 595
Plant Disease Biocontrol and Induced Resistance via Fungal Mycoparasites 141
the control of a constitutive viral promotor, and to fungal pathogens. Transgenic apple plants ex-
p35SR2, a marker for selection after transforma- pressing T. atroviride endochitinase and exochiti-
tion, encoding for acetamidase. Two transformants nase, singly or in combination, were produced and
showed increased constitutive chitinase activity screened for resistance to Venturia inaequalis, the
(specific activity 11 and 5 times higher than the causal agent of apple scab (Bolar et al. 2001). Plants
recipient; Fig. 8.4), and excreted a protein of expressing both enzymes at the same time were
ca. 58 kDa, the expected size of S. marcescens more resistant, demonstrating for the first time in
chitinase, when grown on synthetic medium. planta synergism between the two enzymes. Con-
Antagonistic activity of the transformants was stitutive expression of Trichoderma endochinase
significantly higher than that of the wild-type can be exploited to enhance resistance to fungal
T. harzianum, as evaluated by testing their ability pathogens in important forest tree species. The
to overgrow the plant pathogen S. rolfsii in dual ech42 from T. harzianum was introduced into for-
culture (Haran et al. 1993). The major advantage of est trees, black spruce (Picea mariana) and hy-
such genetic manipulations is the ability to isolate brid poplar (Populus nigra x Populus maximow-
genes from one strain and introduce them into iczii), by Agrobacterium-mediated transformation.
other varieties of fungi, bacteria, or plants. This In vitro assays demonstrated that the transgenic
enhances the potency of biocontrol agents and poplars had increased resistance to the leaf rust
makes a single strain consistently effective against pathogen Melampsora medusae. Seedlings of trans-
more than one plant pathogenic fungus, without genic spruce lines showed an increased resistance
the hazardous effects of chemical pesticides. to the spruce root pathogen Cylindrocladium flori-
This approach was taken by Broglie et al. (1991) danum (Noël et al. 2006).
who, in a pioneering work, produced seedlings con-
stitutively expressing a bean chitinase gene under
the control of the cauliflower mosaic virus 35S pro-
moter. The timing of the natural host defense mech- V. Conclusions
anism was modified to produce fungus-resistant
plants with increased ability to survive in soil in- Mycoparasitism is a quite common, and yet excit-
fested with the fungal pathogen R. solani, delaying ing phenomenon. It appears to play an important
the development of disease symptoms. role in biological control, even though it should be
Since then, genetic manipulations of valuable pointed out that mycoparasitism is only one spe-
crop plants with one or more cell wall-degrading cific aspect in the whole complex system of bio-
enzymes from mycoparasitic fungi have been con- logical control of plant diseases. For example, the
sidered a potent tool for improving plant resistance direct effects of root-colonizing Trichoderma spp.
on plants are at least as important as the direct
effects on pathogens – or perhaps more so. These
fungi have profound impacts on plant growth and
development, and they also induce resistance to
a variety of classes of plant pathogens.
Mycoparasitism is a complex process that
includes the following steps: (1) chemotrophic
growth of the antagonist toward the host;
(2) recognition of the host by the mycoparasite;
(3) attachment; (4) excretion of extracellular
enzymes; and (5) lysis and exploitation of the host.
Mycoparasitism occurs under appropriate eco-
logical conditions. The population and activity of
the mycoparasite can be increased by relatively spe-
cific substances, such as chitin. The gene coding for
Fig. 8.4. Chitinase-specific activity of crude enzyme (units chitinase is only one example of genes with myco-
per mg protein) excreted by the wild-type T. harzianum (wt) parasitic activity. Other potential genes are those
coding for β-1,3-glucanase, protease, and lipase.
and transformants (401 and 402), after 5 days on synthetic
medium. Columns headed by different letters are signifi-
cantly different (p = 0. 05) according to Duncan’s multiple Engineering various chitinases together with
range test (Haran et al. 1993) other genes that may act as antifungal agents
596 Wolf Prize in Agriculture
may lead to better protection of plants against Baker R, Griffin GJ (1995) Molecular strategies for biolog-
pathogenic fungi. It may therefore be possible ical control of fungal plant pathogens. In: Reuveni R
to improve mycoparasitism, and to enhance the (ed) Novel approaches to integrated pest management.
Lewis, CRC Press, Boca Raton, FL, pp 153–182
plants resistance response by integrating cloned Barak R, Elad Y, Mirelman D, Chet I (1985) Lectins: a pos-
chitinase with different lytic enzymes and other sible basis for specific recognition in Trichoderma-
available antifungal polypeptides. By understand- Sclerotium rolfsii interaction. Phytopathology 75:458–
ing the mode of action of biocontrol mycoparasitic 462
Barak R, Elad Y, Chet I (1986) The properties of L-fucose
fungi, we should be able to manipulate the fungal binding agglutinin associated with the cell wall of Rhi-
agent, the plant, and their interactions to achieve zoctonia solani. Arch Microbio1 144:346–349
more effective and safer plant resistance to various Barnett HL, Binder FL (1973) The fungal host-parasite re-
biotic and abiotic stresses. lationship. Annu Rev Phytopathol 11:273–292
Barondes SH (1981) Lectins: their multiple endogenous cel-
Acknowledgements. This work was supported by The Dr. lular functions. Annu Rev Biochem 50:207–231
Alexander and Myrna Strelinger Endowment Fund, The Ju- Benhamou N, Chet I (1993) Hyphal interactions between
dith and Avraham Goldwasser Foundation, and a United Trichoderma harzianum and Rhizoctonia solani: ultra-
States-Israel Binational Agricultural Research and Devel- structure and cytochemistry of the antagonistic pro-
opment Fund (BARD) (US-3507-04). cess. Phytopathology 83:1062–1071
Benitez T, Rincon AM, Limon MC, Codon AC (2004) Biocon-
trol mechanisms of Trichoderma strains. Int Microbiol
7:249–60
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Baldur Rosmund Stefansson
Department of Plant Science
Faculty of Agricultural and Food Sciences
University of Manitoba, Winnipeg, Canada
k
1917–2001
CURRICULUM VITAE
Place and Date of Birth: Veatfold (near Lundar), Manitoba, 1917
Marital Status: Married, 3 children
Positions Held:
1952-1966 Research Associate, Department of Plant Science, University of
Manitoba
1966-1974 Associate Professor, Department of Plant Science, University of
Manitoba
601
602 Wolf Prize in Agriculture
VARIETIES RELEASED:
Soybeans
Portage 1964
Altona 1966
Rape
Tanka 1963
Target 1966
Turret 1970
Tower 1974
Regent 1977
Reston 1982
Pivot 1985
Turnip rape
Polar 1970
Baldur Rosmund Stefansson 603
LIST OF PUBLICATIONS
Stefaneson, B.R. 1959. A method for estimating the percentage of hybrids in seedlots
of first generation advance sunflowers by means of a seedling characteristic.
Can. J. Plant Sci. 39: 71-74.
Howell, R.W., C.J. Wargel, C.A. Brim, E.E. Hartwig, J.W. Lambert, I.R. Torpson,
B.R. Stefansson, J.K. Park, W.E. Seigler and B.K. Webb. 1960. Response of
soybeans to seed treatment with gibberellin under simulated commercial
conditions. Agron. J. 52: 144-146.
Stefansson, B.R., F.W. Hougen and R.K. Downey. 1961. Note on the isolation of
rape plants with seed oil free from erucic acid. Can. J. Plant Sci. 41: 218-219.
Stefansson, B.R. and F.W. Hougen. 1964. Selection of rape plants (Brassica napus)
with seed oil practically free from erucic acid. Can. J. Plant Sci. 44: 359-364.
Kondra, Z.P. and B.R. Stefansson. 1965. Inheritance of erucic and eicosenoic
acid content of rapeseed oil (Brassica napus). Can. J. Genetics and Cytol. 7:
505-510.
Gross, A.T.H. and B.R. Stefansson. 1966. Effect of planting date on protein oil and
fatty acid content of rapeseed and turnip rape. Can. J. Plant Sci. 46: 389-395.
Stefansson, B.R. 1966. Registration of Portage soybeans. Crop. Sci. 6: 612.
Stefansson, B.R. 1966. Altona, a new variety of soybeans. Can. J. Plant Sci. 46: 693.
Stefansson, B.R. 1966. Target, a new variety of summer rape. Can. J. Plant Sci. 46:
694.
Stefaneson, B.R. 1968. Registration of Altona soybeans. Crop Sci. 8: 777.
Stefansson, B.R. 1969. Registration of Target rape. Crop. Sci. 9: 394-395.
Stefansson, B.R. and A.K. Storgaard. 1969. Correlations involving oil and fatty
acids in rapeseed. Can. J. Plant Sci. 49: 573-580.
Kondra, Z.P. and B.R. Stefanseon. 1970. A maternal effect on the fatty acid
composition of rapeseed oil (Brassica napus). Can. J. Plant Sci. 50: 345-346.
Kondra, Z.P. and B.R. Stefansson. 1970. Inheritance of the major glucosinolates of
rapeseed (Brassica napus) meal. Can. J. Plant Sci. 50: 643-647.
Stefansson, B.R. 1971. Registration of Polar summer turnip rape. Crop Sci. 11:
134.
Stefanason, B.F. 1971. Registration of Turret summer rape. Crop Sci. 11: 135.
Fowler, D.B. and B.R. Stefansson. 1972. Effects of the mutagenic agent ethyl
methanesulfonate on the M1 generation of rape (Brassica napus). Can. J.
Plant Sci. 52: 53-62.
Stefansson, B.R. and Z.P. Kondra. 1974. Tower Summer Rape. Can. J. Plant Sci. 54:
343-344.
Fowler, D.B. and B.R. Stefansson. 1975. Ethyl-methanesulfonate-induced mutations
in rape (Brassica napus). Can. J. Plant Sci. 55: 817-821.
Grami, Bahram and B.R. Stefansson. 1977. Gene action for protein and oil content
in summer rape. Can. J. Plant Sci. 57: 625-631.
606 Wolf Prize in Agriculture
Grami, Bahram, RJ. Baker and B.R. Stefansson. 1977. Genetics of protein and
oil content in summer rape: Heritability, number of effective factors, and
correlations. Can. J. Plant Sci. 57: 937-943.
Grami, Bahram and B.R. Stefansson. 1977. Paternal and maternal effects on protein
and oil content in summer rape. Can. J. Plant Sci. 57: 945-949.
Sernyk, J.L. and B.R. Stefansson. 1982. White flower color in rape (Brassica
napus L.) associated with a radish (Raphanus sativus L.) chromosome. Can. J.
Genetics and Cytology 24:729-734.
Sernyk, J.L. and B.R. Stefanesson. 1983. Heterosis in summer rape (Brassica
napus L.). Can. J. Plant Science 63:407-413.
Fan, Z., S.R. Rimmer and B. R. Stefansson. 1983. Inheritance of resistance to
Albugo candida in rape (Brassica napus L.). Can. J. Genetics and Cytology.
25:420-424.
Fan, Z., W. Tai and B.R. Stefansson. 1984. Male sterility in Brassica napus L.
associated with an extra chromosome. Can. J. of Genetics and Cytology.
27:467-471.
Fan, Z. and B.R. Stefansson. 1986. Influence of temperature on sterility of two
cytoplasmic male sterility systems in rape (Brassica napus L.) Can. J. Plant Sci.
66: 221-227.
Fan, Z., B.R. Stefansson and J.L. Sernyk. 1986. Maintainers and restorers of three
male-sterility-inducing cytoplasms in rape (Brassica napus L.). Can. J. Plant
Sci. 66: 229-234.
Li, S., Y. Qian, Z. Wu and B.R. Stefansson. 1988. Genetic male sterility in rape
(Brassica napus L.) conditioned by interaction of genes at two loci. Can. J.
Plant Sci. 68: 1115-1118.
Contributions to Books
Stefansson, B.R. 1973. Oilseeds: production, marketing and processing. In Grains
and Oilseeds, section D-11, International Grains Institute, Winnipeg, Manitoba.
pp. 1-29.
Stefansson, B.R. 1974. Rapeseed Breeding in Western Canada. In The Story of
Rapeseed in Western Canada, edited by D.A. McLeod, Saskatchewan Wheat
Pool. pp. 22-25.
Stefansson, B.R. 1974. Rapeseed. In Principles and Practices of Commercial
Farming, Faculty of Agriculture, University of Manitoba, Winnipeg, Manitoba.
pp. 162-165.
Stefansson, B.R. et al. 1975. Breeding Rapeseed and Mustard Crops. In Oilseed and
Pulse Crops in Western Canada, Western Co-operative Fertilizers Ltd., Calgary,
Alberta. pp. 165-183.
Stefansson, B.R. 1975. Oilseeds: Production. In Grains and Oilseeds, Handling,
Marketing, Processing. International Grains Institute, Winnipeg, Manitoba.
pp. 607-616.
Baldur Rosmund Stefansson 607
Stefansson, B.R. 1976. Tower rapeseed, a major step in the development of high
quality rapeseed. In Fats and Oils in Canada, Department of Industry, Trade
and Commerce, Ottawa. pp. 2-7.
Hougen, F.W. and B.R. Stefansson. 1982. Rapeseed. Chap. 6, in Advances in Cereal
Science and Technology. Vol. V. Edited by Y. Pomeranz. Amer. Assoc. of Cereal
Chemists, Inc., St. Paul, Minnesota. pp. 261-289.
Stefansson, B.R. 1983. The development of improved rapeseed cultivars. Chap. 6,
in High and Low Erucic Acid Rapeseed Oils, edited by J.K.G. Kramer,
F.D. Sauer and W.J. Pigden. Academic Press Canada, Toronto. pp. 143-159.
Stefansson, B.R. 1983. Oilseeds. In The Canadian Encyclopedia, Hurtig Pub. Ltd.,
Edmonton, Alberta. pp. 1314-1315.
Conference Proceedings
B.R. Stefaneson and F.W. Hougen. Breeding for Low Erucic Acid Content in Rape.
Proc. Canadian Barley and Oil Seeds Conf., Winnipeg, February 1960. p. 27.
B.R. Stefanseon. Reply to Question re Soybeans. Proc. Man. Agronomists’ Ann.
Conf., 1967. p. 26.
Stefansson, B.R. Expected Changes in Rapeseed Varieties in the Future. Proc. Man.
Agronomists’ Ann. Conf., 1969. pp. 23-24.
Stefansson, B.R. Plant Breeding and New Varieties of Rapeseed. Proc. 2nd Ann.
Meeting, Rapeseed Assoc. of Canada, Saskatoon, Mar. 1969. pp. 44-46.
Stefansson, B.R. Fatty Acid Composition of Fats and Oils. Proc. Man. Agronomists’
Ann. Conf., 1970. pp. 49-51.
Stefansson, B.R. Influence of Temperature and Soil on Seed Composition. Proc. Int.
Conf. on the Science, Technology and Marketing of Rapeseed and Rapeseed
Products. Ste. Adele, Quebec, Sept. 20-23, 1970. pp. 86-91.
Stefansson, B.R. Rapeseed Varieties Present and Future. Proc. Man. Agronomists’
Ann. Conf., 1971. pp. 59-61.
Stefansson, B.R. Progress Report on “Double Zero” Rapeseed Varieties. Proc.
Canadian Barley and Oil Seeds Conf., Winnipeg, Feb. 1972. p. 103.
Stefansson, B.R. What’s New in 0ilseed Varieties – Rapeseed. Proc. Canadian Barley
and Oil Seeds Conf., Winnipeg February l974. pp. 90-93.
Stefansson, B.R. Rapeseed Varieties. Proc. 7th Ann. Meeting, Rapeseed Assoc. of
Canada, Calgary, March 1974. pp. 46-47.
Stefansson, B.R. Rapeseed for 1975. Proc. Man. Agronomists’ Ann. Conf. 1974.
pp. 74-76.
Stefansson, B.R. Summer Rape (Argentine Type) Varieties Proc. 9th Ann. Meet.
Rapeseed Assoc. of Canada, Winnipeg, March 1976. pp. 37-39.
Stefansson, B.R. Canadian Rapeseed — The Best in the World and Getting Better.
Proc. Manitoba Agronomists’ Ann. Conf., 1978. pp. 39-43.
Stefansson, B.R. Selection for Oil and Protein in Rapeseed Proc. 5th Int. Rapeseed
Conf. Vol. 1, Malmo, Sweden, June 12-16, 1978. pp. 113-114.
608 Wolf Prize in Agriculture
Stefansson, B.R. Rapeseed Varieties of the Future. Proc. Canadian Barley and Oil
Seeds Conf., Winnipeg, Feb. 1980. pp. 75-80.
__________ Rapeseed in China. Report on Canola Technical Seminars People’s
Republic of China, Nov. 9-22, 1980. pp. 14-17.
__________ Canola/Rapeseed Variety Development. Proc. 14th Ann. Conf., Canola
Council of Canada, Vancouver, Mar. 1981. pp. 47-50.
__________ Report on Canada/Japan Canola Consultation Tokyo, Japan, Nov
7-11, 1983.
Gurdev S. Khush
International Rice Research Institute
Makati City, Philippines
k
CURRICULUM VITAE
Date and Place of Birth: August 22, 1935, Rurkee, Punjab, India
Nationality: Indian
Business Address: Adjunct Professor
Dept. of Plant Sciences
University of California, Davis, CA 95616
EDUCATION
(1) Matriculation Punjab University 1951 (topped the list of successful candidates
of the school in first division)
(2) B.Sc. (Agri) Punjab Agricultural University, Ludhiana, Punjab, India 1955 (First
division)
(3) Ph.D. (Genetics) University of California, Davis, Davis, California 1960
609
610 Wolf Prize in Agriculture
(3) Plant Breeder, International Rice Research Institute (IRRI), August 1967 to
June 1972
(4) Plant Breeder and Head, Plant Breeding Dept., (IRRI), July 1972 to December
1985
(5) Principal Plant Breeder and Head, Division of Plant Breeding, Genetics and
Biochemistry, (IRRI), January 1986 to February 2002
(6) Adjunct Professor University of California, Davis 2002-to date
NATIONAL AWARDS
(1) Best all round student award of Punjab Agricultural University, 1955
(2) Achievement Award by the Bureau of Plant Industry, Manila, Philippines,
1983
(3) K. Ramiah Medal, National Academy of Agricultural Sciences, 1997
(4) Medal for the “Cause of Agriculture and Rural Development” of Vietnam,
1998
(5) International Agricultural Cooperation Prize, Ministry of Agriculture,
Government of China, 1999
(6) Friendship Award, Government of China, 1999
(7) B.P. Pal Memorial Award, Indian Science Association, 2000
(8) Padma Shri Award by President of India, 2000
(9) Gold Medal, Ministry of Agriculture, Islamic Republic of Iran, 2000
(10) Gopalan Oration Award and Gold Medal 2000
(11) International Cooperation Award, Government of Egypt, 2001
(12) Citation by President, Government of Philippines, 2002
(13) Plaque of Appreciation by the President of Indonesia, 2002
(14) Amrik Singh Cheema Award by Young Farmer’s association of Punjab, 2006
(15) Swaminathan Award for leadership in agriculture by TAAS, 2006
(16) Golden Sickle Award, Government of Thailand, 2007
(17) NCCPB Genetics and Plant Breeding Award, 2007
(18) CV Raman Medal, Indian National Science Academy, 2007
INTERNATIONAL AWARDS
(1) Borlaug Award for Achievements in Plant Breeding, 1977
(2) Japan Prize, Science and Technology Foundation of Japan, 1987
(3) International Agronomy Award, American Society of Agronomy, USA, 1989
(4) Emil M. Mrak International Award, University of California, Davis, USA, 1990
(5) World Food Prize, World Food Prize Foundation, USA, 1996
(6) Rank Prize, Rank Prize Foundation, United Kingdom, 1998
(7) Wolf Prize in Agriculture, Wolf Prize Foundation, Israel, 2000
Gurdev S. Khush 611
HONORARY DEGREES
(1) Doctor of Science (Honoris Causa), Punjab Agricultural University, India
1987
(2) Doctor of Science (Honoris Causa), Tamil Nadu Agricultural University, India
1995
(3) Doctor of Science (Honoris Causa), C.S. Azad University of Agriculture and
Technology, India 1995
(4) Doctor of Science (Honoris Causa), G.B. Pant University of Agriculture and
Technology, India 1996
(5) Doctor of Science (Honoris Causa), De Montfort University, Leicester, England
1998
(6) Doctor of Science (Honoris Causa), Assam Agricultural University, India,
2000
(7) Doctor of Science (Honoris Causa), Cambridge University, United Kingdom,
2000
(8) Doctor of Science (Honoris Causa) N.D. University of Agriculture and
Technology, India 2003
(9) Doctor of Science (Honoris Causa) Ohio State University, USA, 2006
(10) Doctor of Science (Honoris Causa) Guru Nanak Dev University, India, 2007
in rural trade, transport, and construction activities. The economic miracle underway
in many Asian countries was triggered by the growth in agricultural income and
its equitable distribution, which helped expand the domestic market for non farm
goods and services.
1991 Rice Biotechnology (with G.H. Toenniessen). Wallingford, UK. 320 pp.
1992 Nodulation and Nitrogen Fixation in Rice (With J. Bennett). Manila
Philippines Int. Rice Research Institute, Los Banos 136 pp.
1994 Apomixis: Exploiting Hybrid Vigor in Rice. Int. Rice Research Institute, Los
Banos, Philippines. 76 pp.
1996 Rice Genetics 111. Proceedings of Third International Rice Genetics
Symposium, Manila Philippines, Int. Rice Rice Research Institute, Los Banos
1011 pp.
2000 Aromatic Rices (with R.K. Singh and U.S. Singh). Enfield (USA) and Plymouth
(UK), 292 pp.
2001 Rice Genetics 1V (D.S. Brar and B. Hardy). Proceedings of Fourth
International Rice Genetics Symposium, Manila, Philippines. Int. Rice
Research Institute, Los Banos. 488 pp.
12. 1967. Khush, G. S., and C. M. Rick. Novel compensating trisomics of the
tomato: Cytogenetics, monosomic analysis, and other applications. Genetics
56:297-307.
13. 1967. Khush, G. S. and C. M. Rick. Studies on the linkage map of chromosome
4 of the tomato and on the transmission of induced deficiencies. Genetica
38:74-94.
14. 1967. Khush, G. S., and C. M. Rick. Tomato tertiary trisomics: origin,
identification, morphology, and use in determining position of centromeres
and arm location of markers. Canad. J. Genet. Cytol. 9:610-631.
15. 1967. Khush, G. S., and C. M. Rick. Haplo-triplo-disomics of the tomato:origin,
cytogeneticsand utilization as a source of secondary trisomics. Biol. Zbl.
86:257-265.
16. 1968. Khush, G. S., C. M. Rick. Tomato telotrisomics: origin, identification,
and use in linkage mapping. Cytologia 33:137-148.
17. 1968. Khush, G. S., and C. M. Rick. Cytogenetic analysis of tomato genome
by means of induced deficiencies. Chromosoma 23:452-484.
18. 1968. Reeves, A. F., G. S. Khush, and C. M. Rick. 1968. Segregation
and Recombination of Trisomics: A reconsideration. Canad. J. Genet. Cytol.
10:937-940.
19. 1969. Khush, G. S., and C. M. Rick. Tomato secondary trisomics: Origin,
identification, morphology and use in cytogenetic analysis of the genome.
Heredity 24:129-146.
20. 1969. Suneson, C. A., K. O. Rachie, and G. S. Khush. A dynamic population
of weedy rye. Crop Science 9:121-124.
21. 1969. Beachell, H. M., and G. S. Khush. Objectives of the IRRI breeding
program. SABRAO Newsletter 1:69-80.
22. 1969. Rick, C. M., and G. S. Khush. Cytogenetic exploration in the tomato
genome. pp. 145-168 In R. Bogart, ed. Genetic Lectures. Oregon State
University Press.
23. 1970. Khush, G. S. Breeding rice for resistance to grassy stunt. Paper presented
at the 13th Session of the International Rice Commission Working Party on
Rice Production and Protection, Teheran, Iran, December 1970.
24. 1971. Khush, G. S. Rice Breeding for disease and insect resistance at IRRI.
Oryza 8:111-119.
25. 1971. Khush, G. S., E. Torres, and R. Aquino. Exploiting wild germplasm of
Oryza for improving cultivated rices. In Crop Science Society of the
Philippines. Proceedings of the second annual scientific meeting, May
4-6, 1971. College, Laguna, Philippines. Pages 311-320.
26. 1972. Khush, G. S., and H. M. Beachell. Breeding for disease and insect
resistance at IRRI. In International Rice Research Institute, Rice Breeding. Los
Banos, Philippines. pp. 309-322.
620 Wolf Prize in Agriculture
27. 1972. Murty, V. V. S., and G. S. Khush. Studies on the inheritance of resistance
to bacterial leaf blight in rice varieties. In International Rice Research Institute.
Rice Breeding. Los Banos, Philippines. pp. 301-307.
28. 1972. Beachell, H. M., G. S. Khush, and R. C. Aquino. IRRI’s international
breeding program. In International Rice Research Institute. Rice Breeding,
Los Banos, Philippines. pp. 89-106.
29. 1972. Beachell, H. M., G. S. Khush, and B. O. Juliano. Breeding for high
protein content in rice. In International Rice Research Institute. Rice Breeding.
Los Banos, Philippines. pp. 419-430.
30. 1972. Beachell, H. M. and G. S. Khush. Rice varietal improvement in Asia. In
International Conference on Tropical and Subtropical Agriculture. Conference
Papers, American Society of Agricultural Engineers. Pages 156-160.
31. 1973. Khush, G. S. Cytogenetics of Aneuploids. Academic Press, New York,
301 pp.
32. 1973. Murty, V. V. S., G. S. Khush, and N. F. Jensen. Inheritance of resistance
to bacterial Xanthomonas oryzae (Uyeda et Ishiyama) Dowson in rice.
Allelic relationships of resistance genes in donor varieties. Jap. Journ. Breed.
23:325-328.
33. 1974. Khush, G. S., W. R. Coffman and T. T. Chang. Current status of
the varietal development program at IRRI. Paper presented during the
Annual Rice Research Review Meeting, WARDA, July 15-20, 1974, Monrovia,
Liberia.
34. 1974. Martinez, C. R. and G. S. Khush. Sources and inheritance of resistance
to brown planthopper in some breeding lines of rice Oryza saliva L. Crop
Science 14:264-267.
35. 1974. Khush, G. S. and K. C. Ling. Inheritance of resistance to grassy stunt
virus and its vector in rice. Jour. Hered. 65:135-137.
36. 1975. Pathak, M. D., and G. S. Khush. Control of upland rice insects through
varietal resistance. In International Rice Research Institute. Major Research
in Upland Rice. Los Banos, Philippines. pp. 117-125.
37. 1975. Ou, S. H., K. C. Ling, H. E. Kauffman, and G. S. Khush. Diseases of
upland rice and their control through varietal resistance. In International
Rice Research Institute. Major Research in Upland Rice. Los Banos, Philippines.
pp. 126-135.
38. 1975. Khush, G. S. Rice. In Handbook of Genetics, R. C. King Edit. Plenum
Press, New York. Vol. 2, 31-58 pp.
39. 1976. Librojo, V., H. E. Kauffman, and G. S. Khush. Genetic analysis of
bacterial blight resistance in four varieties of rice. SABRAO J. 8:105-110.
40. 1976. Khush, G. S. Breeding for resistance in rice. In The Genetic Basis of
Epidemics in Agriculture. Annals the New York Academy of Sciences,
287:296-308.
Gurdev S. Khush 621
41. 1976. Siwi, B. H. and G. S. Khush. New genes for resistance to the green
leafhopper in rice. Crop Science 17:17-20.
42. 1976. Lakshminarayana, A., and G. S. Khush. New genes for resistance to
brown planthopper in rice. Crop Science 17:96-100.
43. 1977. Khush, G. S., K. C. Ling, R. C. Aquino and V. M. Aguiero. 1977.
Breeding for resistance to grassy stunt in rice. In Proceedings 3rd Int. Congr.
SABRAO, Canberra, Australia. Plant Breeding Papers l:4(b):3-9.
44. 1977. Khush, G. S., and W. R. Co man. Genetic Evaluation and Utilization
(GEU) Program — The Rice Improvement Program of the International Rice
Research Institute. Theor. Appl. Genet. 51:97-110.
45. 1977. Khush, G. S. Disease and insect resistance in rice. Adv. Agron.
29:265-341.
46. 1977. Olufowote, J. 0., G. S. Khush, and H. E. Kauffman. Genetics of bacterial
blight resistance in rice. Phytopathology 67:772-775.
47. 1977. Petpisit, V., G. S. Khush, and H. E. Kau man. Inheritance of resistance
to bacterial blight in rice. Crop Science 17:551-554.
48. 1977. Sujadi, S. and G. S. Khush. Studies on linkage relations of genes
controlling disease and insect resistance and nature of endosperm in rice.
Euphytica 26:337-342.
49. 1977. Sidhu, G. S., and G. S. Khush. Dominance reversal of a bacterial blight
resistance gene in some varieties of rice. Phytopathology 67:461-463.
50. 1978. Khush, G. S. Breeding methods and procedures employed at IRRI for
developing rice germplasm with multiple resistance to diseases and insects.
In International Symposium on Methods of Crop Breeding. Tropical Agr. Res.
Center, Japan. Series II:69-76.
51. 1978. Siwi, B. H., G. S. Sidhu, and G. S. Khush. Genetic analysis of rice
variety Ptb 8 for resistance to green leafhopper and brown planthopper.
Contr. Centr. Res. Inst. Agric. Bogor. 47:7-10.
52. 1978. Sidhu, G. S., G. S. Khush and T. W. Mew. Genetic analysis of bacterial
blight resistance in seventy-four varieties of rice, Oryza sativa L. Theor.
Appl. Genet. 53:105-111.
53. 1978. Sidhu, G. S. and G. S. Khush. Genetic analysis of brown planthopper
resistance in twenty varieties of rice, Oryza saliva L. Theor. Appl. Genet.
53:199-204.
54. 1979. Ikehashi, H. and G. S. Khush. Breeding for blast resistance in IRRI. In
International Rice Research Institute. Rice Blast Workshop. Los Banos,
Philippines. pp. 69-80.
55. 1979. Khush, G. S. Genetics of and breeding for resistance to brown
planthopper. In International Rice Research Institute. Brown Planthopper:
A Threat to Rice Production in Asia. Los Banos, Laguna, Philippines.
pp. 321-332.
622 Wolf Prize in Agriculture
56. 1979. Pathak, M. D., and G. S. Khush. Studies on varietal resistance to the
brown planthopper at IRRIIn International Rice Research Institute. Brown
Planthopper: A Threat to Rice Production in Asia. Los Banos, Philippines.
pp. 285-302.
57. 1979. Gallun, R. L., and G. S. Khush. Genetic factors affecting expression
and stability of resistance. In F.G. Maxwell and P.R. Jennings, eds. Breeding
Plants Resistant to Insects. John Wiley & Sons, New York. pp. 63-85.
58. 1979. Khush, G. S. Breeding for multiple disease and insect resistance in
rice. In M. K. Harris, ed. Biology and Breeding for resistance to Arthropods
and Pathogens in Agricultural Plants. Texas A&M University, College Station,
Texas. pp. 341-354.
59. 1979. Khush, G. S., C. M. Paule, and N. M. dela Cruz. Rice grain quality
evaluation and improvement at IRRI. In. International Rice Research Institute.
Proc. of Workshop on Chemical Aspects of Rice Grain Quality. Los Banos,
Laguna, Philippines. pp. 21-31.
60. 1979. Ikehashi, H., and G. S. Khush. Methodology of assessing appearance
of the rice grain including chalkiness and whiteness. In International Rice
Research Institute. Proc. of Workshop on Chemical Aspects of Rice Grain
Quality. Los Banos, Laguna, Philippines. pp. 223-229.
61. 1979. Crill, J. P. and G. S. Khush. Effective and stable control of rice blast
with monogenic resistance. Food and Fertilizer Technology Center, Taipei
City, Taiwan. Extension Bulletin No. 128.
62. 1979 . Sidhu, G. S. and G. S. Khush. Linkage relationships of some genes
for disease and insect resistance and semidwarf stature in rice. Euphytica
28:233-237.
63. 1979. Sidhu, G. S., G. S. Khush and F. G. Medrano. A dominant gene for
resistance to whitebacked planthopper in rice and its association with some
other traits. Euphytica 28:227-232.
64. 1980. Khush, G. S. Genetics and breeding of photoperiod sensitive rice
varieties. In Proceedings of the Photoperiod-Sensitive Transplanted Rice. Oct.
1977. Bangladesh Rice Research Institute, Joydebpur. pp. 37-44.
65. 1980. Sidhu, G. S., G. S. Khush and T. W. Mew. Genetic analysis
of bacterial blight resistance in seventy cultivars of rice, Oryza sativa L. from
Indonesia. Crop Improvement 6:19-25.
66. 1981. Khush, G. S. and R. C. Chaudhary. Role of resistant varieties in integrated
pest management of rice. Food and Fertilizer Technology Center, Taipei City,
Taiwan. Extension Bulletin No. 162.
67. 1981. Khush, G. S. Breeding rice for multiple disease and insect resistance.
In Rice Improvement in China and other Asian Countries. International Rice
Research Institute, Los Banos, Philippines. pp. 220-237.
Gurdev S. Khush 623
83. 1984. Khush, G. S. IRRI breeding program and its worldwide impact on
increasing rice production. In J.P. Gustafson, ed. Gene Manipulation in Plant
Improvement. Plenum Press, New York. pp. 61-94.
84. 1984. Khush, G. S. and B. O. Juliano. Rice varietal improvement for protein
content at the International Rice Research Institute. In Cereal Grain Protein
Improvement. International Atomic Energy Agency, Vienna. pp. 199-202.
85. 1984. Khush, G. S. Terminology for rice growing environments. In
Terminology for Rice Growing Environments. International Rice Research
Institute, Los Banos, Philippines. pp. 5-10.
86. 1984. Khush, G. S. Plant Breeding Lectures (in Chinese) China National Rice
Research Institute, Hangzhou 98 pp.
87. 1984. Chaudhary, R. C., G. S. Khush and E. A. Heinrichs. Varietal resistance
to rice stemborers in Asia. Insect Sci. and Its Application. 5:447-463.
88. 1984. Khush, G. S., R. J. Singh, S. C. Sur and A. L. Librojo. Primary trisomics
of rice:origin, morphology, cytology and use in linkage mapping. Genetics
107:141-163.
89. 1984. Khush, G. S. Breeding rice for resistance to insects. Protection Ecology
7:147-165.
90. 1984. Avesi, G. M. and G. S. Khush. Genetic analysis for resistance to the
green leafhopper, Nephotettix virescens (Distant), in some cultivars of rice,
Oryza saliva L. Crop Protection 3:41-51.
91. 1984. Yoshimura, A., T. W. Mew, G. S. Khush and T. Omura. Genetics
of bacterial blight resistance in a breeding line of rice. Phytopathology
74:773-777.
92. 1985. Khush, G. S. and S. S. Virmani. Breeding rice for disease resistance.
In G. E. Russell, ed. Progress in Plant Breeding. Butterworths, London.
pp. 239-279.
93. 1985. Yoshimura, A., T. Omura, T. W. Mew and G. S. Khush. Genetic behavior
of resistance to bacterial blight in differential rice cultivars in the Philippines.
Bull. Inst. Trop. Agr. Kyushu Univ., Japan 8:1-54.
94. 1985. Miah, M.A.A., E. D. Earle, and G. S. Khush. Inheritance of callus
formation ability in anther culture of rice, Oryza saliva L. Theor. Appl.
Genet. 70:113-226.
95. 1985. Mohanty, H. K. and G. S. Khush. Diallel analysis of submergence
tolerance in rice, Oryza saliva L. Theor. Appl. Genet. 70:467-473.
96. 1985. Wu, C. F. and G. S. Khush. A new dominant gene for resistance to
whitebacked planthopper in rice. Crop Sci. 25:505-509.
97. 1985. Kaw, R. N. and G. S. Khush. Heterosis in traits related to low temperature
tolerance in rice. Phil. J. Crop Sci. 10(2)93-105.
98. 1985. Kaw, R. N. and G. S. Khush. Heterosis in traits related to low temperature
tolerance in rice. Phil. J. Crop. Sci. 10:93-105.
Gurdev S. Khush 625
140. 1989. Sahu, R.K. and G.S. Khush. Genetics of resistance to four races of
Xanthomonas campestris pv. oryzae in some rice cultivars. Plant Breed.
102:232-236.
141. 1989. Jena, K.K. and G. S. Khush. Monosomic alien addition lines of
rice: production, morphology, cytology and breeding behavior. Genome
32:449-455.
142. 1989. Dela Cruz, Normita, I. Kumar, R.P. Kaushik and G.S. Khush. Effect of
temperature during grain development on stability of cooking quality
components in rice. Japan. J. Breed. 39:299-306.
143. 1989. Khush, G.S. and K.K. Jena. Biosystematic status of Oryza indandamanica
Ellis. In Proceedings of the 6‘h International Congr. SABRAO, Tsukuba, Japan.
pp. 179-198.
144. 1989. Abdullah, R., J.A. Thompson, G.S. Khush, R.P. Kaushik and E.C. Cocking.
Protoclonal variation in the seed progeny of plants regenerated from rice
protoplasts. Plant Science 65:97-101.
145. 1989. Tang, S.X., G. S. Khush and B.O. Juliano. Diallele analysis of gel
consistency in rice (Oryza sativa L.). Sabrao Journal 21:135-142.
146. 1989. Tang, S.X., G.S. Khush and B.O. Juliano. Variation and correlation
of four cooking and eating quality indices of rices. Philipp. J. Crop Sci.
14:45-49.
147. 1990. Khush, G.S. Rice breeding - accomplishments and challenges. Plant
Breeding Abstracts 60:461-467.
148. 1990. Khush, G.S., S.S. Virmani and D.S. Brar. Expectations of plant
breeders from tissue culture. In S.S. Bhojwani (Ed.) Plant Tissue Culture:
Applications and Limitations. Elsevier Amsterdam - Oxford - New York -
Tokyo. pp. 412-423.
149. 1990. Ogawa, T., T. Yamamoto, G.S. Khush and T.W. Mew. Genetics of
resistance in rice cultivars, Chugoku 45 and Java 14 to Philippine and Japanese
races of bacterial blight pathogen. Japan. J. Breed. 40:77-90.
150. 1990. Ogawa, T., T. Yamamoto, G.S. Khush and T.W. Mew. Genetics of
resistance in rice cultivars, Zenith and Cempo Selak to Philippine and Japanese
races of bacterial blight pathogen. Japan. J. Breed. 40:183-192.
151. 1990. Ogawa, T., T. Yamamoto, G.S. Khush and T.W. Mew. Genetics of
resistance in rice cultivar Sateng to Philippine and Japanese races of bacterial
blight pathogen. Japan. J. Breed. 40:329-338.
152. 1990. Ogawa, T., T. Yamamoto, G.S. Khush and T.W. Mew. Genetics of
resistance in rice cultivar, IR20 and Semora Mangga to Philippine and Japanese
races of bacterial blight pathogen. Japan. J. Breeding 40:435-447.
153. 1990. Jena, K.K. and G.S. Khush. Introgression of genes from Oryza
officinalis Well ex Watt to cultivated rice, O. saliva L. Theor. Appl. Genet.
80:737-745.
Gurdev S. Khush 629
154. 1990. Glaszmann, J.C., R.N. Kaw and G.S. Khush. Genetic divergence among
cold tolerant rices (Oryza saliva L.). Euphytica 45:95-104.
155. 11990. Juliano, B.O., C.M. Perez, R. Kaushik and G.S. Khush. Grain properties
of 1R36-based starch mutants. Starch 42:256-260.
156. 1990. Ishii, T., O. Panaud, D. S. Brar and G. S. Khush. Use of non radioactive
digoxigenin-labelled DNA probes for RFLP analysis in rice. Plant Molecular
Biology Reporter 8:167-171.
157. 1990. Chaudhary, R.C. and G.S. Khush. Breeding rice varieties for resistance
against Chilo spp. of stem borers in Asia and Africa. Insect Sci. Applic.
11:659-669.
158. 1990. Mir, G.N. and G.S. Khush. Genetic analysis of some rice cultivars for
resistance to bacterial blight. Indian J. Mycol. Plant Pathol. 20(2):130-134.
159. 1991. Khush, G.S. and D.S. Brar. Genetics of resistance to insects in crop
plants. Adv. in Agron. 45:223-274.
160. 1991. Khush, G. S. and T. Kinoshita. Rice karyotype, marker genes, and
linkage groups. In G.S. Khush and G.H. Toenniessen (Eds) Rice Biotechnology.
C.A.B. International Wallingford, England and International Rice Research
Institute, P.O. Box 933, Manila, Philippines. pp. 83-108.
161. 1991. Khush, G.S. and R.J. Singh. Chromosome architecture and aneuploids
in rice. In P.K. Gupta and T. Tsuchiya, Eds. Chromosome Engineering
in Plants: Genetics, Breeding, Evolution Part A. Elsevier, Amsterdam.
pp. 577-598.
162. 1991. Ogawa, T., G.A. Bustos, R.E. Tabien, G.O. Romero, N. Endo and
G.S. Khush. Grouping of rice cultivars based on reaction pattern to Philippine
races of bacterial blight pathogen (Xanthomonas campestris pv. oryzae).
Japan J. Breed. 41:109-119.
163. 1991. Ogawa, T., T. Yamamoto, G.S. Khush and T.W. Mew. Resistance and its
inheritance to bacterial blight of 1R8 rice cultivar group. Japan. J. Breed.
41:211-222.
164. 1991. Endo, N., G.A. Bustos, T. Ogawa and G.S. Khush. Rice cultivar
groups in Myanmar based on reaction to bacterial blight. Japan. J. Breed.
41:289-300.
165. 1991. Ogawa, T., T. Yamamoto, G. S. Khush and T.W. Mew. Breeding of near
isogenic lines of rice with single genes for resistance to bacterial blight pathogen
(Xanthomonas campestris pv. oryzae). Japan. J. Breed. 41:523-529.
166. 1991. Tang, S.X., G.S. Khush and B.O. Juliano. Genetics of gel consistency in
rice. J. Genet. 70:69-78.
167. 1991. Kaushik, R.P. and G. S. Khush. Genetic analysis of endosperm mutants
in rice, Oryza sativa L. Theor. Appl. Genet. 83:146-152.
168. 1991. Kaushik, R.P. and G.S. Khush. Endosperm mutants in rice: gene expression
in Japonica and indica backgrounds. Cereal Chemistry 68:487-491.
630 Wolf Prize in Agriculture
169. 1991. Brar, D.S., B.G. delos Reyes, O. Panaud, A. Sanchez, and G.S. Khush.
Genetic mapping in rice using isozymes and RFLP markers. In Rice Genetics
II. International Rice Research Institute, P.O. Box 933, Manila, Philippines.
pp. 137-145.
170. 1991. McCouch, S.R., G. S. Khush and S.D. Tanksley. Tagging genes for
disease and insect resistance via linkage to RFLP markers. In Rice Genetics II.
International Rice Research Institute, P.O. Box 933, Manila, Philippines.
pp. 443-449.
171. 1991. Mir, G.N. and G. S. Khush. Genetics of resistance to bacterial blight in
rice (Oryza sativa L.). Indian J. Genet. 51:72-78.
172. 1992. Khush, G.S. Selecting rice for simply inherited resistances. In H.S.
Stalker and J.P. Murphy, Eds. Plant Breeding in 1990’s. C.A.B. International
Wallingford, England. pp. 303-323.
173. 1992. Khush, G.S. and G.H. Toenniessen. Biotechnology and pest management
in 2000 AD. In A. Aziz, S.A. Kadir and H.S. Barlow, Eds. Pest Management
and the Environment in 2000. C.A.B. International, Wallingford, England.
pp. 348-359.
174. 1992. Khush, G. S. and D. S. Brar. Overcoming barriers in hybridization. In
G. Kallo and J.B. Chaudhury, Eds. Distant hybridization of crop plants. Theor.
Appl. Genet. Monograph Ser. 16:47-61.
175. 1992. Bonman, J.M., G.S. Khush, and R.J. Nelson. Breeding rice for resistance
to pests. Ann. Rev. Phytopath. 30:485-506.
176. 1992. Pooni, H.S., Ish Kumar and G.S. Khush. A comprehensive model for
disomically inherited metrical traits expressed in triploid tissues. Heredity
69:166-174.
177. 1992. Taura, S., T. Ogawa, R.E. Tabien, G.S. Khush, A. Yoshimura and
T. Omura. Resistance gene of rice cultivar, Taichung Native I to Philippine
races of bacterial blight pathogen. Jap. J. Breed. 42:195-202.
178. 1992. Endo, N., T. Ogawa and G.S. Khush. Genetic analysis of Myanmar rice
cultivars for resistance to bacterial blight. Jap. J. Breed. 42:341-352.
179. 1992. Jena, K.K., G.S. Khush and G. Kochert. RFLP analysis of rice (Oryza
saliva L.) introgression lines. Theor. Appl. Genet. 84:608-616.
180. 1993. Brar, D.S. and G.S. Khush. Application of biotechnology in integrated
pest management. J. Insect Sci. 6:7-14.
181. 1993. Khush, G.S. Biotechnology in Agriculture. In M. Nazim and
N. Polunin, eds. Environmental challenges: From Stockholm to Rio and beyond.
Published by Energy and Environment Society of Pakistan and Foundation
for Environmental Conservation, Geneva, Switzerland. pp. 117-148.
182. 1993. Khush, G.S. Varietal needs for different environments and breeding
strategies. In K. Muralidharan and E.A. Siddiq, Eds. New Frontiers in Rice
Research, Directorate of Rice Research, Hyderabad, India. pp. 68-75.
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183. 1993. Khush, G.S., D.S. Brar, F.J. Zapata, R. Nelson, S. McCouch and
D.G. Bottrell. In B.C. Imrie and J.B. Hacker, Eds. Focused Plant Improvement:
Towards Responsible and Sustainable Agriculture. Proceedings Tenth
Australian Plant Breeding Conference. Published by Conference Organizing
Committee, Canberra, Australia. Biotechnology for rice improvement
Vol. I. pp. 258-279.
184. 1993. Khush, G. S. Breeding rice for sustainable agricultural systems. In
D.R. Buxton, R. Shibles, R.A. Forsberg, B.L. Blad, K.H. Asay, G.M. Paulson,
R.F. Wilson, Eds. International Crop Science I. Crop Science Society of America,
Madison, Wisconsin, USA. pp. 189-199.
185. 1993. Bennett, J, and G. S. Khush. In McGraw-Hill Yearbook of Science and
Technology. McGraw-Hill Inc., New York. pp. 321-324.
186. 1993. Pooni, H.S., I. Kumar and G.S. Khush. Genetical control of amylose
content in a diallel set of rice crosses. Heredity 71:603-613.
187. 1994. Abenes, M.L.P., R.E. Tabien, S.R. McCouch, R. Ikeda, P. Ronald,
G.S. Khush and N. Huang. Orientation and integration of the classical and
molecular map of chromosome I 1 in rice. Euphytica 76:81-87.
188. 1994. Fukui, K., N. Ohmido and G.S. Khush. Variability in rDNA loci in the
genus Oryza detected through fluorescence in situ hybridization. Theor. Appl.
Genet. 87:893-899.
189. 1994. Ishii, T., D. S. Brar, D. S. Multani and G. S. Khush. Molecular tagging of
genes for brown planthopper resistance and earliness introgressed from Oryza
australiensis into cultivated rice, O. saliva. Genome 37:217-221.
190. 1994. Jena, K.K., G.S. Khush and G. Kochert. Comparative RFLP mapping
of a wild rice, Oryza officinalis and cultivated rice, O. saliva. Genome
37:382-389.
191. 1994. Multani, D. S., K.K. Jena, D. S. Brar, B. G. delos Reyes, E.R. Angeles, and
G. S. Khush. Development of monosomic alien addition lines and introgression
of genes from Oryza australiensis Domin. to cultivated rice, O. saliva L.
Theor. Appl. Genet. 88:102-109.
192. 1994. Pooni, H. S., Ish Kumar and G. S. Khush. A general method of detecting
additive dominance, and epistatic variation for metrical traits. V. Triple test
cross analysis of disomically inherited traits expressed in triploid tissues.
Heredity 72:563-569.
193. 1994. Sanchez, A.C. and G.S. Khush. Chromosomal location of some marker
genes in rice using primary trisomics. J. Hered. 85:297-300.
194. 1994. Virmani, S.S., G.S. Khush and P.L. Pingali. Hybrid rice for tropics:
Potentials, research priorities and policy issues. In R. S. Paroda and Mangala
Rai, Eds. Hybrid research and development needs in major cereals in the
Asia-Pacific region. Food and Agriculture Organization of the United Nations,
Regional Office for Asia and Pacific, Bangkok. pp. 61-86.
632 Wolf Prize in Agriculture
195. 1994. Khush, G.S. and R.C. Aquino. Breeding tropical japonicas for hybrid
rice production. In Virmani S.S., Ed. Hybrid rice technology: New developments
and future prospects. International Rice Research Institute, P.O. Box 933,
Manila 1099, Philippines. pp. 33-36.
196. 1994. Khush, G.S., D.S. Brar, F.J. Zapata, R. Nelson, S. McCouch, and
D.G. Bottrell. Rice biotechnology at IRRI. In Towards enhanced and sustainable
agricultural productivity in the 2000’s: Breeding research and biotechnology.
Taichung District Agricultural Improvement Station and SABRAO. Pages
387-414.
197. 1994. Khush, G.S. Increasing the genetic yield potential of rice: prospects
and approaches. International Rice Commission Newsletter 43:1-8.
198. 1994. Brar, D. S. and G. S. Khush. Cell and tissue culture for plant
improvement. In A.S. Basra, ed. Mechanisms of plant growth and improved
productivity. Modern Approaches, Marcel Dekker, Inc., New York.
pp. 229-278.
199. 1994. Khush, G. S. Rice Genetics and Breeding. Encyclopedia of Agricultural
Science 3:607-616.
200. 1995. Panda, N. and G. S. Khush. Host Plant Resistance to Insects. C.A.B.
International, Wallingford, England and International Rice Research Institute,
P.O. Box 933, Manila, Philippines. 431 pp.
201. 1995. Khush, G. S. Modern varieties - their real contribution to food supplies
and equity. Geo Journal 335(3):275-284.
202. 1995. Khush, G.S. Breaking the yield barrier of rice. Geo Journal
35(3):329-332.
203. 1995. Alejar M, F.J. Zapata, D. Senadhira, G.S. Khush and S.K. Datta SK.
Utilization of anther culture as a breeding tool in rice improvement.
In M. Terzi et al., eds. Current issues in plant molecular and cellular biology.
Kluwer Academic Publishers, Netherlands. Pages 137-143.
204. 1995. Zheng, K., N. Huang, J. Bennett, and G.S. Khush. PCR-based marker-
assisted selection in rice breeding, International Rice Reserach Institute, Manila
Philippines. IRRI Discussion Paper Series No 12.
205. 1995. Brar, D. S. and G. S. Khush. Wide hybridization for enhancing resistance
to biotic and abiotic stresses in rainfed lowland rice. In Fragile lives in fragile
ecosystems. Proc. International Rice Research Conference. IRRI, P.O. Box 933,
Manila, Philippines. Pages 901-910.
206. 1995. Hittalmani, S, T. Mew, G.S.Khush and N. Huang. DNA marker-assisted
breeding for disease resistance in rice. In Fragiles lives in fragile ecosystems.
Proc. International Rice Research Conference, IRRI, P.O. Box 933, Manila,
Philippines. pp. 947-961.
207. 1995. Katiyar, S.K., Y. Tan, Y. Zhang, B. Huang, Y. Xu, L. Zhao, N. Huang,
G.S. Khush and J. Bennett. Molecular tagging of gall midge resistance genes
Gurdev S. Khush 633
219. 1996. Kohli, A., B. Ghareyazie, H. S. Kim, G. S. Khush and J. Bennett. Cytosine
methylation implicated in silencing of B-glucuronidase genes in transgenic
rice. In G.S. Khush, ed. Rice Genetics III. International Rice Research Institute,
P.O. Box 933, Manila, Philippines. pp. 825-828.
220. 1996. Brar, D. S., R. Dalmacio, R. Elloran, R. Aggarwal, ER. Angeles and
G. S. Khush. Gene transfer and molecular characterization of introgression
grom wild Oryza species into rice. In G. S. Khush, ed. Rice Genetics III.
International Rice Research Institute, P.O. Box 933, Manila, Philippines.
pp. 477-486.
221. 1996. Khush, G. S., K. Singh, T. Ishii, A. Parco, N. Huang, D. S. Brar and
D. S. Multani. Centromere mapping and orientation of the cytological, classical,
and molecular linkage maps of rice. In G.S. Khush, ed. Rice Genetics III.
International Rice Research Institute, P.O. Box 933, Manila, Philippines.
pp. 57-75.
222. 1996. Robeniol, J.A., S.V. Constantino, A.P. Resurreccion, C.P. Villareal,
B. Ghareyazie, B.R. Lu, S.K. Katiyar, N. Huang, C.A. Menguito, E.R. Angeles,
H.Y. Fu, S. Reddy, W. Park, S.R. McCouch, G.S. Khush and J. Bennett. Sequence-
tagged sites and low-cost DNA marker technology for rice. In G.S. Khush,
ed. Rice Genetics III. International Rice Research Institute, P.O. Box 933,
Manila, Philippines. pp. 293-306.
223. 1996. Khush, G.S. Biotechnology for rice improvement. In R. Ishii and
T. Horie, eds. Crop Research in Asia: Achievements and Perspectives. Proceedings
of 2nd Asian Crop Science Congress. “Towards Improvement of Food
Production under Steep Population Increase and Global Environment Change”
Crop Science Society of Japan, Tokyo. pp. 320-325.
224. 1996. Khush, G. S. Genetic improvement of rice for weed management.
In R. Naylor, ed. Herbicide in Asian rice: transition in weed management.
Palo Alto (California): Institute for International Studies, Stanford University,
and Manila (Philippines): International Rice Research Institute. pp. 201-207.
225. 1996. Khush, G.S. and S. Peng. Breaking the yield frontier of rice. In
M.P. Reynolds, S. Rajaram and A. McNab, eds. Increasing the yield potential
in wheat: Breaking the barriers. CIMMYT D.F. Mexico. pp. 36-51.
226. 1996. Datta, S.K., K. Datta, L. Torrizo, M.F. Alam, J. Tu, Y. Fan, I. Altosaar,
J. Bentur, and G. S. Khush. Production of efficient Bt rice and application. In
Proceedings of 2"d Pacific Rim Conference on Biotechnology of Bacillus
thuringiensis and its impact on the enviroment. The Pacific Rim Bt Conference,
Chiang Mai, Thailand. Pages 401-411.
227. 1996. Singh, K., T. Ishii, A. Parco, N. Huang, D. S. Brar and G. S. Khush.
Centromere mapping and orientation of the molecular linkage map of rice
(Oryza sativa L.). Proc. Natl. Acad. Sci. USA 93:6163-6168.
Gurdev S. Khush 635
228. 1996. Singh, K., D.S. Multani and G.S. Khush. Secondary trisomics and
telotrisomics of rice: Origin, characterization and use in determining the
orientation of chromosome map. Genetics 143:517-529.
229. 1996. Tan, Y.J., Y. Zhang, B.C. Huang, Y.K. Xu, L.X. Zhao, S.K. Katiyar,
N. Huang, G.S. Khush, and J. Bennett. Analysis of resistance genetics and
mapping of resistance gene in a rice variety Duokang 1 to gall midge. Acta.
Phytophylacica Sinica 23:315-320.
230. 1996. Zhang, G., E.R. Angeles, M.L.P. Abenes, G.S. Khush and N. Huang.
RAPD and RFLP mapping of bacterial blight resistance gene xa-13 in rice.
Theor. Appl. Genet. 93:65-70.
231. 1996. Huang, N. , B. Courtois, G. S. Khush, H. Lin, G. Wang, P. Wu and
K. Zheng. Association of quantitative trait loci for plant height with major
dwarfing genes in rice. Heredity 74:130-137.
232. 1996. Mukhopadhay, K., N.K. Garrison, D.M. Hinton, C.W. Bacon, G.S. Khush,
H.D. Peck and N. Datta. Identification and characterization of bacterial
endophytes of rice. Mycopathologia 134:151-159.
233. 1997. Khush, G.S. Origin, dispersal, cultivation and variation of rice. Plant
Molecular Biology 35:25-34.
234. 1997. Brar, D. S. and G. S. Khush. Alien introgression in rice. Plant Molecular
Biology 35:35-47.
235. 1997. Bennett, J., M.B. Cohen, S. K. Katiyar, B. Ghareyazie and G. S. Khush.
Enhancing insect resistance in rice through biotechnology. In N. Carozzi and
M. Koziel, eds. Advances in insect control: The role of transgenic plants.
Francis and Taylor, London. pp. 75-93.
236. 1997. Brar, D.S. and G.S. Khush. Wide hybridization for rice improvement:
Alien gene transfer and molecular characterization of introgression.
In M.P. Jones, M. Dingkuhn, D.E. Johnson and S.O. Fagade, eds. Interspecific
hybridization: Progress and Prospects. WARDA, 01 BP 2551, Bouake, Cote
d’Ivoire. pp. 21-29.
237. 1997. Aggarwal, P.K., M.J. Kropff, P.S. Teng and G.S. Khush. The challenge
of integrating systems approach and limitations. In M.J. Kropff, P.S. Teng,
P.K. Aggarwal, J. Bouma, B.A.M. Bouman, J.W. Jones, H.H. Van Laar, eds.
Application of Systems Approaches at Field Level Vol. 2. Kluwer Academic
Publishers, Netherlands. pp. 1-23.
238. 1997. Datta, K., L. Torrizo, N. Oliva, M.F. Alam, C. Wu, E. Abrigo, A. Vasquez,
J. Tu, C. Quimio, M. Alejar, Z. Nicola, G.S. Khush and S.K. Datta. Production
of transgenic rice by protoplast, biolistic and Agrobacterium systems. In
Proceedings, Fifth International Symposium on Rice Molecular Biology.
Yi Hsien Pub. Co. Taipei, Taiwan. Pages 159-167.
239. 1997. Endo, N., T. Ogawa and G. S. Khush. Isozyme classification of Myanmar
rice cultivars resistant to bacterial blight. Breed. Sci. 47:27-32.
636 Wolf Prize in Agriculture
240. 1997. Aggarwal, R.K., D.S. Brar and G.S. Khush. Two new genomes in the
Oryza complex identified on the basis of molecular divergence analysis using
total genomic DNA hybridization. Mol. Gen. Genet. 254:1-12.
241. 1997. Huang, N., A. Parco, T. Mew, G. Magpantay, S. McCouch, E. Guiderdoni,
J. Xu, P. Subudhi, R. Angeles, and G. S. Khush. RFLP mapping of isozymes,
RAPD and QTLs for grain shape, brown planthopper resistance in a doubled
haploid rice population. Molecular Breeding 3:1-8.
242. 1997. Huang, N., E.R. Angeles, J. Domingo, G. Magpantay, S. Singh,
G. Zhang, N. Kumaravadivel, J Bennett, G. S. Khush. Pyramiding of bacterial
blight resistance genes in rice: marker-aided selection using RFLP and PCR.
Theor. Appl. Genet. 95:313-320.
243. 1997. Ghareyazie, B., F. Alinia, C.A. Menguito, L. Rubia, J.M. de Palma,
E.A. Liwanang, M.B. Cohen, G. S. Khush and J. Bennett. Enhanced resistance
to two stem borers in an aromatic rice containing a synthetic crylA(b) gene.
Molecular Breeding 3:401-414.
244. 1998. Khush, G. S. and S. Sarkarung. New plant type and breeding strategies
for rainfed lowland rice. In Rainfed rice for sustainable food security. Central
Rice Research Institute, Cuttack, Orissa, India. pp. 44-52.
245. 1998. Khush, G.S. Strategies for increasing crop productivity. In V.L. Chopra,
R.B. Singh and Anupam Verma, eds. Crop productivity and sustainability —
shaping the future. Proceedings of 2nd International Crop Science Congress,
Oxford & IBH Publishing Co. Pvt. Ltd. New Delhi. pp. 19-43
246. 1998. Khush, G. S. and P. S. Baenziger. Crop improvement: emerging trends
in rice and wheat. In V.L. Chopra, R.B. Singh and Anupam Verma, eds. Crop
productivity and sustainability — shaping the future. Proceedings of 2nd
International Crop Science Congress, Oxford & IBH Publishing Co. Pvt. Ltd.
New Delhi. pp. 113-125.
247. 1998. Khush, G. S. Innovative approaches for improving the yield and grain
quality of rice. In H.Y. Lu, J.M. sung and C.H. Kao, eds. Asian Crop Science.
Proceedings of the 3rd Asian Science Conference. Chinese Society of Agronomy,
Taichung, Taiwan. Pages 436-450.
248. 1998. Khush, G. S. and D. S. Brar. The application of biotechnology to rice.
In C.L. Eves and B.M. Bedford, eds. Agricultural Biotechnology in International
Development. CABI Publishing, Wallingford, U.K. Pages 97-121.
249. 1998. Khush, G. S., D. S. Brar nad J. Bennett. Apomixis in rice and prospects
for its use in heterosis breeding. In Virmani, S.S., E.A. Siddiq and
K. Muralidharan, eds. Advances in hybrid rice technology. Proceedings of the
3rd International Symposium on Hybrid Rice. 14-16 Nov. 1996. International
Rice Research Institute, P.O. Box 933, Manila, Philippines. Pages 297-309.
250. 1998. Khush, G.S., S.B. Peng and S.S. Virmani. Improving the yield
potential by modifying plant type and exploiting heterosis. In J.C. Waterlow,
D.G. Armstrong, L. Fowden and R. Riley, eds. Feeding a world population of
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263. 1999. Peng, S., R.C. Laza, R.M. Visperas, A.L. Sanico, K.G. Cassman and
G.S. Khush. Grain yield of rice cultivars and lines developed in the Philippines
since 1966. Crop Science 40:307-314.
264. 1999. Peng S., K. G. Cassman, S. S. Virmani, J. Sheehy and G. S. Khush. Yield
potential trends of tropical rice since the release of IR8 and the challenge of
increasing rice yield potential. Crop Science 39:1552-1559.
265. 1999. Jalodar N.B., N.W. Blackhall, T.P.W. Hartman, D.S. Brar, G.S. Khush,
M.R. Davey, E.C. Cocking, and J.B. Power. Intergeneric somatic hybrids of
rice [Oryza saliva L. (+) Porleresia coarctala (Roxb.) Tateoka]. Theor. Appl.
Genet. 99:570-577.
266. 2000. Fischer, K.S., J. Barton, G.S. Khush, H. Leung and R. Cantrell.
Collaborations in rice. Science 290:279-280.
267. 2000. Khush, G. S. New plant type of rice for increasing the yield potential
of rice. In J.S. Nanda, ed. Rice breeding and genetics. Science Publishers, Inc.,
P.O. Box 699, Enfield, New Hampshire 03748, USA. Pages 99-108.
268. 2000. Jena K.K. and G.S. Khush. Exploitation of alien species in rice
improvement — opportunities, achievements and future challenges. In J.S.
Nanda, ed. Rice breeding and genetics. Science Publishers, Inc., P.O. Box 699,
Enfield, New Hampshire 03748, USA. Pages 271-285.
269. 2000. Singh, R.J. and G. S. Khush. Cytogenetics of rice. In J. S. Nanda, ed.
Rice breeding and genetics. Science Publishers Inc. P.O. Box 699, Enfield,
New Hampshire 03748, USA. Pages 287-311.
270. 2000. Dela Cruz N, and G. S. Khush. Rice grain quality evaluation procedures.
In R.K. Singh and G. S. Khush, eds. Aromatic Rices. Science Publishers, Inc.
Enfield U.S.A. and Plymouth, U.K. Pages 15-28.
271. 2000. Khush, G.S. Taxonomy and origin of rice. In R.K. Singh, U.S. Singh,
G.S. Khush, eds. Aromatic Rices. Science Publishers, Inc. Enfield, U.S.A. and
Plymouth, U.K. Pages 5-13.
272. 2000. Hossain M, J. Bennett, S.K. Datta, H. Leung and G. S. Khush.
Biotechnology research in rice for Asia: Priorities, focus and directions.
In M. Qaim, A.F. Krattiger and J. von Braun, eds. Agricultural Biotechnology
in Developing Countries: Towards Optimizing the Benefits for the Poor. Kluwer
Academic Publishers. Pages 99-120.
273. 2000. Khush G.S. and H. Leung. Plant genome research and breeding strategies
for sustainable food production in 21st century. In L. Esaki, ed. New frontiers
of science and technology. Proceedings of the International Conference on
Science Frontier Tsukuba 999, November 17-19, 1999 at Tsukuba Center,
Japan. Universal Academy Press Inc., Tokyo, Japan. Pages 15-27.
274. 2000. Khush G.S. Strategies for increasing the yield potential of rice. In
Sheehy JE, Mitchell PL and Hardy B (eds). Redesigning rice photosynthesis to
increase yield. International Rice Research Institute, Manila, Philippines. Pages
207-211.
Gurdev S. Khush 639
275. 2000. Tu J., K. Datta, G.S. Khush, Q. Zhang, and S.K. Datta. Field performance
of Xa21 transgenic indica rice (Oryza saliva L.), IR72. Theor. Appl. Genet.
101:15-20.
276. 2000. Tu J., G. Zhang, K. Datta, C. Xu, Y. He, Q. Zhang, G. S. Khush and S.K.
Datta. Field performance of transgenic elite commercial hybrid rice expressing
Bacillus thuringiensis S-endotoxin. Nature Biotechnology 18:1101-1104.
277. 2000. Tsunematsu, H., M.J. Yanoria, L.A. Ebron, N. Hayashi, 1. Endo,
H. Kato, T. Imbe and G. S. Khush. Development of monogenic lines of rice
for blast resistance. Breeding Science 50:229-234.
278. 2000. Sripongpangkul, K., G.B. T. Posa, D. Senadhira, D. S. Brar, N. Huang,
G. S. Khush, and Z. Li. Genes/QTLs affecting flood tolerance in rice. Theor.
Appl. Genet. 101:1074-1081.
279. 2000. Angeles, E.R. and G.S. Khush. Genetics of resistance to green leafhopper
in five cultivars of rice, Oryza saliva L. SABRAO Journal 32:1-4.
280. 2000. Sanchez, A.C., D.S. Brar, N. Huang, Z. Li and G.S. Khush. Sequence
tagged site marker-assisted selection for three bacterial blight resistance genes
in rice. Crop Sci. 40:792-797.
281. 2000. Zhongchao, Y, J. Chen, L. Zeng, M. Goh, H. Leung, G.S. Khush, and
G.L. Wang. Characterizing rice lesion mimic mutants and identifying mutant
with broad spectrum resistance to rice blast and bacterial blight. MPMI
13:869-876.
282. 2000. Angeles, E.R. and G.S. Khush. Genetic analysis of resistance to green
leafhopper, Nephotettix virescens (Distant) in three varieties of rice. Plant
Breeding 119:446-448.
283. 2000. Sanchez, A.C. and G.S. Khush. Chromosomal localization of five mutant
genes in rice, Oryza sativa L. using primary trisomics. Plant Breeding
119-84-86.
284. 2000. Imbe, T., H. Tsunematsu, H. Kato and G. S. Khush. Genetic analysis of
blast resistance in IR varieties and resistant breeding. In D. Tharreau
et al., eds. Advances in Rice Blast Research. Kluwer Academic Publishers.
Netherlands. Pages 1-8.
285. 2000. Sheehy, J., P. Mitchell, J. Dionora, T. Tsukaguchi, S.B. Peng and
G.S. Khush. Unlocking the yield barrier in rice through a nitrogen-led
improvement in the radiation conversion factor. Plant Prod. Sci. 3(4):
372-374.
286. 2001. Khush, G. S. Green revolution: the way forward. Nature Reviews
Genetics 2: 815-822.
287. 2001. Khush G. S. Challenges for meeting the global food and nutrient needs
in the new millennium. Proc. Nutrition Society 60:15-26.
288. 2001. Khush G.S. Super rice for increasing the genetic yield potential. In
Reeves, E.C.R. ed. Encyclopedia of Genetics. Fitzroy Dearborn Publishers,
London, Chicago. Pages 663-667.
640 Wolf Prize in Agriculture
J.S. Nanda and S.D. Sharma eds. Science Publishers Inc. Enfield (NH) USA,
Plymouth, USA.
301. 2003. Khush, G.S., D.S. Brar, P.S. Virka, S.X. Tang, S.S. Malik, G.A. Bastos,
Y.T. Lee, R. McNally, L.N. Trinh. Y. Jiang and M.A.M. Shata. Classifying rice
germplasm by isozyme polymorphism and origin of cultivated rice. Discussion
paper series No 46. Los Banos (Philippines): International Rice Research
Institute. 279 P.
302. 2003. Kobayashi, S. Y. Fukua, S. Morita, T. Sato, M. Osaki. G.S. Khush
Quantitative trait loci affecting flag leaf development in rice (Oryza SativaL.)
Breeding Science 53: 255-262.
303. 2003. Kobayashi, S., Y. Fukula, T. Sato, M. Osaki and G. S. Khush. Molecular
Marker dissection of rice (Oryza Sativa) plant architecture under temperate
and tropical climates. Theor. Appl. Genet. 107-1350-1356.
304. 2003. Lee, K. S., S. Rasabandith, E. R. Angeles and G. S. Khush. Inheritance of
resistance to bacterial blight of rice. Phytopathology 93: 147-152.
305. 2003. Multani, D. S., G. S. Khush, B. G. delos Reyes, D. S. Brar. Alian
gene introgression and development of monosomic alien addition lines
from Oryza latifolia Desv. to rice, Oryza Sativa L. Theor. Appl. Genet. 107:
395-405.
306. 2003. Peng, S. and G. S. Khush. Four decades of breeding for varietal
improvement of irrigated lowland rice in the International Rice Research
Institute. Plant Production Science 6-157-164.
307. 2004. Ebron, L. A., Y. Fukuta, T. Imbe, H. Kato, J.M.T. Yanoria, H. Tsunematsu,
G. S. Khush and M. Yokoo. Estimation of genes for blast resistance in elite
indica type rice (Oryza sativa L). Varieties bred at the international Rice
Research Institute. Breeding Science. 53: 381-387.
308. 2004. Kabayashi, S., Y. Fukuta, T. Yagif T. Sato, M. Osaki, G. S. Khush.
Identification and characterization of quantitative trait loci affecting
spikelet number per panicle in rice (Oryza sativa L) Field Crops Research 89:
253-262.
309. 2004. Khush, G. S., E. Angeles, P. S. Virk and D. S. Brar. Breeding rice for
resistance to tungro virus at IRRI. SABRAO Journal 36: 101-106.
310. 2004. Nematzadeh, Gh. A., N. Huang and G. S. Khush. Mapping the gene for
aroma in rice (Oryza sativa L.) by bulk segregation analysis via RAPD markers.
J. Agric. Sci. Technol. 6:129-137.
311. 2004. Peng, S., J. Huang, J. E. Sheehy, R. C. Laza, R. M. Visperas, X. Zhong,
G. S. Centano, G.S. Khush and K. G. Cassman. Rice yields decline with higher
night temperature from global warming. Proc. Natl. Acad. Sciences (USA)
101(27): 9971-9975.
312. 2005. Ebron, L. A., Y. Fukuta, T. Imbe, H. Kato, M. J. T. Yanoria, H. Tsunematsu,
G. S. Khush, N. Kobayashi and M. Yokoo. Identification of blast resistance
642 Wolf Prize in Agriculture
genes in elite indica type varieties of rice (Oryza sativa L). SABRAO Journal
37: 19-31.
313. 2005. Khush, G. S. Green Revolution: challenges ahead. Pages 37-51.
In R. Tuberosa, R. L. Phillips and M. Gale (eds.) In the wake of the double
helix. From green revolution to the gene revolution. University of Bologna,
Italy.
314. 2005. Khush, G. S. Taxonomy, ecology and agronomy of rice cultivation
vis-a-vis genetics engineering of rice. Pages 26-37. In V.L. Chopra,
S. Shantaram and R. P. Sharma eds. Biosafetu of transgenic rice. National
Academy of Agricultural Sciences, New Delhi.
315. 2005. Khush, G. S. What it will take to feed 5.0 billion rice consumers in
2030? Plant Molecular Biology 59: 1-6.
316. 2005. Khush, G. S. and P.S. Virk IR varieties and their impact. Los Banos
(Philippines): International Rice Research Institute. 163 P.
317. 2005. Wu, J. L., C. Wu, C. Lei, M. Baraoidan, A. Bordeos, M. R.S. Madamba,
M. Ramos-Pamplona, R. Mauleon, A. Portugal, V. J. Ulat, R. Brukiewich,
G. Wang, J. Leach, G. S. Khush and Hei Leung. Chemical- and irradiation-
induced mutants of indica rice IR64 for forward and reverse genetics. Plant
Molecular Biology. 59: 85-97.
318. 2006. Baisakh, N., S,. Rehana, M. Rai, N. Oliva, J. Tan, D. J. MacKill,
G. S. Khush, K. Datta and S.K. Datta.. Marker free transgenic (MFT) near-
isogenic introgression lines (NILS) of golden indica rice (Cv. IR64) with
accumulation of provitamin A in the endosperm tissue. Plant Biotechnology
Journal 4: 467-475.
319. 2006. Brar, D. S. and G. S. Khush. Cytogenetic manipulation and germplasm
enhancement of rice (Oryza sativa L). Pages 115-158. In R. J. Singh and
P.P. Jauhar eds. Genetic resources, chromosome engineering and crop
improvement. Taylor and Francis, Boca Raton, London, New York.
320. 2006. Kobayashi, S., D. Araki, M. Osaki, G. S. Khush, Y. Fukuta. Localization,
validation and characterization of plant type QTLs on chromosome 4 and 6
in rice (Oryza sativa L) Field Crops Research. 96: 106-112.
321. 2006. Subudhi, P. K., T. Sasaki, G. S. Khush. Rice. Pages 1-78 In Kole C. (ed)
Genome mapping and molecular breeding in plants. Springer: Berlin,
Heidelberg, New York.
322. 2007. Khush, G.S., E.R. Angeles, and D.S. Brar. Genetics analysis of resistance
to green leafhopper, Nephotettix virescens (Distant), in IR rice varieties.
SABRAO Journal 39(2):79-88.
Roger N. Beachy
Danforth Plant Science Center
St. Louis, Missouri, USA
k
Roger Beachy, Ph.D., founding president of the Donald Danforth Plant Science
Center, a non-profit private research institute in St. Louis, MO, earned his Ph.D. in
Plant Pathology at Michigan State Univ.; he conducted post-doctoral positions at
the Univ. of Arizona and at Cornell Univ., NY. He previously held academic positions
at Washington Univ., St. Louis, and The Scripps Research Institute, La Jolla, CA,
where he was co-founder of the International Laboratory for Tropical Agricultural
Biotechnology. He is a member of the U.S. National Academy of Sciences and a
Fellow of the American Academy of Microbiology, and the American Assoc. for the
Advancement of Science. He has received numerous awards including the Wolf
Prize in Agriculture, the D. Robert Hoagland Award from the Society of Plant
Biologists and Ruth Allen Award from the American Phytopathological Society.
Beachy serves as Chair-Elect of the AAAS Section on Agriculture, Food and
Renewable Resources and is President of the International Assoc. of Plant
Biotechnology, among other activities. He is recognized for his work in molecular
virology and gene expression in plants, and for pioneering research in developing
transgenic plants that are resistant to virus infection. His research includes: studies
of mechanisms of transgenic virus resistance including, in rice and sweet potato;
characterizing functional activities of transcription factors; and developing a
chemical gene switching system for use in plants. Beachy has been a vocal advocate
643
644 Wolf Prize in Agriculture
CURRICULUM VITAE
President Professor
Donald Danforth Plant Science Center Department of Biology
975 North Warson Road Washington University
St. Louis, Missouri St. Louis, Missouri
Education:
1966 B.A. - Biology, Goshen College, Goshen, Indiana
1973 Ph.D. - Plant Pathology, Michigan State University (advisor H. H. Murakishi),
East Lansing, Michigan
Post-doctoral:
1973 Department of Agricultural Biochemistry, University of Arizona,
Tucson, Arizona
1973-1976 Department of Plant Pathology (advisor M. Zaitlin), Cornell University,
Ithaca, New York
1976-1978 U.S.D.A. Nutrition Laboratory, (advisor J. F. Thompson), Cornell
University, Ithaca, New York
PROFESSIONAL EXPERIENCE:
Jan. 1999 to present President and Director, Donald Danforth Plant Science
Center, 975 North Warson Road, St. Louis, Missouri
Jan. 1999 to present Professor, Department of Biology, Washington Univer-
sity, St. Louis, Missouri
June 1991 to Jan. 1999 Professor and Scripps Family Chair in Cell Biology, Head,
Division of Plant Biology, Co-Director, Inter-national
Laboratory for Tropical Agricultural Bio-technology
(ILTAB), The Scripps Research Institute, La Jolla, California
1992 to 2005 Adjunct Professor, Department of Biology, Peking
University, Beijing, China
1997 to 1999 Adjunct Professor, Department of Plant Pathology,
University of California, Riverside, California
Jan. 1986 to June 1991 Professor of Biology, Director of the Center for Plant
Science and Biotechnology, Washington University,
St. Louis, Missouri
Roger N. Beachy 645
HONORS:
Betty Klepper Crop Science Society of America Endowed Lecture Award, 2006
Wolf Prize in Agriculture, 2001
Dennis Robert Hoagland Award, American Society of Plant Biologists, 2000
Fellow, St. Louis Academy of Science, 2000
Fellow, American Academy of Microbiology, 2000
Honorary Doctor of Science, Michigan State University, 2001
Culture for Service Award, Goshen College, 2001
Scientist of the Year, R&D Magazine, 1999
Member, National Academy of Sciences, U.S.A., 1997
Commonwealth Award for Science and Industry, Bank of Delaware, 1991
Ruth Allen Award, American Phytopathological Society, 1990
Fellow, American Association for the Advancement of Science, 1989
LOCAL RECOGNITION:
Agri-Business Leader of the Year, St. Louis Agri-Business Club, 2004
Honoree, Huntington’s Disease Society of America-St. Louis Chapter, 2002
Entrepreneur of the Year, Ernst & Young, 2001
Wm. D. Phillips Technology Advancement Award, St. Louis County Economic
Council, 2000
Headliner of the Year Award, San Diego Press Club, 1995
646 Wolf Prize in Agriculture
Past:
Member, American Society for Gravitational and Space Biology, 2004-present
Councilor, National Academy of Sciences, 2003-2006
Member, National Academy of Sciences Council Committee on Budget and Internal
Affairs, 2003-2006
Member, UC-Davis Office of Research External Research Advisory Board,
2003-2006
Member, Editorial Board, Current Opinion in Plant Biology, 2002-present
Member, Portland State University Project Advisory Group for “Public Goods and
University-Industry Relationships in Agricultural Biotechnology”, 2002-2006
Member, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT);
Governing Board of Directors; Pantancheru, India, 2001-2007
Member, DOE Biomass Research and Development Technical Advisory Committee,
2003-2005
Member, NASA’s Biological and Physical Research Maximization and Prioritization
(REMAP) Task Force, 2002
Member, National Academy of Sciences/National Research Council Opportunities
in Agriculture Committee, 2001-2002
Member, Burrill and Company; Board of Advisors; San Francisco, CA, 2000-2007
Member, German American Academic Council, National Academy of Sciences,
1999-2000
Roger N. Beachy 647
Member, Board of Trustees, The Academy of Science of St. Louis; St. Louis, MO,
2000-present
Member, Board of Trustees, St. Louis Science Center; St. Louis, MO, 2003-present
Member, Science Advisory Board Chlorogen, Inc.; St. Louis, MO, 2003-2007
St. Louis Regional Chamber & Growth Association (RCGA), St. Louis, MO
2002-present
Member, Technology Board, Midwest Bancshares; St. Louis, MO, 2001-present
The World Affairs Council of St. Louis; St. Louis, MO, 2000-2005
NAMED LECTURES:-
Storer Life Sciences Endowment Lecture Series, University of California-Davis, 2008
National Academy of Sciences Lectureship Series, University of Minnesota, 2007
The Horning Endowment Lecture, Oregon State University, 2007
President’s Circle, NAS, Washington, D.C., 2006
Sackler Colloquium, Organizer, Washington, D.C., 2006
Betty Klepper Endowed Lecturer, Crop Science Society of America, Washington,
DC, 2006
Stadler Genetics Symposium, Columbia, MO 2006
Boyer Symposium: Marine and Terrestrial Molecular Bioscience-New Frontiers,
Philadelphia, PA, 2005
Roger N. Beachy 649
Regional Commerce and Growth Association’s Missouri Life Sciences Summit, Lake
of the Ozarks, MO, 2002
University of California-Irvine 15th Annual Chief Executive Roundtable Retreat,
2002
University of Minnesota Plant Disease & Pest Management Symposium, 2002
University of California-Irvine Virology Seminar, 2002
North Carolina State University Emerging Issues Forum, 2002
International Rice Genome Meeting, Forum X, Japan, 2002
Michigan State University Plant Breeding & Genetics Symposium, 2001
American Farm Bureau Annual Meeting, Orlando, FL, 2001
University of Minnesota Conference Series, Minneapolis, MN, 2001
Seminar Series for University of California at Davis, 2001
Knowledge Millennium II: Biotechnology – The New World, New Delhi, India,
2001
Forum Agrosante, Paris, France, 2001
National Academy of Sciences 2001 Symposium, Washington, DC, 2001
World Agricultural Forum World Congress, St. Louis, MO, 2001
2001 Congress on In Vitro Biology, St. Louis, MO, 2001
International Seminar on Future of Wheat and Wheat Research, Paris, France,
2001
2001 Cold Spring Harbor Laboratory Summer Course, Cold Spring Harbor, NY,
2001
Global Consortium of Higher Education & Research for Agriculture, San Francisco,
CA, 2001
Society for Industrial Microbiology Annual Meeting, St. Louis, MO, 2001
Seminar on Agricultural Biotechnology at Academia Sinica in Taipei, Taiwan, 2001
American Chemical Society, Chicago, IL, 2001
Pharmacia/Monsanto Annual Symposium, 2001
Keynote Speaker, Iowa State University Symposium, 2001
National Association of State University and Land Grant Colleges, 2001
National Association of Conservation Districts Symposium, Colorado Springs, CO,
2000
Princeton University, Department of Molecular Biology, Princeton, NJ, 2000
Bio 2000, Boston, MA, 2000
U.S.-India High Level Science and Technology Dialogue, 2000
Missouri Venture Forum, St. Louis, MO2000
Michigan State University, East Lansing, MI, 2000
John Innes Centre, Norwich, United Kingdom, 2000
Goshen College, Goshen, IN, 2000
Max-Planck-Institut, Cologne, Germany, Symposium, 2000
Great Plains Cereals Biotechnology Symposium, Manhattan, KS, 2000
652 Wolf Prize in Agriculture
LIST OF PUBLICATIONS
INVITED ARTICLES, BOOKS - 1974 TO PRESENT:
1. Zaitlin, M. and R.N. Beachy. 1974. Protoplasts and separated cells: Some new
vistas for plant virology. In: “Proc. of the 3rd Intern. Congress for Plant
Tissues and Cell Culture.” Leicester, pp. 265-286.
2. Zaitlin, M. R.N. Beachy, G. Bruening, C.P. Romaine, and R. Scalla. l976.
Translation of tobacco mosaic virus RNA. In “Animal Virology,” eds.
D. Baltimore, A. Huang, and C.F. Fox, Academic Press, pp. 567-58l.
3. Bruening, G., R.N. Beachy, and M. Zaitlin. l979. Replication of RNA plant
viruses. In “Molecular Biology of Plants,” eds. I. Rubenstein, R.L. Phillips,
C.E. Green, and B.G. Gengenbach, Academic Press pp. 24l-272.
4. Beachy, R.N., J.F. Thompson, and J.T. Madison. l979. Isolation and
characterization of messenger RNAs that code for the subunits of soybean
seed protein. In “The Plant Seed: Development, Preservation, and
Germination,” eds., Rubenstein, Green, Phillips and Gengenbach, Academic
Press, New York, pp. 67-84.
5. Beachy, R.N., K.A. Barton, J.F. Thompson, and N.P. Jarvis. l980. The mRNAs
that code for soybean seed proteins. In “Genome Organization and Expression
in Plants,” C. Leaver, ed., Plenum Press, N.Y., pp. 273-28l.
6. Walbot, V., R.N. Beachy, and M.C. Yao. l980. Molecular techniques applied
to polyploids. In “Polyploidy: Biological Relevance,” W.H. Lewis, ed., Plenum
Publ. Corp., N.Y., pp. 529-535.
7. Beachy, R.N. Molecular aspects of seed storage protein biosynthesis. 1981. In
“Critical Reviews in Food Science and Nutrition”, CRC Press, Vol. 16(2):
187-198.
8. Beachy, R.N., J.J. Doyle, B.F. Ladin, and M.A. Schuler. 1983. Structure and
expression of genes encoding the soybean 7S seed storage proteins. In
“Structure and Function of the Plant Genome”, Plenum Press, N.Y.
9. Beachy, R.N., J. Bryant, J.J. Doyle, K. Kitamura, and B.F. Ladin. 1983. Molecular
characterization of a soybean variety lacking a subunit of the 7S seed storage
protein. ed. R.B. Goldberg. Plant Mol Biol A.R. Liss, Inc., N.Y. pp. 413-422.
Roger N. Beachy 653
10. Doyle, J.J., R.N. Beachy, and W.H. Lewis. 1984. Evolution of rDNA in Claytonia
polyploid complexes. In: “Plant Biosystematics,” ed. W.G. Grant, Academic
Press, N.Y., pp. 321-342.
11. Beachy, R.N. 1984. Toward an understanding of gene expression in plants.
In: “Proceedings of the Stadler Genetics Symposium,” ed. P. Gustafson, Plenum
Press, Inc., pp. 605-626.
12. Beachy, R.N. and R. Fraley. 1985. Potentials for applications of genetic
engineering technology to soybeans. In “New Protein Foods,” eds. A.M. Altschul
and H. Wilke, Academic Press, N.Y., pp. 89-105.
13. Beachy, R.N., P. Abel, M.J. Oliver, B. De, R.T. Fraley, S.G. Rogers, and
R.B. Horsch. 1985. Potential for applying genetic transformation to studies of
viral pathogenesis and cross-protection. In: “Biotechnology in Plant Sciences:
Relevance to Agriculture in the 1980’s”, eds. M. Zaitlin, P. Day, and
A. Hollaender, Academic Press, pp. 265-276.
14. Beachy, R.N., P.P. Abel, R.S. Nelson, S.G. Rogers, and R.T. Fraley. 1987.
Transgenic plants that express the coat protein gene of TMV are resistant to
infection by TMV. In: “Molecular Strategies for Crop Protection,” eds. C.S.
Arntzen and C.A. Ryan. A.R. Liss, Inc., N.Y., pp. 205-213.
15. Bray, E.A., and R.N. Beachy. February, 1986. A modulation by abscisic acid of
genes encoding bð-ðconglycinin in developing soybean cotyledons. In:
“Proceedings of UCLA Symposium on Plant Growth Regulators,” Lake Tahoe,
CA.
16. Jaworski, E.G., R.T. Fraley, S.G. Rogers, R.B. Horsch, R.N. Beachy, and N.-H.
Chua. 1987. Genetic transformation of plants. In: “Protein Engineering:
Applications in Science, Medicine, and Industry,” eds. M. Inouye and
R. Sarma. Academic Press, Inc., pp. 383-391.
17. Beachy, R.N., D.M. Stark, C.M. Deom, M.J. Oliver, and R.T. Fraley. 1987.
Expression of sequences of tobacco mosaic virus in transgenic plants and
their role in disease resistance. In: “Tailoring Genes for Crop Improvement,”
eds. G. Bruening, J. Harada, T. Kosuge and J. Hallaender, Plenum Publ. Corp.,
N.Y., pp. 169-180.
18. Beachy, R.N., S.G. Rogers and R.T. Fraley. 1987. Genetic transformation to
confer resistance to plant virus disease. In: “Genetic Engineering,” Vol. 9,
eds. J. Setlow, Plenum Press, N.Y., pp. 229-247.
19. Beachy, R.N., P. Powell Abel, R.S. Nelson, J.C. Register III, N. Tumer, and
R.T. Fraley. 1987. Genetic engineering of plants for protection against virus
diseases. In: “Plant Resistance to Viruses,” Ciba Foundation Symposium, eds.
D. Evered and S. Harnett, Wiley and Sons, pp. 151-169.
20. Beachy, R.N. 1988. Virus cross-protection in transgenic plants. In: “Plant
Gene Research. Temporal and Spatial Regulation of Plant Genes,” eds. D.P.S.
Verma and R.B. Goldberg, Springer-Verlag, N.Y., pp. 313-327.
654 Wolf Prize in Agriculture
21. Hemenway, C., N. Tumer, P.A. Powell and R.N. Beachy. 1989. In: “Genetic
Engineering of Plants for Viral Disease Resistance.” eds. I. Vasil and J. Schell.
22. Register III, J.C., P.A. Powell, and R.N. Beachy. 1989. Genetically engineered
cross protection against TMV interferes with initial infection and long distance
spread of the virus. In: “Molecular Biology of Plant-Pathogen Interactions,”
eds. B. Staskowicz, P. Ahlquist, and O.C. Yoder, Alan R. Liss, N.Y.
pp. 269-281.
23. Clark, W. G., J.C. Register III, and R.N. Beachy. Application of genetic
engineering to agriculture. In: “Recombinant DNA Technology and
Application,” Chapter 6. Prokop, Editor. McGraw-Hill.
24. Clark, W.G., J.C. Register III, and R.N. Beachy. Engineering virus resistance
in transgenic plants. In: “Horticultural Biotechnology; Plant Biology,”
Vol. 11:1-11. A.B. Bennett and S.D. O’Neill, Editors. Wiley-Liss, New York.
25. Lucas, W.J., S. Wolf, C.M. Deom, G.M. Kishore, and R.N. Beachy. 1990.
Plasmodesmata-virus interaction. In: “Parallels in Cell-to-Cell Junctions in
Plants and Animals, eds., A.W. Robards, H. Jongsma, W.J. Lucas, J. Pitts,
D. Spray, NATO ASI series H; Cell Biology Vol. 46:261-274, Springer-Verlag,
Berlin.
26. Fauquet, C.M., and R.N. Beachy. 1990. Coat protein mediated resistance, a
new type of resistance to control plant viruses. In: Molecular Methods for
Potato Improvement, Proc. Planning Conference “Application of molecular
techniques to potato germplasm enhancement.” Ed. CIP, pp 72-84, Lima.
27. Fauquet, C.M., and R.N. Beachy. 1990. Virus resistance and genetic
engineering, concepts, efficacy, and stability. In: Monograph. “Enhancing
production of tropical crops with biotechnology.” Ed. Thotthapilly, Ibadan,
Nigeria.
28. Fauquet, C.M., and R.N. Beachy. 1990. Cassava viruses and genetic
engineering. In: Monograph. “Enhancing production of tropical crops with
biotechnology,” ed. Thotthapilly, Ibadan, Nigeria.
29. Fauquet, C.M., and R.N. Beachy. 1990. “Constructing genes for virus resistance
in tropical plants,” ed. Gamborg, San Jose, Costa Rica.
30. Beachy, R.N. 1990. Plant transformation to confer resistance against virus
infection. In: “Gene Manipulation in Plant Improvement II,” ed. J.P. Gustafson,
Plenum Press, N.Y.
31. Moore, P., C.M. Deom, and R.N. Beachy. 1991. The p30 movement protein of
TMV alters plasmodesmata structure and function. In: “Cell-Cell Interactions
in Early Development,” pp 273-282. Wiley-Liss, Inc.
32. Beachy, R.N. 1991. The very structure of scientific research does not mitigate
against developing products to help the environment, the poor, and the hungry.
In: “Journal of Agricultural & Environmental Ethics,” Vol. 4, No. 2.
Roger N. Beachy 655
33. Beachy, R.N. 1992. Coat protein mediated protection and the potential for its
application in agriculture. In: “Biotechnology and Environmental Science:
Molecular Approaches,” eds. S. Mongkolsuk et al., Plenum Press, N.Y.
34. Sturtevant, A.P., and R.N. Beachy. 1992. Virus resistance in transgenic plants:
Coat protein-mediated resistance. In: “Transgenic Plants.” pp 93-112. Marcel
Dekker, Inc.
35. Beachy, R.N. 1993. Virus resistance through expression of coat protein genes.
In: “Biotechnology in Plant Disease Control,” ed. Ilan Chet, pp 89-104,
Wiley-Liss, N.Y.
36. Fitchen, J. and R.N. Beachy. 1993. Genetically engineered protection
against viruses in transgenic plants. In: “Annu. Rev. Microbiol,” Vol. 47:
739-763.
37. Schöpke, C., Franche, C., Bogusz, D., Chavarriaga, P., Fauquet, C., and
R.N. Beachy. 1993. Transformation in cassava (manihot esculenta Crantz).
In: “Biotechnology In Agriculture and Forestry,” 23:273-289 Plant Protoplasts
and Genetic Engineering IV. ed. Y.P.S. Bajaj. Springer Verlag, Heidelberg.
38. Rochester, D.E., R.N. Beachy, and C.M. Fauquet. 1993. Geminivirus
Nomenclature: The need to set taxonomic standards. Virology Division News
132:221-224.
39. Reimann-Philipp, U., and R.N. Beachy. 1993. Plant resistance to virus diseases
through genetic engineering: Can a similar approach control plant-parasitic
nematodes? In: “Journal of Nematology,” 25(4):541-547.
40. Holt, C.A., and R.N. Beachy. 1993. Detection of localization of plant virus
antigens. In: “Tissue Printing: Tools for the Study of Anatomy, Histochemistry,
and Gene Expression,” eds. Philip D. Reid and Raphael F. Pont-Lezica.
Academic Press, Inc., Orlando, FL.
41. Beachy, R.N. 1993. Introduction: Transgenic resistance to plant viruses.
In: “Seminars in Virology,” 4:(6):327-328.
42. Barefoot, S.F., R.N. Beachy, and M.S. Lilburn. 1994. Labeling of food-plant
biotechnology products. In: CAST Issue Paper No. 4:1-8.
43. Beachy, R.N., E. Schell-Frederick, and J. Schell. 1995. La biotechnologie végétale
au service de la santé. In: “Diogènes,” No. 172, 43(4): 97-110, ed. Françoise
Héritier, Imprimerie Alenconnaise, Paris.
44. Beachy, Roger N. 1996. Virus-resistant transgenic plants. In: “Biotechnology
and Integrated Pest Management,” ed. G.J. Persley. CAB International,
pp 226-232.
45. Fenczik, C.A., B.L. Epel, and R.N. Beachy. 1996. Role of plasmodesmata and
virus movement proteins in spread of plant viruses. In: “Signal Transduction
in Plant Development.,” pp. 249-272, ed. D.P.S. Verma. Springer-Verlag, New
York.
656 Wolf Prize in Agriculture
46. Beachy, R.N., J.H. Fitchen, and M.B. Hein. 1996. Use of plant viruses for
delivery of vaccine epitopes. In: “Engineering Plants for Commercial Products
and Applications,” eds. Glenn B. Collins and Robert J. Shepherd, Annals of
the New York Academy of Sciences, New York 792:43-50.
47. Beachy, R.N., J.H. Fitchen. 1996. Pathogen-derived resistance to plant viruses
and plant viruses as vaccines. In: “Microbe Hunters – Then and Now,” eds.
H. Koprowski and M. Oldstone, Medi-Ed Press, Bloomington. pp. 295-305.
48. Beachy, R.N. and C.M. Fauquet. 1996. La cooperation scientifique nord-sud
en biotechnologie vegetale: l’ILTAB un modele parmi d’autres. In “Les Sciences
Hors D’Occident Au XXE Siecle,” 7:277-283. ORSTOM Editions, Paris.
49. Beachy, R.N. 1997. Mechanisms and applications of pathogen-derived
resistance in transgenic plants. In Current Opinion in Biotechnology.
8:215-220.
50. Voloudakis, A.E., Y. Yin and R. N. Beachy. 1999. Recombinant protein
expression in plants. In: “Gene Expression Systems: Using Nature for the Art
of Expression. Academic Press. pp. 429-461.
51. Beachy, R.N., J. Bennetzen, B. Chassy, M. Chrispeels, J. Chory, J. Ecker, J. Noel,
S. Kay, C. Dean, C. Lamb, J. Jones, C. Santerre, J. Schroeder, J. Umen,
M. Yanofsky, S. Wessler, Y. Zhao, and W. Parrott. 2002. Letter to the Editor:
Divergent perspectives on GM food. In “Nature Biotechnology,” 20:1-2.
52. Beachy, R.N. 2003. IP Policies and Serving the Public. Science 299:473.
53. Frommer, W.B. and R. N. Beachy. 2003. Editorial overview. Plant biotechnology:
A future for plant biotechnology? Naturally! In “Current Opinion in Plant
Biology,” 6:147-149.
54. Beachy, R.N. 2003. Foreward. In Encyclopedia of Plant and Crop Science; 1st
Ed; Goodman, R.M., Ed.; Taylor & Francis Group, LLC: New York, NY.
55. Yadav, J. S., R. N. Beachy, and C. M. Fauquet. 2005. Control of plant virus
diseases. In Encyclopedia of Plant and Crop Science; 1st Ed; Goodman, R.M.,
Ed.; Taylor & Francis Group, LLC: New York, NY. pp. 1-6. DOI: 10.1081./E-
EPCS-120019914.
56. Berg, R. Howard, R.N. Beachy. 2006. Fluorescent Protein Applications in
Plants. In Fluorescent Proteins, Second Edition, Volume 87 of Methods in
Cell Biology. Elsevier Inc. In print.
1. Murakishi, H.H., J.X. Hartmann, L.E. Pelcher, and R.N. Beachy. 1970. Improved
inoculation of cultured plant cells resulting in high virus titer and crystal
formation. Virology 41:365-367.
2. Murakishi, H.H., J.X. Hartmann, R.N. Beachy, and L.E. Pelcher. 1971. Growth
curve and yield of tobacco mosaic virus in tobacco callus cells. Virology
43:62-68.
Roger N. Beachy 657
3. Beachy, R.N. and H.H. Murakishi. 1971. Local lesion formation in tobacco
tissue culture. Phytopathology 61:877-878.
4. Beachy, R.N. and H.H. Murakishi. 1973. Effect of cycloheximide on TMV
synthesis in callus from hypersensitive tobacco. Virology 55:320-328.
5. Zaitlin, M. and R.N. Beachy. 1974. The use of protoplasts and separated cells
in plant virus research. Adv Virus Res 19:1-37.
6. Beachy, R.N. and M. Zaitlin. l975. Replication of tobacco mosaic virus VI.
Replicative intermediate and TMV-RNA- related RNAs associated with
polyribosomes. Virology 63:84-97.
7. Beachy, R.N. and H.H. Murakishi. l976. Changes in soluble proteins in callus
cells of hypersensitive tobacco inoculated with tobacco mosaic virus. In Vitro
12:5l7-520.
8. Bruening, G., R.N. Beachy, R. Scalla, and M. Zaitlin. l976. In vitro and in vivo
translation of the ribonucleic acids of a cowpea strain of tobacco mosaic
virus. Virology 7l:498-5l7.
9. Beachy, R.N., M. Zaitlin, G. Bruening, and H.W. Israel. l976. A genetic map
for the cowpea strain of TMV. Virology 73:498-507.
10. Zaitlin, M., G. Bruening, R.N. Beachy, and R. Scalla. l976. The cowpea strain
of TMV is a pseudo-multicomponent virus. Ann Microbiol l27A:37-38.
11. Beachy, R.N. and M. Zaitlin. l977. Characterization and in vitro translation
of the RNAs from less-than-full-length, virus-related, nucleoprotein rods
present in tobacco mosaic virus preparations. Virology 8l:l60-l69.
12. Zaitlin, M., R.N. Beachy, and G. Bruening. 1977. Lack of molecular
hybridization between RNAs of two strains of TMV: a reconsideration of the
criteria for strain relationships. Virology 8:237-24l.
13. Beachy, R.N., J.F. Thompson, and J.T. Madison. l978. Isolation of polyribosomes
and messenger RNA active in in vitro synthesis of soybean seed proteins.
Plant Physiol 6l:l39-l44.
14. Beachy, R.N., K. Barton, J.F. Thompson and J.T. Madison. l980. In vitro synthesis
of the α and α’ subunits of the 7S storage protein (conglycinin) of soybean
seeds. Plant Physiol 65:990-994.
15. Beachy, R.N., N.P. Jarvis, and K.A. Barton. 1981. Biosynthesis of subunits of
the soybean 7S storage protein. J Mol Appl Genet 1:19-27.
16. Meinke, D.W., J. Chen, and R.N. Beachy. 1981. Expression of storage protein
genes during soybean seed development. Planta 153:130-139.
17. Bernal-Lugo, I., R.N. Beachy, and J.E. Varner. 1981. The response of barley
aleurone layers to gibberellic acid includes the transcription of new sequences.
Biochim Biophys Acta 102:617-623.
18. Barton, K.A., J.F. Thompson, J.T. Madison, R. Rosenthal, N.P. Jarvis, and R.N.
Beachy. The biosynthesis and processing of high molecular weight precursors
of soybean glycinin subunits. J Biol Chem 257:6089-6095.
658 Wolf Prize in Agriculture
19. Schuler, M.A., B.F. Ladin, J.C. Polacco, G. Freyer, and R.N. Beachy. 1982.
Structural sequences are conserved in the genes coding for the a, a’,
and a-subunits of the soybean 7S seed storage protein. Nucleic Acids Res
10:8245-8261.
20. Schuler, M.A., E.S. Schmidt, and R.N. Beachy. 1982. Closely related families
of genes code for the α and α’-subunits of the soybean 7S storage proteins.
Nucleic Acids Res 10:8225-8244.
21. Foard, D.E., P.A. Gutag, B.F. Ladin, R.N. Beachy, and B.A. Larkins. 1982. In
vitro synthesis of Bowman-Birk and related soybean protease inhibitors.
Plant Mol Biol 1:227-243.
22. Schuler, M.A., J.J. Doyle and R.N. Beachy. 1983. Nucleotide homologies between
the glycosylated seed storage proteins of Glycine max and Phaseolus vulgaris.
Plant Mol Biol 2:119-127.
23. Ladin, B.F., J.J. Doyle and R.N. Beachy. 1984. Molecular characterization of a
deletion mutation affecting the α’-subunit of β-conglycinin of soybean.
J Mol App Genet 2:372-380.
24. Vance, V.S. and R.N. Beachy. 1984. Detection of genomic-length soybean
mosaic virus RNA on polyribisomes of infected soybean leaves. Virology
132:271-281.
25. Vance, V.S. and R.N. Beachy. 1984. Cloning of soybean mosaic virus RNA:
Detection of a subgenomic mRNA in infected leaves. Virology 138:26-36.
26. Doyle, J.J., B.F. Ladin, and R.N. Beachy. 1985. Antigenic relationship of legume
seed proteins to the 7S seed storage protein of soybean. Biochem Syst Ecol
13:123-132.
27. Doyle, J.J. and R.N. Beachy. 1985. Ribosomal gene variation in soybean
(Glycine) and its relatives. Theor Appl Genet 70: 369-376.
28. Shattuck-Eidens, D.M. and R.N. Beachy. 1985. Degradation of β-conglycinin
in early stages of soybean embryogenesis. Plant Physiol 78:895-898.
29. Bray, B.A. and R.N. Beachy. 1985. Regulation by ABA of β-conglycinin
expression in cultured developing soybean cotyledons. Plant Physiol
79:746-750.
30. Beachy, R.N., Z.-L. Chen, R.B. Horsch, S.G. Rogers, N.J. Hoffmann, and
R.T. Fraley. 1985. Accumulation and assembly of soybean β-conglycinin in
seeds of transformed petunia plants. EMBO J 4:3047-3053.
31. Reinero, A., and R.N. Beachy. 1986. Association of TMV coat protein with
thylakoid membrane in virus infected leaves. Plant Mol Biol 6:291-301.
32. Doyle, J.J., Schuler, M.A., J. Slightom, W.D. Goddette, V.E. Zenger, and
R.N. Beachy. 1986. The glycosylated seed storage proteins of Glycine max
and Phaseolus vulgaris: structural homologies of genes and proteins. J Biol
Chem 261:9228-9238.
Roger N. Beachy 659
33. Powell-Abel, P.A., R.S. Nelson, B. De, N. Hoffmann, S.G. Rogers, R.T. Fraley,
and R.N. Beachy. 1986. Delay of disease development in transgenic
plants that express the tobacco mosaic virus coat protein gene. Science
232:738-743.
34. Oliver, M.J., C.M. Deom, B.K. De, and R.N. Beachy. 1986. In vitro transcription
and translation of cloned cDNA’s encoding the 30 kD protein gene of TMV.
Virology 155:277-283.
35. Chen, Z.L., M.A. Schuler, and R.N. Beachy. 1986. Functional analysis of
regulatory elements in a plant embryo-specific gene. Proc Natl Acad Sci USA
83:8560-8564.
36. Ladin, B.F., M. L. Tierney, D.W. Meinke, P. Hosángadi, M. Vieth, and
R.N. Beachy. 1987. Developmental regulation of β-conglycinin in soybean
axes and cotyledons. Plant Physiol 84:35-41.
37. Nelson, R.E., P. Powell Abel, and R.N. Beachy. 1987. Lesions and virus
accumulation in inoculated transgenic tobacco plants expressing the coat
protein gene of tobacco mosaic virus. Virology 158:126-132.
38. Tumer, N.E., K.M. O’Connell, R.S. Nelson, P.R. Sanders, and R.N. Beachy.
1987. Expression of alfalfa mosaic virus coat protein gene confers cross-
protection in transgenic tobacco and tomato plants. EMBO J 6:1181-1188.
39. Bray, E.A., S. Naito, N.-S. Pan, E. Anderson, P. Dubé, and R.N. Beachy. 1987.
Expression of the β-subunit of β-conglycinin in seeds of transgenic plants.
Planta 172:364-370.
40. Tierney, M.L., E.A. Bray, R.D. Allen, R.E. Droug, Y. Ma, J. Slighton, and
R.N. Beachy. 1987. Isolation and characterization of a genomic clone encoding
the β-subunit of β-conglycinin. Planta 172:356-363.
41. Della-Cioppa, G., G.M. Kishore, R.N. Beachy, and R.T. Fraley. 1987. Protein
trafficking in plant cells. Plant Physiol 84:965-968.
42. Deom, C.M., M.J. Oliver, and R.N. Beachy. 1987. The 30-kilodalton gene
product of tobacco mosaic virus potentiates virus movement. Science
237:389-394.
43. Lawton, M.A., M.L. Tierney, I. Nakamura, E. Anderson, Y. Komeda,
P. Dubé, N. Hoffmann, R.T. Fraley, and R.N. Beachy. 1987. Expression of a
soybean β-conglycinin gene under the control of the cauliflower mosaic
virus 35S and 19S promoters in transformed petunia tissues. Plant Mol Biol
9:315-324.
44. Nelson, R.S., S.M. McCormick, X. Delannay, P. Dubé, J. Layton, E.J. Anderson,
M. Kaniewska, R.K. Proksch, R.B. Horsch, S.G. Rogers, R.T. Fraley, and
R.N. Beachy. 1988. Virus tolerance, plant growth, and field performance of
transgenic tomato plants expressing coat protein from tobacco mosaic virus.
Bio/technology 6:403-409.
660 Wolf Prize in Agriculture
45. Chen, Z.-L., N.-S. Pan, and R.N. Beachy. 1988. Identification of a
developmentally regulated enhancer element in a soybean embryo-specific
gene. EMBO J 7:297-302.
46. Naito, S., P.H. Dubé, and R.N. Beachy. 1988. Differential expression
of developmentally regulated genes in transgenic plants. Plant Mol Biol
11:109-123.
47. Register III, J.C., and R.N. Beachy. 1988. Resistance to TMV in transgenic
plants results from interference with an early event in infection. Virology
166:524-532.
48. Reinero, A., and R.N. Beachy. 1989. Inhibition of Photosystem II and
accumulation of viral coat protein in thylakoids of leaves infected by tobacco
mosaic virus. Plant Physiol 89:111-116.
49. Chen, Z.-L., S. Naito, I. Nakamura, and R.N. Beachy. 1989. Regulated
expression of genes encoding soybean β-conglycinins in transgenic plants.
Dev Genet 10:112-122.
50. Eggenberger, A.L., D.M. Stark, and R.N. Beachy. 1989. The nucleotide sequence
of a soybean mosaic virus coat protein-coding region and its expression in
Escherichia col, Agrobacterium tumefaciens and tobacco callus. J Gen Virol
70:1853-1860.
51. Carr, J.P., R.N. Beachy, and D.F. Klessig. 1989. Are the PR1 proteins of
tobacco involved in genetically engineered resistance to TMV? Virology
169:470-473.
52. Anderson, E.J., D.M. Stark, R.S. Nelson, N.E. Tumer, and R.N. Beachy.
1989. Transgenic plants that express the coat protein genes of TMV or
AlMV interfere with disease development of some non-related viruses.
Phytopathology 12:1284-1290.
53. Hodgson, R.A.J., R.N. Beachy, and H.B. Pakrasi. 1989. Selective inhibition of
photosystem II in spinach by tobacco mosaic virus: An effect of the viral coat
protein. FEBS Lett. 245(1,2):267-270.
54. Powell, P.A., D.M. Stark, P. Sanders, and R.N. Beachy. 1989. Protection against
tobacco mosaic virus in transgenic plants that express TMV antisense RNA.
Proc Natl Acad Sci USA 86:6949-6952.
55. Allen, R.D., F. Bernier, P.A. Lessard, and R.N. Beachy. 1989. Nuclear
factors interact with a soybean β-conglycinin enhancer. Plant Cell
1:623-631.
56. Stark, D.M., and R.N. Beachy. 1989. Protection against potyvirus infection in
transgenic plants: Evidence for broad-spectrum resistance. Bio/technology
7:1257-1262.
57. Nejidat, A., and R.N. Beachy. 1989. Decreased Levels of TMV Coat Protein in
Transgenic Tobacco Plants at Elevated Temperatures Reduce Resistance to
TMV Infection. Virology 173:531-538.
Roger N. Beachy 661
58. Osbourn, J.K., J.W. Watts, R.N. Beachy, and T.M.A. Wilson. 1989. Disassembly
of pseudovirus particles is inhibited in electroporated protoplasts from
transgenic tobaccos expressing tobacco mosaic virus coat protein. Evidence
that virus replication is also inhibited at a later stage of infection. Virology
172:370-373.
59. Register III, J.C., and R.N. Beachy. 1989. A transient protoplast assay for
capsid protein-mediated protection: Effect of capsid protein aggregation state
on protection against tobacco mosaic virus. Virology 173:656-663.
60. Wolf, S., C.M. Deom, R.N. Beachy, and W.J. Lucas. 1989. Movement protein
of tobacco mosaic virus modified plasmodesmatal size exclusion limit. Science
246:377-379.
61. Powell, P.A., P.R. Sanders, N. Tumer, and R.N. Beachy. 1990. Protection
against tobacco mosaic virus infection in transgenic plants requires
accumulation of capsid protein rather than coat protein RNA sequences.
Virology 175:124-130.
62. Deom, C.M., K.R. Schubert, S. Wolf, C.A. Holt, W.J. Lucas, and R.N. Beachy.
1990. Molecular characterization and biological function of the movement
protein of tobacco mosaic virus in transgenic plants. Proc Natl Acad Sci USA
87:3284-3288.
63. Wisniewski, L.A., P.A. Powell, R.S. Nelson, and R.N Beachy. 1990. Local and
systemic movement of tobacco mosaic virus (TMV) in tobacco plants that
express the TMV coat protein gene. Plant Cell 2:559-567.
64. Franche, C., D. Bogusz, C. Fauquet, C. Schöpke, C. Jobe, J. Pease, and
R.N. Beachy. 1990. A la frontière de la balistique et de la biologie végéale: le
canon à particules. ORSTOM Actualités, 29:8-10.
65. Nejidat, A., and R.N. Beachy. 1990. Transgenic tobacco plants expressing a
coat protein gene of tobacco mosaic virus are resistant to some other
tobamoviruses. Mol Plant Microbe Interact 3:247-251.
66. Sebastiani, F.L., L.B. Farrell, M.A. Schuler, and R.N. Beachy. 1990. Complete
sequence of cDNA of a-subunit of soybean β-conglycinin. Plant Mol Biol
15:197-201.
67. Nejidat, A., W.G. Clark, and R.N. Beachy. 1990. Engineered resistance against
plant virus diseases. Physiol Plant 80:662-668.
68. Beachy, R.N., S. Loesch-Fries, and N.E. Tumer. 1990. Coat protein-mediated
resistance against virus infection. Annu Rev Phytopathol 28:451-474.
69. Rochester, D.E., W. Kositratana, and R.N. Beachy. 1990. Systemic movement
and symptom production following agroinoculation with a single DNA of
tomato yellow leaf curl virus (Thailand) geminivirus. Virology 178:520-526.
70. Nejidat, A., F. Cellier, C.A. Holt, R. Gafny, A.L. Eggenberger, and R.N. Beachy.
1990. Transfer of the movement protein gene between two tobamoviruses:
Influence on local lesion development. Virology 180:318-326.
662 Wolf Prize in Agriculture
71. Clark, W.G., J.C. Register, III, A Nejidat, D.A. Eichholtz, P.R. Sanders,
R.T. Fraley, and R.N. Beachy. 1990. Tissue-specific expression of the
TMV coat protein in transgenic tobacco plants affects the level of coat protein-
mediated virus protection. Virology 179:640-647.
72. Holt, C.A., R.A.J. Hodgson, F.. Coker, R.N. Beachy, and R.S. Nelson. 1990.
Characterization of the masked strain of tobacco mosaic virus: Identification
of the region responsible for symptom attenuation by analysis of an infectious
cDNA clone. Mol Plant Microbe Interact 3(6):17-423.
73. Farrell L.B., and R.N. Beachy. 1990. Manipulation of β-glucuronidase for use
as a reporter in vacuolar targeting studies. Plant Mol Biol 15:821-825.
74. Deom, C.M., S. Wolf, C.A. Holt, W.J. Lucas, and R.N Beachy. 1991. Altered
function of the tobacco mosaic virus movement protein in a hypersensitive
host. Virology 180:251-256.
75. Atkins, D., R. Hull, B. Wells, K. Roberts, P. Moore, and R.N. Beachy. 1991. The
TMV 30K movement protein in transgenic tobacco plants is localized to
plasmodesmata. J Gen Virol 72:209-211.
76. Wu, X., R.N. Beachy, T.M.A. Wilson, and J G. Shaw. 1990. Inhibition
of uncoating of tobacco mosaic virus particles in protoplasts from
transgenic tobacco plants that express the viral coat protein gene. Virology
179:893-895.
77. Lessard, P.A., R.D. Allen, F. Bernier, J.D. Crispino, T. Fujiwara, and R.N. Beachy.
1991. Multiple nuclear factors interact with upstream sequences of
differentially regulated β-conglycinin genes. Plant Mol Biol 16:397-413.
78. Jones, M., K. Gough, I. Dasgupta, B.L. Subba Rao, J. Cliffe, R. Qu, P. Shen,
M. Kaniewska, M. Blakebrough, J.W. Davies, R.N. Beachy, and R. Hull. 1991.
Rice tungro disease is caused by an RNA and a DNA virus. J Gen Virol
72:757-761.
79. Holt, C.A., and R.N. Beachy. 1991. In vivo complementation of infectious
transcripts from mutant tobacco mosaic virus cDNAs in transgenic plants.
Virology 181:109-117.
80. Berna, A., R. Gafny, S. Wolf, W.J. Lucas, C.A. Holt and R.N. Beachy. 1991. The
TMV movement protein: Role of the C-terminal 73 amino acids in subcellular
localization and function. Virology 182:682-689.
81. Franche, C., D. Bogusz, C. Schöpke, C. Fauquet, and R.N. Beachy. 1991.
Transient gene expression in cassava using high-velocity microprojectiles.
Plant Mol Biol 17:493-498.
82. Sebastiani, F.L., L.B. Farrell, M. Vasquez, and R.N. Beachy. 1991. Conserved
amino acid sequences among plant proteins sorted to protein bodies and
plant vacuoles: can they play a role in protein sorting?. Eur J Biochem
199:441-450.
Roger N. Beachy 663
83. Wolf, S., C.M. Deom, R.N. Beachy, and W.J. Lucas. 1991. Plasmodesmatal
function is probed using transgenic tobacco plants that express a virus
movement protein. Plant Cell 3:593-604.
84. Niblett, C.L., L.A. Calvert, T.L. Kendall, D.M. Stark, K.R. Zagula, C.E. Smith,
R.N. Beachy, and S.A. Lommel. 1991. cDNA Cloning and Nucleotide
Sequence of the Wheat Streak Mosaic Virus Capsid Protein Gene. J Gen Virol
72:499-504.
85. Qu, R., M. Bhattacharyya, G.S. Laco, A. de Kochko, B.L. Subba Rao,
M.B. Kaniewska, J.S. Elmer, D.E. Rochester, C.E. Smith, and R.N. Beachy.
1991. The complete sequence of rice tungro bacilliform virus DNA —
comparison with commelina yellow mottle virus and caulimoviruses. Virology
185:354-364.
86. Jain, R.K., N.M. McKern, S.A. Tolin, J.H. Hill, O.W. Barnett, M. Tosic, R.E.
Ford, R.N. Beachy, M.H. Yu, C.W. Ward, and D.D. Shukla. 1992. Confirmation
that fourteen potyvirus isolates from soybean are strains of one virus by
comparing coat protein peptide profiles. Phytopathology. 82:294-299.
87. Gafny, R., M. Lapidot, A. Berna, C. Holt, C.M. Deom, and R.N. Beachy. 1992.
Effects of terminal deletion mutations on function of the movement protein
of tobacco mosaic virus. Virology 187:499-507.
88. Fujiwara, T., P.A. Lessard, and R.N. Beachy. 1992. Seed-specific repression
of GUS activity in tobacco plants by antisense RNA. Plant Mol Biol
20:1059-1069.
89. Perl, M., R. Gafney, and R.N. Beachy. 1992. Phosphodiesterase activities in
transgenic tobacco plants associated with the movement protein of tobacco
mosaic virus. Theor Appl Genet 84:730-734.
90. Deom, C.M., M. Lapidot, and R.N. Beachy. 1992. Plant virus movement
proteins. Cell 69:221-224.
91. Ding, B., J.S. Haudenshield, R.J. Hull, S. Wolf, R.N. Beachy, and W.J. Lucas.
1992. Secondary plasmodesmata are specific sites of localization of the
tobacco mosaic virus movement protein in transgenic tobacco plants. Plant
Cell 4:915-928.
92. Moore, P.J., C.A. Fenczik, C.M. Deom, and R.N. Beachy. 1992. Developmental
changes in the structure of plasmodesmata in transgenic tobacco expressing
the movement proteins of tobacco mosaic virus. Protoplasma 170:115-127.
93. Huttner, S.L., C. Arntzen, R.N. Beachy, G. Bruening, L. DeFrancesco,
E. Nester, C. Qualset, and A. Vidaver. 1992. Revising oversight of genetically
modified plants. Bio/technology 10(9):967-971.
94. Fujiwara, T., and R.N. Beachy. 1993. Expression of soybean seed storage
protein genes in transgenic plants: Their effects on expression of a neighboring
gene and position dependency. Plant Cell Physiol 34(1):13-20.
664 Wolf Prize in Agriculture
95. Fujiwara, T., P.A. Lessard, and R.N. Beachy. 1993. Inactivation of the nopaline
synthase gene by double transformation: reactivation by segregation of the
induced DNA. Plant Cell Rep 12:133-138.
96. Shen, P., M. Kaniewska, C. Smith, and R.N. Beachy. 1993. Nucleotide
sequence and genomic organization of rice tungro spherical virus. Virology
193:621-630.
97. Li, L., R. Qu, A deKochko, C. Fauquet, and R.N. Beachy. 1993. An improved
rice transformation system using the biolistic approach. Plant Cell Rep
12:250-255.
98. Padgett, H.S., and R.N. Beachy. 1993. Nucleotide sequence and mutational
analysis of an infectious cDNA clone of Ob, a tobamovirus that overcomes
the N resistance gene of Nicotiana. Plant Cell 5:577-586.
99. Lucas, W.J., A. Olesinski, R.J. Hull, J.S. Haudenshield, C.M. Deom, R.N. Beachy,
and S. Wolf. 1993. Influence of the tobacco mosaic virus 30-kDa movement
protein on carbon metabolism and photosynthate partitioning in transgenic
tobacco plants. Planta 190:88-96.
100. Nelson, R.S., G. Li, R.A.J. Hodgson, R.N. Beachy, and M. H. Shintaku. 1993.
Impeded phloem-dependent accumulation of the masked strain of tobacco
mosaic virus. Mol Plant Microbe Interact 6(1):45-54.
101. Reimann-Philipp, U. and R.N. Beachy. 1993. Coat protein-mediated resistance
in transgenic tobacco expressing the TMV coat protein from tissue specific
promoters. Mol Plant Microbe Interact 6: 323-330.
102. Mathews, H., C. Schöpke, R. Carcamo, P. Chavarriaga, C. Fauquet, and
R.N. Beachy. 1993. Improvement of somatic embryogenesis and plant recovery
in cassava. Plant Cell Rep 12:328-333.
103. Bhattacharyya-Pakrasi, M., J. Peng, J.S. Elmer, G. Laco, P. Shen, M.B. Kaniewska,
H. Konowicz, F. Wen, T.K. Hodges, and R.N. Beachy. 1993. Specificity of a
promoter from the rice tungro bacilliform virus for expression in phloem
tissues. Plant J 4:71-79.
104. Lessard, P.A., R.D. Allen, T. Fujiwara, and R.N. Beachy. 1993. Upstream
regulatory sequences from two β-conglycinin genes. Plant Mol Biol 22:
873-885.
105. Nakamura, I., P.H. Dube, and R.N. Beachy. 1993. Accumulation of the
products of β-conglycinin a’-subunit gene constitutively expressed in seeds
and non-seed tissues of the transgenic petunia plants. Plant Cell Physiol
34(6):865-872.
106. Lapidot, M., R. Gafny, B. Ding, S. Wolf, W.J. Lucas, and R.N. Beachy. 1993. A
dysfunctional movement protein of tobacco mosaic virus that partially
modifies the plasmodesmata and limits virus spread in transgenic plants.
Plant J 4(6): 959-970.
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107. Reimann-Philipp, U., and R.N. Beachy. 1993. The mechanism(s) of coat
protein-mediated resistance against tobacco mosaic virus. Semin Virol 4(6):
349-356.
108. Fujiwara, T., and R.N. Beachy. 1993. Tissue-specific and temporal regulation
of a β-conglycinin gene: roles of the RY repeat and other cis-acting elements.
Plant Mol Biol 24:261-272.
109. Rance, I., W. Tian, H. Mathews, A. de Kochko, R.N. Beachy, and C.M. Fauquet.
1994. Partial desiccation of mature embryo-derived calli, a simple treatment
that dramatically enhances the regeneration ability of Indica rice. Plant Cell
Rep 13:647-651.
110. Ngon a Yassi, M. C. Ritzenthaler, C. Brugidou, C. Fauquet, and R.N. Beachy.
1994. Nucleotide sequence and genome characterization of rice yellow mottle
virus RNA. J Gen Virol 75:249-257.
111. Marcos, J.F., and R.N. Beachy. 1994. In vitro characterization of a cassette to
accumulate multiple proteins through synthesis of a self-processing
polypeptide. Plant Mol Biol 24:495-503.
112. Rochester, D.E., J.J. DePaulo, C.M. Fauquet, and R.N. Beachy. 1994. Complete
nucleotide sequence of the geminivirus, tomato yellow leaf curl virus
(Thailand isolate). J Gen Virol 75: 477-485.
113. Wenzhong, T., I. Rance, E. Sivamani, C. Fauquet, and R.N. Beachy. 1994.
Improvement of plant regeneration frequency in vitro in indica rice. Chin J
Genet 21(3):1-9.
114. Laco, G., and R.N. Beachy. 1994. Rice tungro bacilliform virus encodes reverse
transcriptase, DNA polymerase, and ribonuclease H activities. Proc Natl Acad
Sci USA 91:2654-2658.
115. De Kochko, A., R. Qu, P. Marmey, I. Rance, C. Fauquet, and R.N. Beachy.
1994. Protein slot blotting; an easy, rapid and reliable technique to
identify the expression of a protein in transgenic plants. Plant Mol Biol Rep
12(3):274-278.
116. Deom, C.M., Z.H. Xian, R.N. Beachy, and A.K. Weissinger. 1994. Influence of
heterologous tobamovirus movement protein and chimeric-movement protein
genes on cell-to-cell and long-distance movement. Virology 205:198-209.
117. Rance, I.M., W. Tian, H. Mathews, A. deKochko, R.N. Beachy, and
C. Fauquet. 1994. Partial desiccation of mature embryo-derived calli, a simple
treatment that dramatically enhances the regeneration ability of indica rice.
Plant Cell Rep 13:647-651.
118. Cooper, B., M. Lapidot, J.A. Heick, J.A. Dodds, and R.N. Beachy. 1995. A
defective movement protein of TMV in transgenic plants confers resistance
to multiple viruses whereas the functional analog increases susceptibility.
Virology 206:307-313.
666 Wolf Prize in Agriculture
119. Brugidou, C., C. Holt, M. Ngon a Yassi, S. Zhang, R.N. Beachy, and C. Fauquet.
1995. Synthesis of an infectious full-length cDNA clone of rice yellow mottle
virus and mutagenesis of the coat protein. Virology 206:108-115.
120. Padidam, M., R.N. Beachy, and C.M. Fauquet. 1995. Sequence comparison
can be used to identify and classify geminiviruses. J Gen Virol 76:249-263.
121. Padidam, M., R.N. Beachy, and C.M. Fauquet 1995. Tomato leaf curl
geminivirus from India has abipartite genome and coat protein is not essential
for infectivity. J Gen Virol 76:25-35.
122. Yin, Y. and R.N. Beachy. 1995. The regulatory regions of the rice tungro
bacilliform virus promoter and interacting nuclear factors in rice (Oryza
sativa L.). Plant J 7(6):969-980.
123. Laco, G.S., S.B.H. Kent, and R.N. Beachy. 1995. Analysis of the proteolytic
processing and activation of the rice tungro bacilliform virus reverse
transcriptase. Virology 208:207-214.
124. Zahalak, M., R.N. Beachy, and T. Thiel. 1995. Expression of the movement
protein of tobacco mosaic virus in the cyanobacterium Anabaena sp. strain
PCC 7120. Mol Plant Microbe Interact 8(2):192-199.
125. Clark, W.G., J.H. Fitchen, and R.N. Beachy. 1995. Studies of coat protein-
mediated resistance to TMV: I. The PM2 assembly defective mutant confers
resistance to TMV. Virology 208:485-491.
126. Arce-Johnson, P., T.W. Kahn, U. Reimann-Philipp, R.R. Bustamante, and
R.N. Beachy. 1995. The amount of movement protein produced in transgenic
plants influences the establishment, local movement, and systemic spread of
infection by movement protein-deficient tobacco mosaic virus. Mol Plant
Microbe Interact (3):415-423.
127. Clark, W. Gregg, J. Fitchen, A. Nejidat, C. M. Deom, and R.N. Beachy. 1995.
Studies of coat protein-mediated resistance to TMV: II. Challenge by a mutant
with altered virion surface does not overcome resistance conferred by TMV
CP. J Gen Virol 76(10):2613-2617.
128. Fitchen, J., R.N. Beachy, and M.B. Hein. 1995. Plant virus expressing hybrid
coat protein and added murine epitope elicits autoantibody response. Vaccine
13(12):1051-1057.
129. Heinlein, M., B.L. Epel, H.S. Padgett, and R.N. Beachy. 1995. Interaction
of tobamovirus movement proteins with the plant cytoskeleton. Science
270:1983-1985.
130. Fenczik, C.A., H.S. Padgett, C.A. Holt, S.J. Casper, and R.N. Beachy. 1995.
Mutational analysis of the movement protein of odontoglossum ringspot
virus to identify a host-range determinant. Mol Plant Microbe Interact
8(5):666-673.
131. Bryant, J.L., P. Hosangadi, and R.N. Beachy. 1995. The β-conglycinins of
soybean remain assembled in a 7S conformation while undergoing proteolytic
Roger N. Beachy 667
cleavage during germination and early seedling growth. Plant Cell Physiol
36:1059-1065.
132. Marcos, J.F., R.N. Beachy, R.A. Houghten, S.E. Blondelle, and E. Perez-Paya.
1995. Inhibition of a plant virus infection by analogs of melittin. Proc Natl
Acad Sci USA 92:12466-12469.
133. Sivamani, E., P. Shen, N. Opalka, R.N. Beachy, and C.M. Fauquet. 1995.
Selection of large quantities of embryogenic subcultured calli from Indica
rice seeds for production of fertile transgenic plants using the biolistic method.
Plant Cell Rep 15:322-327.
134. Cooper, Bret, I. Schmitz, A.L.N. Rao, R.N. Beachy, and J.A. Dodds. 1996. Cell-
to-cell transport of movement-defective cucumber mosaic and tobacco mosaic
viruses in transgenic plants expressing heterologous movement protein genes.
Virology 216:208-213.
135. Schopke, C., N. Taylor, R. Carcamo, K.K. N’Da, P. Marmey, G.G. Henshaw,
R.N. Beachy, and C.M. Fauquet. 1996. Regeneration of transgenic cassava
plants (Manihot esculenta Crantz) from microbombarded embryogenic
suspension cultures. Nat Biotechnol 14:731-735.
136. Epel, B.L., H.S. Padgett, M. Heinlein, and R.N. Beachy. 1996. Plant virus
movement protein dynamics probed with a gfp-protein fusion Gene
173(1):75-79.
137. Aleman, M.-E., J.F. Marcos, C. Brugidou, R.N. Beachy, and C.M. Fauquet.
1996. The complete nucleotide sequence of yam mosaic virus (Ivory Coast
isolate) genomic RNA. Arch Virol 141:1259-1278.
138. Verdaguer, B., A. de Kochko, R.N. Beachy, and C.M. Fauquet. 1996. Isolation
and expression in transgenic plants of the cassava vein mosaic virus (CVMV)
promoter. Plant Mol Biol 31:1129-1139.
139. Zhang, S., L. Chen, R. Qu, P. Marmey, R.N. Beachy, and C.M. Fauquet. 1996.
Regeneration of fertile transgenic Indica (Group 1) rice plants following
microprojectile-transformation of embryogenic suspension culture cells. Plant
Cell Rep 15:465-469.
140. Padidam, M., R.N. Beachy, and C.M Fauquet. 1996. The role of AV2
(“precoat”) and coat protein in viral replication and movement in tomato
leaf curl geminivirus. Virology 224:390-404.
141. Padgett, H.S., B.L. Epel, M.H. Heinlein, Y. Watanabe, and R.N. Beachy. 1996.
Distribution of tobamovirus movement protein in infected cells and
implications for cell-to-cell spread of infection. Plant J 10:1079-1088.
142. Qu, R., A. de Kochko, L. Zhang, P. Marmey, L. Li, W. Tian. S. Zhang,
C.M. Fauquet, and R.N. Beachy. 1996. Analysis of a large number of
independent transgenic rice plants produced by the biolistic method. In vitro
Cell Dev Biol Plant 32:233-240.
668 Wolf Prize in Agriculture
143. Beachy, R.N., H.S. Padgett, T. Kahn, M. Bendahmane, J.H. Fitchen, M. Heinlein,
Y. Watanabe, and B.L. Epel. 1996. Cellular and molecular mechanisms of
coat protein and movement protein mediated resistance against TMV. Molec
Plant Microbe Interact 259-264.
144. Kahn, T.W., R.N. Beachy, and M. M. Falk. 1997. Cell-free expression of
a GFP fusion protein allows quantitation in vitro and in vivo. Curr Biol
7:R207-R208.
145. Beachy, R.N. 1997. The 2’,5’A antiviral system in plants: A dose of mammalian
medicine. Nat Biotechnol 14:1538-39.
146. Arce-Johnson, P., U. Reimann-Philipp, H.S. Padgett, R. Rivera-Bustamante,
and R.N. Beachy. 1997. Requirement of the movement protein for long distance
spread of tobacco mosaic virus in grafted plants. Mol Plant Microbe Interact
10:691-699.
147. Marcos, J.F. and R.N. Beachy. 1997. Transgenic accumulation of two
plant virus proteins on a single self-processing polypeptide. J Gen Virol
78:1771-1778.
148. Aleman-Verdaguer, M.-E., C. Goudou-Urbino, J. Dubern, R.N. Beachy, and
C.M. Fauquet. 1997. Analysis of the sequence diversity of the P1, HC, P3,
Nib and CP genomic regions of several yam mosaic potyvirus isolates:
implications for the intraspecies molecular diversity of potyviruses. J Gen
Virol 78:1253-1264.
149. Padgett, H.S., Y. Watanabe, and R.N. Beachy. 1997. Identification of the
tobamovirus protein that activates the N gene-mediated hypersensitive
response. Mol Plant Microbe Interact 10:709-715.
150. Yin, Y., Q. Zhu, S. Dai, C. Lamb, and R.N. Beachy. 1997. RF2a, a bZIP
transcriptional activator of the phloem-specific rice tungro bacilliform virus
promoter, functions in vascular development. EMBO J 16:5247-5259.
151. Yin, Y., L. Chen, and R.N. Beachy. 1997. Promoter elements required for
phloem-specific gene expression from the RTBV promoter in rice. Plant J
12:1179-1188.
152. Bendahmane, M., J.H. Fitchen, G. Zhang, and R.N. Beachy. 1997. Studies of
coat protein mediated resistance to TMV: Correlation between assembly of
mutant coat proteins and resistance. J Virol 71:7942-7950.
153. Ceriani, M.F., J.F. Marcos, H.E. Hopp, and R.N. Beachy. 1997. Simultaneous
accumulation of multiple viral coat proteins from a TEV-NIa based expression
vector. Plant Mol Biol 36:239-248.
154. Oparka, K.J., D.A.M. Prior, S. Santa Cruz, H.S. Padgett, and R.N. Beachy.
1997. Gating of epidermal plasmodesmata is restricted to the leading edge
of expanding infection sites of tobacco mosaic virus. Plant J 12:781-789.
155. Ares, X., G. Calamante, S. Cabral, P. Hemenway, R.N. Beachy and
A. Mentaberry. 1998. Transgenic plants expressing PVX ORF2 protein (p24)
are resistant to TMV and Ob tobamoviruses. J Virol 72:24-31.
Roger N. Beachy 669
169. Umaharan, P., M. Padidam, R.H. Phelps, R.N. Beachy and C.M. Fauquet.
1998. Distribution and diversity of geminiviruses in Trinidad and Tobago.
Phytopathology 88(12):1262-1268.
170. Karrer, E.E., R.N. Beachy, and C.A. Holt. 1998. Cloning of tobacco genes that
elicit the hypersensitive response. Plant Mol Biol 36:681-690.
171. Ceriani, M.F., J.F. Marcos, H.E. Hopp, and R.N. Beachy. 1998. Simultaneous
accumulation of multiple viral coat proteins from a TEV-Nia based expression
vector. Plant Mol Biol 36:239-248.
172. Mas, P. and R.N. Beachy. 1998. Distribution of TMV movement protein in
single living protoplasts immobilized in agarose. Plant J 15:835-842.
173. Zhang, S., W-Y Song, L. Chen, D. Ruan, N. Taylor, P. Ronald, R.N. Beachy and
C. M. Fauquet. 1998. Transgenic elite Indica rice varieties, resistant to
xanthomonas oryzae pv. oryzae molecular breeding. Mol Breed 4:551-558.
174. Marmey, P., B. Bothner, E. Jacquot, A. deKochko, C. A. Ong, P. Yot,
G. Siuzdak, R.N. Beachy, and C. M. Fauquet, 1999. Rice tungro bacilliform
virus open reading frame 3 encodes a single 37-kDa coat protein. Virol
253:319-326.
175. Beachy, R.N. 1999. Coat-protein-mediated resistance to tobacco mosaic
virus: discovery mechanisms and exploitation. Philos Trans R Soc Lond B
B 354:659-664.
176. Szécsi, J., X.S. Ding, C.O. Lim, M. Bendahame, M.J. Cho, R.S. Nelson, and
R.N. Beachy. 1999. Development of TMV infection sites in Nicotiana
benthamiana. Mol Plant Microbe Interact 12:143-152.
177. Padidam, M., V.S. Reddy, R.N. Beachy and C. M. Fauquet. 1999. Molecular
characterization of a plant mitochondrial chaperone GrpE. Plant Mol Biol
39:871-881.
178. Padidam, M., R.N. Beachy and C. M. Fauquet. 1999. A phage single-stranded
DNA (ssDNA) binding protein complements ssDNA accumulation of a
geminivirus and interferes with viral movement. J Virol 73:1609-1616.
179. Lazarowitz, S.G. and R. N. Beachy. 1999. Viral movement proteins as
probes for intracellular and intercellular trafficking in plants. Plant Cell
11:535-548.
180. Bendahmane, M. and R. N. Beachy. 1999. Control of tobamovirus infections
via pathogen-derived resistance. Adv Virus Res 53:369-386.
181. Bendahame, M., J.C. Koo, E. Karrer and R. N. Beachy. 1999. Display of
epitopes on the surface of tobacco mosaic virus: Impact of charge and
isoelectric point of the epitope on virus-host interaction. J Mol Biol 290:
9-20.
182. Koo, M., M. Bendahmane, G. A. Lettieri, A. D. Paoletti, T.E. Lane, J.H. Fitchen,
M.J. Buchmeier, and R.N. Beachy. 1999. Protective immunity against murine
hepatitis virus (MHV) induced by intranasal or subcutaneous administration
Roger N. Beachy 671
of hybrids of tobacco mosaic virus that carries an MHV epitope. Proc Natl
Acad Sci USA 96:7774-7779.
183. Pickard, B.G. and R. N. Beachy. 1999. Intercellular connections are
developmentally controlled to help move molecules through the plant. Cell
98:5-8.
184. Kawakami, S., H.S. Padgett, D. Hosokawa, Y. Okada, R. N. Beachy, and
Y. Watanabe, 1999. Phosphorylation and/or presence of serine 37 in the
movement protein of tomato mosaic tobamovirus is essential for intracellular
localization and stability in vivo. J Virol 73:6831-6840.
185. Reichel, C., P. Más, and R. N. Beachy. 1999. The role of the ER and cytoskeleton
in viral trafficking. Trends Plant Sci 4:458-462.
186. Chatterji, A., M. Padidam, R. N. Beachy and C.M. Fauquet. 1999. Identification
of replication specificity determinants in two strains of tomato leaf curl
virus from New Delhi. J Virol 73:5481-5489.
187. Más, P. and R. N. Beachy. 1999. Replication of tobacco mosaic virus on
endoplasmic reticulum and role of the cytoskeleton and virus movement
protein in intracellular distribution of viral RNA. J Cell Biol 147:945-958.
188. Staczek, J., M. Bendahmane, L. B. Gilleland, R. N. Beachy, and H. E. Gilleland
Jr. 2000. Immunization with a chimeric tobacco mosaic virus containing an
epitope of outer membrane protein F of Pseudomonas aeruginosa provides
protection against challenge with P. aeruginosa. Vaccine 18:2266-2274.
189. Gilleland, Jr., H.E., L. B. Gilleland, J. Staczek, B. Harty, A. Garcia-Sastre,
P. Palese, F. R. Brennan, W.D.O. Hamilton, M. Bendahmane, and R .N. Beachy.
2000. Chimeric animal and plant viruses expressing epitopes of outer
membrane protein F as a combined vaccine against Pseudomonas aeruginosa
lung infection. FEMS Immunol and Med Microbiol 27:291-297.
190. Brill, L.M., R.S. Nunn, T.W. Kahn, M. Yeager, and R. N. Beachy. 2000.
Recombinant tobacco mosaic virus movement protein is an RNA-binding, α-
helical, polytopic membrane protein. Proc Natl Acad Sci USA 97:7112-7117.
191. Reichel, C. and R. N. Beachy. 2000. Degradation of tobacco mosaic virus
movement protein by the 26S proteasome. J Virol 74:3330-3337.
192. Más, P. and R. N. Beachy. 2000. Role of microtubules in the intracellular
distribution of tobacco mosaic virus movement protein. Proc Natl Acad Sci
USA 97:12345-12349.
193. Fondong, V.N., J.S. Pita, M.E.C. Rey, A. deKochko, and R.N. Beachy. 2000.
Evidence of synergism between African cassava mosaic virus and a new
double-recombinant geminivirus infecting cassava in Cameroon. J Gen Virol
81:287-297.
194. Opalka, N., M. Tihova, C. Brugidou, A. Kumar, R. N. Beachy, C.M. Fauquet, and
M. Yeager. 2000. Structure of native and expanded sobemoviruses by electron
cryo-microscopy and image reconstruction. J Mol Biol 303:197-211.
672 Wolf Prize in Agriculture
195. Beachy, R. N. and M. Heinlein. 2000. Role of P30 in replication and spread
of TMV. Traffic 1:540-544.
196. Qu, C., L. Liljas, N. Opalka, C. Brugidou, M. Yeager, R. N. Beachy,
C. M. Fauquet, J.E. Johnson, and T. Lin. 2000. 3D domain swapping of a
molecular switch for quasi-equivalent symmetry modulates the stability of
an icosahedral virus. Structure 8:1095-1103.
197. Dai, S., P. Zheng , P. Marmey , S. Zhang, W. Tian, S. Chen , R. N. Beachy, and
C. M. Fauquet. 2001. Comparative analysis of transgenic rice plants obtained
by Agrobacterium-mediated transformation and particle bombardment. Mol
Breed 7: 25-33.
198. Petruccelli, S., S. Dai, R. Carcamo, Y. Yin, S. Chen, and R. N. Beachy. 2001.
Transcription factor RF2a alters expression of the rice tungro bacilliform
virus promoter in transgenic tobacco plants. Proc Natl Acad Sci USA
98:7635-7640.
199. Kotlizky, G., A. Katz, J. van der Laak, V. Boyko, M. Lapidot, R.N. Beachy,
M. Heinlein, and B. Epel. 2001. A dysfunctional movement protein of tobacco
mosaic virus interferes with targeting of wild-type movement protein to
microtubules. Mol Plant Microbe Interact 14:895-904.
200. Dai, S., R. Carcamo, Z. Zhang, S. Chen, and R. N. Beachy. 2001. Bacterial
cytosine deaminase gene used as a conditional negative selection marker in
transgenic rice plants. Plant Cell Rep 20:738-743.
201. Bendahmane, M., J. Szecsi, I. Chen, R.H. Berg, and R.N. Beachy.
2002. Characterization of mutant tobacco mosaic virus coat protein
that interferes with virus cell-to-cell movement. Proc Natl Acad Sci USA
99:3645-3650.
202. Zhu, Q., M. Ordiz, T. Dabi. R.N. Beachy, and C. Lamb. 2002. Rice TATA
binding protein interacts functionally with transcription factor IIB and the
RF2a bZIP transcriptional activator in an enhanced plant in vitro transcription
system. Plant Cell 14: 795-803.
203. Ordiz, M.I., C.F. Barbas III, and R. N. Beachy. 2002. Regulation of transgene
expression in plants with polydactyl zinc finger transcription factors. Proc
Natl Acad Sci USA 99:13290-13295. DOI 10.1073/pnas.202471899.
204. Beachy, R.N. and M.I. Ordiz. 2002. The fingers of gene regulation in plants.
ISB News Report: December 2002:5-6.
205. Stege, J.T., X. Guan, T. Ho, R.N. Beachy, and C.F. Barbas III. 2002. Controlling
gene expression in plants using synthetic zinc finger transcription factors.
Plant J 32:1077-1086. DOI 10.1046/j.1365-313X.2007.01492.X
206. Brugidou, C. N. Opalka, M. Yeager, R.N. Beachy, and C.M. Fauquet. 2002.
Stability of Rice Yellow Mottle Virus and cellular compartmentalization during
the infection process in Oryza sativa (L.). Virology 297:98-108.
207. Atkinson, R.C., R.N. Beachy, G. Conway, F.A. Cordova, M.A. Fox, K.A. Holbrook,
D.F. Klessig, R. L. McCormick, P.M. McPherson, H.R. Rawlings IIII, R. Rapson,
Roger N. Beachy 673
L.N. Vanderhoef, J.D. Wiley, and C.E. Young. 2003. Public Sector Collaboration
for Agricultural IP Management. Science 301:174-175.
208. Dai, S., S. Petruccelli, M.I. Ordiz, Z. Zhang, S. Chen, and R.N. Beachy.
2003. Functional analysis of RF2a, a rice transcription factor. J Biol Chem
278:36396-36402.
209. Whitelaw, C.A., W.B. Barbazuk, G. Pertea, A.P. Chan, F. Cheung, Y. Lee,
L. Zheng, S. van Heeringen, S. Karamycheva, J.L. Bennetzen, P. SanMiguel,
N. Lakey, J. Bedell, Y. Yuan, M.A. Budiman, A. Resnick, S. Van Aken,
T. Utterback, S. Riedmuller, M. Williams, T. Feldblyum, K. Schubert,
R. Beachy, C. M. Fraser, J. Quackenbush. 2003. Enrichment of gene-coding
sequences in maize by genome filtration. Science 302:2118-2120.
210. Dai, S., Z. Zhang, S. Chen, R.N. Beachy. 2004. RF2b, a rice bZIP transcription
activator, interacts with RF2a and is involved in symptom development of
rice tungro disease. Proc Natl Acad Sci USA 101:687-692.
211. Asurmendi, S., H. Berg, J. Koo, and R.N. Beachy. 2004. Coat protein regulates
formation of replication complexes during TMV infection. Proc Natl Acad
Sci USA 101:1415-1420.
212. Koo, J.C., S. Asurmendi, J. Bick, T. Woodford-Thomas, and R.N. Beachy.
2004. Ecdysone agonist-inducible expression of a coat protein gene from
Tobacco Mosaic Virus confers viral resistance in transgenic Arabidopsis.
Plant J 37:439-448.
213. Brill, L.M., S. Dechongkit, B. DeLaBarre, J. Stroebel, R.N. Beachy, and
M. Yeager. 2004. Dimerization of recombinant TMV movement protein.
J Virol 78:3372-3377.
214. Voloudakis, A.E., C.A. Malpica, M-E. Aleman-Verdaguer, D.M. Stark,
C.M. Fauquet, and R.N. Beachy. 2004. Structural characterization of Tobacco
Etch Virus coat protein mutants. Arch Virol 149:699-712.
215. Hossain, T., I. Rosenberg, J. Selhub, G. Kishore, R.N. Beachy, and K. Schubert.
2004. Enhancement of folates in plants through metabolic engineering. Proc
Natl Acad Sci USA 101:5158-5163.
216. Kawakami, S., Y. Watanabe, R.N. Beachy. 2004. Tobacco mosaic virus infection
spreads cell to cell as intact replication complexes. Proc Natl Acad Sci USA
101:6291-6296.
217. Beachy, R.N. 2004. Continuing the Effort. AgBioForum 7(1&2):1-1.
218. Marmey, P., A. Rojas-Mendoza, A. de Kochko, R.N. Beachy, and
C.M. Fauquet. 2005. Characterization of the protease domain of rice tungro
bacilliform virus responsible for the processing of the capsid protein from
the polyprotein. Virol J 2:33-45.
219. Voloudakis, A.E., M-E. Aleman-Verdaguer, H.S. Padgett, and R.N. Beachy.
2005. Characterization of resistance in transgenic Nicotiana benthamiana
encoding N-terminal deletion and assembly mutants of the tobacco etch
potyvirus coat protein. Arch Virol 150:2567-2582.
674 Wolf Prize in Agriculture
220. Dai, S., Z. Zhang, J. Bick, and R. N. Beachy. 2006. Essential role of box II cis
element and cognate host factors in regulating the promoter of rice tungro
bacilliform virus. J Gen Virol 87:715-722. DOI 10.1099/vir.0.81488-0.
221. Bazzini, A., S. Asurmendi, H. E. Hopp, and R. N. Beachy. 2006. TMV and PVX
coat proteins confer heterologous interference to PVX and TMV infection,
respectively. J Gen Virol 87:1005-1012. DOI 10.1009/vir.0.81396-0.
222. Petruccelli S., M. Otegui, F. Lareu, O. Tran Dinh, A.-C. Fitchette, A. Circosta,
M. Rumbo, M. Bardor, R. Carcamo, V. Gomord, and R.N. Beachy. 2006.
A KDEL-tagged monoclonal antibody is efficiently retained in the endoplasmic
reticulum in leaves, but is both partially secreted and sorted to protein storage
vacuoles in seeds. Plant Biotechnol J 4:511-517. DOI: 10.1111/j.1467-
7652.2006.00200.x
223. Kouassi, N.K., L. Chen, C. Siré, M. Bangratz-Reyser, R.N. Beachy,
C.M. Fauquet, and C. Brugidou. 2006. Expression of rice yellow mottle virus
coat protein enhances virus infection in transgenic plants. Arch Virol.
151: 2111-2122. DOI 10.1007/s00705-006-0802-3.
224. Fujiki, M., S. Kawakami, R.W. Kim, and R.N. Beachy. 2006. Domains of
Tobacco mosaic virus movement protein essential for its membrane
association. J Gen Virol 87:2699-2707.
225. Bazzini, A., H.E. Hopp, R.N. Beachy, and S. Asurmendi. 2006.
Posttranscriptional gene silencing does not play a significant role in PVX coat
protein mediated resistance. Phytopathology, Vol. 96, No. 11. 1175-1178.
226. Liu, Y., S. Dai, R.N. Beachy. 2007. Role of the C-terminal domains of rice
bZIP proteins RF2a and RF2b in regulating transcription. Biochem J. 405:
243-249. DOI 10.1042/BJ2006/375.
227. Bazzini, A., HE. Hopp, R.N. Beachy and Sebastian Asuremendi. 2007. Infection
and co-accumulation of Tobacco Mosaic Virus proteins alter microRNA
accmumulation, correlating with symptom on plant development. PNAS. 104
(29) 12157-12162. DOI 10.1073/pnas 0705/14104.
228. Asurmendi, S., R.H. Berg, T.J. Smith, M. Bendahmane, and R.N. Beachy. 2007.
Aggregation of TMV CP plays a role in CP functions and in coat-protein
mediated resistance. Virology. 366: 98-106. DOI 10.1016/j.viol.2007.03.014.
229. Bendahmane, M., I.Chen, S. Asurmendi, A. Bazzini, J. Szecsi, and R.N. Beachy.
2007. Coat protein-mediated resistance to TMV infection of Nicotiana
tabacum involves multiple modes of interference by coat protein. Virology.
366: 107-116.
230. Berg, H. and R.N. Beachy. 2008. Fluorescent Protein Applications
in Plants. Methods in Cell Biology, 85: 153-177. DOI: 10.1016/S0091-
679X(08)85008-X.
Roger N. Beachy 675
The above article is reprinted with permission from Macmillan Publishers Ltd., Bio/Technology,
Vol. 6, pp. 403-409 (April 1988).
676 Wolf Prize in Agriculture
CURRICULUM VITAE
PRESENT POSITION AND ADDRESS:
Distinguished Professor of Veterinary Pathobiology and Genetics
Director, Center for Animal Biotechnology and Genomics
Department of Veterinary Pathobiology
College of Veterinary Medicine and Biomedical Sciences
Texas A&M University
College Station, Texas 77843-4467
EDUCATION:
Degree Conferring Institute Field Year
B.S. Abilene Christian College Mathematics Ed 1964
Ph.D. Oregon State University Genetics 1968
683
684 Wolf Prize in Agriculture
PROFESSIONAL ORGANIZATIONS:
American Genetic Association, Member - 1981-
President - 1985
Executive Vice President - 1996-
Genetics Society of America - 1968-
American Association for the Advancement of Science - 1969-
Fellow - 1999-
The American Society for Human Genetics - 1972-
Texas Genetics Society - 1978
Board of Directors - 1982-1987
President - 1989
International Society for Animal Genetics - 1988-
Chairman, Committee on Gene Mapping - 1988-1996
President - 2000-2006
Human Genome Organization (HUGO) - 1993-
Steering Committee, NIH Bovine Genome Sequencing Project - 2004-
Technical Committee, NIH Bovine Genome Sequencing Project - 2004-
686 Wolf Prize in Agriculture
EDITORIAL BOARDS:
Genomics, - 1987-1995
The Journal of Heredity - 1987-
Biochemical Genetics - 1987-
Animal Biotechnology - 1989-
Mammalian Genome - 1990-
Genome Research - 1995-2004
Animal Genetics - 1991-1999
POSTDOCTORAL FELLOWS:
Name Ph.D. Institution Dates
Eric Hallerman The Hebrew University 1986-1987
Duke Rogers University of California, Berkeley 1987-1989
Daniel Gallagher Texas A&M University 1989-1993
Claude Schelling Univ. of Berne, Switzlerland 1993-1995
Jorg Schlapher Univ. of Berne, Switzerland 1995-1996
Prapti Mahyuddin Institute for Animal Production, Indonesia 1998-2001
Tom Goldammer Univ. of Rostock, Germany 1999-2001
Edward Cargill Texas A&M University 2005-2006
Christopher Seabury Texas A&M University 2006-
I was honored to share the 2001 Wolf Prize in Agriculture with Roger Beechy for
our respective work in animal and plant genomics. My award reads “for the use of
recombinant DNA technology to revolutionize animal sciences, paving the way for
applications to neighboring fields.” I’ll try to recount my career path to this highest
honor in agricultural research which was anything but a straight line. In fact, the
winding path of my academic training and research career is marked by relatively
688 Wolf Prize in Agriculture
few tracks pointing in the direction of livestock genomics. There are subtle
hoofprints, however, which are largely overshadowed by chance encounters with a
variety of scientific mentors, colleagues, friends and family who unknowingly
shaped my path.
I was born in Anson, Texas in 1941 and didn’t range far in the first 23 years of
my life except for a couple of years when my dad was stationed in Michigan before
the end of WWII. My parents purchased a small stock farm near Dad’s homeplace
after the war and built a modest house while Dad taught at a country school and
completed his education on the G.I. bill at Abilene Christian College. My mother
had only a high school education but could have and should have been an engineer.
She could design, construct or repair anything from a baby dress to a cattle barn.
Dad was offered a job with the Hawley Independent School District in the fall of
1946, the year I started first grade. He taught all the high school English courses,
coached all the boys and girls athletic teams, and drove the school bus that traveled
a 50 mile route to pick up about 30 students which when added to the locals
produced a student body of just under 100 students spread over 12 grades. Less
than a month into the school year, the Superintendent resigned and Dad was
tapped for that job, a position he held until his retirement 33 years later. Over
those years, he never gave up coaching basketball and retired with a national
record for career wins and was inducted into the Texas High School Basketball
Hall of Fame in 2005. My two sisters and brother also became teachers and
needless to say, we were all exposed to basketball. In addition to basketball, I was
exposed to farm life. Our homestead was several hundred acres and we raised beef
cattle and horses, partly to supplement Dad’s teaching salary but mostly, I think for
the educational experience it provided the children. I made my spending money
from a few steers we pasture finished each year and by breaking and selling raw
horses we bought each summer from a dealer in Mexico. I enjoyed 4H and FFA
and assumed I would someday have a small ranch of my own. I finished all 12
grades of public education at Hawley, graduating with a senior class of 16 students.
Despite being less than six feet tall, I had a number of basketball scholarship
offers, including a couple from major universities. Still a homeboy, I chose to
follow Dad’s footsteps at nearby Abilene Christian College.
I entered college with the intent of becoming a high school math and science
teacher and basketball coach. My main interest, however, was playing basketball. I
had a good career that actually spanned 5 years, thanks to a “redshirt” sophomore
year. Although not a prolific scorer as in high school, I started two years, one year
as team captain and played in two NCAA postseason tournaments. My closest
friends to this day are some of my basketball teammates. I met Raby Jean Beakley
during my freshman year but we didn’t start dating until a couple of years later.
She came from a ranching family and a small school and we were seated next to
each other in one class by a professor who knew us both and thought we should
James E. Womack 689
get to know each other. He was right. She finished her teaching degree in 1962
and taught for a year before we married in June of 1963. I still didn’t have my
degree and had another year of basketball ahead of me. I had taken a number of
education courses along with chemistry, physics, and math and was still planning
a career in teaching and coaching. I had never taken a biology course. At some
point it struck me that most public school teachers couldn’t afford the luxury of
my life long dream of living in the country, owning some land, and raising horses
and cattle. Raby and I spent a lot of time on a ranch owned by her dad and with a
dentist cousin who also had a nice cattle ranch. I came to the conclusion that my
quickest route to land and cattle was through dentistry, not through teaching. The
requirements for dental school included courses in embryology and comparative
anatomy both of which had lab requirements and one other biology course. I
chose genetics as my other biology course since it was the only course offered in
the fall that didn’t include a laboratory. I was already pushing my limit with labs
that conflicted with basketball practice. All three courses required permission of
the instructor since I had none of the prerequisite biology courses. Probably the
biggest single moment in shaping my scientific career was my request for permission
to take introductory genetics. Dr. Jim Throneberry was a cynical sort who apparently
had some bad experiences with athletes in his classes. He let me know that
I was welcome to give it a try but he was pretty sure I couldn’t pass his course. I
took his expectation of my failure as a personal challenge and immediately went to
the bookstore and purchased the textbook, an introductory genetics text by Irwin
Herskowitz. By the first day of class, I had read much of the text and was fascinated
by the supplemental material I had discovered at the back of the book. The
Supplement consisted of part of Mendel’s 1867 letter to Nageli, and the Nobel
Prize lectures of Morgan (1934), Muller (1946), Beadle (1958), Tatum (1958),
Kornberg (1959) and Lederberg (1958). Although dental school was still my goal,
I was convinced that genetics would provide most of the scientific excitement of
the last half of the century.
After the first exam, Jim Throneberry became a friend and mentor and he
along with Drs. Tommy McCord (organic chemistry) and Alvie Davis (biochemistry)
began to try to convince me that dental school would be a waste of my talents and
interest. Nonetheless, I applied to the Baylor College of Dentistry, and received my
letter of acceptance for the fall class of 1964. I was still on track to graduate with
a degree in Math Education, but now with a good background in biology and still a
strong interest in genetics. I had broken my foot in the first basketball game of the
season and had not worked my way back into the starting lineup until near the end
of the season. Raby was already establishing herself as an outstanding elementary
school teacher and loved her job at Dyess Elementary. It is hard to believe now but
Texas schools had just come to grips with integration and the Dyess school was the
first in Abilene to open their doors to both black and white students. Raby’s love
690 Wolf Prize in Agriculture
for children lessened the challenges of integration for both her and her students.
As with every first or second grade she has taught, she still gets appreciative phone
calls and emails from her former students. It was difficult for her to resign her job
in Abilene but she applied, was interviewed, and quickly hired for the Fall of 1964
in Richardson, near Dallas and the Baylor College of Dentistry. Three days in
August of 1964 marked another sharp turn in my career path. I was invited to
work in the office of a dentist friend who suspected my interest in dentistry might
be waning if in fact, it had ever been genuine. After the first day, I came home
excited about all the new things I’d seen and learned. After the second day, I told
Raby it was pretty much a repeat of the day before. After the third day, I told her I
couldn’t do that for the rest of my life. Her reaction was totally unselfish and left
me no room to ever doubt that she loved me. She said “we’ll do what makes you
happy.” I couldn’t shake my interest in genetics. I talked to Dr. Throneberry who
suggested I teach biology labs at ACC in the fall while applying to graduate programs
in genetics. Raby resigned the job in Richardson and was quickly welcomed back
in Abilene. We forfeited our $100 deposit at Baylor and I settled into an academic
career, understanding fully that we would probably never have our little ranch
with cows and horses.
I enjoyed teaching general biology labs and began to envision a career as a
professor of genetics. In retrospect, I know I would have found a research career
even if I had gone to dental school but now my interest was graduate school and
genetics. Without the benefit of the internet, selection of a graduate school in
1964 was by word of mouth and Peterson’s Guide. My love for the outdoors,
including hunting and fishing, pulled me toward the Pacific Northwest and
eventually to Oregon State University where Dr. Ralph Bogart, an Animal Scientist,
directed a large, well funded, interdisciplinary Genetics Institute. I accepted an
offer from Abilene Christian for support of $200 per month with the agreement
that I would return to their faculty upon receiving my PhD. This was a nice
supplement to the NASA fellowship Ralph provided. We arrived in Corvallis in
August of 1965 pulling all our belongings in a small covered trailer I had made
over the summer. Raby was pregnant with our first child and I was excited about
the breadth of course offerings in genetics at the University. My mother had been
diagnosed with cancer just a few months before we left making the decision to
move so far away a difficult one. My family assisted us with the move and enjoyed
some time in Oregon while helping us move into our new apartment. Watching
them drive away brought tears to our eyes. Our daughter Wendy was due in
December but didn’t arrive until January of 1966. We tried to induce labor with
weekend trips following streams up the rough logging roads or climbing over the
massive timbers drifted up on the beautiful Oregon beaches. But like everything
else she has done in her life, Wendy arrived when she chose, not at our convenience.
Raby’s dad had passed away in 1964 and her mom spent quite a bit of time with
James E. Womack 691
us over the next three years, providing greatly appreciated day care as Raby was
able to teach between the birth of our two children. I spent a lot of time in the
graduate student cubicles in the basement of Withycombe Hall with people like
Jerry Reeves, Paul Humes, Carol Nix, Prentice Schilling, and Joe Templeton, who
with his wife Jamie had moved up with us from Abilene Christian. I had never
doubted the quality of my education from a small public school and a small
college in West Texas and found my courses in genetics and radiation biophysics
challenging but not overwhelming and totally delightful. Ralph was a quantitative
geneticist by training but had incorporated considerable physiological genetics
into his research program. When he suggested I might find a PhD research project
in the wealth of data he was collecting from selection experiments in cattle,
I distinctly recall telling him I didn’t really want to study cattle genetics or animal
breeding. I was more interested in laboratory animals and biomedical applications.
I had done some reading on mutagenesis and we agreed on a radiation mutagenesis
project in mice with emphasis on quantitative traits. I measured selection response
for body weight and fertility traits in irradiated populations and published two
papers from my dissertation research. More importantly, I learned about mammalian
mutagenesis and became fascinated with mouse genetics. Ralph taught me a lot
about mentoring although I’m sure I never practiced it like he did. He had an
enormous garden and orchard that was planted with poor starving graduate students
in mind. Of course, we spent some of our weekend time tilling the soil. He and
Fran became adopted grandparents to his students’ children. He guided us into our
postdoctoral work and he was disappointed that I had made a commitment to
Abilene Christian. He thought I should look for a postdoc in a medical school or
research institute but he accepted my decision and encouraged me to grow and
develop as a teacher.
Our second child was already overdue when I got my PhD degree in May of
1968. Our good friends from church, Gail and Chuck Woosley, escorted Raby to
the graduation ceremony and had parked a car near Gill Coliseum for Raby if she
needed a quick exit. She didn’t. Louis and Janice Stone, who along with Pat and
Carolyn Agnew, and Larry and Ella Rogers, have been lifelong friends from basketball
days at ACC, had come to Corvallis to help us move. Raby, Wendy, the baby, and
her mother would fly back to Texas, and the Stones would alternate with me in
driving the U-Haul truck (we had outgrown the home made trailer) and our car.
We waited and waited and waited. As before, the doctor had to induce labor and
this was not an easy one for Raby….or Jimmy who was born with some ugly
forceps bruises which fortunately disappeared before he and his mother, sister and
grandmother arrived back in Texas. I would arrive a few days later with the Stones,
our belongings and two black lab puppies.
The summer of 1968 was both a happy and sad one, but memorable. Thanks
to a gift of stocks from Raby’s parents, and her teaching salary from the beginning
692 Wolf Prize in Agriculture
of our marriage, we had managed to buy 160 acres of farm/pasture land near
Hawley, only 12 miles from where I would begin work in the fall and only 6 miles
from my parents. Mom was very sick and I am thankful for the time she had with
our two children before her death that fall. They made her smile. The property had
an old and very small house in which we were determined to live. My grandpa
Hollums, a carpenter by trade, helped me add a room and refurbish the house that
summer, an experience I treasure. We built what we thought was a rattlesnake
proof fence around the back yard but were proved wrong on several occasions.
Despite the snakes, the sandstorms, the red mud, and constant concern about a
half mile of river and a large pond, it was a nice place to raise young children.
They had wonderful pets including a young skunk which is a story in itself. Raby
was quickly employed by the Abilene schools and I began teaching genetics, cell
biology and zoology in the fall of 1968 at what was now Abilene Christian
University. I even bought that small herd of cattle I had long coveted although the
ranching venture was a financial failure, thanks to cattle thieves who knew we left
the property unoccupied for most of the daylight hours.
My most vivid memories of my first teaching experience include the high
quality of the undergraduate students at ACU. The Biology department offered a
Master’s degree and I took on a few grad students, including Loren Skow who is
now a colleague on the Veterinary faculty at Texas A&M. Dr. Clark Stevens was a
wonderful department head and we were blessed with teachers like Mike Kemp,
Kenneth Williams, and Roy Shake. Although I enjoyed teaching, my graduate school
experience had kindled a burning desire for research that wouldn’t go away. I
wanted to know more about mutagenesis in mammals and wrote a proposal to the
National Foundation March of Dimes to develop a biochemical genetic assay of
mutation in mice using allozymes as an endpoint. I was familiar with mutagenesis
studies of Tom Roderick and Earl Green at the Jackson Laboratory and had contacted
Tom who agreed to be a collaborator in my proposal. If funded, I would come to
Bar Harbor to study in the summers and continue my teaching on a nine month
basis with ACU. The work was funded and we made our first of several trips to Bar
Harbor in the summer of 1970. Although we didn’t discover any radiation induce
allozyme variants in one summer of starch gels, we developed the concept into a
germinal mutation assay the NIEHS would eventually use on a grand scale in their
chemical mutagenesis program. I also developed a lasting professional and personal
friendship with Tom Roderick and a number of other scientists at The Jackson
Laboratory such as Murriel Davisson, Eva Eicher and Ben Taylor, who along with
visitors like John Hutton and Verne Chapman had begun to use allozyme variants
as markers for populating the mouse linkage map. Enzyme markers were especially
exciting to me because the human gene map was in its infancy and was being
developed through somatic cell genetics and biochemical markers. By mapping the
orthologous gene products in mice we had the first opportunity to do focused
James E. Womack 693
their work was a simple matter of staying out of their way. Skippy Lane kept us
busy mapping new spontaneous mutations in the mouse, the most notable being
ashen and muscle deficient, and we were able to use the growing list of biochemical
genetic variants to map induced inversions that Norm and Muriel uncovered. But
mostly, I was interested in watching the human synteny map develop and building
a comparative linkage map of the mouse. I was fascinated by the cluster of
parologous esterase genes on mouse chromosome 8 and the evolution of gene
families as could be revealed by linkage mapping with biochemical markers. Along
with Tom, Muriel, Eva, Ben and of course, Margaret Green, I became part of a
formidable team of mouse gene mappers with an eye toward comparing the genomes
of the two most important mammals in the world of genetics. The first International
Workshop of Human Gene Mapping held in New Haven in the summer of 1973
recognized the role of comparative mapping in understanding the human genome
and was organized as individual workshops for each human chromosome with
special workshops on topics like nomenclature and comparative mapping. These
workshops which met around the world at two year intervals became the focal
point of the comparative mapping community which consisted entirely of mouse
geneticists at that time.
Earl Green, Director of The Jackson Laboratory, forced me into the toughest
decision of my professional life in the Spring of 1975 when he offered me a Staff
Scientist position and my own laboratory when Tom returned from Washington. I
missed ACU and the undergraduate students and I really hated to disappoint Clark
Stevens who had invested heavily in me. However, our children enjoyed Bar Harbor,
Raby was enjoying her work where she was gaining a reputation as an outstanding
reading specialist, and I was doing science that rewarded me with the discovery of
new knowledge, almost on a weekly basis. I resigned my position at ACU with
great sadness but welcomed the opportunity to pursue mammalian genetics full
time.
Scientific meetings and conferences have been extremely important to me and I
made it a point to attend and present as often as possible early in my career,
sometimes totally or partially at my personal expense. I tried to attend the annual
meetings of the Genetics Society of America, the previously mentioned Human
Gene Mapping Workshops, and the annual Biochemical Genetics workshops
organized by Ken Paigen and attended mostly by JAX and Roswell Park scientists.
Of course, JAX ran its summer short courses that brought in outstanding human
and mammalian geneticists on an annual basis. The people I met at meetings were
probably more important to me than staying on the cutting edge of my science,
something I could have done through the literature. I could easily list 50 names of
people who influenced my career that I met at meetings, workshops or seminars
during my tenure at JAX. I’ll keep it to a handful. JAX attracted people like Jim
Crow, Frank Ruddle, Victor McKusick, Mary Lyon, Tony Searle, Susumu Ohno, and
James E. Womack 695
Charles Ford. Of course, daily contact with Tibby Russell and George Snell influenced
every young scientist who came through The Jackson Laboratory. One on one
discussions with the academic giants were inspirational. But it was young scientists
from the mapping workshops like Steve O’Brien, Pete Lalley, Mike Sicilliano,
and Sue Naylor who were in the trenches mapping genes and with whom I
communicated on a regular basis and developed professional and personal
friendships that continue to the present. I became keeper of the mouse map
(bioinformatics on 3x5 index cards) when Margaret Green retired, a job Muriel
Davisson would later do much better immediately before conversion to electronic
databases. I contributed both mouse and rat gene maps to O’Brien’s “Genetic
Maps”, a valuable compilation of genome maps, again before the advent of electronic
databases. I always had my eye on the mapping of genetic orthologues in mouse
and man and drew the original “Oxford grid” for Mouse Newsletter, a valuable
publication of mouse mutants and linkages read cover to cover by practicing
mouse geneticists. By the fall of 1976 I felt like a card carrying mouse geneticist
fortunate to work in the Mecca of mouse genetics. Although I had several excellent
summer students such as Debra Kendall and Philip Giampietro, I missed teaching. I
don’t remember thinking about it a lot but it all came in focus one day when I got
a call from my undergraduate and graduate school buddy, Joe Templeton.
After receiving his PhD at Oregon State, Joe had taken a position in
immunogenetics with the University of Oregon Medical School in Portland. He had
recently been recruited to the Baylor College of Medicine in Houston to head up
the genetics component of a state funded Institute of Comparative Medicine, a
joint venture between Baylor College of Medicine and the College of Veterinary
Medicine at Texas A&M, 90 miles away in College Station. The Institute was looking
to fill a new faculty position in comparative genetics at A&M and Joe suggested I
apply if interested. I was surprised and also worried when I found myself interested
in the position. The same person who had difficulty settling on an undergraduate
major hadn’t kept his family in one location for more than five years since
graduating. I discussed it with Raby and her reply was the same as before, “we’ll
do what makes you happy”. But this time her poker face failed her and I could see
a little smile with the thought of moving back to Texas and away from the Maine
winters. I applied for the job, made an interview visit, and in June of 1977 joined
the Department of Veterinary Pathology at Texas A&M. A farewell gesture from
mother nature was a little snow flurry as we departed Bar Harbor in early June.
Most of my concerns about my professional restlessness are now gone as I approach
the end of my 31st year at Texas A&M.
The College of Veterinary Medicine had very few non-DVM faculty and the
Department of Pathology had none. Charlie Bridges and later Ken Pierce, as heads
of the Department, were willing to take a chance on a so-called “naked PhD”. I
was attracted to the University’s interdisciplinary faculty in Genetics that would
696 Wolf Prize in Agriculture
had not embraced gene mapping, the USDA and animal breeders were interested
in the concept of genetic engineering in the early 1980’s and became especially
excited when Palmiter and Brinster produced transgenic mice expressing and
responding to introduced growth hormone genes. I did everything within my
power to tie gene mapping to the genetic engineering bandwagon. Dewey Kraemer
led a strong embryo physiology group at A&M and he appreciated the eventual
role of genome mapping in the success of genetic engineering of farm animals.
My first external funding for the cattle map came from the Hillcrest Foundation of
Dallas after a fascinating meeting between Dewey, myself and Mr. W. C. Caruth of
the Hillcrest Board of Trustees. Mr. Caruth was intrigued by a popular article he
had read predicting that genetic engineering could produce wooly pigs, transgenic
pigs producing wool in addition to meat. We didn’t promise Mr. Caruth a wooly
pig but we promised significant advances in the genetic mapping and genome
manipulation of cattle in exchange for a generous $100,000 grant from the
Foundation that Dewey and I would divide between our laboratories. The cattle
map was off the ground and we published our first cow map in Steve’s “Genetic
Maps” in 1984 and the first cow-human comparative map in the Journal of
Heredity in 1986. The cattle genome proved to be more conserved with human
than the mouse genome but not to the extent of the cat-human comparison.
I came to know Morris Soller of the Hebrew University through my appreciation
for his far-sighted projections of “genome genetics” in livestock and the use of
gene markers in selection for quantitative traits. He was clearly the theoretical
leader of the genomic era of animal breeding. We teamed up to write a BARD
grant in 1985, proposing to significantly expand the cattle somatic cell map using
DNA markers and also to build a RFLP map for linkage analysis of quantitative
traits. We got our grant although RFLPs were soon to be replaced by microsatellites
as markers of choice for linkage mapping. We got another BARD grant in 1990
and I have treasured our collaboration and friendship. Several small grants from
the USDA to my laboratory were forthcoming in the 1980’s, mostly through Animal
Health and with ties to genes for disease resistance. My first competitive grant
from the USDA purely for genome mapping in cattle came in 1986. I also received
some support from the Kleberg Foundation. I spent much of the 1980’s in the dual
role of trying to get my own mapping efforts funded and in trying to convince
funding agencies of the value of genome maps for livestock species. The idea of a
human genome initiative was gaining traction in the early 80’s and we were
beginning to imagine having access to the complete sequence of the human genome
and several model organism genomes including the mouse. With the human genome
sequence projected to cost $3.5 billion, it was difficult to imagine that the genomes
of livestock species would ever be sequenced. My pitch was for funding of
comparative gene mapping which would identify regions of conservation between
cattle (or pigs or sheep) and humans and mice on a fine scale. The mouse and
698 Wolf Prize in Agriculture
human sequence could then be reasonably well exploited to find important livestock
genes. This idea complemented proposals by Soller, Jacques Beckmann and others
to develop markers for building linkage maps and mapping quantitative trait loci
(QTL). I always assumed that finding the genes underlying QTL would come from
mining the sequenced human and mouse genomes at the addresses suggested by
the comparative livestock maps.
The concept of an international bovine genome project was discussed in a
workshop setting for the first time to my knowledge at the opening of the
Internatioinal Trypanotolerance Center in the Gambia in March of 1987 followed
by a meeting of scientists from the European Economic Community in Brussels in
December of 1987. Mapping the bovine genome received considerable attention at
the XXI Conference on Animal Blood Groups and Biochemical Polymorphisms in
Torino in July of 1988. I was an invited speaker at that conference and remember
it well, having popped a couple of stitches from my triple bypass surgery two
months earlier. This group was to soon become the International Society for Animal
Genetics (ISAG) and would play a major role in animal gene mapping over the
next 20 years through its workshop structure at biennial meetings. I had the
privilege of serving as President of ISAG a few years later. I also had a wonderful
opportunity visit Australia as a McMaster Fellow with Jay Hetzel’s CSIRO group in
Rockhampton in 1990, doing a little lab work but mostly helping campaign for
funding of their mapping program and I participated in the organization of the
Canadian animal mapping program in a workshop in Ottawa later that year. These
and many other international experiences were important to me professionally but
more important to me personally for the wonderful friends Raby and I discovered
among animal gene mappers.
Several events in the early 1990’s were important to the US effort in bovine
genomics (the term “genomics” was coined by Tom Roderick in 1986 for the title
of the journal Ruddle and McKusick proposed to accompany the human genome
initiative). Charlie Arntzen had taken the position of Director of the Texas
Agricultural Experiment Station and the Institute of Biosciences and Technology at
Texas A&M. He continued the support of my mapping laboratory that Neville had
begun and also supported and helped organize a Banbury Conference on Mapping
the Genomes of Agriculturally Important Animals at Cold Spring Harbor in the
summer of 1990. This conference brought representatives of the human genome
project ( Jim Watson, Victor McKusick, David Housman, and Ray White) together
with some of the animal mapping community (Michel Georges, Jay Hetzel, Brian
Kirkpatrick, Harris Lewin, Joan Lunney, Larry Schook, Loren Skow, Morris Soller,
Alan Teale and myself) in the presence of administrators from the USDA and
several of the land grant universities. The conference was followed in the fall by
the first Allerton Conference organized by Lewin, Schook and others at the University
of Illinois. These two workshops led to the organization of a USDA National Animal
James E. Womack 699
Genome Research Progam in 1993 for which I have served as Cattle Coordinator
for the last 15 years.
Most of the ideas and essentially all of the data credited to me over the years
have come from the excellent students, postdocs and technical staff who have
blessed my laboratory. More than 40 graduate students have completed their degrees
with me at Texas A&M and I can’t really name one without naming them all
(which I’ve done in my c.v). These students and staff developed the hybrid somatic
cell panel into a valuable resource for comparative mapping, beginning with
biochemical markers and evolving through Southern blotting and PCR genotyping
of DNA markers, always with an eye toward the comparative maps of homologous
genes in humans and mice. Dan Gallagher, an exceptional postdoctoral fellow,
brought the cytogenetics expertise to the lab to assign syntenic groups to
chromosomes, not an easy chore with 29 acrocentric autosomes in the bovine
karyotype. Dan played a critical role in standardizing the karyotype of domestic
cattle which became the prototype for chromosome identification in other
artiodactyls. The construction of radiation hybrid panels allowed us to order markers
in the cattle genome and define conservation and disruption of gene order within
conserved syntenic groups. These panels, especially when genotyped with ESTs
and BAC end sequences in our collaboration with Harris Lewin’s laboratory, became
valuable tools for fine scale mapping, for mining gene loci underlying traits of
economic and biological interest, and ultimately for assembly of the cattle genome
sequence. The citation for my election to the National Academy of Sciences USA in
1999 reads “Womack launched the discipline of livestock genomics with his initial
map of the bovine genome and the demonstration of extensive chromosomal
conservation in cattle, humans, and mice. His international leadership resulted in
a comprehensive bovine linkage map and the first genetic mapping of economically
important traits in cattle.” It is obvious to me, if not to everyone familiar with my
work, that my students and collaborators are equally deserving of this and other
recognitions I have received. Perhaps I am considered a leader in the discipline
because I got an early start and couldn’t work fast enough to get out of the way. A
highlight of my career was in April 2007 when a group of my laboratory alumni
organized a symposium in my honor entitled Advances in Applied and Comparative
Genetics. The two-day scientific program was comprised entirely of presentations
from my former students, postdocs and collaborators.
Early in this decade, it became obvious that the genomes of human and selected
model organisms would be sequenced ahead of schedule and under budget by the
National Human Genome Research Institute. A call went out for white paper
proposals for sequencing of additional genomes that could aid in informing human
genetics. I was able to organize a National Academy of Science workshop in early
2002 to discuss the status of domestic animal genomics and to help prioritize
different species for genome sequencing. Shortly thereafter, Richard Gibbs and
700 Wolf Prize in Agriculture
George Weinstock of the Baylor College of Medicine Human Genome Center, Steve
Kappes of the USDA, Larry Schook of the University of Illinois, and Loren Skow
and myself of Texas A&M wrote a white paper proposal for sequencing the bovine
genome. In September of that year the cattle and dog genomes were approved with
high priority for sequencing. Our work had only begun, however. The cost for
7-8x coverage of a 3000 megabase genome had come down over the years to
about $50 million. While appreciating the comparative information the bovine
genome could provide medical genetics, the NIH was well aware that there were,
or should be, significant agricultural interest in sequencing this genome. They
agreed to fund half the total cost and, in all fairness, asked us to come up with the
other $25 M. Despite pledges from the USDA, the dairy and beef industries, and
international agricultural agencies, the funding came together very slowly. As of
March, 2003 we had only $15 M committed. In fact, we were within a week of
losing our high priority status when Texas Governor Rick Perry committed the
final $10 M. This would have never happened without the efforts of a wonderful
lady from South Texas. Mrs.Anne Armstrong, former U.S. Ambassador to Great
Britain and a member of the Board of Regents at Texas A&M, learned about the
bovine sequencing project. She and her husband, Tobin, owned and operated the
Armstrong Ranch in Armstrong, Texas and were influential in the beef cattle
industry as well as in State and National politics. She recognized the potential of
genome sequencing to the improvement of beef cattle health and productivity and
used her influence to persuade the Governor’s office to insure its initiation. I
suppose the bovine genome would have ultimately been sequenced without the
help of Anne Armstrong, but it would have certainly been delayed by several years.
I am planning to reap some of the benefits of the cattle genome sequence for a
few years before I retire. I have aggressively avoided administrative responsibilities
at Texas A&M and have maintained a laboratory totally with external funding. Jan
Elliot, Elaine Owens and Mary Jewell are still the nucleus of my research team. We
continue to train graduate students and postdocs and are focusing our efforts on
finding diversity in the bovine genome that translates to differential susceptibility
to pathogens. Raby and I plan to retire to our little ranch in Robertson County
where we currently relax (and work) on weekends and have the occasional barbecue
with family, friends and guests. We enjoy our little cattle herd (with one bison)
and I’ve (gently) broken and trained a young filly who has become more of a
pasture pet than a working horse. I can imagine being very happy spending my
retirement years with the animals on our ranch and I’m sure I’ll have questions
about the composition of their genomes, the same kind of questions that have
excited me about going to work every day for the past 40 years. In the early days of
human gene mapping, Frank Ruddle responded to the question of “why map
genes” with the reply “ because mapping genes is good for you.” I kept that quote
on my office wall for a number of years and can now summarize my own career
James E. Womack 701
with the statement “mapping genes has not only been good for me, mapping genes
has been good to me.”
COMMENCEMENT ADDRESS:
President Gates, Regent Jones, Provost Prior, Dean Watson Mr. Taylor, General Van
Alstyne, and others on the stage, I am deeply honored to be here among you and to
address this 2006 graduating class of Fightin’ Texas Aggies.
Howdy Ags! Congratulations on a job well done. And congratulations to your
families who have supported you in every step of this long and difficult journey.
Graduates, I’m here today as a representative of your faculty to thank you for
the privilege of working with you these last four..(or five)…(or six) years. I’m here
to wish you well in the many endeavors that tomorrow holds. And I’m here to give
you one last lecture before you leave. Ags, along with your family, we the faculty
have invested our lives in you, we care about your future, and we love you. So,
bear with me for a few minutes.
A new world awaits you outside the doors of this arena. A world full of
excitement and opportunities, and challenges. A world my generation inherited,
and molded and shaped for you, improving it in some areas and really messing it
up in others.
I sat where you are in the early 1960s, not at this great university that adopted
me 29 years ago, but at a fine little school in West Texas. I had a wonderful
undergraduate experience. I went through college on a basketball scholarship
(I was taller then). I played in a couple of NCAA tournaments. I found and married
my life-long partner. I could solve mathematical equations with the best of them
and I had read and absorbed just about everything that had been written at that
time about a wonderful new molecule called DNA. But what I got that day was a
diploma, my education was only beginning.
My diploma did not prepare me to deal with a war being fought half-way
around the world that was taking the lives of some of my classmates and splitting
our great country down the middle. We haven’t improved the world much for you
in 40 years, have we?
But we’ve done some good things, especially in science and technology. We got
you into space, we connected you to the world through the internet, we gave you
Dolly the sheep and we sequenced your genome. You will undoubtedly figure out
what to do with these gifts, then continue to advance technology in ways I cannot
even imagine. My new world of the 60’s and your new world today are not that
different, otherwise I wouldn’t be so presumptuous as to offer you a few words of
advice.
James E. Womack 703
So, what can I say to help you succeed in a world that is both ever changing
and never changing. I’m certainly not here to tell you how to become rich or
famous, and if I were it wouldn’t be from first hand experience. However, if you
can look up from what you are doing 30 years from today and say “I have
contributed some little something to making this world a better place, I have great
friends and family who love me, and I wouldn’t trade with anyone what I do for a
living”, you are a success.
That’s my definition of success, now let me give you three keys to finding it.
And let me tell you from the beginning, these aren’t original ideas.
1. Work Hard. (Have you heard that before?) Don’t allow yourself to fail at
anything for lack of effort. Don’t be ashamed to be an overachiever, in fact, strive
for it. All of you are talented or you wouldn’t be here, some of you are blessed
with exceptional talents. For you gifted Aggies who managed to get your diploma
despite spending more of the last 6 years in the Dixie Chicken than in Sterling
Evans, I say “shame on you”. But now, it’s time to get with it. If you don’t push
your talents to their limits, then a step beyond, you will look back on your life with
serious regrets. Be an overachiever in your job..and that begins with finding work
that you enjoy doing. If you love your job you will go the extra mile.
My admonition to work hard extends beyond your job, however. A successful
marriage or any worthwhile relationship requires hard work. And nothing requires
more effort than parenthood. Hard work does not guarantee success in any endeavor,
but less than your best effort is surely an invitation for failure. We would all do
well to heed the words of King Solomon who wrote in the book of Ecclesiastes
“Whatever you find to do, do it with all your might.”
2. Live Smart. Don’t ever, ever, ever stop learning. Learn from your mistakes, you
can turn your wounds into wisdom. Choose your friends wisely. Surround yourself
with the kind of people you would like to emulate. You are Aggies, you are going
to be bosses…don’t be afraid to hire people who are smarter than you. I’ve never
taken a graduate student into my laboratory who wasn’t smarter than me. They
haven’t made me rich or famous, but I wouldn’t trade my job of teaching and
research at this university for any in the world. (There went any hope I had for
getting a raise this year.) Select your life-long mate very carefully. Marry up, as the
saying goes, and I’m not talking about money. It worked for me. The person you
will learn the most from the rest of your life is probably the person you wake up
with every morning. And besides that, your spouse will contribute the DNA that
makes your children smarter than you… and they will be…if you don’t believe
me, ask them when they are 16.
Elbert Hubbard, American essayist from some100 yrs ago said “The recipe for
perpetual ignorance is to be satisfied with your opinion and content with your
knowledge.”
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Stay informed, and weigh conflicting information wisely. Keep up with the
technical and scientific advances of the world, whether you are trained in science
or not. Your new world is a technical one and you are going to be a juror in the
court of public opinion on many issues outside the discipline named in your
diploma.
You will decide whether stem cell research is allowed to proceed with its life
saving potential.
You will determine whether starving children in developing countries will eat
food that has been genetically modified to grow in harsh environments.
You will make political decisions to stop environmental deterioration and restrain
global warming.
Don’t make your decisions on the basis of my obvious biases. Read and learn,
and always weigh the motivation behind the source of your information.
And above all, don’t let other people do your thinking for you. I encourage
loyalty to your religion, to your country and to this great university. But blind
loyalty is wasted loyalty. Participate in shaping the future of the institutions you
hold dear, but be an informed participant.
3. Be a team player. Regardless of how hard you work, or how smart you become,
your success and happiness is going to boil down to relationships. The world has
become far too complex to go it alone. Globalization is a reality. You may spend
your working hours alone with a computer, but on the other end of that broad
band or satellite beam are real people. Whether in our professional lives or in our
personal lives, our relationships with people are the keys to our success. Of course,
developing business and personal relationships begins with our choices of associates.
But it doesn’t end there, it requires putting the success of the team ahead of
personal gratification, and that’s an Aggie tradition. In the end, team play translates
to unselfishness. It requires taking our work seriously without taking ourselves
seriously. There is no room for an overbearing ego in a team endeavor, or in any
successful relationship. Team play is a big part of your heritage as Aggies, take it
with you.
So, my three keys to success for you today are pretty simple, Work Hard, Work
Smart, and Work Together. It’s perfectly legitimate in our academic world to use
the ideas of other people, so long as we give proper credit. I told you earlier I am
an old jock and most of my personal philosophy has at one time or another been
posted on a locker room wall. Some of you may recognize my keys to success as the
motto of our outstanding basketball coach Billy Gillispie, who demands, and I
emphasize demands, that his Aggies “Play hard, Play smart, and Play together”.
I recommend Coach Gillispie’s motto as a sound rule for a successful life. We’ve
seen it work miracles here on this floor, I believe it will work for you outside the
doors of this arena.
Gig em’ Ags, and God Bless You.
James E. Womack 705
LIST OF PUBLICATIONS
(Selected Publications from a total of 331 in Refereed Journals):
Womack, J.E., Yan, D.L.S., and Potier, M.: Gene for neuraminidase activity on mouse
chromosome 17 near H-2: pleiotropic effects on multiple hydrolases. Science
212:63-65, 1981.
O’Brien, S.J., Moore, J.L., Martin, M.S., and Womack, J.E.: Evidence for the horizontal
acquisition of murine AKR virogenes by recent horizontal infection of the
germ line. J. Exptl. Med. 155:1120-1132, 1982.
Womack, J.E. and Moll, Y.D.: A gene map of the cow: Conservation of linkage with
mouse and man. J. Hered. 77:2-7, 1986.
O’Brien, S.J., Seuanez, H.M., and Womack, J.E.: Mammalian genome organization:
an evolutionary view. Ann. Rev. Genet. 22:223-251, 1988.
Threadgill, D.S., Krause, J.P., Krawetz, S.A., and Womack, J.E.: Evidence for the
evolutionary origin of human chromosome 21 from comparative gene mapping
in the cow and mouse. Proc. Natl. Acad. Sci. USA. 88:154-158, 1991.
Gallagher, D.S. and Womack, J.E.: Chromosome conservation in the bovidae.
J. Hered. 83:287-298, 1992.
O’Brien, S.J., Womack, J.E., Lyons, L.A., Moore, K.J., Jenkins, N.A., and Copeland,
N.G.: Anchored reference loci for comparative genome mapping in mammals.
Nature Genetics 3, 103-112, 1993.
Georges, M., Dietz, A.B., Mishra, A., Nielsen, D., Sargeant, L., Sorensen, A., Steele,
M.R., Zhao, X., Leipold, H., and Womack, J.E.: Microsatellite mapping of the
gene causing weaver disease in cattle will allow the study of an associated
quantitative trait locus. Proc. Natl. Acad. Sci. USA 90:1058-1062, 1993.
Georges, M., Drinkwater, R., King, T., Mishra, A., Moore, S.S., Nielsen, D., Sargeant,
L.S., Sorensen, A., Steele, M.R., Zhao, X., Womack, J.E. and Hetzel, J:
Microsatellite mapping of a gene affecting horn development in Bos taurus.
Nature Genetics 4:206-210, 1993.
Barendse, W., Armitage, S.M., Kossarek, L.M., Shalom, A., Kirkpatrick, B.W., Ryan,
A.M., Clayton, D., Li, L., Neibergs, H.L., Zhang, N., Grosse, W.M., Weiss, J.,
Creighton, P., McCarthy, F., Ron, M., Teale, A.J., Fries, R., McGraw, R.A., Moore,
S.S., Georges, M., Soller, M., Womack, J.E., and Hetzel, D.J.S.: A genetic linkage
map of the bovine genome. Nature Genetics 6:227-235, 1994.
Womack, J.E. and Kata, Srinivas R.: Bovine genome mapping: evolutionary inference
and the power of comparative genomics. Current Opinion in Genetics &
Development 5:725-733, 1995.
Lyons, L.A., Laughlin, T.F., Copeland, N.G., Jenkins, N.A., Womack, J.E. and O’Brien,
J.O.: Comparative anchor tagged sequences (CATS) for integrative mapping of
mammalian genomes. Nature Genetics 15:47-56, 1997.
Womack, J.E., Johnson, J.S., Owens, E.K., Rexroad, C.E. III, Schläpfer, J., and Yang,
Y.-P.: A whole-genome radiation hybrid panel for bovine gene mapping.
Mammalian Genome 8:854-856, 1997.
706 Wolf Prize in Agriculture
Yang, Y.-P. and Womack, J.E.: Parallel radiation hybrid mapping: A powerful
tool for high-resolution genomic comparison. Genome Research 8:731-736,
1998.
O’Brien, S.J., Menotti-Raymond, M., Murphy, W.J., Nash, W.G., Wienberg, J.,
Stanyon, R., Copeland, Neal G., Jenkins, N.A., Womack, J.E., and Graves, J.A.M.:
The promise of comparative genomics in mammals. Science 286:458-481,
1999.
Winter, A., Kramer, W., Werner, F.A., Kollers, S., Kata, S., Durstewitz, G., Buitkamp,
J., Womack, J.E., Thaller, G., and Fries, R.: Association of a lysine-232/alanine
polymorphism in a bovine gene encoding acyl-CoA:diacylglycerol
acyltransferase (DGAT1) with variation at a quantitative trait locus for milk
fat content. Proc. Natl. Acad. Sci. USA 99-9300-9305, 2000.
Ozawa, A., Band, M.R., Larson, J.H., Donovan, J., Green, C.A., Womack, J.E., and
Lewin, H.A.: Comparative organization of cattle chromosome 5 revealed by
comparative mapping by annotation and sequence similarity and radiation
hybrid mapping. Proc. Natl. Acad. Sci. USA 97:4150-4155, 2000.
Goldammer, T., Kata, S.R., Brunner, R.M., Dorroch, U., Sanftleben, H., Schwerin,
M., and Womack, J.E.: A comparative radiation hybrid map of bovine
chromosome 18 and homologous chromosomes in human and mice. Proc.
Natl. Acad. Sci. USA 99:2106-2111, 2002.
Takeda, H., Takami, M., Oguni, T., Tsuji, T., Yoneda, K., Sato, H., Ihara, N., Itoh, T.,
Kata, S.R., Mishina, Y., Womack, J.E., Moritomo, Y., Sugimoto, Y. and Kunieda, T.:
Positional cloning of the gene LIMBIN responsible for bovine chondrodysplastic
dwarfism. Proc. Natl. Acad. Sci. USA 99:10549-10554, 2002.
Winter, A., Krämer, W., Werner, F.A., Kollers, S., Kata, S., Durstewitz, G., Buitkamp,
J., Womack, J.E., Thaller, G., and Fries, R.: Association of a lysine-232/alanine
polymorphism in a bovine gene encoding acyl-CoA:diacylglycerol
acyltransferase (DGAT1) with variation at a quantitative trait locus for milk
fat content. Proc. Natl. Acad. Sci. USA 99:9300-9305, 2002.
White, S.N., Taylor, K.H., Abbey, C.A., and Gill, C.A., and Womack, J.E.: Haploytype
variation in bovine Toll-like receptor 4 and computational predication of a
positively selected ligand-binding domain. Proc. Natl. Acad. Sci. USA
100:10364-10369, 2003.
Larkin, D.M., der Wind, A.E., Rebeiz, M., Schweitzer, P.A., Bachman, S., Green, C.,
Wright, C.L., Campos, E.J., Benson, L.D., Edwards, J., Liu, L., Osoegawa, K.,
Womack, J.E., de Jong, P., and Harris, L.A.: A cattle-human comparative map
built with cattle BAC-ends and human genome sequence. Genome Research
13:1966-1973, 2003.
Murphy, W.J,. Larkin D.M., Everts-van der Wind A., Bourque G., Tesler G., Auvil L.,
Beever J.E., Chowdhary B.P., Galibert F., Gatzke L., Hitte C., Meyers S.N., Milan
D., Ostrander E.A., Pape G., Parker G.H., Raudseep T., Rogatcheva M.B., Schook
James E. Womack 707
L.B., Skow L.C., Welge M., Womack J.E., O’Brien S.J., Pevzner P.A., and
Lewin H.A.: Dynamics of mammalian chromosome evolution inferred from
multispecies comparative maps. Science 309, 613-617, 2005.
Everts-van der Wind , A., Larkin, D.M., Green, C.A., Elliott, J.S., Olmstead, C.,
Chiu, R., Schein, J.E., Marra, MA, Womack, J.E., Lewin, H.A.: A high-resolution
whole-genome cattle-human comparative map reveals details of mammalian
chromosome evolution. Proc Natl Acad Sci USA; 102:18526-18531, 2006.
Sugimoto, M., Fujikawa A., Womack, J.E., Sugimoto Y.: Evidence that bovine forebrain
embryonic zinc finger-like gene influences immune response associated with
mastitis resistance. Proc Natl Acad Sci USA 103:6454-6459, 2006.
On a shelf in Dr. James Womack’s modest office at Texas A&M University is a gray
book, published in 1962. Its edges are worn and its 543 pages are yellowed now,
but if not for that book and a skeptical Abilene Christian University biology professor,
the world of genetics might be very different today.
Womack (’63), a former Hawley (Texas) High School basketball star who at
5’10” made the all-state team twice and averaged 30 points a game, now chuckles
as he recalls how his career path took a sharp turn.
Planning to teach and coach, Womack entered ACU in 1959 on a basketball
scholarship, lettered four years, and captained the 1962-63 Wildcat team. By his
junior year, he decided he wanted to be a dentist instead, but lacked the biology
prerequisites. He visited Dr. James Throneberry, who taught Genetics, the only
course that didn’t interfere with basketball workouts.
“He gave me a challenge that I took very personally,” Womack says. “He said in
the first place, normally athletes didn’t pass his class, and second, that without the
prerequisites, I didn’t have a chance. But if I wanted to fail his course, I was
welcome to sign up. With a challenge like that, I went to the bookstore and bought
the book, and to make a long story short, I just read the textbook before the first
class.”
James E. Womack 711
Despite Womack’s success, “he’s never forgotten where he came from, and
things that were important to him before he was successful still are,” says Templeton.
Church and family are two of those things. Womack and his wife have been
members of the A&M Church of Christ since moving to College Station in 1977,
and he has served as a deacon for 20 years. The couple lives in College Station and
maintains a 100-acre farm near Wheelock with 30 cows, one bison and two
horses. He and Raby, a retired second-grade teacher, built a log cabin there they
plan to expand and eventually move into when he retires. It will be close to their
daughter, Wendy (Womack ’90) Faltys, a nurse in Franklin, and their son, James
Michael Womack (’90), a business owner in Austin.
Throughout his years at A&M, Womack has not only influenced the future of
genetics, but also the lives of future geneticists. More than 30 graduate students
such as Rexroad did their doctoral studies under Womack’s tutelage and have gone
on to successful careers in genetics.
In late April, his former graduate students will hold a symposium at A&M
sharing how Womack’s influence, in and out of the lab, benefited their careers. It
will be a time to celebrate, once again, the spark ignited in a young man by a book
and a challenge.
Among his key discoveries, Bazer isolated a uterine protein called uteroferrin and
identified that it is a hematopoietic growth factor that influences the survival
of the neonate and may be useful in treating diseases, such as leukemia and
osteoporosis.
Bazer determined that estrogen in pigs and interferon-t in ruminant species,
are the signals for pregnancy maintenance. The ability of interferon-t to suppress
transcription of the estrogen receptor gene, provides a model for potential treatment
of estrogen-dependent tumors.
F.W. Bazer exemplifies how devotion to basic research in agriculture can lead
to practical outcomes that impact both animal production, as well as human
health and well-being.
715
716 Wolf Prize in Agriculture
1979 Bazer FW, Roberts RM, Basha SMM, Zavy MT, Caton D, Barron DH. Study
of ovine uterine secretions obtained from unilaterally pregnant ewes.
J Anim Sci 49:1522-1527.
1986 Bazer FW, Vallet JL, Roberts RM, Sharp DC, Thatcher WW. Role of conceptus
secretory products in the establishment of pregnancy. J Reprod Fertil
76:841-850.
1989 Vallet JL, Bazer FW. The effect of ovine trophoblast protein-1, oestrogen
and progesterone on oxytocin-induced phosphatidylinositol turnover in
endometrium of sheep. J Reprod Fertil 87:755-761.
1990 Mirando MA, Henry JP, Beers S, Pontzer CH, Torres BA, Johnson HM, Bazer
FW. Onset of secretion of protein with antiviral activity in pig conceptuses.
J Reprod Fertil 88:197-203.
1991 Bazer FW, Worthington-White D, Fliss MFV, Gross S. Uteroferrin: A
progesterone-induced hematopoietic growth factor of uterine origin.
J Exp Hematol 1991; 19:910-915.
1994 Harney JP, Smith LC, Simmen RCM, Fliss AE, Bazer FW. Retinol-binding
protein immunolocalization of protein and abundance of messenger
ribonucleic acid in conceptus and maternal tissues during pregnancy in
pigs. Biol Reprod 50:1126-1135.
1996 Spencer TE, Bazer FW. Ovine interferon tau suppresses transcription of the
estrogen receptor and oxytocin receptor genes in the ovine endometrium.
Endocrinology 137:1144-1147.
1996 VanHeeke G, Ott TL, Strauss A, Ammaturo D, Bazer FW. High yield
expression and secretion of the pregnancy recognition hormone
ovine-τ interferon by Pichia pastoris. J Interferon Res 16:119-126.
1997 Laurenz JC, Hadjisavas M, Chovanic GW, Bazer FW. Myelosuppression in
the pig (Sus scrofa): Uteroferrin reduces the myelosuppressive effects of
5-flourouracil in young pigs. Comp Biochem Physiol 116A:369-377.
1998 Spencer TE, Ott TL, Bazer FW. Expression of interferon regulatory factors
one and two in the ovine endometrium: Effects of pregnancy and ovine
interferon tau. Biol Reprod 58:1154-1162.
1998 Tuo W, Brown WC, Rogers E, Zhu D, Lin G, Smith R, Bazer FW. Trophoblast
IFN-τ differentially induces lymphopenia and neutropenia in lambs.
J Interferon Cytokine Res 18:731-737.
1999 Johnson GA, Spencer TE, Burghardt RC, Bazer FW. 1999. Ovine osteopontin:
I. Cloning and expression of mRNA in the uterus during the peri-
implantation period. Biol Reprod 61: 884-891.
1999 Johnson GA, Burghardt RC, Spencer TE, Newton GR, Ott TL, Bazer FW.
1999. Ovine osteopontin: II. Osteopontin and αvβ3 integrin expression in
the uterus and conceptus during the peri-implantation period. Biol Reprod
61:892-899.
Fuller W. Bazer 717
Originally published in
Fuller W. Bazer 719
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Fuller W. Bazer 723
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Fuller W. Bazer 725
The above article is reprinted with permission from the Society for the Study of Reproduction, Inc.
726 Wolf Prize in Agriculture
Stainsall Detection of OPN Protein One clone, found to be large enough to potentially contain
Endometrium was thawed and homogenized, and the con- the entire coding sequence, was sequenced in both direc-
centration of protein was determined as described for Western tions and the nucleotide sequence for ovine OPN cDNA
blotting. Protein (120 g) in endometrial extracts was dena- (clone 10.2.1) is shown in Figure 2. The inferred amino
tured in Laemmli buffer and separated in 10% gels by two- acid sequence predicts that ovine OPN is a single chain
dimensional (2D) SDS-PAGE. After electrophoresis, gels protein of 278 amino acids with a hydrophobic leader se-
were fixed (10% acetic acid:45% methanol:45% H2O) for 1 quence of 16 amino acids characteristic of secreted pro-
h at room temperature, and then soaked in 50% methanol: teins. Ovine OPN contains 1) a potential calcium phosphate
50% H2O for 4 h with two changes of this solution to remove apatite binding region of consecutive Asp residues begin-
acetic acid. Phosphoproteins were detected by staining over- ning at amino acid 86, 2) a cell-attachment Gly-Arg-Gly-
night at room temperature (in the dark) with Stainsall [24]: 5 Asp-Ser (GRGDS) sequence (residues 151–155), 3) a
mg Stainsall (1-ethyl-2-[3-(1-ethylnaphtho[1,2-D]thiazolin-2- thrombin KS cleavage site (residues 161 and 162), and 4)
ylidene)-2-methylpropenyl]naphtho-[1,2-D]thiazolium bro- two glutamines that are recognized substrates for transglu-
mide; Sigma, St. Louis, MO) in 5 ml formamide, 25 ml iso- taminase in other species.
propanol, 0.5 ml 3 M Tris-HCl (pH 8.8), and 69.5 ml H2O. Nucleotide and amino acid similarities of ovine OPN
Gels were then soaked in 40% methanol/60% H2O, placed on with bovine, rat, mouse, and porcine OPN are summarized
in Table 1. Sequences that are highly conserved in all of
a gel light-box, and photographed through transmitted white
these species include 1) four of the 9 or 10 residues in the
light.
poly-Asp region, 2) the GRGDS, 3) 15 serines that include
an SSEEK sequence (residues 26–30), 4) three threonines
Statistical Analysis (residues 131, 140, and 145), 5) an NES sequence (residues
Data were subjected to least-squares analysis of variance 79–81), and 6) glutamines at positions 50 and 52. A feature
(ANOVA) using the general Linear Models (GLM) proce- of the ovine OPN sequence, shared with bovine OPN only,
dures of the Statistical Analysis System (Users guide 1990, is deletion of 22 amino acids that would otherwise be in-
Ver. 6; SAS Institute, Cary, NC). Slot blot hybridization serted between residues 196 and 197. Also, ovine and bo-
data (total counts) were adjusted for differences in sample vine OPN have a KS, rather than RS putative thrombin
loading using the 18S rRNA data as a covariate [25]. All cleavage site.
tests of statistical significance were performed using the The OPN cDNA detected a ⬃1-kilobase (kb) mRNA in
appropriate error terms according to the expectation of Northern blot analysis of ovine endometrial total RNA (Fig.
mean squares. Data are presented as least-square means 3A). The amount of OPN mRNA increased in pregnant
(LSM) with standard errors (SE). ewes. Steady-state levels (Fig. 3B) did not increase in cy-
clic ewes but did increase in pregnant ewes after Day 17
RESULTS (P ⬍ 0.05).
In situ hybridization analysis of cyclic and pregnant uteri
Western blot analyses of ovine uterine flushings from revealed two distinct patterns of OPN mRNA expression
Day 15 cyclic and pregnant ewes indicated the presence of (Fig. 4). During the estrous cycle and early pregnancy, ex-
70-kDa and 45-kDa immunoreactive proteins (Fig. 1). The pression of transcript was observed in a small percentage
amount of immunoreactive 70-kDa and 45-kDa proteins of cells scattered throughout the endometrium. The cells
was greater in uterine flushings from pregnant as compared that expressed OPN mRNA became concentrated in the
to cyclic ewes. stratum compactum immediately beneath the LE on Days
Hybridization and screening of the ovine endometrial 1, 5, and 7 of the estrous cycle. This expression was inter-
cDNA library with the 291-bp ovine OPN cDNA fragment mittent, and extended along only a small percentage of the
resulted in isolation of 20 positive plaques. Several plaques total circumference of the uterine wall. In pregnant ewes,
were purified and cDNA inserts recovered by self excision. OPN mRNA was present in the GE, where expression was
728 Wolf Prize in Agriculture
DISCUSSION
This is the first report of OPN expression and secretion
by the ovine uterus. Prior to these studies, it was hypoth-
esized that OPN was the IFN-regulated acidic 70-kDa se-
first detected in some ewes on Day 13 of pregnancy. Ex- FIG. 3. Detection of OPN mRNA in ovine endometrium. A) Northern
pression of OPN mRNA by GE increased markedly in all blot analyses of OPN mRNA (20 g/lane) in ovine endometrium from
cyclic (C) and pregnant (P) ewes. Endometrial OPN mRNA (⬃1 kb) in-
ewes between Days 15 and 19 of pregnancy. However, the creased during early pregnancy. B) Steady-state levels of OPN mRNA in
temporal rise in glandular OPN mRNA varied between ovine endometrium during the estrous cycle and pregnancy. There was a
ewes. Day 19 conceptuses did not express OPN mRNA day ⫻ pregnancy status interaction (P ⬍ 0.05) and, within pregnant ewes,
(Fig. 4). there was an effect of Day (P ⬍ 0.05).
Fuller W. Bazer 729
FIG. 4. In situ hybridization analysis of OPN mRNA in ovine endometrium during the estrous cycle (this page, C panels) and pregnancy (opposite
page, P panels). C panels) Corresponding brightfield and darkfield images of endometrium in cycling animals. Note OPN transcript in scattered cells
immediately beneath LE on Days 1, 5, and 7 of the estrous cycle (1C, 5C, 7C) and the decline in number of these cells later in the cycle. A representative
section from Day 1 hybridized with radiolabeled sense cRNA probe (1CS) serves as a negative control. P panels) Brightfield and darkfield images of
endometrium in pregnant animals. Note that OPN transcript appears in GE and increases during pregnancy but is absent in the Day 19 conceptus (19P,
TR). A representative section from Day 19 of pregnancy hybridized with radiolabeled sense cRNA probe (19PS) serves as a negative control. All sections
represent examples of maximal OPN mRNA expression for each day observed. LE, Luminal epithelium; GE, glandular epithelium; S, stroma; TR,
trophectoderm. ⫻60.
730 Wolf Prize in Agriculture
FIG. 4. Continued.
cretory protein. This hypothesis was reinforced by the pres- flushings, steady-state levels of endometrial OPN mRNA
ence of increased OPN protein in uterine flushings from increased during pregnancy. In situ hybridization analysis
pregnant ewes. However, when endometrial protein extracts indicated an increase in OPN mRNA in the GE of some
from Day 17 pregnant ewes were examined by 2D-PAGE, pregnant ewes as early as Day 13, and all ewes by Day 19.
the 70-kDa OPN protein exhibited a number of isoelectric In cyclic and pregnant ovine uteri, OPN mRNA was local-
variants ranging in pI from 5–6. Because the IFN-regu- ized to different cell types. In cyclic ewes, OPN mRNA
lated 70-kDa protein has a single pI of 4, identification of was detected in scattered immune cells that represent a
this protein as OPN is questionable. small percentage of total cells in the endometrium. Secre-
The cDNA sequence for ovine OPN shares high homol- tion of OPN by these cells may be responsible for the pres-
ogy with those for other mammalian species [28–31] and ence of the 70- and 45-kDa OPN forms in uterine flushings
is most similar to bovine OPN (91.7% amino acid sequence from cyclic ewes. In pregnant ewes, there was significant
similarity) [21]. Features of OPN are that it 1) is a glyco- induction of OPN gene transcript in GE. This increase of
sylated phosphoprotein, 2) contains an RGD peptide inte- GE expression is likely responsible for increased OPN pro-
grin binding sequence, 3) is secreted, and 4) is susceptible tein in uterine flushings from pregnant ewes.
to cleavage by proteases [9]. The inferred OPN amino acid Pregnancy-specific expression of OPN mRNA in ovine
sequence is consistent with these properties. Ovine OPN uterine epithelial cells is not surprising. In humans, OPN
has a 16 amino acid hydrophobic signal peptide, GRGDS mRNA has been localized to luminal epithelial cells of the
cell attachment and KS thrombin cleavage sites, 41 serines gastrointestinal tract, gall bladder, pancreatic ducts, distal
of which 13 phosphoserines are found in the same location tubules of the kidney, lung, thyroid, GE of the breast, mu-
in all species, and three threonine O-glycosylation sites as cinous epithelium of the endocervix, and fallopian tubes
well as a potential N-glycosylation (NES) site. [13]. The secretory phase glands of both nonpregnant and
Consistent with the presence of OPN protein in uterine pregnant human uteri express OPN [13, 32]. In mice, the
uterine epithelium, adjacent to deciduoma elicited in pseu- 4. Vallet JL, Barker PJ, Lamming GE, Skinner N, Huskisson NS. A low
dopregnant females, expresses OPN, while the epithelium molecular weight endometrial secretory protein which is increased by
ovine trophoblast protein-1 is 2 microglobulin-like protein. J Endo-
in the contralateral horn does not [12]. Results of the pres- crinol 1991; 130:R1-R4.
ent study indicate that OPN mRNA is not present in LE or 5. Austin KJ, Ward SK, Teixeira MG, Dean VC, Moore DW, Hansen
the conceptus but restricted to the endometrial glands of TR. Ubiquitin cross-reactive protein is released by the bovine uterus
Day 13–19 pregnant ewes. in response to interferon during early pregnancy. Biol Reprod 1996;
The role of OPN during the estrous cycle is unknown. 54:600–606.
The OPN-positive cells in uterine stroma are probably im- 6. Teixeira MG, Austin KJ, Perry DJ, Dooly VD, Johnson GA, Francis
BR, Hansen TR. Bovine granulocyte chemotactic protein-2 is secreted
mune cells because 1) OPN/Eta-1 has been cloned and se- by the endometrium in response to interferon-tau (IFN-). Endocrine
quenced from activated T helper lymphocytes [33] and may 1997; 6:31–37.
be the most abundant protein secreted by various T lym- 7. Ott TL, Spencer TE, Lin JY, Kim H-T, Gerami B, Bartol FF, Wiley
phocyte populations [11, 34, 35]; 2) OPN/Eta-1 is ex- AA, Bazer FW. Effects of the estrous cycle and early pregnancy on
pressed by monocytes and macrophages after tissue injury uterine expression of Mx protein in sheep (Ovis aries). Biol Reprod
[36]; 3) osteoclasts, which deposit OPN/Eta-1 into lacunae 1998; 59:784–789.
8. Short EC, Geisert RD, Helmer SD, Zavy MT, Fulton RW. Expression
of resorbing bone, are specialized tissue macrophages [37]; of antiviral activity and induction of 2⬘,5⬘-oligoadenylate synthetase
and 4) OPN/Eta-1 has been detected in activated CD8⫺ by conceptus secretory proteins enriched in bovine trophoblast pro-
CD4⫺ natural killer cells, including granulated metrial tein-1. Biol Reprod 1991; 44:261–268.
gland cells [39]. The immunological role of OPN/Eta-1 9. Butler WT, Ridall AL, McKee MD. Osteopontin. In: Bilezikian JP,
may be significant since it binds the CD44 receptor on lym- Raisz LG, Rodan GA (eds), Principals of Bone Biology. New York:
phocytes and monocytes to induce chemotaxis of these cells Academic Press, Inc.; 1996: 167–181.
10. Weber GF, Cantor H. The immunology of Eta-1/osteopontin. Cytokine
out of the bloodstream and into sites of inflammation [10]. Growth Factor Rev 1996; 7:241–248.
The OPN/Eta-1 gives rise to 24- and 45-kDa proteins that 11. Fresno M, McVay-Boudreau L, Nabel G, Cantor H. Antigen-specific
bind antigen and suppress T-helper cells, respectively [11]. T-lymphocyte clones. II. Purification and biological characterization
And, the 45-kDa fragment increases immunoglobulin pro- of an antigen-specific suppressive protein synthesized by cloned T-
duction by B lymphocytes [39]. cells. J Exp Med 1981; 153:1260–1274.
During early pregnancy, OPN may act as an adhesion 12. Nomura S, Wills AJ, Edwards JK, Heath JK, Hogan BLM. Devel-
opmental expression of 2ar (osteopontin) and SPARC (osteonectin)
molecule. It binds primarily to ␣v3 integrin heterodimer RNA as revealed by in situ hybridization. J Cell Biol 1988; 106:441–
on tissues via its GRGDS sequence to promote cell-cell 450.
attachment, cell spreading, and cell-extracellular matrix 13. Brown LF, Berse B, Van DeWater L, Papadopoulos-Sergiose A, Pe-
communication [40, 41]. In women [42], expression of ␣v3 ruzzi CA, Manseau EJ, Dvorak HF, Senger DR. Expression and dis-
is unique to the ‘‘implantation window’’ and increases in tribution of osteopontin in human tissues: widespread association with
response to progesterone in uteri of baboons [43]. Temporal luminal epithelial surfaces. Mol Biol Cell 1992; 3:1169–1180.
14. Senger DR, Perruzzi CA, Papadopoulos A, Tenen DG. Purification of
and spatial alterations in expression of extracellular matrix a human milk protein closely similar to tumor secreted phosphopro-
proteins (ECMs) and integrins by the uterus and concep- teins and osteopontin. Biochim Biophys Acta 1989; 996:43–48.
tuses of pigs during the periimplantation period have also 15. Khori K, Nomura S, Kitamura Y, Nagata T, Yoshioka K, Iguchi M,
been reported [44]. Thus, our working hypothesis is that Yamate T, Umekawa T, Suzuki Y, Shinohara H. Structure and expres-
modulation of expression of OPN by progesterone and/or sion of the mRNA encoding urinary stone protein (osteopontin). J Biol
sheep trophoblast interferons may induce expression and Chem 1993; 268:15180–15184.
16. Mesrogli M, Schneider J, Maas DHA. Early pregnancy factor as a
secretion of OPN by uterine epithelium. This OPN then marker for the earliest stages of pregnancy sera. J Biol Chem 1988;
binds ␣v3 integrin heterodimer expressed by trophecto- 264:2266–2271.
derm and/or uterus to 1) stimulate changes in morphology 17. Morton H, High V, Clunie GJA. Immunosuppression detected in preg-
of trophectoderm and extra-embryonic endoderm that result nant mice by rosette inhibitor test. Nature 1974; 249:459–460.
in elongation of the conceptus and 2) induce adhesion be- 18. Clark IQ, Orosco C, Cock IE, Clark FM. The ‘early pregnancy factor’
tween luminal epithelium and trophectoderm essential for revisited: the effect of ammonium sulfate on the capacity pf pregnant
attachment and superficial implantation. Experiments de- mouse sera to express activity in the rosette inhibitor assay. J Reprod
Fertil 1994; 100:279–289.
signed to determine if OPN expression is regulated in vivo 19. Weise DW, Newton GR, Emish GC. The effects of day of the estrous
by progesterone and/or IFN are in progress. cycle or pregnancy on protein secretion by caprine endometrial tissue.
Biol Reprod 1993; 49: 522–527.
ACKNOWLEDGMENTS 20. Fisher LW, Stubbs III JT, Young MF. Antisera and cDNA probes to
human and certain animal model bone matrix noncollagenous pro-
The authors thank Dr. Marian F. Young for the rabbit polyclonal anti- teins. Acta Orthop Scand 1995; 66:61–65.
bodies to recombinant human OPN (LF-123 and LF-124); Dr. Mary C. 21. Kerr JM, Fisher LW, Termine JP, Young MF. Cloning and RNA dis-
Farach-Carson of the University of Delaware Biological Sciences Depart- tribution of bovine osteopontin. Gene 1992; 108:227–234.
ment for advice concerning Stainsall detection of OPN; and Dr. Shawn W. 22. Ausebel FM, Brent R, Kinston RE, Moore DD, Seidmen JG, Smith
Ramsey and Mr. Todd Taylor of the Texas A&M University Sheep & Goat HA, Struhl KA. Current Protocols in Molecular Biology. New York:
Center for care and management of ewes. Photomicrographs were prepared John Wiley and Sons; 1992.
using facilities in the College of Veterinary Medicine Image Analyses 23. Spencer TE, Ing NA, Ott TL, Mayes JS, Becker WC, Watson GH,
Laboratory, which is supported, in part, by NIH Grant P30 ES09106. Mirando MA, Bazer FW. Intrauterine injection of ovine interferon-tau
(oIFN-) alters oestrogen receptor and oxytocin receptor expression in
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31. Wrana JL, Zhang Q, Sodek J. Full length cDNA sequence of porcine tegrin stimulates immediate cell signals in osteoclasts. J Biol Chem
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Fuller W. Bazer 733
Center for Animal Biotechnology and Genomics (H.K., L.A.J., G.A.J., T.E.S., F.W.B.) and Departments
of Animal Science (H.K., G.A.J., T.E.S., F.W.B.) and Veterinary Anatomy and Public Health (L.A.J.),
Texas A&M University, College Station, Texas 77843-2471
2303
734 Wolf Prize in Agriculture
2304 REGULATION AND FUNCTION OF KGF IN THE PORCINE UTERUS Endo • 2001
Vol. 142 • No. 6
estrogens and/or P4 may be responsible for the increased charcoal-stripped serum, antibiotics, 2 mm glutamine (Sigma), and 0.1
KGF expression in the porcine uterine endometrium during U/ml bovine insulin (Sigma).
early pregnancy.
During early pregnancy, pig conceptuses undergo dra- [3H]Thymidine incorporation assay
matic changes in morphology and differentiation in prepa-
Effect of KGF on proliferation of pTr cells was determined as de-
ration for implantation and placentation (13, 14). It is well scribed previously (19). Briefly, pTr cells were plated at 20,000 cells/cm2
known that KGF stimulates the proliferation and migration in DMEM/F-12 containing 5% FBS, then serum-starved for 24 h in
of various epithelia and also affects cellular differentiation serum-free DMEM/F-12, containing 2 mm glutamine and 0.1% BSA.
processes (1). The biological activity of KGF is achieved Cells were then treated with recombinant rat KGF (rKGF; 0, 1, 10, or 100
ng/ml) for 24 h at 37C in serum-free DMEM/F-12 containing 5 Ci/ml
through intracellular signaling activated by KGFR, and KGF [3H]thymidine, precipitated in 10% trichloroacetic acid for 30 min on ice,
activates phosphorylation of KGFR and the mitogen-acti- and fixed in cold methanol. The fixed cells were solubilized in 0.6 ml
vated protein kinase (MAPK) pathway (15). Given that KGF 0.05% trypsin/0.1% SDS for 30 min at 37 C. [3H]Thymidine incorpora-
is a component of histotroph, it may stimulate the prolifer- tion was counted using an LS 3801 liquid scintillation counter (Beckman
ation and differentiation of the conceptus. Therefore, the Coulter, Inc., Palo Alto, CA). The total DNA content was determined
using Picogreen (Molecular Probes, Inc., Eugene, OR) as described by the
objectives of this study were to determine: 1) the effects of manufacturer. Data are expressed as disintegrations per min/g total
estrogens and P4 on KGF expression in the porcine uterine DNA.
endometrium, and 2) the effects of KGF on proliferation and
differentiation of porcine conceptus trophectoderm in vitro.
Northern and slot blot hybridization analysis
Materials and Methods Total cellular RNA was isolated from endometrial explant tissues and
cultured pTr cells using TRIzol reagent (Life Technologies, Inc.). Ex-
Animals and tissue collection pression of KGF in explant tissues and of urokinase-type plasminogen
Experimental and surgical procedures involving animals were ap- activator (uPA) in pTr cells was determined by Northern blot and slot
proved by the agricultural animal care and use committee of Texas A&M blot hybridization analyses as described previously (16). Twenty mi-
University (Animal Use Protocol 2000 –120). Sexually mature, cross-bred crograms of total cellular RNA were hybridized with 32P-radiolabeled
gilts were observed daily for estrous behavior and were either bred or antisense complementary RNA probes generated against a linearized
allowed to continue cycling. For endometrial explant culture, gilts (n ⫽ 690-bp porcine KGF partial complementary DNA (cDNA) (4), 511 bp
3) were hysterectomized on day 9 postestrus, and uteri were transported bovine uPA partial cDNA (provided by Dr. A. R. Menino, Jr., Oregon
on ice directly to the laboratory and processed. For collection of cyclic State University, Corvallis, OR), or 18S ribosomal RNA (pT718S, Am-
and pregnant endometrium, gilts (n ⫽ 3 gilts/day) were hysterecto- bion, Inc., Austin, TX). Autoradiographs of Northern blots to determine
mized on days 9, 12, and 15 of the estrous cycle and on days 9, 10, 12, the size of the uPA transcript were prepared using Kodak X-OMAT
15, 30, and 60 of pregnancy as described previously (16). Conceptuses x-ray film (Eastman Kodak Co., Rochester, NY). The radioactivity in each
from days 9, 12, and 15 of pregnancy were obtained at hysterectomy by slot was quantified using a Packard Instant Imager (Packard, Meriden,
flushing the uterine horns with 40 ml Hanks’ Balanced Salt Solution CT) and is expressed as total counts.
(Sigma, St. Louis, MO). Tissue samples for paraffin sections were fixed
in 4% paraformaldehyde in PBS (pH 7.2) as described previously (16).
RT-PCR
Explant culture Expression of KGFR and interferon-␦ (IFN␦) by pTr was determined
by RT-PCR as described previously (20). Five micrograms of total RNA
Endometrium was dissected from myometrium and placed into from pTr cells were reverse transcribed to obtain cDNAs using Super-
warm phenol red-free DMEM/F-12 culture medium (DMEM/F-12; script II reverse transcriptase (Life Technologies, Inc.). Newly synthe-
Sigma) containing penicillin G (100 IU/ml), streptomycin (0.1 mg/ml), sized cDNA was acid-ethanol precipitated, resuspended in 20 l water,
and amphotericin (0.25 g/ml; Life Technologies, Inc., Grand Island, and stored at ⫺20 C. The cDNAs were then diluted (1:10) with sterile
NY), as described previously (17). Endometrium was then minced with water, and templates were amplified by PCR using AmpliTaq DNA
scalpel blades into small pieces (2–3 mm3), and aliquots of 500 mg were polymerase (Perkin-Elmer Corp., Foster City, CA) and specific primers
placed into culture dishes (100 ⫻ 15 mm) with serum-free modified based on the human KGFR (GenBank accession no. M80637; forward,
DMEM/F-12 containing 10 g/ml insulin (Sigma, catalogue no. I5500), 5⬘-TCTGTTCAATGTGACCGAGG; reverse, 5⬘-GTTTTGGCAGGACA-
10 g/ml transferrin (Sigma, catalogue no. T1428), and 10 ng/ml hy- GTGAGC) or the porcine IFN␦ (GenBank accession no. Z22706; forward,
drocortisone (Sigma, catalogue H0396). Endometrial explants were cul- 5⬘-ATGGATTGTCCCCATGTAGG; reverse, 5⬘-CTGAGCTACCAGG-
tured immediately after mincing in the presence of E2 (0, 0.05, 0.5, 5, and GTTACCG). PCR conditions were 35 cycles of 95 C for 30 sec, 55 C for
50 ng/ml), P4 (0, 0.03, 0.3, 3, and 30 ng/ml), catechol estrogens [5 ng/ml KGFR or 58 C for IFN␦ for 30 sec, and 72 C for 1 min. PCR products (104
2-hydroxy-E2 (2OH-E2) or 5 ng/ml 4OH-E2], estrogen receptor (ER) bp for KGFR and 296 bp for IFN␦) were separated in 2% agarose gels
antagonist [50 ng/ml ICI 182,780 (ICI)], or E2 (5 ng/ml) plus P4 (3 ng/ml) and visualized by ethidium bromide staining. The identity of each am-
for 48 h with rocking under an atmosphere of 45% nitrogen, 5% carbon plified PCR product was verified by sequence analysis after cloning into
dioxide, and 50% oxygen. E2 (catalogue no. E8875), P4 (catalogue no. the pCRII vector (Invitrogen, Carlsbad, CA).
P0130), 2OH-E2 (catalogue no. H3131), and 4OH-E2 (catalogue no.
H4637) were obtained from Sigma, and ICI was purchased from Tocris
(Ballwin, MO). Explant tissues were then harvested, and RNA was Immunohistochemistry
extracted for slot blot analysis of KGF expression. These experiments
were conducted using endometrium from three individual gilts. Treat- Expression of immunoreactive proliferating cell nuclear antigen
ments were performed in triplicate using tissues obtained from each gilt. (PCNA) was evaluated in conceptus and paraformaldehyde-fixed, par-
affin-embedded, uterine tissue cross-sections (5 m) using 2 g/ml of
Porcine trophectoderm cells a monoclonal antibody to PCNA (Santa Cruz Biotechnology, Inc., Santa
Cruz, CA; catalogue no. sc-56) and a Super ABC Mouse IgG Kit
Porcine trophectoderm cells were isolated using nonenzymatic dis- (Biomeda, Foster City, CA), and procedures described previously (16).
persion of trophoblast from conceptuses collected on day 12 of gestation A boiling citrate buffer antigen retrieval protocol was used to reveal the
(18). A trophectoderm cell line (pTr) was established by repeated pas- PCNA according to the manufacturer’s recommendations. Purified nor-
sage and culture of the cells on Primaria tissue culture plastic (Falcon, mal mouse IgG (Sigma) at 2 g/ml was substituted for mouse anti-
Lincoln Park, NJ). Cells were maintained in DMEM/F-12 containing 5% PCNA and served as a negative control.
Fuller W. Bazer 735
2306 REGULATION AND FUNCTION OF KGF IN THE PORCINE UTERUS Endo • 2001
Vol. 142 • No. 6
extracellular signal-regulated kinases 1 and 2 (ERK1/2), was assessed by to ERK1/2 (1:400 dilution in 2% milk-TBST; catalogue no. sc-94-g) were
immunoprecipitation and Western blot analysis. Briefly, monolayer cul- obtained from Santa Cruz Biotechnology, Inc. Blots were then incubated
tures of pTr cells were grown to 75% confluence on 75-cm2 tissue culture with rabbit antigoat IgG or goat antimouse IgG conjugated to peroxidase
flasks and then incubated in serum-free DMEM/F-12 containing 0.1% (Kirkegaard & Perry Laboratories, Bethesda, MD) for 1 h at room tem-
BSA for 48 h. Whole cell extracts were prepared as previously described perature, and immunoreactive proteins were detected using enhanced
(22). The protein concentration of the lysate supernatant was determined chemiluminescence (Amersham Pharmacia Biotech, Arlington Heights,
by Bradford assay (Bio-Rad Laboratories, Inc., Burlingame, CA) using NY) according to the manufacturer’s recommendations.
BSA as the standard and 1 mg of each extract used for immunoprecipi-
tation. Five micrograms of KGFR antibody (Santa Cruz Biotechnology, Effect of KGF on pTr cell differentiation
Inc., catalogue no. sc-122) or normal rabbit IgG were added to each
extract, and bound proteins were purified using protein A/G plus aga- To determine whether KGF affects functional cell differentiation, pTr
rose as described previously (22). Immunoprecipitated proteins were cells were treated with increasing doses of rKGF (0, 1, 10, and 100 ng/ml)
separated by SDS-PAGE and analyzed by Western blotting (as described in serum-free DMEM/F-12 for 24 h. Expression of uPA was used as a
below) with antibody to phosphotyrosine (Santa Cruz Biotechnology, marker for pTr cell differentiation by Northern and slot blot analyses.
Inc.; catalogue no. sc-7020) diluted 1:100 in 5% BSA-TBST (Tris-buffered
saline/0.1% Tween-20). Statistical analysis
In similar experiments pTr cells were serum-starved and treated with
rKGF as described above. In addition to KGF treatment, some cells were All quantitative data were subjected to least squares ANOVA using
pretreated with 0 or 50 m of the MAPK/ERK kinase 1 (MEK1) inhibitor, the general linear models procedures of the Statistical Analysis System
PD98059 (New England Biolab, Beverly, MA; catalogue no. 9900L) for (SAS Institute, Inc., Cary, NC) (23). Data from dose-response studies on
1 h, then treated with 0 or 10 ng/ml rKGF for 60 min. Twenty micro- KGF and uPA expression were analyzed by least squares regression
grams of whole cell extract protein from each sample were separated by analysis. Slot blot data (total counts) were analyzed using the 18S ri-
SDS-PAGE and transferred to nitrocellulose as described previously bosomal RNA data as a covariate to correct for differences in sample
(21). Blots were blocked for 4 h at 4 C with either 5% BSA-TBST for loading. Preplanned contrasts (control vs. E2 plus ICI; E2 vs. E2 plus ICI;
phospho-specific antibodies or 5% nonfat milk-TBST for other antibod- control vs. catechol estrogens; control vs. E2; P4 vs. E2 plus P4) were used
ies, and then incubated with primary antibodies overnight at 4 C. Mono- to test for effects of treatments in slot blot analyses. All tests of statistical
clonal antibodies to phospho-ERK1/2 (pERK1/2; 1:400 dilution in 5% significance were performed using the appropriate error terms accord-
BSA-TBST; catalogue no. sc-7383) and phospho-tyrosine (1:100 dilution ing to the expectation of mean squares. Data are presented as least
in 5% BSA-TBST; catalogue no. sc-7020), and goat polyclonal antibody squares means with ses.
Fuller W. Bazer 737
Results
Effects of estrogens on KGF expression
KGF expression was increased (quadratic, P ⬍ 0.05) by E2
(Fig. 1A). The increase in KGF expression by E2 was blocked
by addition of the ER antagonist, ICI (Fig. 1B; E2 vs. E2 plus
ICI, P ⬍ 0.05). Catechol estrogens, 2OH-E2 and 4OH-E2, did
not affect KGF expression in endometrium (Fig. 1C; control
vs. 2OH-E2 and control vs. 4OH-E2, P ⬎ 0.05).
2308 REGULATION AND FUNCTION OF KGF IN THE PORCINE UTERUS Endo • 2001
Vol. 142 • No. 6
mediated decrease in KGF expression is not known. Inter- alter conceptus trophectoderm cell differentiation in this
estingly, E2 increased KGF expression in the presence of P4, study. In various cell types, uPA, aromatase, surfactant pro-
a situation to which the endometrial epithelium would be tein A and D, syndecan-1, and Na⫹/K⫹-adenosine triphos-
exposed in vivo during pregnancy. As P4 down-regulates phatase are increased by KGF (1, 37, 38). In particular, KGF
endometrial ER␣ in ovine uterus (16, 35, 36), the decreased increases uPA expression and activity in human uterine exo-
expression of KGF in endometrial explants treated with P4 cervical epithelial cells (39) and keratinocytes (40). In this
alone and E2 plus P4 may be the result of a P4-mediated study KGF increased uPA expression in pTr cells. Differen-
decrease in ER and an attenuated response to E2. We cannot tiating pig conceptuses produce uPA from trophoblast (41),
rule out the effects of any residual levels of estradiol on and pig conceptus trophectoderm produces uPA in a bipha-
decreased KGF expression by P4 in this system. It is also sic manner between days 10 and 12 and between days 14 and
possible that P4 inhibits KGF expression through a direct 16 of early pregnancy (42), coinciding with estrogen pro-
mechanism or indirectly by modulation of other unknown duction by conceptuses. Therefore, increased expression of
factors which then down-regulate KGF expression. uPA by KGF in pTr cells suggests that KGF expression in-
KGF affects epithelial cell proliferation in various tissues creases within endometrial LE in response to estrogen, is
(37). In the present study KGF increased DNA synthesis of secreted into the uterine lumen, and may be an important
pTr cells, which are of epithelial cell origin and express regulator of uPA secretion by conceptus trophectoderm.
KGFR, and IFN␦, a porcine trophectoderm cell-specific Like most receptor tyrosine kinase-activating growth fac-
marker (24). Porcine conceptuses undergo dramatic mor- tors, KGFR signals through the MAPK pathway (15). In the
phological changes during early pregnancy (13, 14). Between present study phosphorylation of KGFR and ERK1/2, mem-
days 10 and 12 of pregnancy there is a rapid transition from bers of the MAPK family, was detected in KGF-treated pTr
spherical (9 –10 mm in diameter) to tubular (10 –50 mm in cells, suggesting that effects on trophectoderm proliferation
length) and elongated filamentous forms (⬎100 mm long) and differentiation were mediated by interaction of KGF
(14). In the present study trophectoderm cell proliferation with the KGFR. Among the several FGFR isoforms, KGF
was detected between days 9 and 15 of conceptus develop- recognizes only KGFR with high affinity and biological ac-
ment as reported previously (14). Thus, in vitro results of the tivity (43), precluding the possibility that ERK1/2 is acti-
present study suggest that KGF of endometrial epithelial vated by other members of the FGFR family. The precise
origin may increase the proliferation of conceptus trophec- mechanisms of intracellular KGF signaling for proliferation
toderm during the periimplantation phase of development. and differentiation in pTr cells remain to be determined.
This hypothesis is supported by results from PCNA staining In summary, the results of the present study indicate that
of conceptuses in vivo between days 9 –15 of pregnancy,
E2, a pregnancy recognition signal in pigs, increases KGF
indicating high intensity PCNA staining from day 10 of preg-
expression in the uterine endometrium, and that KGF in-
nancy and thereafter. The PCNA staining in the conceptuses
creases the proliferation of conceptus trophectoderm and
is coordinate with increasing KGF expression in the endo-
stimulates the expression of uPA, a marker for differentia-
metrium between days 10 and 15 of pregnancy (4). The lack
tion. Thus, KGF of endometrial origin affects both the growth
of detection of PCNA in the LE and GE during early preg-
and differentiation of trophectoderm during this crucial
nancy suggests that epithelial cells do not proliferate to any
phase of conceptus development in pigs.
great degree at this time. Therefore, although KGF is present
in the uterine lumen, and KGFR is expressed in LE and GE,
it is unlikely that KGF affects epithelial cell proliferation in Acknowledgments
the endometrium during early pregnancy. However, it is
The authors thank Dr. Robert C. Burghardt for assistance with immuno-
possible that KGF affects differentiation of endometrial ep- fluorescence image capture and use of microscopy and imaging facilities in
ithelial cells. the Image Analysis Laboratory of College of Veterinary Medicine. We also
In addition to effecting proliferation, KGF was found to thank Dr. Wallace L. McKeehan for providing recombinant rat KGF.
740 Wolf Prize in Agriculture
2310 REGULATION AND FUNCTION OF KGF IN THE PORCINE UTERUS Endo • 2001
Vol. 142 • No. 6
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Fuller W. Bazer 741
A&M University, College Station, Texas 77843-2471, USA; and 2Department of Veterinary
Anatomy and Public Health, Texas A&M University, College Station, Texas 77843-4458,
USA
Endometrial glands are necessary for conceptus epithelium with no detectable differences between normal
implantation and growth. In the ovine uterine gland and UGKO ewes. Uterine flushes from pregnant ewes, but
knockout (UGKO) model, blastocysts hatch normally but not cyclic or UGKO ewes, contained abundant
fail to survive or elongate. This peri-implantation defect in immunoreactive interferon τ and the cell adhesion
UGKO ewes may be due to the absence of endometrial proteins, osteopontin and glycosylation-dependent cell
glands or, alternatively, to the lack of certain epithelial adhesion molecule one. In study two, UGKO ewes were
adhesion molecules or the inability of the endometrium to fitted with uterine catheters 5 days after oestrus, infused
respond to signals from the conceptus. Two studies were with recombinant ovine interferon τ or control proteins
performed to examine these hypotheses. In study one, from 11 to 15 days after oestrus, and underwent
normal (n = 8) and UGKO (n = 12) ewes were mated at hysterectomy 16 days after oestrus. Expression of several
oestrus (day 0) with intact rams and their uteri were interferon τ-stimulated genes (ISG17, STAT1, STAT2 and
flushed 14 days after oestrus. Normal ewes (n = 4) were IRF-1) was increased in the endometrium from interferon
also flushed on 14 days after oestrus. Uterine flushes τ-infused UGKO ewes. These results support the
from bred normal ewes contained filamentous con- hypothesis that the defects in conceptus elongation and
ceptuses (n = 7 of 8), whereas those from UGKO ewes survival in UGKO ewes are due to the absence of
contained no conceptus (n = 5 of 12), a growth-retarded, endometrial glands and their secretions rather than to
tubular conceptus (n = 6 of 12), or a fragmented, alterations in expression of anti-adhesive or adhesive
filamentous conceptus (n = 1 of 12). In all groups, molecules on the endometrial luminal epithelium or to the
expression of mucin 1 and integrin αv, α5, β3 and β5 was responsiveness of the endometrium to the conceptus
localized at the apical surface of the endometrial luminal pregnancy recognition signal.
other Müllerian duct-derived female reproductive tract 1999a,b, 2001a) and glycosylation-dependent cell adhesion
structures (Gray et al., 2000b, 2001b). Despite repeated molecule 1 (GlyCAM-1) (Spencer et al., 1999b), are
matings with fertile rams, adult UGKO ewes do not secreted by the endometrial GE and are thought to bind to
establish pregnancy (Gray et al., 2000a, 2001b,c). Transfer trophectoderm and LE to stimulate elongation, adhesion
of hatched blastocysts recovered from superovulated and attachment of the ovine conceptus (Johnson et al.,
normal donor ewes into the uteri of UGKO ewes failed to 2001a).
ameliorate the pregnancy defect (Gray et al., 2001c). The available evidence supports the hypothesis that
Normal hatched blastocysts were found in the uterine the failure of peri-implantation conceptus survival and
flushings of mated UGKO ewes 6 and 9 days, but not 14 elongation in UGKO ewes is due to an absence of
days, after mating (Gray et al., 2001b,c). Uterine flushings endometrial glands and, by default, their secretory
taken 14 days after mating from mated UGKO ewes products. Alternatively, the pregnancy defect in UGKO
contained either no conceptus or a severely growth- ewes could be attributed to deficient expression of adhesion
retarded conceptus that had failed to elongate from a molecules on the LE or inability of the endometrium to
tubular to filamentous form (Gray et al., 2001c). Therefore, respond to conceptus signals such as IFN-τ. Two studies
UGKO ewes exhibit a peri-implantation pregnancy defect, were conducted to test these hypotheses by determining
the timing of which correlates with most of the embryo loss differences in normal and UGKO ewes on: (1) distribution
that occurs during pregnancy in livestock and humans of Muc-1 and integrin subunit expression in the uterine LE
(Bazer, 1975; Kane et al., 1997). 14 days after mating; (2) abundance of osteopontin and
Implantation in ruminants is a highly co-ordinated GlyCAM-1 in the uterine lumen 14 days after mating; and
process that involves apposition, attachment, and adhesion (3) endometrial expression of IFN-τ-stimulated genes in
of the conceptus trophectoderm to the endometrial luminal response to intrauterine administration of recombinant
epithelium (LE) (Guillomot et al., 1981; Guillomot, 1995). ovine IFN-τ.
In sheep, the peri-implantation period is marked by rapid
elongation of the conceptus from a tubular to filamentous
Materials and Methods
form on days 12–13 of gestation and the production of large
amounts of interferon τ (IFN-τ), a type I interferon that is the Animals
signal for maternal recognition of pregnancy (Bazer et al.,
Experimental and surgical procedures complied with the
1997). Elongation of the conceptus is critical for de-
Guide for Care and Use of Agriculture Animals and were
velopmentally regulated production of IFN-τ (Farin et al.,
approved by the Institutional Agricultural Animal Care and
1989) and requires the uterus, as hatched blastocyts fail to
Use Committee of the Texas A&M University System
elongate in vitro unless transferred to the uterus (Heyman et
Agricultural Experiment Station (Animal Use Protocol 7-
al., 1984). Apposition of conceptus trophectoderm and
286).
endometrial LE is initiated on day 14, adhesion occurs on
UGKO ewes were produced as described by Spencer et
day 15, and firm attachment on days 16–18 of gestation
al. (1999a) and Gray et al. (2000a) by implanting crossbred
(Guillomot et al., 1981). Adhesion of conceptus tro-
Rambouillet ewe lambs with a single Synchromate B®
phectoderm to the LE is temporally regulated by non-
(Sanofi, Overland Park, KS) implant within 12 h of birth and
adhesive and adhesive factors on the apical surface of the
every 2 weeks thereafter for a total of 8 weeks. Implants
endometrial LE (Burghardt et al., 1997, in press; Johnson
were inserted s.c. into the periscapular area and released
et al., 2001a). It is hypothesized that, initially, non-
approximately 6 mg norgestomet (17α-acetoxy-11β-
adhesive factors, such as mucin 1 (Muc-1), sterically impair
methyl-19-norpreg-4-ene-3,20-dione), a potent synthetic
interactions between adhesive glycoproteins expressed on
19-norprogestin, over 14 days (Bartol et al., 1988). Normal
the apical surfaces of conceptus trophectoderm and LE
control ewes did not receive implants.
by means of their extensive glycosylation and extended
extracellular structure (Carson et al., 2000; Johnson et al.,
Study one
2001a). In sheep, immunoreactive Muc-1 expression on
the LE decreases progressively between day 9 and day 17 Adult UGKO (n = 18) and normal ewes (n = 12) were
of early pregnancy, presumably to unmask adhesive given two i.m. injections (at 07:00 and 17:00 h) of 10 mg
glycoproteins on the LE for interaction with the tro- prostaglandin F2α (Lutalyse, Upjohn, Kalamazoo, MI) 9 days
phectoderm (Johnson et al., 2001a). Integrins are thought to apart to synchronize oestrus. Ewes were monitored each
be the dominant glycoproteins that regulate trophectoderm day for oestrous behaviour using vasectomized rams. All
adhesion. During the peri-implantation period in ewes, UGKO and some normal control ewes (n = 8) were mated
integrin subunits αv, α4, α5, β1, β3 and β5 were con- at oestrus (day 0) and at 12 and 24 h after oestrus with intact
stitutively expressed on the conceptus trophectoderm as rams of proven fertility. The remaining normal control ewes
well as the apical surface of the endometrial LE (Johnson et (n = 4) were assigned to cyclic status, and oestrus was
al., 2001a). During the peri-implantation period, in addition determined using vasectomized rams.
to constitutive expression of integrins, two molecules Fourteen days after oestrus or mating, all ewes were
involved in cell adhesion, osteopontin (Johnson et al., subjected to mid-ventral laparotomy, and their uterine
Fuller W. Bazer 743
lumina were flushed with 20 ml sterile saline. Uterine black, cytoplasm and muscle fibres red, and extracellular
flushes were analysed under a dissecting microscope to matrix (ECM) components blue. For this stain, uterine
recover conceptuses, if any, and determine their morphology. sections were deparaffinized in CitraSolv (Fisher Scientific;
Conceptuses were fixed in 4% paraformaldehyde in PBS Fairlawn, NJ) and rehydrated through a graded alcohol
(pH 7.2). Uterine flushes were clarified by centrifugation series to distilled water. Tissues were then incubated for 1 h
(2000 g for 30 min at 4⬚C), and aliquots were snap-frozen at 55⬚C in Bouin’s solution (71% (v/v) picric acid, 24%
in liquid nitrogen and stored at –80⬚C. Several sections formaldehyde (40%), and 5% (v/v) glacial acetic acid) and
(1–1.5 cm) from the middle of each uterine horn were rinsed in water. Slides were incubated sequentially at room
snap–frozen in Tissue-Tek OCT compound (Miles, temperature for 5 min each in Weigert’s iron haematoxylin
Oneonta, NY). Several sections from the middle region of (50% (v/v) ethanol (95%), 4% (v/v) ferric chloride (29%
each uterine horn were also fixed in 4% paraformaldehyde aqueous), 1% (v/v) hydrochloric acid, and 1% (w/v)
in PBS (pH 7.2). After 24 h, fixed tissues were changed haematoxylin), biebrich scarlet-acid fuchsin solution (90%
to 70% ethanol for 24 h and then dehydrated and (v/v) biebrich scarlet (1% aqueous), 9% (v/v) acid fuchsin
embedded in Paraplast-Plus (Oxford Labware, St Louis, (1% aqueous), and 1% (v/v) glacial acetic acid),
MO). The endometrium was physically dissected from phosphomolybdic–phosphotungstic acid solution (2.5%
myometrium from the remainder of each uterine horn (v/v) phosphomolybdic acid, 2.5% (w/v) phosphotungstic
ipsilateral to the ovary bearing the corpus luteum and then acid), aniline blue solution (2.5% (w/v) aniline blue, 2%
snap–frozen in liquid nitrogen and stored at –80⬚C for RNA (v/v) glacial acetic acid), and then in 1% glacial acetic acid
extraction. (v/v) for 5 min. Slides were then dehydrated through alcohol
to xylene, and coverslips fixed with Permount (Fisher
Study two Scientific, Fair Lawn, NJ). Photomicrographs of stained
tissues were captured using a Zeiss Axioplan2
Fourteen days after mating, another group of UGKO
photomicroscope (New York, NY) fitted with a Hamamatsu
ewes (n = 8) was subjected to mid-ventral laparotomy, their
chilled 3CCD colour camera (Hamamatsu, Hamamatsu
uterine lumina flushed with 20 ml sterile saline, and their
City).
uterine horns fitted with catheters as described by Spencer
et al. (1995). Ewes were given two i.m. injections (07:00
and 17:00 h) of 10 mg prostaglandin F2α (Lutalyse, Upjohn, Western blot analyses
Kalamazoo, MI) to synchronize oestrus, and were
Uterine flushes (2 ml) from day 14 cyclic, pregnant and
monitored twice a day for oestrous behaviour using a
UGKO ewes were concentrated using Centricon-3 columns
vasectomized ram, and then assigned randomly to receive
(Amicon, Beverly, MA). Protein content was determined
daily intra-uterine injections (at 07:00 and 19:00 h each
using a Bradford protein assay (Bio-Rad, Hercules, CA) with
day) of control proteins (6 mg serum proteins per day) or
bovine serum albumin (BSA) as the standard. Uterine flush
recombinant ovine IFN-τ (roIFN-τ; 2 ⫻ 107 antiviral units
proteins (30 µg) were denatured, separated by SDS-PAGE
per day) from day 11 to day 15 after mating. Recombinant
using 12% acrylamide gels, and transferred to nitrocellulose
oIFN-τ was produced from a synthetic gene construct in
membranes as described by Spencer et al. (1999b).
Pichia pastoris and purified as described by Van Heeke et
Membranes were blocked for 1 h at room temperature with
al. (1996). Preparation of control proteins and roIFN-τ was
5% (w/v) milk–TBST (20 mmol Tris l–1 (pH 7.5), 137 mmol
performed as described by Spencer et al. (1995). Blood
NaCl l–1, 0.05% (v/v) Tween 20) and then incubated with
samples were collected on days 11–15 after mating, via
mouse anti-ovine IFN-τ (HL129; 4 µg ml–1) (Swann et al.,
jugular venepuncture into Vacutainer evacuated blood
1999), rabbit anti-human osteopontin (LF-123 and LF-124;
collection tubes with sodium heparin (Becton-Dickinson,
1 : 2500 each) (Johnson et al., 1999a), or rabbit anti-rat
Franklin Lakes, NJ), and plasma was stored at –20⬚C. All
GlyCAM-1 (CAM02; 1 µg ml–1; kindly provided by S. D.
ewes were hysterectomized 16 days after mating. At
Rosen (University of California) (Singer and Rosen, 1996) in
hysterectomy, portions (approximately 1.0 cm) from the
5% milk-TBST as described by Spencer et al. (1999b).
middle region of each uterine horn were fixed in fresh 4%
Negative control blots were performed in which primary
paraformaldehyde in PBS for 24 h and embedded in
antibody was replaced by mouse IgG (IFN-τ), rabbit IgG
Paraplast-Plus (Oxford Labware). From the remainder of
(GlyCAM-1) or normal rabbit serum (osteopontin) at the
each uterine horn, endometrium was dissected from
same concentration used for the respective primary anti-
myometrium, frozen separately in liquid nitrogen, and
bodies. After an overnight incubation at 4⬚C, membranes
stored at –80⬚C.
were washed for 30 min with TBST and then incubated with
either goat anti-mouse or goat anti-rabbit IgG–horseradish
Histological analyses
peroxidase-conjugated secondary antibody (KPL, Bethesda,
Conceptus tissues were sectioned (5 µm) and stained MD) for 1 h at room temperature. Membranes were again
with haematoxylin and eosin as described by Gray et al. washed with TBST for 30 min before detection by chemi-
(2000a). Uteri were sectioned (5 µm) and stained with luminescence using a Super Signal West Pico kit (Pierce;
Masson’s trichrome stain. This procedure stains nuclei Rockford, IL) and Kodak X-OMAT AR film.
744 Wolf Prize in Agriculture
(a) (b)
sc SC
L
v
ss
SS
G
(c) (d)
(e) (f )
Fig. 1. Representative photomicrographs of the uterus from cyclic (a,b), pregnant (c,d) and uterine gland knockout
(UGKO) ewes (e,f) 14 days after mating, using a Masson’s trichrome stain. This procedure stains nuclei black,
cytoplasm and muscle fibres red, and extracellular matrix (ECM) components blue. The left column (a,c) from normal
ewes depicts intercaruncular endometrial regions and the right column (b,d) depicts caruncular endometrial areas. No
distinct caruncular or intercaruncular regions are distinguishable in uteri from UGKO ewes (e,f). L: luminal epithelium;
G: glandular epithelium; SC: stratum compactum; SS: stratum spongiosum; M: myometrium; V: blood vessel. Scale bar
represents 200 µm.
Myometrial histoarchitecture did not appear different contrast, growth-retarded conceptuses were observed in
between uteri from UGKO and control ewes. the uterine flush of some UGKO ewes. A representative
degenerating tubular conceptus found in the uterine flush of
an UGKO ewe is shown (Fig. 2c).
Study one: IFN-τ, osteopontin and GlyCAM-1 in uterine
Immunoreactive IFN-τ was absent in uterine flushes
flushes
obtained from cyclic ewes (Fig. 2b, lanes 1–4), but detected
Filamentous conceptuses were present in most uterine in uterine flushes obtained from pregnant ewes (Fig. 2b,
flushes from normal ewes 14 days after mating (Fig. 2a). In lanes 5–8). The amount of IFN-τ present in the uterine
746 Wolf Prize in Agriculture
17.4
7.2
1 2 3 4 5 6 7 8
(c) (d) None Tubular Fil
32
17.4
7.2
1 2 3 4 5 6
Fig. 2. Conceptus morphology and uterine flush content of interferon τ (IFN-τ) from cyclic, pregnant and
uterine gland knockout (UGKO) ewes. Photomicrographs (a,c) are haematoxylin and eosin-stained
conceptuses that were present in the uterine flushes. (a) Elongating filamentous conceptus
representative of conceptuses found in the uterine flushes from normal bred ewes 14 days after mating.
(c) Growth-retarded, or tubular, conceptus detected in the uterine flush of an UGKO ewe 14 days after
mating. Photomicrographs are shown at the same magnification (⫻ 164). (b,d) Western blot analyses of
IFN-τ in concentrated uterine flushes obtained from (b) normal ewes on day 14 of oestrus (Cyclic, lanes
1–4) or 14 days after mating (Pregnant, lanes 5–8) and (d) UGKO ewes 14 days after mating. (d) Uterine
flushings from UGKO ewes were loaded depending upon the morphology of the conceptus present in
the uterine flush. No conceptus (lanes 1–2), a growth-retarded or degenerating tubular conceptus (lanes
3–5), or a fragmented filamentous conceptus (Fil; lane 6) was detected in the uterine flushes from UGKO
ewes. Each lane in the blots represents proteins from a single flush from an individual ewe (30 µg per
lane). Immunoreactive protein was detected using mouse monoclonal antibody directed against ovine
IFN-τ. Positions of pre-stained molecular weight standards (⫻ 10–3) are indicated.
flushes corresponded to the number of conceptuses present (Johnson et al., 1999a). The 70 kDa form of osteopontin was
in the uterine flush. The uterine flush from the ewe present in only one uterine flush (Fig. 3a, lane 5), which
represented in lane 5 contained three filamentous contained three filamentous conceptuses. Immunoreactive
conceptuses, whereas those in lanes 6 and 7 were from osteopontin was not detectable in any uterine flush
ewes with two conceptuses, and lane 8 represents a flush obtained from UGKO ewes, regardless of the presence of a
that contained a single conceptus. Western blot analyses of degenerating conceptus (Fig. 3b, lanes 1–6).
uterine flushes that contained a conceptus from UGKO Immunoreactive GlyCAM-1 was detectable in two of the
ewes is presented (Fig. 2d). Immunoreactive IFN-τ was not uterine flushes obtained from cyclic ewes (Fig. 4a, lanes 2
detected in uterine flushes from UGKO ewes containing no and 4), but abundant in all uterine flushes obtained from
conceptus (lanes 1 and 2), degenerating tubular pregnant ewes (Fig. 4a, lanes 5–8). Small amounts of
conceptuses (lanes 3 and 4), or a fragmenting filamentous GlyCAM-1 were detected in uterine flushings from three
conceptus (lane 6). Immunoreactive IFN-τ was within UGKO ewes (Fig. 4b, lanes 1, 3 and 5). However, the
detection limits in the uterine flush of one UGKO ewe that amount of GlyCAM-1 in uterine flushings from UGKO ewes
contained a degenerating, tubular conceptus (lane 5). was not abundant and appeared similar to that in normal
Immunoreactive osteopontin was absent in uterine cyclic ewes.
flushings from cyclic ewes (Fig. 3a, lanes 1–4), but present
in uterine flushings from pregnant ewes (Fig. 3a, lanes 5–8).
Study one: expression of Muc-1 and integrin subunits in
Several molecular mass forms (70, 45, and 25 kDa) of
the endometrium
immunoreactive osteopontin were detected in the uterine
flushings from pregnant ewes. The 45 and 25 kDa forms of Immunoreactive Muc-1 was restricted to the apical
osteopontin are cleavage products of the 70 kDa form surface of uterine LE and GE (Fig. 5). Apical staining of
Fuller W. Bazer 747
v
5
3
5
MUC-1
Fig. 5. Immunofluorescent localization of integrins αv, α5, β3, β5 and MUC-1 in the endometrium of normal ewes on day 14
of oestrus (Cyclic; left column) or 14 days after mating (Pregnant; middle column), or uterine gland knockout (UGKO) ewes
after mating (UGKO; right column). Frozen cross-sections of the uterine wall from control and UGKO ewes were fixed in
methanol and blocked in 5% normal goat serum. Sections were incubated overnight at 4⬚C with 20 µg ml–1 rabbit IgG
against αv, α5, β3, β5 or MUC-1, and then incubated with fluorescein-conjugated goat anti-rabbit IgG. Photomicrographs
were taken using a fluorescein filter. Scale bar represents 200 µm.
Fuller W. Bazer 749
conceptus or a growth-retarded conceptus on day 14 after in agreement with previous studies in normal cyclic ewes
mating. Taken together, the evidence indicates that infused with IFN-τ (Spencer et al., 1999c; Johnson et al.,
endometrial glands and, by default, their secretions are not 2000; Choi et al., 2001). Taken together, available results
required for development of the conceptus to the hatched indicate that the endometrium of UGKO ewes can respond
blastocyst state, but are crucial for blastocyst elongation. properly to the pregnancy recognition signal produced by
Normal conceptuses by day 14 after mating have de- the ovine conceptus, but is unable to produce the proper
veloped from a tubular to an elongated filamentous form, secretions to stimulate conceptus development.
secreted large amounts of IFN-τ, and transiently contacted The process of superficial implantation in sheep involves
the maternal epithelium in preparation for implantation a sequence of events, including removal of anti-adhesion
(Guillomot, 1995). molecules on the LE, expression of receptors on the LE and
In the present study, uterine flushes from UGKO ewes on trophectoderm, and secretion of adhesive factors by the
day 14 after mating had either no detectable or very low endometrial LE and GE (Bowen and Burghardt 2000;
amounts of IFN-τ associated with severely growth retarded Johnson et al., 2001a; Burghardt et al., in press). Muc-1 is a
conceptuses. Secretion of IFN-τ by the trophectoderm heavily glycosylated mucin thought to project above the
serves as a marker for conceptus health and state of apical surface of LE cells to sterically block cell–cell and
development. IFN-τ is secreted by the ovine conceptus cell–ECM adhesion (Wesseling et al., 1995) and, thereby,
between days 8 and 21 after mating, with maximum trophectoderm access to the uterine LE. In both humans and
amounts secreted on days 14–16 as the conceptus develops rodents, the expression pattern of glycoproteins, such as
from a tubular to a filamentous form (Ashworth and Bazer, MUC-1, on the uterine LE may control the accessibility
1989; Farin et al., 1989). In the present study, the uterine of integrin receptors to their ligands and provide a barrier
flush of UGKO ewes did not contain the high concen- to invasiveness (Carson et al., 2000). The implantation ad-
trations of immunoreactive IFN-τ characteristic of uterine hesion cascade in rodents is initiated after downregulation
flushings from normal ewes on day 14 after mating. The of MUC-1 (Carson et al., 1998). In sheep, MUC-1 is also
lack of appreciable IFN-τ in the uterine flush of UGKO ewes expressed on the apical surface of the LE and is down-
is correlated directly with the lack of a conceptus or the regulated during implantation to allow interactions of
presence of a growth-retarded tubular conceptus in the uteri integrins and their ligands (Johnson et al., 2001a). Integrins
of UGKO ewes. comprise a family of intrinsic membrane proteins of non-
In ewes, IFN-τ acts on the endometrium to induce or covalently linked α and β subunit heterodimers. Integrins
increase the expression of several inteferon-stimulated genes. serve as receptors and bind ECM and other ligands to aid in
IFN-τ-stimulated genes include ISG17/ubiquitin cross- cellular adhesion, reorganization of cytoskeletal molecules
reactive protein (Johnson et al., 1999b, 2000), 2⬘,5⬘- or signal transduction (Miyamoto et al., 1995; Burghardt et
oligoadenylate synthetase (OAS) (Mirando et al., 1991; al., 1997, in press). In sheep, integrin expression on the
Johnson et al., 2001b; Stewart et al., 2001a), STATs 1 and 2 apical surface of the LE is constitutive (Johnson et al.,
(Choi et al., 2001; Stewart et al., 2001a,b), and IRF-1 and -9 2001a). The ovine endometrial LE and trophectoderm
(Choi et al., 2001; Stewart et al., 2002). Despite retarded express integrin subunits αv, α4, α5, β1, β3 and β5 (Johnson
conceptus development and absence of IFN-τ production et al., 2001a). The present study examined expression of the
by UGKO conceptuses, findings from the present study glycoprotein MUC-1 and integrin subunits αv, α5, β3 and β5
indicate that the endometrium of UGKO ewes is responsive by the endometrial epithelium of normal and UGKO ewes
to IFN-τ in terms of induction of or increases in expression 14 days after mating. There were no differences between
of IFN-τ-stimulated genes. The ability of IFN-τ to stimulate normal and UGKO ewes in the patterns of Muc-1 and
increases in IFN-τ-stimulated genes in the UGKO uterus is integrin subunit expression on the endometrial LE. Previous
750 Wolf Prize in Agriculture
studies have demonstrated similar results for temporal L-selectin which, in turn, activates integrins and promotes
expression of steroid hormone receptors (Gray et al., fibronectin adhesion to aid in extravasation of lymphocytes
2000a) and LE-specific genes (Gray et al., 2001c) in the in lymph nodes (Rosen, 1993; Hwang et al., 1996; Giblin et
endometrium of UGKO compared with normal ewes. al., 1997). In pregnant ewes, GlyCAM-1 in the LE and GE
Taken together, the results of these studies support the was low on days 11 and 13, increased on day 15, and
contention that the endometrial LE of UGKO ewes is not was abundant on days 17 and 19 after mating (Spencer
abnormal in phenotype and is not the underlying cause of et al., 1999b). In pregnant ewes, the relative amount of
conceptus growth retardation and mortality. immunoreactive GlyCAM-1 in uterine flushings was low on
In several species, conceptus implantation involves days 11 and 13, but high on days 15 and 17 after mating.
molecules that are secreted by the endometrial glands Similar to findings for osteopontin, the abundance of
during pregnancy. In rodents, the endometrial glands GlyCAM-1 was low or undetectable in uterine flushings
secrete leucocyte inhibitory factor and calcitonin, which act from cyclic and UGKO ewes, but was abundant in uterine
on the uterine LE or conceptus to promote conceptus flushings from pregnant ewes 14 days after mating. Thus,
development, establishment of uterine receptivity and the absence of GlyCAM-1 in the uterine environment of
implantation (Carson et al., 2000). Available evidence from UGKO ewes could also impede conceptus elongation and
the ovine UGKO model supports the contention that the implantation. It is likely that several other molecules are
endometrial glands also secrete molecules that support also lacking in the UGKO uterus, resulting in conceptus
conceptus survival and development. The endometrial mortality and growth retardation during the peri-
glands of the pregnant ovine uterus synthesize and secrete implantation period. The lack of a closed uterine lumen
osteopontin, which is hypothesized to play a role in with discreet protruding caruncles may also contribute to
conceptus attachment, adhesion and elongation during defects in conceptus survival and elongation observed in
peri-implantation (Johnson et al., 1999a,b, 2001a). UGKO ewes. However, available results support the
Osteopontin is a 70 kDa acidic glycoprotein component of hypothesis that defects in peri-implantation conceptus
the ECM (Butler et al., 1996) that gives rise to 25 and 45 kDa survival and growth in UGKO ewes are not the result of
fragments upon treatment with proteases and freezing or alterations in the endometrial expression of either anti-
thawing (Weber and Cantor, 1996). Osteopontin binds to adhesion molecules or integrin receptors, but are rather the
cell surface integrins present on both trophectoderm and LE, result of an absence of endometrial glands and their
via its RGD sequence present on the 70 and 45 kDa forms, secretions, including molecules involved in cell–cell
to promote cell–cell attachment during the period of adhesions such as osteopontin and GlyCAM-1. A functional
implantation (Johnson et al., 1999b). Osteopontin engages a genomics and proteomics approach will be necessary to
number of receptors, including the integrins αv(β1, β3 or determine which uterine factors are absent in UGKO ewes
β5) and (α4, α5, α8 or α9)β1 (Denhardt et al., 2001). In the as compared with normal ewes during early pregnancy.
present study, osteopontin was found to be absent in uterine
flushings from cyclic and UGKO ewes and present in all These studies were supported by Grants 1998-35203-6322
three forms in uterine flushings from pregnant ewes 14 days and 2001-35203-10700 from the NRI Competitive Grants
after mating. This pattern of osteopontin expression in Program/CSREES/USDA and, in part, by P30 ES09106. The authors
normal ewes is similar to that reported by Johnson et al. appreciate the gift of reagents by T. Hansen (University of
Wyoming) and D. Carson (University of Delaware).
(1999a). If osteopontin is essential for normal conceptus
elongation and implantation in sheep, then its absence in
the uterine milieu of UGKO ewes would be detrimental to References
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of the integrin receptors for osteopontin on the endometrial endometrial function following asynchronous embryo transfer or
LE. The process of conceptus elongation and implantation administration of progesterone Biology of Reproduction 40 425–433
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Burghardt et al., in press). Results from the present study Bartol FF, Wiley AA, Floyd JG, Ott TL, Bazer FW, Gray CA and Spencer TE
concur with this hypothesis and provide new insight (1999) Uterine differentiation as a foundation for subsequent fertility
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752 Wolf Prize in Agriculture
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Fuller W. Bazer 753
Cathepsins (CTS) are peptidases that have biological roles in conceptus trophectoderm, and pro-CTSL was detected in
degrading extracellular matrix, catabolism of intracellular uterine flushings from ewes between d 12 and 16 of pregnancy.
proteins, and processing of prohormones. Expression of In ovariectomized and catheterized ewes, CTSL mRNA in the
CTSB, CTSD, CTSH, CTSK, CTSL, CTSS, and CTSZ genes was endometrium was increased by progesterone and intrauter-
detected in the endometria of cyclic and early pregnant ewes ine injections of ovine interferon (IFN). Other endometrial
with distinct temporal and spatial expression patterns. In the CTS genes were also regulated by progesterone alone (CTSB,
d 18 and 20 conceptus, expression of CTSB, CTSD, CTSL, and CTSK, CTSS, and CTSZ) or progesterone and IFN (CTSH,
CTSZ mRNA was detected in the trophectoderm. Of particular CTSK, CTSS, and CTSZ). These results indicate that CTS of
note, CTSL mRNA was the most abundant CTS mRNA in the endometrial and conceptus origin may regulate endometrial
ovine endometrium and detected only in the luminal epithe- remodeling and conceptus implantation, endometrial CTS
lium and superficial glandular epithelium of cyclic and preg- genes are regulated by ovarian and placental hormones, and
nant ewes. CTSL mRNA increased 8-fold between d 10 and 18 CTSL is a novel IFN-stimulated gene expressed only in lu-
in endometria of pregnant ewes, whereas it declined between minal epithelium and superficial glandular epithelium of the
d 14 and 16 in cyclic ewes. CTSL protein was also detected in endometrium. (Endocrinology 146: 4825– 4833, 2005)
4825
754 Wolf Prize in Agriculture
4826 Endocrinology, November 2005, 146(11):4825– 4833 Song et al. • Progesterone and IFN Regulate Endometrial CTSL
Materials and Methods TABLE 1. Sequences of primers used for RT-PCR and cloning
Animals
Sequence (5⬘–3⬘): GenBank Product
Gene
Mature crossbred Suffolk ewes (Ovis aries) were observed daily for forward and reverse accession no. size (bp)
estrus in the presence of vasectomized rams and used in experiments CTSB GCAAAACACCACTTGGAAGG L06075 581
only after they had exhibited at least two estrous cycles of normal AGGAACTGCATCCAAAATGC
duration (16 –18 d). All experimental and surgical procedures were in CTSD ACCTTCGACATCCACTACGG AF164143 520
compliance with the Guide for the Care and Use of Agriculture Animals GTAGCTCTCGCACCTCATCC
and approved by the University Laboratory Animal Care and Use Com- CTSH GGTCAGAGCCTCAGAATTGC NM_004390 420
mittee of Texas A&M University. CATCGTTCAGTGTGATGTTGG
CTSK GGGTCTCACGGTTCTACTGC NM_000396 504
Experimental design CAGTCCACCAGGTTCTGAGG
CTSL AATGGAGGAGAGCAGTGTGG X91755 579
Study 1. At estrus (d 0), ewes were mated to either an intact or CCTTCATAAGGGCCTTCTCC
vasectomized ram as described previously (22) and then hysterecto- CTSS CCTGGAAGCACAAGTGAAGC X62001 330
mized (n ⫽ 5 ewes/d) on d 10, 12, 14, or 16 of the estrous cycle or d GAATGGCTCGCGTCTATACC
10, 12, 14, 16, 18, or 20 of pregnancy. Pregnancy was confirmed on d CTSZ GGCCTCATGAGTACCTGTCC NM_001336 501
10 –16 after mating by the presence of a morphologically normal TTGCCATTATGCCACAGC
conceptus(es) in the uterus. At hysterectomy, several sections (⬃0.5
cm) from the midportion of each uterine horn ipsilateral to the corpus
luteum were fixed in fresh 4% paraformaldehyde in PBS (pH 7.2). CTSH, CTSS, CTSD and CTSZ : 1) 95 C for 5 min; 2) 95 C for 45 sec; 59.1
After 24 h, fixed tissues were changed to 70% ethanol for 24 h and then C (for CTSB and CTSH), 56.5 C (for CTSD, CTSK, CTSL, and CTSZ), or 64.5
dehydrated and embedded in Paraplast-Plus (Oxford Labware, St. C (for CTSS) for 1 min; and 72 C for 1 min for 35 cycles; and 3) 72 C for 10
Louis, MO). Several sections (1–1.5 cm) from the middle of each min. Partial cDNAs of the correct size were cloned into pCRII using a T/A
uterine horn were embedded in Tissue-Tek OCT compound (Miles, cloning kit (Invitrogen) and their sequences verified by sequencing.
Oneonta, NY), frozen in liquid nitrogen vapor, and stored at ⫺80 C.
The remaining endometrium was physically dissected from myome-
Slot blot hybridization analyses
trium, frozen in liquid nitrogen, and stored at ⫺80 C for subsequent
RNA or protein extraction. In monovulatory pregnant (PX) ewes, Steady-state levels of mRNA in ovine endometria were assessed by
uterine tissue samples were marked as either contralateral or ipsi- slot blot hybridization as described previously (28, 29). Radiolabeled
lateral to the ovary bearing the corpus luteum. No tissues from the antisense and sense cRNA probes were generated by in vitro tran-
contralateral uterine horn were used for study. Uterine flushes were scription using linearized plasmid template, RNA polymerases, and
clarified by centrifugation (3000 ⫻ g for 30 min at 4 C) and frozen at [␣-32P]uridine 5-triphosphate. Denatured total endometrial RNA (20
⫺80 C for Western blot analysis. g) from each ewe in studies 1 and 2 was hybridized with radiola-
beled cRNA probes. To correct for variation in total RNA loading, a
Study 2. Cyclic (C) ewes (n ⫽ 20) were checked daily for estrus and duplicate RNA slot membrane was hybridized with radiolabeled
then ovariectomized and fitted with indwelling uterine catheters on antisense 18S cRNA (pT718S; Ambion, Austin, TX). After washing,
d 5 as described previously (23). Ewes were then assigned randomly the blots were digested with ribonuclease A and radioactivity asso-
(n ⫽ 5 per treatment) to receive daily im injections of progesterone ciated with slots quantified using a Typhoon 8600 MultiImager (Mo-
and/or a progesterone receptor (PR) antagonist (ZK 136,317; Schering lecular Dynamics, Piscataway, NJ). Data are expressed as relative
AG, Berlin, Germany) and intrauterine infusions of control serum units.
proteins and/or recombinant ovine IFN protein as follows: 1) 50 mg
progesterone (P, d 5–16) and 200 g control (CX) serum proteins (d
11–16) (P⫹CX); 2) P and 75 mg ZK 136,317 (d 11–16) and CX proteins In situ hybridization analyses
(P⫹ZK⫹CX); 3) P and IFN (2 ⫻ 107 antiviral units, d 11–16) (P⫹IFN); Location of mRNA expression in sections (5 m) of the ovine
or 4) P and ZK and IFN (P⫹ZK⫹IFN). Steroids were administered uterus was determined by radioactive in situ hybridization analysis
daily in corn oil vehicle. Both uterine horns of each ewe received as described previously (28, 29). Radiolabeled antisense and sense
twice-daily injections of either CX proteins (50 g/horn per injection) cRNA probes were generated by in vitro transcription using linear-
or IFN (5 ⫻ 106 antiviral units/horn per injection). Recombinant ized plasmid template, RNA polymerases, and [␣-35S]uridine
ovine IFN was produced in Pichia pastoris and purified as described 5-triphosphate. Deparaffinized, rehydrated, and deproteinated uter-
previously (24). Proteins were prepared for intrauterine injection as ine tissue sections were hybridized with radiolabeled antisense or
described previously (23). This regimen of progesterone and recom- sense cRNA probes. After hybridization, washing, and ribonuclease
binant ovine (ro)IFN mimics the effects of progesterone and the A digestion, slides were dipped in NTB-2 liquid photographic emul-
conceptus on endometrial expression of hormone receptors and IFN- sion (Kodak, Rochester, NY) and exposed at 4 C for 2 wk. Slides were
stimulated genes during early pregnancy in ewes (25–28). All ewes developed in Kodak D-19 developer, counterstained with Gill’s he-
were hysterectomized on d 17, and the uteri and endometria pro- matoxylin (Fisher Scientific, Fairlawn, NJ), and then dehydrated
cessed as described in study 1. through a graded series of alcohol to xylene. Coverslips were then
affixed with Permount (Fisher Scientific). Images of representative
RNA isolation fields were recorded under bright-field or dark-field illumination
using an Eclipse 1000 photomicroscope (Nikon Instruments Inc.,
Total cellular RNA was isolated from frozen ipsilateral endometrium Lewisville, TX) fitted with a Nikon DXM1200 digital camera.
(studies 1 and 2) using Trizol reagent (Life Technologies, Inc.-BRL,
Bethesda, MD) according to manufacturer’s recommendations. The
quantity and quality of total RNA was determined by spectrometry and
Immunohistochemistry
denaturing agarose gel electrophoresis, respectively. Immunocytochemical localization of immunoreactive CTSL protein
in the ovine uterus was performed as described previously (22) in
Cloning of partial cDNAs for ovine CTSB, CTSK, CTSL, uterine tissue cross-sections from studies 1 and 2 using rabbit antihuman
CTSH, CTSS, CTSD, and CTSZ CTSL polyclonal antibody (catalog no. 3192-100; BioVision, Mountain
View, CA) at a final concentration of 1 g/ml. Antigen retrieval was
Partial cDNAs for ovine CTSB, CTSD, CTSK, CTSL, CTSH, CTSS, and performed by using boiling citrate buffer as described previously (30).
CTSZ mRNAs were amplified by RT-PCR using total RNA from endo- Negative controls included substitution of the primary antibody with
metrial tissues from d 16 –18 of pregnancy using specific primers (Table 1). nonimmune rabbit IgG (Sigma Chemical Co., St. Louis, MO) at the same
PCR amplification was conducted as follows for ovine CTSB, CTSK, CTSL, final concentration.
Fuller W. Bazer 755
Song et al. • Progesterone and IFN Regulate Endometrial CTSL Endocrinology, November 2005, 146(11):4825– 4833 4827
Western blot analyses corrected for differences in sample loading using the 18S rRNA data as
a covariate. Data from study One were analyzed for effects of day,
Protein concentrations of uterine flushes were determined using the pregnancy status (C or PX), and their interaction. Effects of day were
Bradford protein assay (Bio-Rad Laboratories, Hercules, CA) with BSA determined by least squares regression analysis. Data from study 2 were
as the standard. Proteins were denatured and separated by 12% SDS- analyzed using preplanned orthogonal contrasts (P⫹CX vs. P⫹IFN,
PAGE and Western blot analysis conducted as described previously (22) P⫹CX vs. P⫹ZK⫹CX, and P⫹IFN vs. P⫹ZK⫹IFN). Data are presented
by using enhanced chemiluminescence (SuperSignal West Pico, Pierce, as least squares means with overall se values.
Rockford, IL) and X-OMAT AR x-ray film (Kodak) according to the
manufacturer’s recommendations. Immunoreactive CTSL protein was
detected using rabbit antihuman CTSL polyclonal antibody (catalog no. Results
3192-100; BioVision) at 0.5 g/ml.
Effects of estrous cycle and pregnancy on expression of CTS
mRNAs in ovine endometrium (study 1)
Statistical analyses
Data from slot blot hybridization analyses were subjected to least Steady-state levels of ovine CTSB, CTSD, CTSH, CTSK, CTSL,
squares ANOVA using the general linear models procedures of the CTSS, and CTSZ mRNAs in endometria from C and PX ewes
Statistical Analysis System (Cary, NC). Slot blot hybridization data were were determined by slot blot hybridization analyses (Fig. 1).
FIG. 1. Steady-state levels of CTSB, CTSD, CTSH, CTSK, CTSL, CTSS, and CTSZ mRNAs in endometria from C and PX ewes as determined
by slot blot analysis. See text for description of effects of day of the C or PX on mRNA levels in the endometrium.
756 Wolf Prize in Agriculture
4828 Endocrinology, November 2005, 146(11):4825– 4833 Song et al. • Progesterone and IFN Regulate Endometrial CTSL
Expression of CTSB mRNA was lowest on d 10 and increased however, abundant CTSD mRNA was detected in the tro-
to d 16 or 20 in C and PX ewes, respectively (linear effect of day, phectoderm of the conceptus. CTSH mRNA was expressed
P ⬍ 0.01). Endometrial levels of CTSD mRNA did not change at moderate levels in the endometrial LE and GE, particularly
in C ewes but increased from d 10 –20 in PX ewes (linear effect on d 18 and 20 in PX ewes. In C and PX ewes, CTSK mRNA
of day, P ⬍ 0.01). CTSH mRNA levels increased from d 10 –14 was expressed at moderate levels in the endometrial LE and
in C ewes and from d 10 –20 in PX ewes (linear effect of day, P ⬍ stroma as well as cells within the stroma that appeared to be
0.01). In contrast, CTSK mRNA did not change (P ⬎ 0.10) in immune cells based on their morphology and location.
endometria of C and PX ewes. CTSL mRNA was affected (P ⬍ CTSL mRNA was the most abundant CTS genes expressed
0.05) by day, status, and their interaction. In C ewes, CTSL in the endometrium, and it was detected only in endometrial
mRNA increased from d 10 –14 and then declined to d 16 LE and sGE (Fig. 3). Furthermore, CTSL mRNA was ex-
(quadratic effect of day, P ⬍ 0.05). In PX ewes, CTSL mRNA pressed by conceptus trophectoderm on d 18 and 20 of PX.
increased about 8-fold between d 10 and 18 (linear effect of day, CTSS mRNA was detected at low levels in the endometrial
P ⬍ 0.01). Furthermore, CTSL mRNA levels in the endometrium LE and cells within the stroma that appeared to be immune
were greater on d 14 and 16 in PX than C ewes (day ⫻ status, cells based on their morphology and distribution. The num-
P ⬍ 0.05). Endometrial CTSS and CTSZ mRNA levels were not ber of CTSS mRNA-positive immune-like cells increased be-
affected by pregnancy status or day or their interaction (P ⬎ tween d 14 and 16 of pregnancy. CTSZ mRNA was detected
0.10). at low levels specifically in the endometrial LE and sGE as
In situ hybridization analyses determined the location of well as conceptus trophectoderm on d 18 and 20 of preg-
CTS gene expression in endometria. In C and PX ewes, CTSB nancy. No differences in expression of CTS mRNAs in the LE
mRNA was detected in the endometrial LE, ductal sGE, or stroma of the intercaruncular endometria were found
stratum compactum stroma and cells distributed throughout when compared with the caruncular endometria in the
the stroma that appeared to be immune cells based on their uterus of either C or PX ewes (data not shown).
morphology (Fig. 2). Abundant CTSB mRNA was detected Collectively, results of slot blot and in situ hybridization
in the trophectoderm of the conceptus. CTSD mRNA was analyses indicated that CTSL mRNA was the most abundant
expressed at low levels in the endometrial LE and sGE; CTS gene expressed in the endometrium and the only CTS
FIG. 2. In situ hybridization analyses of CTSB, CTSD, CTSH, and CTSK mRNAs in uteri of C and PX ewes. Cross-sections of the uterine wall
from C and PX ewes were hybridized with radiolabeled antisense or sense ovine CTS cRNA probes. C, Conceptus; S, stroma. Scale bar, 10 m.
Fuller W. Bazer 757
Song et al. • Progesterone and IFN Regulate Endometrial CTSL Endocrinology, November 2005, 146(11):4825– 4833 4829
4830 Endocrinology, November 2005, 146(11):4825– 4833 Song et al. • Progesterone and IFN Regulate Endometrial CTSL
ceiving P alone (P⫹CX vs. P⫹IFN, P ⬍ 0.01) or P⫹ZK ovine endometrium may play an important role in estab-
(P⫹ZK⫹CX vs. P⫹ZK⫹IFN, P ⬍ 0.01). lishing a regulatory network of multiple proteolytic enzymes
responsible for ECM remodeling during implantation and
Discussion placentation. Although decidualization of the endometrial
Similar to endometria of other mammals, expression of stroma does not occur in sheep, the endometrium undergoes
many CTS genes was detected in endometria of C and early dramatic remodeling after pregnancy recognition and estab-
PX ewes. The CTS family of cysteine and aspartyl proteases lishment between d 12 and 20 of early pregnancy. In the
as well as other proteases, including MMPs and serine pro- intercaruncular endometrium, the endometrial epithelium is
teases, are implicated in the degradation of ECM required for removed by the trophoblast giant binucleate cells during
uterine remodeling during decidualization, implantation, synepitheliochorial placentation, the stroma becomes very
and placentation (11–14). In rodents, for example, it has been compact and begins to express new genes such as osteopon-
hypothesized that CTS play a crucial role in digestion of tin, and the glands undergo hypertrophy followed by hy-
matrix molecules and activation of other proenzymes re- perplasia (33–36). In the caruncular endometrium, the pla-
sponsible for intracellular breakdown of molecules that are cental cotyledons attach to the maternal caruncles and
phagocytosed by cells (4). The dynamic and differential ex- develop into placentomes (35). These morphogenetic and
pression of CTS genes between C and PX ewes suggests differentiation events undoubtedly involve regulation by
functional diversity in mechanisms responsible for expres- CTS and extensive remodeling of the ECM.
sion of CTS genes that may be responsible for optimization The present studies found that CTSL mRNA was partic-
of a uterine environment that supports conceptus implan- ularly abundant in the endometrial LE and sGE and up-
tation and placentation during establishment and mainte- regulated during early pregnancy in association with con-
nance of pregnancy (12). In the present study, cysteine pro- ceptus elongation and implantation (16). CTSL is normally
teases CTSB, CTSH, CTSK, CTSL, CTSS, and CTSZ and localized in lysosomes, in which it plays a major role in
aspartyl protease CTSD were found to be expressed in the intracellular protein catabolism. In the present studies, the
ovine endometrium, and expression of CTSB, CTSD, CTSH, 38- to 40-kDa latent pro-CTSL form of CTSL protein was
CTSL, and CTSZ mRNA increased between d 10 and 20 of abundant in uterine flushings from d 12, 14, and 16 PX ewes.
early pregnancy. Consistent with above results, CTSL pro- This latent pro-CTSL must be cleaved by proteases, such as
tein in the porcine uterus was observed in endometrial GE MMPs, to generate the active two-chain form made up of 21-
as well as the uterine lumen and induced by progesterone and 5-kDa subunits (1). The presence of the pro-CTSL in
during the periods of implantation and placentation (9). uterine flushings from PX ewes between d 12 and 16 of
Interestingly, the ovine placenta expresses large numbers pregnancy suggests that CTSL is secreted by the endometrial
of aspartic proteinase inhibitor genes, termed pregnancy- LE and/or conceptus. Indeed, the synthesis and secretion of
associated glycoproteins (31), and the endometrial glands the 39-kDa pro-CTSL has been demonstrated for many tu-
express large amounts of serine protease inhibitors, termed mors, including cancers of the kidney, lung, colon, breast,
serpins or uterine milk proteins (32), that could regulate the and ovary (37). In rodents, interactions of CTSB, CTSL, and
activity of endometrial CTS identified in the present study. cystatin C, a CTSL inhibitor, are important for implantation
Therefore, the molecular control of expression of CTS in the and placentation because inhibition of endometrial CTSL
Fuller W. Bazer 759
Song et al. • Progesterone and IFN Regulate Endometrial CTSL Endocrinology, November 2005, 146(11):4825– 4833 4831
FIG. 6. Steady-state levels of CTSB, CTSD, CTSH, CTSK, CTSS, and CTSZ mRNA in the ovine endometrium from ewes in study 2. Steady-state
levels of mRNA in endometria from treated ewes were determined by slot blot analysis. See text for description of effects of treatment on mRNA
levels in the endometrium. The asterisk (*) denotes an effect of treatment (P ⬍ 0.10).
and CTSB results in abnormal embryonic development and pregnancy in sheep as well as many other mammals. CTSL
uterine decidualization during the periimplantation period is capable of degrading ECM proteins, suggesting a role in
(4). Invasion by the ectoplacental cone of mouse trophoblast conceptus attachment by altering the composition of the
was prevented by cysteine proteinase inhibitors in vitro (38). ECM present on the apical surfaces of the endometrial LE
Recently Cheon et al. (39) found that cytotoxic T lymphocyte and/or trophoblast.
antigen-2, a cysteine protease inhibitor, was up-regulated In the present study, temporal changes in expression of
by progesterone in the decidua and proposed to regulate endometrial CTSL mRNA in C and PX ewes supported the
blastocyst implantation by neutralizing the activities of one hypothesis that ovarian progesterone regulates transcription
or more proteases, including CTSL, generated by the prolif- of the CTSL gene in the endometrial LE. Similarly, an increase
erating trophoblast. CTSL has been studied in uteri of cats in CTSB, CTSD, CTSH, and CTSZ was also observed in the
(6 – 8), pigs (9), and mice (4, 40). In cats, CTSL is localized to endometrium during early pregnancy. The increase in CTSL
the GE and can be detected in the uterine lumen, in which and CTSZ mRNAs in LE and sGE, between d 10 and 12 after
it is implicated in blastocyst invasion (6). In pigs, CTSL was estrus/mating, and CTSH mRNA in LE and GE, between d
also found to be expressed in the endometrial GE and as a 14 and 16 after mating, is coincident with the disappearance
progesterone-regulated component of the uterine lumen of PR mRNA and protein in these epithelia (41). Similarly, the
during implantation and placentation (9). Thus, available decrease in CTSL and CTSZ mRNAs between d 14 and 16 of
results suggest that CTSL may be an essential regulator of the cycle is coincident with the reappearance of PR protein
endometrial remodeling and conceptus implantation during in endometrial LE. In study 2, CTSL mRNA was detected in
760 Wolf Prize in Agriculture
4832 Endocrinology, November 2005, 146(11):4825– 4833 Song et al. • Progesterone and IFN Regulate Endometrial CTSL
Song et al. • Progesterone and IFN Regulate Endometrial CTSL Endocrinology, November 2005, 146(11):4825– 4833 4833
binant ovine interferon on the endometrium of cyclic ewes. Biol Reprod significance of the binucleate giant cells of the placenta of ruminants. J Anat
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29. Choi Y, Johnson GA, Burghardt RC, Berghman LR, Joyce MM, Taylor KM, 37. Nomura T, Katunuma N 2005 Involvement of cathepsins in the invasion,
Stewart MD, Bazer FW, Spencer TE 2001 Interferon regulatory factor-two metastasis and proliferation of cancer cells. J Med Invest 52:1–9
restricts expression of interferon-stimulated genes to the endometrial 38. Babiarz BS, Romagnano LC, Kurilla GM 1992 Interaction of mouse ectopla-
stroma and glandular epithelium of the ovine uterus. Biol Reprod 65:1038 – cental cone trophoblast and uterine decidua in vitro. In Vitro Cell Dev Biol
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30. Taylor KM, Gray CA, Joyce MM, Stewart MD, Bazer FW, Spencer TE 2000 39. Cheon Y-P, DeMayo FJ, Bagchi MK, Bagchi IC 2004 Induction of cytotoxic
Neonatal ovine uterine development involves alterations in expression of T-lymphocyte antigen-2, a cysteine protease inhibitor in decidua: a potential
receptors for estrogen, progesterone, and prolactin. Biol Reprod 63:1192– regulator of embryo implantation. J Biol Chem 279:10357–10363
1204 40. Hamilton RT, Bruns KA, Delgado MA, Shim JK, Fang Y, Denhardt DT,
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RM 2000 Pregnancy-associated bovine and ovine glycoproteins exhibit spa- rasHa in the mouse placenta. Mol Reprod Dev 30:285–292
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phylogeny of ovine uterine serpin. Am J Reprod Immunol 45:266 –272 42. Johnson GA, Spencer TE, Burghardt RC, Taylor KM, Gray CA, Bazer FW
33. Johnson GA, Burghardt RC, Bazer FW, Spencer TE 2003 Osteopontin: roles 2000 Progesterone modulation of osteopontin gene expression in the ovine
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Endocrinology is published monthly by The Endocrine Society (http://www.endo-society.org), the foremost professional society serving the
endocrine community.
762 Wolf Prize in Agriculture
Establishment of pregnancy in ruminants results from para- that the basal promoter activity was dependent on the region
crine signaling by interferon (IFNT) from the conceptus to from ⴚ144 to ⴚ4 bp that contained only SP1 sites. IFNT did not
uterine endometrial luminal epithelia (LE) that prevents re- affect activity of the OXTR promoter. In cells transfected with
lease of luteolytic prostaglandin F2␣ pulses. In cyclic and preg- ESR1, E2, and ICI 182,780 increased promoter activity due to
nant ewes, progesterone down-regulates progesterone recep- GC-rich SP1 binding sites at positions ⴚ104 and ⴚ64. Mutation
tor (PGR) gene expression in LE. In cyclic ewes, loss of PGR analyses showed that the proximal SP1 sites mediated ESR1
allows for increases in estrogen receptor ␣ (ESR1) and then action as well as basal activity of the promoter. In response to
oxytocin receptor (OXTR) gene expression followed by oxy- progesterone, progesterone receptor B also increased OXTR
tocin-induced prostaglandin F2␣ pulses. In pregnant ewes, promoter activity. SP1 protein was constitutively expressed
IFNT inhibits transcription of the ESR1 gene, which presum- and abundant in the LE of the ovine uterus. These results
ably inhibits OXTR gene transcription. Alternatively, IFNT support the hypothesis that the antiluteolytic effects of IFNT
may directly inhibit OXTR gene transcription. The 5ⴕ promot- are mediated by direct inhibition or silencing of ESR1 gene
er/enhancer region of the ovine OXTR gene was cloned and transcription, thereby precluding ESR1/SP1 from stimulating
found to contain predicted binding sites for activator protein OXTR gene transcription. (Endocrinology 147: 899 –911, 2006)
1, SP1, and PGR, but not for ESR1. Deletion analysis showed
899
Fuller W. Bazer 763
900 Endocrinology, February 2006, 147(2):899 –911 Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription
Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription Endocrinology, February 2006, 147(2):899 –911 901
⫺104 and ⫺64. Mutated DNAs were confirmed by sequencing, and otide with the same base composition as the SP1 consensus oligonu-
fragments containing only the desired point mutations were direction- cleotide to minimize charge differences, but having a different sequence
ally subcloned into the luciferase vector as described above. (Table 1). Radiolabeled probes (10 fmol per reaction) were added to
reactions that were incubated at room temperature for an additional 15
Transient transfections and luciferase assays min. Binding reactions were electrophoresed in 5% native polyacryl-
amide gels in 1⫻ TBE. Imaging of dried gels was done with a Molecular
Cells were subcultured into 12-well plates (67–75% confluent) and Dynamics Typhoon phosphorimager.
transiently transfected as described previously (23) with the following
modifications. Luciferase constructs (500 ng/well) were cotransfected Immunohistochemistry
with either pEF1-Myc-His-LacZ (500 ng/well; Invitrogen) or expression
plasmids for ovine IRF1, ovine IRF2, human ESR1wt, human ESR1IIc, Mature crossbred Suffolk ewes (Ovis aries) were observed daily for
or human progesterone receptor B (PGR-B) (500 ng/well except where estrus in the presence of vasectomized rams and used in experiments
noted). The ovine IRF1 and IRF2 mammalian overexpression plasmids only after they had exhibited at least two estrous cycles of normal
have been described previously (24). The ESR1wt (wild-type human duration (16 –18 d). All experimental and surgical procedures were in
ESR1) and ESR1IIc (a DNA binding domain mutant construct) overex- compliance with the Guide for the Care and Use of Agriculture Animals
pression plasmids have been described previously (32, 33). The PGR-B and approved by the University Laboratory Animal Care and Use Com-
plasmid, pSV40-hPGR-B that overexpresses the B form of human PGR, mittee of Texas A&M University.
was kindly provided by Dr. M.-j. Tsai (Baylor College of Medicine, At estrus (d 0), ewes were mated to either an intact or vasectomized
Houston, TX). Transfected cells were grown in 10% FBS for approxi- ram as described previously (34) and then hysterectomized (n ⫽ 5 ewes
mately 14 –16 h before treatment for 24 h in serum-free medium. Phenol per day) on either d 10, 12, 14, or 16 of the estrous cycle or d 10, 12, 14,
red-free DMEM-F12 medium and dextran-coated charcoal-stripped FBS 16, 18, or 20 of pregnancy. Pregnancy was confirmed on d 10 –16 after
were substituted in experiments when testing for effects of steroids. mating by the presence of a morphologically normal conceptus(es) in the
Steroid agonists and antagonists were dissolved in 100% ethanol. For the uterus. At hysterectomy, several sections (⬃0.5 cm) from the mid-por-
control, cells were treated with the same volume of 100% ethanol alone. tion of each uterine horn ipsilateral to the CL were fixed in fresh 4%
Luciferase and protein assays were done as described previously (23, 24). paraformaldehyde in PBS (pH 7.2). After 24 h, fixed tissues were
Transfection assays were repeated a minimum of three times. changed to 70% ethanol for 24 h and then dehydrated and embedded in
Paraplast-Plus (Oxford Labware, St. Louis, MO). In monovulatory preg-
EMSA nant ewes, uterine tissue samples were marked as either contralateral or
ipsilateral to the ovary bearing the CL. No tissues from the contralateral
Double-stranded oligonucleotide primers listed in Table 1 were end- uterine horn were used for study.
labeled to high specific activity with [␥-32P]ATP and T4 DNA kinase by Immunoreactive SP1 proteins were localized in cross-sections (5 m)
standard methods. Purified SP1 protein (54 ng/reaction) was incubated of the uterus using a rabbit polyclonal antibody to human SP1 (catalog
in 10 l reactions containing 4% glycerol, 1 mm MgCl2, 0.5 mm EDTA, SC-59; Santa Cruz Biotechnology Inc., Santa Cruz, CA) at a final working
50 mm NaCl, 10 mm Tris-HCl (pH 7.5), 0.5 mm DTT, 0.05 mg/ml concentration of 0.1 g/ml and a VectaStain Rabbit IgG Elite ABC kit
poly(dI-dC) at room temperature for 15 min. A 100-fold excess of un- (Vector Laboratories, Burlingame, CA) using methods described previ-
labeled specific or nonspecific competitor oligonucleotides was included ously (35). Antigen retrieval using boiling citrate buffer was performed
in some reactions. The nonspecific competitor was a 22-nt oligonucle- as described previously (35, 36). The chromagen used for peroxidase
Fuller W. Bazer 765
902 Endocrinology, February 2006, 147(2):899 –911 Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription
localization was 3,3⬘-diaminobenzidine tetrahydrochloride from Sigma sites were found at ⫺748, ⫺543, and ⫺444 relative to the
Chemical Co. (St. Louis, MO). Negative controls were performed in translational start site (ATG ⫹1) in addition to consensus
which the primary antibody was substituted with the same concentra-
tion of purified normal rabbit IgG from Sigma Chemical Co. Multiple
sites at ⫺104 and ⫺64 (Fig. 1). The putative ⫺64 SP1 binding
tissue sections from each ewe were processed as sets within an exper- site is identical to the consensus SP1 binding motif (5⬘-
iment. Sections were not counterstained before affixing coverslips. Rep- CCCCGCCCC-3⬘, sense strand). The ⫺104 putative binding
resentative photomicrographs of uterine tissues were taken using a site (5⬘-CCCCACCCCGCCCC-3⬘, sense strand) also contains
Nikon Eclipse 1000 photomicroscope (Nikon Instruments Inc., Lewis- a consensus binding motif that is immediately adjacent to
ville, TX) fitted with a Nikon DXM1200 digital camera. Digital images
were captured and assembled using Adobe Photoshop (Adobe Systems, and overlaps a sequence that resembles a nonconsensus SP1
Seattle, WA). binding site (5⬘-CCCCACCCC-3⬘) (Fig. 1). Nonconsensus
SP1 sites having the sequence 5⬘-CCCCACCCC-3⬘ have been
Statistical analyses reported for the long terminal repeats of the human endog-
The effects of steroid hormones, ICI 182,780, or IFNT on the activity enous retrovirus H family of human retrovirus-like elements
of promoter reporter constructs in transient transfection assays were (39). Alternatively, part of the sequence of the OXTR ⫺104
analyzed by least squares ANOVA using the General Linear Models SP1 site may contain a CACCC box at the 5⬘ end. The CACCC
procedure of the Statistical Analysis System (Cary, NC). A P value of 0.10 elements bind SP1 and related Sp/Kruppel-like factor tran-
or less was considered statistically significant. Unless denoted, data are
reported as mean with sd. Effects of treatment were determined using
scription factors (40) and are involved in regulation of both
orthogonal or nonorthogonal contrasts using the PDIFF option of Gen- basal (41) and induced levels of transcription in other sys-
eral Linear Models. tems (42). A consensus AP-1 binding site at ⫺680 and non-
consensus AP-1 sites at ⫺574, ⫺329, and ⫺221 were identi-
Results fied, which also mediate hormone actions (38). No full
Isolation and sequence analysis of the OXTR 5⬘ progesterone response elements (PREs) were found, but sev-
upstream region eral putative PRE half sites were detected at ⫺639, ⫺486,
⫺477, and ⫺187. No full EREs or ERE half sites were found
A phagemid DNA containing approximately 9.8 kbp of
by computer-assisted analysis of the ovine OXTR upstream
ovine genomic DNA was isolated from a ovine genomic
region. Furthermore, elements mediating effects of type I or
library by conventional screening. Southern blotting, phys-
II IFNs or STATs, including ISRE, IRFE, or ␥ activation sites
ical mapping, and sequence analysis with several OXTR-
(GAS) (43), were also not found by bioinformatic analyses.
specific primers revealed that the clone contained a 1059-bp
region that included the 5⬘ end of the coding sequence of the
IFNT does not regulate OXTR promoter activity
ovine OXTR gene contiguous to 830 nt of upstream sequence.
The 1059-bp clone was sequenced in both directions and To determine whether IFNT regulated activity of the
deposited into GenBank (accession AY163261). Sequence OXTR promoter, transient transfection assays were con-
comparisons of the ovine upstream region with the previ- ducted in 2fTGH parental and U3A (STAT1 null) cells, which
ously characterized Bos taurus OXTR promoter region and are responsive to IFNT (44 – 46) in a STAT-dependent
adjacent coding regions (GenBank AF100633) indicated that (2fTGH) and STAT1-independent (U3A) manner. These cells
the two sequences were 91% identical. Although the coding have been extensively used by our laboratory as a model for
sequence displayed high homology with the OXTR cDNA the endometrial stroma and LE, respectively (44 – 46). Cells
from other species (human, dog, pig, gorilla, rat, mouse, were transfected with the full-length OXTR(⫺830/⫺4) and
chicken), the promoter region had very little or no homology sequential 5⬘-deletion reporter constructs and treated with
to those of other species except for bovine. different doses of recombinant ovine IFNT (103–106 antiviral
Bioinformatic analyses found that the ovine OXTR pro- units). No dose-dependent effects of IFNT on promoter ac-
moter region contained multiple putative binding sites for tivity were detected (P ⬎ 0.10) with any of the OXTR pro-
the transcription factors SP1 and activator protein 1 (AP-1), moter-reporter constructs in either 2fTGH or U3A cells (data
which regulates target genes by steroid hormone receptors not shown). Cells were also cotransfected with the
(see Refs. 37 and 38 for review). Putative nonconsensus SP1 OXTR(⫺830/⫺4) promoter-reporter construct and either
Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription Endocrinology, February 2006, 147(2):899 –911 903
904 Endocrinology, February 2006, 147(2):899 –911 Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription
Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription Endocrinology, February 2006, 147(2):899 –911 905
of either the 5⬘ nonconsensus SP1 site or the 3⬘ consensus SP1 6), double mutated (constructs 7 and 9), or point mutated
element was performed, because it could alter the integrity (construct 8). Indeed, basal luciferase levels of the ⫺64 SP1
of the overlapping site and affect the ability of the ⫺104 mutants were about 10% of wild-type parental levels. Mu-
sequence to bind SP1. Therefore, in addition to mutations of tation of the 5⬘ consensus SP1 site or both the nonconsensus
the ⫺104 site tested in initial gel shift analyses, some C3 A and consensus sites at ⫺104 and the ⫺64 SP1 site (construct
transversions that mutated either the consensus or noncon- 10) further reduced (P ⬍ 0.10) basal activity to about 2% of
sensus portion of the site, without altering the other over- activity observed for the wt construct.
lapping moiety, were also introduced. The oligonucleotides Estradiol-induced activity, in all cells transfected with the
used to create the mutations were tested initially for their minimal OXTR(⫺220/⫺4) promoter, was lower (P ⬍ 0.10) in
ability to bind SP1 protein by EMSA (Fig. 7). Mutation of the constructs containing a mutation in the ⫺104 and ⫺64 SP1
5⬘ nonconsensus site did not affect SP1 binding regardless of binding sites (Fig. 8). Mutation of both sites at position ⫺104
whether the overlapping 3⬘ consensus portion remained further reduced stimulation by estradiol, and this was con-
wild-type (mut3; lane 5) or was slightly altered (mut2; lane sistent with the EMSA data suggesting that both the left and
4). Mutation of the 3⬘ consensus sequence alone (mut1, lane right moieties are involved in binding SP1 protein. However,
3) substantially reduced SP1 binding compared with the mutation of the ⫺64 SP1 site decreased (P ⬍ 0.10) stimulation
wild-type ⫺104 OXTR oligonucleotide (lane 2), but did not by estradiol, regardless of whether the ⫺104 SP1 site was wt
eliminate binding entirely. Mutation of both 5⬘ and 3⬘ sites or mutated. Overall, these results suggest that both the ⫺104
(mut4) completely prevented SP1 binding (lane 6). In com- and ⫺64 SP1 sites in the proximal OXTR promoter regulates
petition experiments (Fig. 7B), excess unlabeled wt ⫺104 basal levels of OXTR gene transcription, and that both SP1
oligonucleotide and the mut1, mut2, and mut3 oligonucle- binding sites, especially the consensus sequences, regulate
otides reduced or prevented SP1 binding to the radiolabeled E2 stimulation of OXTR gene expression.
wt ⫺104 oligonucleotide (lanes 6, 7, and 8). As expected,
competition with the fully mutated mut4 oligonucleotide did
Progesterone stimulates OXTR promoter activity
not affect SP1 binding to the radiolabeled ⫺104 OXTR wt
oligonucleotide (lane 9). Collectively, these results indicate During metestrus and estrus in cyclic ewes, PGR return to
that SP1 binding to the ⫺104 OXTR sequence depends pri- the OXTR-expressing endometrial epithelia as systemic pro-
marily on the 3⬘ consensus GC-rich element. However, the 5⬘ gesterone declines to undetectable levels (9, 10, 12–14). The
nonconsensus GC-rich element appears to weakly bind SP1 prevailing theory is that PGR do not directly inhibit OXTR
protein, suggesting that the two overlapping motifs at ⫺104 gene expression, but rather the progesterone block to OXTR
may act cooperatively to bind SP1. formation is indirect via progesterone inhibition of ESR1
Transient transfection assays were then conducted to de- expression (4, 8, 28). To determine whether the OXTR pro-
termine the relative contributions of the ⫺104 and ⫺64 SP1 moter is regulated by PGR, 2fTGH cells were cotransfected
binding sites on hormonal responsiveness of the ovine OXTR with human PGR-B and OXTR promoter-LUC constructs
promoter (Fig. 8). Wild-type or mutant OXTR(⫺220/⫺4) and then treated with R5020 (10⫺8 m), a nonmetabolizable
promoter-reporter constructs were cotransfected with form of progesterone (Fig. 9). As observed previously, basal
ESR1IIc into 2fTGH cells and left unstimulated or treated activity of the OXTR promoter constructs increased (P ⬍
with E2 (10⫺8 m). Mutation of either the 5⬘ nonconsensus site 0.05) by deletion of the region from ⫺830 to ⫺711. Treatment
(constructs 2 and 3), the 3⬘ consensus site (construct 4), or with R5020 stimulated (P ⬍ 0.05) the activity of all OXTR
both sites (construct 5) at ⫺104 reduced (P ⬍ 0.05) basal promoter-reporter constructs. Next, the same experiment
luciferase expression in unstimulated cells by 25– 40% com- was repeated with the reporter constructs and PGR-B, but
pared with the wild-type reporter (construct 1) depending on cells were treated with a range of R5020 doses. Dose-depen-
the location of the mutation. Mutation of the ⫺64 SP1 site dent effects of R5020 (10⫺14–10⫺5 m) on activity of the ⫺830
reduced (P ⬍ 0.01) basal luciferase expression to very low and ⫺220 OXTR promoter constructs were observed in
levels, regardless of whether the ⫺104 site was wt (construct 2fTGH cells cotransfected with PGR-B, with concentrations
Fuller W. Bazer 769
906 Endocrinology, February 2006, 147(2):899 –911 Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription
as low as 10⫺10 m R5020 stimulated transactivation in cells The OXTR promoter constructs were then transfected into
transfected with OXTR constructs (data not shown). 2fTGH cells along with ESR1IIc or PGR-B and treated with
E2 (10⫺8 m) or R5020 (10⫺8 m) for 24 h, respectively (Fig. 10).
Deletion of the AP-1 and PRE half site does not affect E2 or Deletion of the AP-1 site and PRE half site did not affect (P ⬎
R5020 stimulation of OXTR promoter activity 0.10) the stimulatory effects of E2 and R5020 on OXTR pro-
The minimal OXTR(⫺220/⫺4) promoter contains an AP-1 moter activity. These results indicate that the SP1 binding
site at ⫺220 and a PRE half site at position ⫺187. To deter- sites at ⫺104 and ⫺64 are the only cis elements required for
mine whether these sites are involved in hormone respon- basal activity of the OXTR promoter and its responsiveness
siveness, the sites were sequentially removed by 5⬘ deletion. to stimulation by ligand-activated ESR1 and PGR-B.
770 Wolf Prize in Agriculture
Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription Endocrinology, February 2006, 147(2):899 –911 907
908 Endocrinology, February 2006, 147(2):899 –911 Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription
a protein kinase C-dependent pathway that can be syner- regulator of OXTR gene expression (27). Similarly in sheep,
gistically augmented by dexamethasone (58). In rabbit am- in vivo administration of estrogen increases OXTR mRNA
nion cells, in vitro treatment with forskolin and/or cortisol and the number of oxytocin binding sites in the endometrium
activates OXTR gene transcription (59). Protein kinase A, (15, 16, 22, 62). The classical pathway for estrogen regulation
protein kinase C, and nerve growth factor-dependent OXTR of target genes involves ligand-activated ESR1/ERE inter-
up-regulation is also observed in MCF-7 and SK-N-SH cells actions or ERE half sites (37, 47). In the rat OXTR gene, a
transfected with the rat OXTR gene promoter (60). In the palindromic ERE was identified about 4 kb upstream of the
human, basal promoter activity is conferred by the 85-bp transcriptional start site (52). This sequence was hormone-
flanking the 5⬘ end of the human OXTR gene. In this se- responsive in transfection studies in human MCF-7 breast
quence, there is a consensus ETS binding sequence. GABP cancer cells, but only when the 3.3-kb fragment between the
(GA binding protein transcription factor), which binds to the ERE and the basal promoter of the rat OXTR gene was trun-
ETS element, cooperates with AP-1 (FOS/JUN) to activate cated. In the wild-type construct, this ERE was not activated
human OXTR promoter transcription in Hs578T cells (61). by estrogen. However, the human OXTR gene does not have
However, elements that regulate activity of the human and a palindromic ERE in the area comparable to that of the rat
rat OXTR promoter/enhancer regions are not present in the gene (27). Therefore, estrogen induction of ovine OXTR gene
ovine and bovine OXTR promoters, suggesting that the iden- transcription may be indirect rather than due to direct ESR1/
tified pathways regulating OXTR expression in other species ERE interactions.
may not be the same for ruminants. The present study found that an estrogen agonist (E2) and
In every studied mammal, estrogen is considered a key antagonist (ICI 182,780) transactivated the ovine OXTR pro-
moter in cells transfected with ESR1. Deletion and mutation
analyses found that induction of ovine OXTR promoter ac-
tivity by E2-activated ESR1 is dependent on GC-rich SP1
binding sites at ⫺104 and ⫺64 in the minimal promoter, and
basal activity of this promoter is also regulated by the same
GC-rich SP1 elements adjacent to the translational start site
of the OXTR gene. One caveat of the present studies is that
human fibrosarcoma cells were used for transfection assays
instead of an ovine endometrial epithelial cell line the en-
dogenous expresses the OXTR gene. Indeed, the GC-rich
motifs in several gene promoters are cis-acting elements for
an increasing number of ligand-activated nuclear and or-
FIG. 9. Comparative activation of OXTR constructs by wild-type hu- phan receptors that interact with SP1 and related proteins
man PGR-B in 2fTGH cells. Cells were cotransfected with constructs (see Refs. 37 and 47). In the present study, SP1 transcription
containing the OXTR promoter fragments and PGR-B, treated with
vehicle as a control or the nonmetabolizable R5020 progestin (10⫺8 M)
factor was constitutively expressed in nearly all cell types of
for 24 h, and luciferase activity was determined as described in Ma- the endometrium from both cyclic and pregnant ewes and
terials and Methods. Significant induction (P ⬍ 0.10) is indicated with was particularly abundant in LE and sGE. Therefore, the
an asterisk, and results are expressed as mean relative light units primary determinant of OXTR gene regulation in the endo-
(RLU) with SD. Four replicate determinations for each treatment
group were conducted in each experiment. A representative experi-
metrial epithelia is expression of the ESR1 gene. Adminis-
ment of three independent experiments with similar results is pre- tration of estrogen on d 11 or 12 postestrus induces structural
sented. and functional luteolysis in cyclic ewes with the following
772 Wolf Prize in Agriculture
Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription Endocrinology, February 2006, 147(2):899 –911 909
temporal events: 1) increase ESR1 mRNA and protein in Development of the endometrial luteolytic mechanism
endometrial epithelia between 12–24 h; 2) moderate increase and uterine release of luteolytic pulses of PGF occur in the
in endometrial OXTR density between 12–36 h; 3) large in- presence of high circulating levels of progesterone. However,
crease in OXTR density between 36 – 48 h; 4) a decline in the endometrial epithelia are not responsive to receptor-
concentrations of progesterone in plasma after 36 h; and 5) dependent actions of progesterone, because they are PGR-
decrease in CL weight by 48 h (15, 17). Similar temporospatial negative between d 11–15 of the estrous cycle. As proges-
alterations in ESR1 and OXTR gene expression occur in the terone levels decrease and estrogen levels increase during
endometrium of cyclic ewes between d 12–16 postestrus (9, proestrus (d 15–17), PGR protein returns to the endometrial
10, 12). Although exogenous estrogen can clearly elicit de- epithelia of the ovine uterus (10, 12). In the present study,
velopment of the endometrial luteolytic mechanism and in- PGR also increased activity of the ovine OXTR promoter
duce OXTR promoter activity, the precise physiological role through GC-rich SP1 sites rather than the AP-1 site or PRE
of estrogen in the luteolytic mechanism in vivo during a half sites also present in the promoter of the OXTR gene.
natural estrous cycle has not been adequately investigated. Progestin-dependent regulation of OXTR through GC-rich
Estrogen may not be needed to increase OXTR gene expres- sites is not unprecedented becausePGR/SP1 activation of
sion, because ESR1 can be activated in a ligand-dependent glycodelin A and CDKN1A (cyclin-dependent kinase inhib-
manner by estrogens or a ligand-independent manner by itor 1A or p21) in endometrial and breast cancer cells, re-
growth factors (63), such as IGF-I and IGF-II that are abun- spectively, is also due to specific GC-rich SP1 binding sites
dant in the ovine endometrial stroma (64). Furthermore, ICI in their promoters (66, 67). Indeed, uterine OXTR mRNA and
182,780, a pure antiestrogen, will not be useful for in vivo protein increases in the endometrial epithelia and stroma
investigations, because it activated ESR1 and induced OXTR after d 15 and is maximal at estrus, which is coincident with
promoter activity in the present study. Similarly, the anties- PGR mRNA and protein as well as SP1 expression in the
trogens 4⬘-hydroxytamoxifen (4-OHT) and ICI 182,780 acti- same cells (10, 12, 13). Therefore, the progesterone block to
vate reporter gene activity in cells transfected with constructs OXTR formation must be due to inhibition of ESR1 gene
containing GC-rich promoters with SP1 and SP3 binding transcription via direct or indirect actions that does not in-
sites (65). In addition to the OXTR, many other endometrial volve SP1.
genes that are regulated by hormone and orphan receptors In the present study, IFNT did not affect OXTR promoter
are likely to involve SP1 and related transcription factors. activity or E2 induction of OXTR promoter activity. These
910 Endocrinology, February 2006, 147(2):899 –911 Flemming et al. • ESR1 Regulates Ovine OXTR Gene Transcription
results were consistent with the lack of classical interferon Ovine interferon- inhibits estrogen receptor up-regulation and estrogen-in-
duced luteolysis in cyclic ewes. Endocrinology 136:4932– 4944
regulatory elements (ISRE, IRFE, or GAS) in the ovine OXTR 16. Spencer TE, Becker WC, George P, Mirando MA, Ogle TF, Bazer FW 1995
promoter and the lack of IRF1 and IRF2 effects on OXTR Ovine interferon- regulates expression of endometrial receptors for estrogen
promoter activity in transient transfection assays. Collec- and oxytocin but not progesterone. Biol Reprod 53:732–745
17. Hixon JE, Flint AP 1987 Effects of a luteolytic dose of oestradiol benzoate on
tively, results of the present and previous studies continue to uterine oxytocin receptor concentrations, phosphoinositide turnover and pros-
support our theory that the antiluteolytic effects of IFNT taglandin F-2␣ secretion in sheep. J Reprod Fertil 79:457– 467
from the conceptus are to inhibit or silence ESR1 gene tran- 18. Vallet JL, Lamming GE, Batten M 1990 Control of endometrial oxytocin
receptor and uterine response to oxytocin by progesterone and oestradiol in
scription, which in turn precludes ESR1 stimulation of OXTR the ewe. J Reprod Fertil 90:625– 634
gene transcription and, therefore, oxytocin-induced release 19. Roberts RM, Ealy AD, Alexenko AP, Han CS, Ezashi T 1999 Trophoblast
of luteolytic pulses of PGF by uterine epithelia. Future ex- interferons. Placenta 20:259 –264
20. Wathes DC, Lamming GE 1995 The oxytocin receptor, luteolysis and the
periments are needed to address how progesterone down- maintenance of pregnancy. J Reprod Fertil Suppl 49:53– 67
regulates PGR gene expression, what factors critically reg- 21. Kim S, Choi Y, Spencer TE, Bazer FW 2003 Effects of the estrous cycle,
ulate PGR inhibition of ESR1 gene transcription in the pregnancy and interferon on expression of cyclooxygenase two (COX-2) in
ovine endometrium. Reprod Biol Endocrinol 1:58
endometrial epithelia of the uterus, and if SP1 and related 22. Spencer TE, Bazer FW 1996 Ovine interferon suppresses transcription of the
transcription factors regulate other endometrial genes in the estrogen receptor and oxytocin receptor genes in the ovine endometrium.
Endocrinology 137:1144 –1147
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ovine estrogen receptor-␣ promoter and functional regulation by ovine inter-
Acknowledgments feron-. Endocrinology 142:2879 –2887
24. Choi Y, Johnson GA, Burghardt RC, Berghman LR, Joyce MM, Taylor KM,
We thank Haijun Gao for assistance with immunohistochemical anal- Stewart MD, Bazer FW, Spencer TE 2001 Interferon regulatory factor-two
ysis of SP1 protein and other members of our laboratories who con- restricts expression of interferon-stimulated genes to the endometrial stroma
tributed to the research presented in this paper. and glandular epithelium of the ovine uterus. Biol Reprod 65:1038 –1049
25. Vallet JL, Bazer FW 1989 Effect of ovine trophoblast protein-1, oestrogen and
progesterone on oxytocin-induced phosphatidylinositol turnover in endome-
Received September 1, 2005. Accepted October 19, 2005. trium of sheep. J Reprod Fertil 87:755–761
Address all correspondence and requests for reprints to: Fuller W. 26. Telgmann R, Bathgate RA, Jaeger S, Tillmann G, Ivell R 2003 Transcriptional
Bazer, Center for Animal Biotechnology and Genomics, 442 Kleberg regulation of the bovine oxytocin receptor gene. Biol Reprod 68:1015–1026
Center, 2471 TAMU, Texas A&M University, College Station, Texas 27. Kimura T, Saji F, Nishimori K, Ogita K, Nakamura H, Koyama M, Murata
77843-2471. E-mail: fbazer@cvm.tamu.edu. Y 2003 Molecular regulation of the oxytocin receptor in peripheral organs. J
Financial support for these studies was obtained from National In- Mol Endocrinol 30:109 –115
stitutes of Health R01 Grant HD32534 and National Institute on Envi- 28. Spencer TE, Johnson GA, Burghardt RC, Bazer FW 2004 Progesterone and
ronmental Health Sciences 5 P30 ES09106-03. placental hormone actions on the uterus: insights from domestic animals. Biol
Reprod 71:2–10
29. Pellegrini S, John J, Shearer M, Kerr IM, Stark GR 1989 Use of a selectable
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Endocrinology is published monthly by The Endocrine Society (http://www.endo-society.org), the foremost professional society serving the
endocrine community.
Fuller W. Bazer 775
Center for Animal Biotechnology and Genomics,4 Departments of Animal Science,5 and Veterinary Integrative
Biosciences,6 Texas A&M University, College Station, Texas 77843
Department of Biological Resources and Technology,7 Yonsei University, Wonju 220-710, South Korea
The above article is reprinted with permission from the Society for the Study of Reproduction, Inc.
776 Wolf Prize in Agriculture
174 KA ET AL.
FIG. 1. Detection of FGF7 mRNA in the endometrium. A) Northern blot analysis of FGF7 mRNA (20 lg) in the endometria of ovariectomized pigs treated
with P4, P4 þ E2 (P4E2), P4 þ ZK (P4ZK), and P4ZKE2. The positions of the 28S (4.7-kb) and 18S (1.8-kb) rRNAs are indicated. A single transcript for FGF7
(;2.4 kb) is detected. B) Steady-state levels of FGF7 mRNAs in CO-, E2-, P4-, P4E2-, P4ZK-, and P4ZKE2-treated pig endometria, expressed as least square
means of the relative units of counts per minute with overall SEM, normalized for differences in sample loading using 18s rRNA and representing 20 lg of
total endometrial mRNA per sample.
immunohistochemistry and in situ hybridization figures were assembled using ESR2. However, although ESR1 was readily detected by RT-
Adobe Photoshop 8.0 (Adobe Systems Inc., San Jose, CA). PCR, ESR2 was barely detected above background in total
endometrium from pigs from all the treatment groups (Fig. 3).
Statistical Analysis In all the endometrial samples, the level of ESR1 was ;8-fold
All quantitative data were subjected to least-squares ANOVA using the
higher than that of ESR2. In addition, ESR2 was not detected
general linear models procedures of the Statistical Analysis System (SAS above the level of the sense cRNA probe or irrelevant IgG
Institute, Cary, NC). Slot blot hybridization data were corrected for differences background using in situ hybridization or immunohistochem-
in sample loading using the 18S rRNA data as a covariate. Orthogonal contrasts istry, respectively (data not shown). In situ hybridization
were used to determine the effects of treatment. Preplanned contrasts were CO analysis of pig endometrium localized FGF7 mRNA to the LE
vs. E2, E2 vs. P4, P4 vs. P4 þ E2, P4 þ ZK vs. P4 þ ZK þ E2, and P4 vs. P4 þ of ovariectomized pigs treated with P4, P4 and E2 or P4, E2,
ZK. All tests of significance were performed using the appropriate error terms
according to the expectation of the mean squares for error. A P value equal to
and ZK. However, FGF7 mRNA was not detected in the
or less than 0.05 was considered statistically significant. Data are presented as endometrial LE of pigs treated with CO vehicle, P4 and ZK or
least-square means (LSM) with standard errors (SEM). E2 alone (Fig. 4). Immunohistochemistry for PGR revealed
expression in the endometrial LE of pigs treated with CO
RESULTS vehicle or E2, whereas PGR expression was absent in the LE of
pigs treated with P4 alone or P4 in combination with ZK and/or
Steady-State Levels of FGF7 mRNA in the Pig Endometrium E2. In contrast, PGR was expressed in the endometrial stromal
and of FGF7 Protein in the Uterine Lumen cells of pigs in all the treatment groups (Fig. 4). Immunohis-
tochemistry for ESR1 revealed expression in the LE, stromal
The FGF7 cRNA detected a single transcript of ;2.4 kb in cells, and GE of all pigs in all treatment groups. In the
Northern blot analysis of pig endometrial total RNA (Fig. 1A). following section, the relationships between treatment, PGR
The steady-state levels of endometrial FGF7 mRNA were not and ESR1 status, and FGF7 mRNA expression are summarized
different between pigs that received CO or E2 (P . 0.10), or by treatment group (Fig. 4).
between pigs that received P4 or P4ZK (P . 0.10). However Ovariectomized pigs injected with corn oil. In the
FGF7 mRNA increased in pigs injected with P4 compared to absence of ovarian steroid hormones following ovariectomy,
those injected with E2 (P , 0.05). The combination of P4 and the absence of P4 and E2 resulted in default expression of both
E2 increased endometrial FGF7 mRNA compared to P4 alone PGR and ESR1 in the endometrial LE, stromal cells, and GE
(P , 0.01), and the addition of E2 (PZKE2) to the P4ZK (Fig. 4). The presence of PGR in the LE is considered to
treatment increased the level of FGF7 mRNA significantly (P preclude the expression of FGF7 mRNA in CO-treated pigs.
. 0.01). The results of the Western blotting analysis to detect Ovariectomized pigs injected with progesterone. Con-
FGF7 protein purified from uterine flushings using heparin- tinuous 9-day exposure of pigs to P4 resulted in the down-
coated agarose beads are shown in Figure 2. An immunore- regulation of PGR in endometrial LE but not in stromal cells,
active FGF7 protein of 17-kDa was detected in the flushings and did not affect the expression of ESR1 in LE, GE or stromal
from pigs treated with P4 and E2, and two molecular mass cells (Fig. 4). The combined effects of P4-induced down-
variants of the FGF7 protein, 17 kDa and 25 kDa, were regulation of PGR in LE and the maintenance of PGR in
detected in the flushings from pigs treated with the combina- stromal cells is considered to enable P4 to interact with PGR in
tion of P4, E2, and ZK, which indicates the secretion of FGF7 stromal cells to induce the synthesis and secretion of a putative
into the uterine lumen of pigs (Fig. 2). progestamedin that induces FGF7 in the LE.
Ovariectomized pigs treated with progesterone and
Relationships Between FGF7 mRNA, PGR, and ESR1 ZK137,316. It appears that 75 mg ZK daily does not
in Pig Endometria recapitulate the previous results obtained for sheep [30] to
block completely the effects of P4 on the pig endometrium.
Previous studies have reported the presence of ESR2 mRNA PGR in the LE and stromal cells are differentially sensitive to
and ESR2 protein in the pig uterus [27–29]. A specific PCR the effects of ZK. Although ZK did not inhibit P4-induced
product was detected in the pig endometrium using primers for down-regulation of PGR in endometrial LE in 5/5 pigs, ZK
ESR2. Sequence analysis indicated that the product was pig bound PGR in the endometrial stromal cells, thereby rendering
778 Wolf Prize in Agriculture
176 KA ET AL.
780 Wolf Prize in Agriculture
FIG. 5. Schematic illustration of the proposed hormonal regulation of FGF7 expression in the porcine pregnant uterine endometrium. During early
pregnancy, continuous exposure of the uterine epithelia to progesterone down-regulates PGR, thereby eliminating PGR-dependent inhibition of
expression of most P4-regulated genes, e.g., those in the LE. Therefore, the endocrine effects of P4 in inducing the expression of FGF7 in endometrial LE
and its secretion into the uterine lumen are mediated by a paracrine-acting factor(s) (progestamedin) produced by the PGR-positive stromal cells.
Furthermore, in PGR-negative epithelial cells, estrogen produced by pig conceptuses binds to ESR1 in the LE to induce FGF7 expression. The combined
endocrine/paracrine effects of ovarian P4 and conceptus estrogens are likely responsible for the high levels of FGF7 mRNA expression in the endometria
and of FGF7 protein in the uterine lumen on Day 12 of pregnancy. During the estrus cycle, FGF7 expression increases in the LE during the P4-dominated
luteal phase, whereas maximal levels of FGF7 are attained on Day 12 of pregnancy after the PGR are down-regulated and when LE is stimulated by high
levels of estrogen released by pig conceptuses for pregnancy recognition and P4 can stimulate the secretion of a progestamedin(s) from uterine stromal
cells. It is hypothesized that secreted FGF7 acts on the conceptus to stimulate the proliferation and differentiation of the trophectoderm.
LE that simultaneously down-regulate PGR and either allow or dissect effectively P4 regulation of gene expression in the LE
stimulate the expression of endometrial genes [30, 46]. In pigs, during the estrus cycle and early pregnancy.
the expression of PGR in endometrial LE and GE is down- Three conclusions can made based on the results of the
regulated by Day 10 of the estrus cycle and pregnancy, whereas present study. First, similar to sheep [23], P4 negatively auto-
the expression of PGR is maintained in stromal cells and the regulates PGR in the LE, but not PGR in the endometrial
stromal cells of pigs. The expression of PGR in the LE and
myometrium [19]. Removal of PGR from LE correlates with stromal cells was detected in pigs that received CO but it was
loss of MUC1 and expression of secreted phosphoprotein 1 down-regulated in the LE of all pigs treated with P4. Second,
(SPP1; also known as osteopontin) on the apical surface of the similar to the results for P4-regulated genes in the endometrial
LE, which exposes integrins to extracellular matrix proteins for epithelia of sheep [23, 30], the presence of PGR in pig LE
trophoblast attachment to the uterus [38, 47]. This is also the precludes the induction of FGF7. All pigs with PGR in the LE,
period during which the endometrium releases many cytokines i.e., those treated with CO alone or E2 alone, failed to express
and growth factors into the uterine lumen of the pig to support FGF7 in LE. Third, the combined effects of P4-induced down-
conceptus development and trophoblast elongation [48]. regulation of PGR in LE and P4 interaction with PGR in
Although the loss of PGR from the endometrial epithelia of stromal cells result in the expression of a progestamedin(s)
from stromal cells that acts on LE to induce FGF7. Pigs treated
pigs is well established [19], the present results are the first to with P4 exhibited down-regulation of PGR in LE concomitant
with induction of FGF7. Furthermore, FGF7 expression
requires functional stromal PGR, since ZK treatment ablated
3 P4-induced FGF7 expression. However, in the absence of
functional PGR (treatment with P4, E2, and ZK), E2 alone
FIG. 4. Interrelationships between FGF7 mRNA, PGR protein, and interacts with ESR1 to induce FGF7 expression.
estrogen receptor a protein in pig endometria. First column: nuclear
localization of ESR1 protein in the luminal epithelia (LE) of corn oil (CO) The mechanisms by which P4 both down-regulates PGR
and E2-treated (E2) pigs, whereas PGR is present in stromal cells (ST) of the and up-regulates the expression of other genes within the
endometria from pigs in all the treatment groups. A section from an E2- uterine LE of pigs are not understood. Induction of stromal-
treated pig stained with nonimmune mouse IgG (IgG) serves as a negative derived progestamedins is one explanation for these phenom-
control. The width of each field is 540 lm. Second column: nuclear ena, although an attractive alternative hypothesis has been put
immunostaining for PGR in the LE, GE, and ST of endometria from pigs in
all treatment groups. A section from an estradiol valerate-treated pig
forward by Geisert and colleagues [46]. They propose that P4
stained with nonimmune rat IgG serves as a negative control. The width of interacts with PGR in LE to stimulate factors that activate
each field is 540 lm. Third and fourth columns: in situ hybridization nuclear factor kappa B (NF-jB), which then functions to
analysis of FGF7 mRNA expression in pig endometria. The left and right inhibit PGR expression and activate transcription of genes that
panels represent corresponding bright-field and dark-field images, are believed to be involved in implantation [46]. The results of
respectively, of endometria from pigs in each treatment group. A the present study are consistent with both of these theories of
representative section from a P4-treated pig hybridized with a radiola-
beled sense cRNA probe (Sense) serves as a negative control. Note that endometrial gene regulation by P4.
FGF7 mRNA is detectable only in the LE, and that the hybridization signal Estrogens secreted by pig conceptuses on Day 12 of
is evident only in the endometria from pigs treated with P4, P4 þ E2 (PE2), gestation comprise the maternal recognition signal that
and P4 þ ZK þ E2 (PZKE2). The width of each field is 690 lm. switches the secretion of endometrial prostaglandin F2a from
Fuller W. Bazer 781
178 KA ET AL.
the endocrine to exocrine direction to prevent CL regression differentiation of conceptus trophectoderm [4, 5] by influencing
[16]. In addition, conceptus estrogens modulate uterine gene DNA synthesis and through motility, differentiation, cytopro-
expression to support uterine secretions and the controlled tective, and antiapoptotic effects on cells [54]. Western blotting
inflammatory-like events that characterize changes in concep- detected an immunoreactive FGF7 protein of about 25 kDa only
tus morphology and uterine remodeling for implantation in pigs in the uterine flushings of pigs treated with E2. Interestingly, a
[49]. Indeed, secreted SPP1 is induced by estrogen in LE, and 17-kDa FGF7 protein was also detected in uterine flushings
it initially localizes to the LE in close proximity to the Day-12 from pigs treated with E2; this protein may be a cleavage
implanting conceptuses [47], whereas conceptus secretion of fragment of the native 25-kDa FGF7 protein. These results
estrogens correlates with conceptus secretion of IL1b and may indicate that E2 regulates the secretion of FGF7 from LE cells. It
modulate uterine responses to this cytokine [50]. The is known that estrogen (of conceptus origin or administered
importance of estrogen for the early survival of pig conceptuses exogenously) induces pseudopregnancy, modulates the redi-
is underscored by pregnancy loss in response to premature rection of prostaglandin F2a from primarily endocrine secretion
exposure of the pregnant uterus to estrogen. Administration of into the underlying vasculature to exocrine secretion into the
estrogen on Days 9 and 10 of pig pregnancy is associated with uterine lumen [16], and increases the secretory activities of
altered expression of SPP1 and cyclooxygenase 1 in LE [46, endometrial epithelia directly or by increasing uterine expres-
51] and degeneration of conceptuses by Day 15 [52]. In pigs, sion of prolactin receptors on uterine epithelia [55]. Therefore,
ESR1 is readily detectable in LE from Day 5 through Day 12 of we hypothesize that FGF7 secretion is regulated by conceptus
the estrus cycle and pregnancy, then decreases, but remains estrogens during pregnancy and by ovarian estrogens during the
detectable until Day 15 [18]. The presence of ESR1 in LE estrus cycle. The detailed cellular mechanism by which estrogen
provides a mechanism by which estrogens secreted by the affects epithelial FGF7 secretion remains to be determined.
elongating pig conceptus can stimulate the changes in uterine In summary, the uterine environment of early pregnancy in
function that are necessary for the maintenance of pregnancy pigs is complex and influenced by the overlapping actions of
[18]. Previous reports, using endometrial explant cultures and conceptus estrogens for pregnancy recognition and the
in vivo steroid replacement experiments, have strongly permissive effects of P4 acting on uterine epithelia to down-
suggested that the induction of FGF7 in uterine LE during regulate PGR and/or via PGR expressed by uterine stromal
early pregnancy in pigs is stimulated primarily by conceptus cells to stimulate the expression of a progestamedin(s) that
estrogens [5, 42]. The results of the present study support these mediates the developmental changes in uterine functions
previous reports and further indicate that FGF7 is induced in necessary for the establishment and maintenance of pregnancy.
the uterine LE of ovariectomized pigs injected with E2, but The dynamic regulation of FGF7 by estrogen and P4 reflects
only when P4 has down-regulated PGR in the LE. The uterine this complexity, and strongly suggests important roles for
LE of pigs treated with E2 alone does not express FGF7 FGF7 in stimulating the proliferation and differentiation of the
mRNA, probably due to the lack of P4 to down-regulate PGR pig conceptus during elongation and implantation. The results
in the LE, whereas pigs treated with a combination of E2 and of the present study provide new insights into the intricate
P4 exhibit FGF7 expression. Furthermore, functional stromal interplay between the actions of estrogen and P4 to modify
PGR, and therefore progestamedins, are not required for E2 gene expression in the peri-implantation pig uterus. A similar
induction of FGF7 because the addition of ZK did not alter the interplay between pregnancy recognition signals and proges-
effects of the combination of P4 and E2. terone has been reported for sheep [56, 57]. However, the
There is a discrepancy between the results of the present magnitude and extent of the influence that conceptus estrogens
study and those in the previous reports with respect to the impart on the early pregnant uterus are remarkable and unique
fining that estrogen in the absence of P4 can increase to the pig. Clearly, further studies are warranted to dissect
endometrial expression of FGF7 [5, 37]. Ka et al. [5] used further the endocrine and paracrine regulation of gene
endometrial explant cultures from Day 9 of the estrus cycle, at expression in the uterus of pregnant pigs.
which time-point the PGR are significantly reduced compared
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R. Michael Roberts
Department of Animal Sciences
University of Missouri, Columbia, Missouri, USA
k
CURRICULUM VITAE
Date and Place of Birth: October 23, 1940; Ilkley, Yorkshire, England
EDUCATION:
Bradford Grammar School, England - 1951-59
B.A. (Botany) Oxford University, England - 1962
D. Phil. Oxford University, England - 1965
APPOINTMENTS:
- Research Associate, State University of New York at Buffalo NY, 1965-67.
- Assistant Professor of Biology, State University of New York at Buffalo, NY,
1967-68.
- Senior Research Fellow, UK Atomic Energy Authority, Radiochemical Centre,
Amersham, England, 1968-69.
- Assistant Professor of Biochemistry, University of Florida, January 1970-September
1972.
785
786 Wolf Prize in Agriculture
- 1st Recipient, Research Award, College of Agri., Food & Natural Resources,
University of Missouri-Columbia, 1995.
- Member, National Academy of Sciences, elected 1996.
- The Alexander vom Humboldt Award for Agriculture, 1996.
- President’s Award for Research and Creativity, University of Missouri system,
1997.
- Foreign Specialist, National Institute of Animal Industry, Japan, 1998.
- Docteur Honoris Causa (Honorary Doctorate) from the University of Liege,
Belgium, 1998.
- Nalbandov Memorial Lecturer, University of Illinois, 2000.
- Wolf Prize for Agriculture 2003; For the discoveries of interferon-tau and other
pregnancy-associated proteins, which clarified the biological mystery of signaling
between the embryo and mother to maintain pregnancy.
- Larry Ewing Memorial Lecturer, John Hopkins University School of Public Health
(2003).
- University of Missouri Faculty/Alumni Award 2005.
- Scientific American Top 50 List (Dec) 2005 Acknowledged for accomplishment
in research and technological leadership.
- Patent Recipient Awardee-University of Missouri-Columbia Technology Transfer
Showcase 2006.
- Carl G. Hartman Award-Recognition of a research career and scholarly activities
in the field of reproductive biology 2006.
- International Federation Placenta Association 2007 Senior Investigator Award.
SOCIETY MEMBERSHIP:
American Society of Biological Chemists
Society for Study of Reproduction
American Society for Advancement of Science
International Society for Interferon and Cytokine Research
American Society for Cell Biology
Endocrine Society
AUTOBIOGRAPHY
I was born in my parents’ home on October 23, 1940, in Menston, a village in
West Yorkshire in the postal district of Ilkley, a few miles north of the industrial
cities of Bradford and Leeds. My father was a wool sorter in Bradford, while my
mother had been employed until my birth in a shirt factory. My one brother, Glyn,
was also a war baby, born in February 1944. I was enrolled in Menston Primary
School from the autumn of 1945 until the summer of 1951 and then in Bradford
(Boys) Grammar School until 1959. The latter was a “Direct Grant Grammar
788 Wolf Prize in Agriculture
subject, but who ensured that classical botany was never much emphasized. Instead
students were exposed to extensive amounts of genetics and plant biochemistry.
Even though my decision to become a plant scientist appeared radically unsound
to my father, I thoroughly enjoyed my remaining years as an undergraduate, and
graduated with an “Upper Second” honors degree in June of 1962.
There are two events during my undergraduate life that are worth remarking
upon, since they have had a profound effect on my future. The first was that I
married Susan Elizabeth Davies, a technician in the department of Botany, in
January 1961. We had our first child, Mark Glyn, in the late summer of that year.
My college was silently aghast, and the addition of a family certainly strained what
financial resources we had available, particularly as it was impossible, even in the
1960s, for a young mother to find child care and to continue to work. Our flat was
also pretty primitive and certainly cheap by Oxford standards; it had an outside
lavatory, a bath tub in the kitchen, and no central heating. In the record, arctic-
like winter of 1963, when Susan was pregnant with our second child, Samantha
Clare, everything seemed to freeze, including the lavatory. Despite this unpropitious
start and impoverished state, I was extremely happy and certainly a more motivated
individual than before. Our marriage has survived for 47 years.
The second noteworthy occurrence was that in March of my final undergraduate
year I was awarded a prestigious Christopher Welch Scholarship in biology, which
would pay me a stipend of £500 (now £13,500) a year to pursue a D.Phil. degree
at Oxford. It was an honor to accept a scholarship previously held by such scientists
as John Eccles, J. Z. Young, Peter Medawar, Anne McLaren, Alec Jeffreys, and John
Gurdon, among others. The examination for this award was competitive and involved
written and oral sections, which required that the applicant map out a research
problem they proposed to tackle. I developed a theoretical approach to studying
the biosynthesis of cell wall polysaccharides in roots through the incorporation of
radioactive precursors and then using autoradiography to follow patterns of
deposition along files of cells from their stem cell origins in the meristem through
the main zone of elongation further along the root. Oddly, this theoretical scheme
worked almost perfectly when I came to perform real experiments during my
graduate training.
My nominal supervisor during my graduate student days was a plant biochemist,
Vernon Butt, a nice man but who was largely absent from the laboratory during
my three years of research as he tried to recover from the premature death of his
wife and to bring up two young children. I worked more closely with Lionel
Clowes, who discovered the quiescent centre within root meristems, and Barrie
Juniper, a microscopist, with whom I published my first paper on the production
of mucus-like, lubricating polysaccharides by the maize root cap ( Juniper and
Roberts 1966). My thesis eventually led to the publication of six papers, but
getting Vernon to even read them (I was in the U.S.A. by then) was like pulling
790 Wolf Prize in Agriculture
teeth. The gambit that eventually worked was to submit and publish a paper
without his authorship (Roberts 1967). I had no trouble after that with getting
him to edit and co-author the subsequent manuscripts.
In the summer of 1964, I attended the International Botanical Congress in
Edinburgh and had the opportunity to meet several plant scientists from North
America, who were recruiting postdoctoral fellows. The post-Sputnik, U.S. science
expansion was in full swing. Although Buffalo New York was not the place of my
dreams, I eventually decided to join Frank Loewus there in the Department of
Biology. America seemed such a potential adventure, and it was. I had never travelled
by air before; I didn’t possess a driver’s license, and fondly believed that we could
get about on bicycles, just as we had in Oxford. That first winter in Buffalo, we had
135 inches of snow, but at least we were warmer than we had been in our frigid
flat on Divinity Road, Oxford. I drove a VW bug illegally (you need an automobile
to survive a Buffalo winter) for six months before taking my test and became an
expert on ice.
My U.S. education began in September 1965. Frank Loewus was best known
for his work in ascorbic acid biosynthesis, but had recently discovered the inositol
pathway in plants in which myo-inositol is oxidatively cleaved to D-glucuronic
acid. I was able to demonstrate that myo-inositol, via this cleavage reaction, is a
precursor of the uronic acids and pentoses incorporated into cells wall pectins and
hemicelluloses (Roberts, Deshusses et al. 1968). I also had the opportunity to study
the formation of apiose, an unusual sugar found in the walls of the pond weed
Lemna gibba, and of phytic acid, the polyphosphorylated form of inositol (Roberts,
Shah et al. 1967). Frank taught me an awful lot, particularly some carbohydrate
chemistry and the use of rigorous methods for identifying metabolites (with no
help from mass spectrometry). The lack of course work associated with an Oxford
D.Phil. and the narrow scope of the prior undergraduate curriculum allows a
motivated student enormous license to follow what interests him, but the down
side can be a profound ignorance of all sorts of other useful stuff, a fact I soon
began to appreciate. Another feature of the Loewus laboratory was that it required
scrupulously annotated note books, another valuable training lesson. I thoroughly
enjoyed my time in Frank’s laboratory and owe him a great deal about how to run
and how not to run a research group. In my third postdoctoral year, I was also
appointed as a temporary Assistant Professor and given the opportunity to lecture
and run a laboratory course, all useful experience for the future, although a low
grade in 1967 could condemn a student to a vacation in Vietnam.
The late 1960s were heady times. As a green card recipient (Brits were a
favored nationality in those days), I was obliged to register for the draft myself, but
my advanced age (24) and two children meant I was unlikely ever to be called up
for war duty. In any case, we were only 20 minutes from the Canadian border. The
Vietnam War was not yet at its peak and popular when we first arrived, but out of
R. Michael Roberts 791
hand and facing massive protests by the time we left in 1968. That year the U.S.
experienced the deaths of Robert Kennedy and Martin Luther King, riots in central
cities, including the predominantly black areas of Buffalo, the Johnson resignation,
and the extraordinary Democratic Convention in Chicago, which we watched from
our hotel room in New York, just before embarking on the great ocean liner,
“France”, for Southampton at the end of August. I look back fondly to those years
in the U.S.A. and the freedom of being a postdoc. Our family were also liberated by
the possibilities of traveling where we wanted by automobile, camping where we
chose, and the seductive opportunities of having little extra money in our pockets.
I returned to England in August 1968 as a Fellow of the Atomic Energy Authority
to work at the Radiochemical Center in Amersham, then a government owned,
commercial enterprise that sold radiochemicals. It was an odd 16 months. I had to
sign the “Official Secrets Act” although, in retrospect, that seems no more bizarre
than signing a Loyalty Oath for my subsequent job at the University of Florida. I
was given a lot of freedom to pursue topics in which I was interested, learned a lot
of enzymology, and was able to make use of the almost limitless supplies of
labeled compounds available for metabolic tracer studies. At the same time, fixed
work hours, the limited availability of equipment, and the constant risk of
radioactive contamination were irksome. My mail was opened, and, I assume,
read, but I benefitted from being surrounded by excellent professionals and a
knowledge that I had a job for life should I have wanted such security, which I
didn’t. After having a series of job interviews in botany departments at U.K.
universities I also decided that I didn’t desire that sort of job either, and I seized
upon the opportunity to interview for a position in the Department of Biochemistry
at the University of Florida in Gainesville, then something of a backwater tucked
away in North Florida. I was offered that position, and the family moved to
Gainesville in January 1970.
At Florida, I was hired as a plant biochemist but my laboratory was located in
the Medical School. I continued to concentrate on plant cell wall formation and on
determining whether plants could utilize glucosamine as a precursor
of glycoproteins (Roberts, Connor et al. 1972). I wrote my first NSF grant application
that first month of 1970 with nothing much in the way of preliminary results, but
by late summer was funded. How the world has changed! I also began to gravitate
towards animal biochemistry through collaborations. I teamed up with Owen
Rennert, a pediatrician, to determine whether there was anything abnormal about
the glycosaminoglycans secreted by cultured dermal fibroblasts from patients with
cystic fibrosis (there wasn’t) (Welch and Roberts 1975), but on the basis of a few
ideas wrote a successful application for a Career Development Award to NIH, a
grant that paid my salary for five years from 1972 through 1977. Also, through a
peculiar series of circumstances, one of which was being the faculty advisor for
the University of Florida cricket team, I became acquainted with the reproductive
792 Wolf Prize in Agriculture
special interest in this regard because iron deficiency in neonatal pigs is a frequently
encountered problem for producers (Roberts and Bazer 1988) .
In the mid-1970s, Bazer and I, with the later collaboration of Bill Thatcher, a
dairy cow physiologist, began to turn our attention to the subject of maternal
recognition of pregnancy and specifically how the conceptus signaled to the mother
that she was pregnant. This problem was important to producers, as a female
breeder that failed to remain pregnant was a costly liability. This research became
possible through a second NIH grant, which still remains active here in Missouri
30 years after it was first awarded. Prior work from Bob Moore’s lab in Cambridge
in the 1960s and later by Jacques Martal in France, had indicated that extracts of
sheep conceptuses infused into the uterus were capable of extending the estrous
cycles of non-pregnant ewes and that the active factor was only present in such
extracts for a few days just prior to the time that placentation was initiated, i.e.
before the conceptus had made firm attachment to the uterine wall. Moreover, all
evidence suggested that this factor was a protein. Our group at Florida decided
that the easiest way to identify the factor was probably not by grinding up embryos.
Instead we predicted that it would be secreted. We therefore cultured intact ovine
conceptuses, recovered by flushing the uterus of pregnant ewes at different days of
pregnancy, in culture medium lacking serum but containing radioactive amino
acids, reasoning that the best source of protein would be the medium recovered at
the end of an extended culture period. Two-dimensional electrophoresis allowed
us to identify a major protein of molecular weight approximately 18,000, consisting
of three to four isoforms, that was produced transiently during early pregnancy
and that proved remarkably easy to purify by using a combination of ion-exchange
chromatography and gel filtration (Godkin, Bazer et al. 1982). Initially called
Protein X, we eventually settled on the name ovine trophoblast protein-1 or oTP-1.
Sufficient purified oTP-1 was eventually accumulated to allow us to determine
whether its infusion into the uteri of non-pregnant ewes could lengthen the estrous
cycle, i.e. induce a short pseudopregnancy, by extending the functional lifespan of
the corpus luteum, the progesterone-producing structure on the ovary that normally
regresses at the end of a non-fertile estrous cycle. Indeed it did. A new hormone of
pregnancy had been discovered and one that might have the potential to improve
pregnancy rates in animals at risk for early pregnancy loss. Parallel studies in cows
revealed a similar protein, although the bovine variety appeared to be glycosylated
and of slightly higher molecular weight (Bartol, Roberts et al. 1985).
In late 1985, I took a position at the University of Missouri, with appointments
in both Biochemistry and Animal Sciences. Importantly, I now had direct access to
the sheep flock. Colleagues in Animal Sciences at Missouri, particularly Duane
Keisler, Alan Garverick, and Mike Smith, crucially helped me harvest and culture
embryos that autumn before my official arrival, so providing a stock of oTP-1 and
RNA. My immediate objective was to clone a cDNA for this protein, although the
794 Wolf Prize in Agriculture
laboratory had no experience with the required technology. Moreover, those were
days before the appearance of “kits” and other aids to neophytes such as me who
wanted to become “molecular biologists”. To accomplish the cloning quickly, I
established a collaboration with Keith Marotti at the UpJohn Co. in Kalamazoo,
Michigan. An energetic postdoctoral fellow, Kaz Imakawa, was dispatched to Keith’s
laboratory to establish cDNA expression libraries in lambda phage with RNA
extracted from ovine embryos and to screen these libraries with an antiserum
prepared against the purified oTP-1. The first sequencing data emerged in late
1986 and immediately revealed that oTP-1 was allied to the type I interferons
(IFN), proteins known for their abilities to protect cells from virus. That phone call
with Keith when he described the results was one of those “wow moments” that
happen infrequently but sufficiently often to make research so rewarding an
experience. Despite the clear signal that we were dealing with an IFN, however, it
was clear that the resemblance to human, mouse, and bovine IFNα and IFNβ, the
best characterized kinds of Type I IFN at the time, was relatively distant (~60 %
and 30 % amino acid sequence identities, respectively). Moreover, the predicted
primary structure of oTP-1 indicated that it possessed an extra sequence of six
amino acids on its carboxyl tail, which was absent on IFNα, which are generally
166 residues in length. On the other hand, oTP1 did bear a closer resemblance
(~70 % identity) to a newly discovered Type I IFN, eventually named IFNω. These
data were published in late 1987 in Nature (Imakawa, Anthony et al. 1987), but
not before the manuscript had been misplaced for several weeks. Soon after this
publication, bTP-1, the bovine equivalent of oTP-1 was also cloned (Imakawa,
Hansen et al. 1989). Both the native proteins and their recombinant forms possessed
an antiviral potency similar to that reported for other Type I IFN. On the basis of
this information, I chose to name the oTP-1 and bTP-1 proteins interferon-tau
(IFNτ), although it required several years for the official nomenclature committee
to affirm this designation, largely because there was concern that this new group
was part of the established IFNω family with which it shared structural
characteristics (Allen and Diaz 1994).
A question immediately arose was whether relatives of the IFNτ serve as signaling
molecules for maternal recognition of pregnancy in other mammals and especially
the human. We were excited to find that antiviral activity was released by murine
and porcine embryos (Cross, Farin et al. 1990). The nature of the former activity
still remains mysterious, whereas that of the pig is due to the secretion of a
mixture of IFN-gamma (Lefevre, Martinat-Botte et al. 1990), a cytokine structurally
unrelated to Type I IFN, and IFN-delta, a novel Type I IFN, most closely related to
IFNβ (Lefevre and Boulay 1993). No antiviral activity could be detected in the
medium in which human blastocysts had been cultured (Roberts, Cross et al.
1991). Ultimately, my group went on to show what we had long anticipated,
namely that the gene family encoding the IFNτ (IFNT) was of recent evolutionary
R. Michael Roberts 795
origin (Leaman and Roberts 1992). We showed that it had diverged from the IFNω
gene (IFNW) lineage approximately 36 million years ago (Roberts, Liu et al. 1997).
As a result of this recent origin, the IFNT were confined to a single sub-order, the
Ruminantia, a grouping that contains among others, cattle and sheep and their
relatives, antelope, deer, and giraffe. The IFNT are absent from rodents, primates,
and even other taxa within the Artiodactyl order, such as swine. The ruminant
ungulates are an especially successful group of mammals with a geographic spread
from equatorial regions, e.g. wildebeest, impala, to the arctic, e.g. musk oxen,
reindeer, and elk, and from wetland, e.g. water buffalo, to desert, e.g. addax. Many,
such as the American bison, once roamed in huge numbers and have only become
diminished as a result of recent human activity. The emergence of the IFNT
correlated in time with the origin of the Ruminantia and with the evolution of the
highly efficient, non-invasive placentation that characterizes this taxon. I like to
believe that the recruitment of this interferon for reproductive purposes has had a
great deal to do with the triumphant rise of the magnificent animals that occupy
this sub-order of mammals.
A second and more persistent question has been whether the IFNτ possess
some unique features that especially equip them for their role in maternal
recognition of pregnancy. In our studies at Missouri, we found that the IFNτ have
an antiviral activity comparable or superior to other type I IFN (Alexenko, Leaman
et al. 1997), that they bind to the same receptor as other Type I IFN (Li and Roberts
1994), and that this receptor is especially concentrated on the surface epithelium
of the uterus targeted by IFNτ (Rosenfeld, Han et al. 2002). My group also showed
that the ability of different forms of IFNτ to cause a pseudopregnancy in sheep is
strongly correlated with their potency as antiviral agents (Niswender, Li et al.
1997). The IFNτ activate the same signal transduction system as other Type I IFN
and regulate most, if not all the same genes (Chen, Antoniou et al. 2007). Conversely,
other Type 1 IFN can extend estrous cycle length in ewes provided they are supplied
in amounts that would evoke an equivalent antiviral response to the IFNτ (Ealy,
Green et al. 1998). As a result of such observations, I had begun, by the late
1990s, to favor the alternative hypothesis, namely that the proteins had not evolved
unusual properties but instead their unique role depended upon their being
produced in the right place, at the right time, and in sufficient quantities to trigger
maternal responses that would allow the pregnancy to be maintained. Accordingly
the attention of the laboratory became focused not so much on the proteins as on
IFNT gene expression itself and specifically on the control elements of these genes.
It was already known that IFNT expression was unlike that of the other known
Type I IFN. The genes were not upregulated in response to viruses and other
pathogens, but instead were expressed constitutively as part of a program associated
with conceptus development (Roberts, Cross et al. 1992). This expression is confined
to a single layer of cells known as trophectoderm, the precursor tissue of the fetal
796 Wolf Prize in Agriculture
days a year of sunshine. Certainly, Lake of the Ozarks wasn’t much compensation,
but I felt energized by the change and relished having full control over the animal
studies. Susan and I enjoyed the courtesy of Midwest. Getting your car fixed or
calling a plumber were somehow pleasanter events than they had been. The
university was smaller and more accessible than that of the University of Florida,
and Columbia was a delightful place in which to re-settle. I was treated well; all
agreements with the administration were kept and generally exceeded. I had
dependable support from my Dean, Roger Mitchell, equivalent to an extra grant,
which has been maintained to this date. That source of funding allowed me to try
new directions and take risks that would have been much more difficult with just
federal grant support. No, there have been no regrets, even though I never imagined
I could spend so long in one place. I began to receive more recognition for my
work, particularly for the research on interferon-tau. In 1989, Fuller Bazer and I
shared the Research Award from the Society for Study of Reproduction, the
professional society I still love best. I continued to maintain at least two grants
from NIH, and in 1992 received one of the best presents of my career, a Merit
Award from the institute at NIH, NIHCD, which has supported me for over 30
years. That Merit Award was not only an honor, it provided an additional stable
source of funding for the next ten years. The research on interferon was also
recognized outside the reproductive biology community when I was awarded the
1995 Milstein Award by International Society for Interferon and Cytokine Research.
I suppose that 1996 has to be my banner year because I was elected to the U.S.
National Academy of Sciences, a huge thrill not just for me but for my laboratory,
and later in the year received the Alexander Von Humboldt Award for Agriculture.
The research that most likely gained me NAS membership and provided the
main basis for my selection as the 2002/2003 Wolf Awardee for Agriculture was
undoubtedly that on interferon-tau, work that can be considered important for
several reasons. First, it described a Type 1 IFN functioning in a constitutive biological
process unrelated to pathogenesis. Second, the find provided the first example of a
cytokine involved in trophoblast signaling, plus an example of how a defined
trophoblast product might intervene in maternal immune responses. Third, the
IFNT would appear to be the most recent mammalian Type 1 gene family to have
arisen and provide a clear illustration of how a control region of a gene can be
commandeered and then refined to provide a radically changed pattern of
expression. Most importantly, IFNτ illustrated why cattle, sheep, and goats are able
to recognize that they are pregnant even before the conceptus has attached to the
uterine wall and why many pregnancies fail at this stage of development. I was
immensely proud to receive the Wolf Prize for this and other work on farm species
with my long-time collaborator, Fuller Bazer, and to accept the prize simultaneously
with three other friends, Oliver Smithies, Mario Capecchi, and Ralph Brinster, who
shared the Prize for Medicine.
R. Michael Roberts 799
References
Alexenko, A. P., D. W. Leaman, et al. (1997). “The antiproliferative and antiviral
activities of IFN-tau variants in human cells.” J Interferon Cytokine Res 17(12):
769-79.
Allen, G. and M. O. Diaz (1994). “Nomenclature of the human interferon proteins.”
J Interferon Res 14(4): 223-6.
Antanaitis, B. C., P. Aisen, et al. (1980). “The novel “g’ = 1.74" EPR spectrum of
pink and purple uteroferrin.” J Biol Chem 255(23): 11204-9.
Bartol, F. F., R. M. Roberts, et al. (1985). “Characterization of proteins produced in
vitro by periattachment bovine conceptuses.” Biol Reprod 32(3): 681-93.
Baumbach, G. A., P. T. Saunders, et al. (1984). “Uteroferrin has N-asparagine-
linked high-mannose-type oligosaccharides that contain mannose 6-
phosphate.” Proc Natl Acad Sci U S A 81(10): 2985-9.
Buhi, W. C., C. A. Ducsay, et al. (1982). “Iron transfer between the purple
phosphatase uteroferrin and transferrin and its possible role in iron metabolism
of the fetal pig.” J Biol Chem 257(4): 1712-23.
Chen, T. T., F. W. Bazer, et al. (1973). “Purification and properties of a progesterone-
induced basic glycoprotein from the uterine fluids of pigs.” J Biol Chem
248(24): 8560-6.
Chen, Y., E. Antoniou, et al. (2007). “A microarray analysis for genes regulated
by interferon-{tau} in ovine luminal epithelial cells.” Reproduction 134(1):
123-35.
Couso, R., L. Lang, et al. (1986). “Phosphorylation of the oligosaccharide
of uteroferrin by UDP-GlcNAc:glycoprotein N-acetylglucosamine-1-
phosphotransferases from rat liver, Acanthamoeba castellani, and Dictyostelium
discoideum requires alpha 1,2-linked mannose residues.” J Biol Chem 261(14):
6326-31.
Cross, J. C., C. E. Farin, et al. (1990). “Characterization of the antiviral activity
constitutively produced by murine conceptuses: absence of placental mRNAs
for interferon alpha and beta.” Mol Reprod Dev 26(2): 122-8.
Das, P., T. Ezashi, et al. (2008). “Combinatorial Roles of Protein Kinase A, Ets2, and
3',5'-Cyclic-Adenosine Monophosphate Response Element-Binding Protein-
Binding Protein/p300 in the Transcriptional Control of Interferon-{tau}
Expression in a Trophoblast Cell Line.” Mol Endocrinol 22(2): 331-43.
Das, P., T. Ezashi, et al. (2008). “Effects of FGF2 and oxygen in the BMP
4-driven differentiation of trophoblast from human embryonic stem cells.”
Stem Cell Research 1(1): 61-74.
Ealy, A., J. Green, et al. (1998). “Differences in biological activities among
recombinant variants of ovine interferon-tau.” Biology of Reproduction
58(566-573).
800 Wolf Prize in Agriculture
Ezashi, T., P. Das, et al. (2008). “The Role of Homeobox Protein Distal-Less 3 and
Its Interaction with ETS2 in Regulating Bovine Interferon-Tau Gene Expression-
Synergistic Transcriptional Activation with ETS2.” Biol Reprod.
Ezashi, T., P. Das, et al. (2005). “Low O2 tensions and the prevention of
differentiation of hES cells.” Proc Natl Acad Sci U S A 102(13): 4783-8.
Ezashi, T., A. D. Ealy, et al. (1998). “Control of interferon-tau gene expression by
Ets-2.” Proc Natl Acad Sci U S A 95(14): 7882-7.
Ezashi, T. and R. M. Roberts (2004). “Regulation of interferon-tau (IFN-tau) gene
promoters by growth factors that target the Ets-2 composite enhancer: a
possible model for maternal control of IFN-tau production by the conceptus
during early pregnancy.” Endocrinology 145(10): 4452-60.
Gaber, B. P., J. P. Sheridan, et al. (1979). “Resonance Raman Scattering from
Uteroferrin, the Purple Glycopprotein of the Porcine Uterus.” J. Biol. Chem.
254: 8340-8342.
Godkin, J. D., F. W. Bazer, et al. (1982). “Purification and properties of a major,
low molecular weight protein released by the trophoblast of sheep blastocysts
at day 13-21.” J Reprod Fertil 65(1): 141-50.
Green, J. A., T. E. Parks, et al. (2005). “The establishment of an ELISA for the
detection of pregnancy-associated glycoproteins (PAGs) in the serum of
pregnant cows and heifers.” Theriogenol. 63: 1481-1503.
Green, J. A., S. Xie, et al. (2000). “Pregnancy-associated bovine and ovine
glycoproteins exhibit spatially and temporally distinct expression patterns
during pregnancy.” Biol Reprod 62(6): 1624-31.
Hunt, D. F., J. R. Yates, et al. (1987). “Sequence homology in the metalloproteins:
purple acid phosphatase from beef spleen and uteroferrin from porcine
uterus.” Biochem. & Biophys. Res. Comm. 144: 1154-1160.
Imakawa, K., R. V. Anthony, et al. (1987). “Interferon-like sequence of ovine
trophoblast protein secreted by embryonic trophectoderm.” Nature 330(6146):
377-9.
Imakawa, K., T. R. Hansen, et al. (1989). “Molecular cloning and characterization
of cDNA’s corresponding to bovine trophoblast protein-1. A comparison with
ovine trophoblast protein-1 and bovine interferon-*II.” Molec. Endocrinol 3:
127-139.
Juniper, B. E. and R. M. Roberts (1966). “Polysaccharide Synthesis and the Fine
Structure of Root Cells.” J. Roy. Microscop. 85: 63-72.
Ketcham, C. M., G. A. Baumbach, et al. (1985). “The type 5, Acid Phosphatase
from Spleen of Humans with Hairy Cell Leukemia.” J. Biol. Chem. 260:
5768-5776.
Lang, L., M. Reitman, et al. (1984). “Recognition of a Protein-dependent Determinant
allows Specific Phosphorylation of Oligosaccharides present on Lysosomal
Enzymes.” J. Biol. Chem. 259: 14663-14671.
R. Michael Roberts 801
Larson, M. A., K. Kimura, et al. (2001). “Sexual dimorphism among bovine embryos
in their ability to make the transition to expanded blastocyst and in the
expression of the signaling molecule IFN-tau.” Proc Natl Acad Sci U S A
98(17): 9677-82.
Leaman, D. W. and R. M. Roberts (1992). “Genes for the trophoblast interferons in
sheep, goat, and musk ox and distribution of related genes among mammals.”
J Interferon Res 12(1): 1-11.
Lefevre, F. and V. Boulay (1993). “A novel and atypical type one interferon gene
expressed by trophoblast during early pregnancy.” J Biol Chem 268(26):
19760-8.
Lefevre, F., F. Martinat-Botte, et al. (1990). “Interferon-gamma gene and protein
are spontaneously expressed by the porcine trophectoderm early in gestation.”
Eur J Immunol 20(11): 2485-90.
Li, J. and R. M. Roberts (1994). “Interferon-tau and interferon-alpha interact with
the same receptors in bovine endometrium. Use of a readily iodinatable form
of recombinant interferon-tau for binding studies.” J Biol Chem 269(18):
13544-50.
Malathy, P. V., K. Imakawa, et al. (1990). “Molecular cloning of the uteroferrin-
associated protein, a major progesterone-induced serpin secreted by the porcine
uterus, and the expression of its mRNA during pregnancy.” Mol. Endocr. 4:
428-440.
Meyer, M. D., P. J. Hansen, et al. (1995). “Extension of corpus luteum lifespan and
reduction of uterine secretion of prostaglandin F2 alpha of cows in response
to recombinant interferon- tau.” J Dairy Sci 78(9): 1921-31.
Meyer, M. D., P. J. Hansen, et al. (1995). “Effect of bovine interferon-tau on body
temperature and plasma progesterone concentrations in cyclic dairy cows.”
J Dairy Sci 78(7): 1470-6.
Niswender, K. D., J. Li, et al. (1997). “Effect of variants of interferon-tau with
mutations near the carboxyl terminus on luteal life span in sheep.” Biol Reprod
56(1): 214-20.
Renegar, R. H., F. W. Bazer, et al. (1982). “Placental Transport and Distribution of
Uteroferrin in the Fetal Pig.” Biol. Reprod. 27: 1247-1260.
Roberts, R., J. Cross, et al. (1991). Trophoblast Interferons. In: Growth Factors in
Reproduction. D. W. Schomberg, Serono Symposia, USA, Springer-Verlag, New
York: 235-245.
Roberts, R. M. (1967). “Incorporation of 14C-labeled D-glucuronate and D-galactose
into Segments of the Root Tip of Corn.” Phytochemistry 6: 525-533.
Roberts, R. M. and F. W. Bazer (1988). “The functions of uterine secretions.”
J Reprod Fertil 82(2): 875-92.
Roberts, R. M., A. B. Connor, et al. (1972). “The Formation of Glycoproteins in
Tissues of Higher Plants. Specific Labeling Using (1-14C) glucosamine.”
Biochem. J. 125: 999-1008.
802 Wolf Prize in Agriculture
REFEREED PUBLICATIONS
1. Juniper, B.E. and Roberts, R.M. (1966) Polysaccharide Synthesis and the Fine
Structure of Root Cells. J. Roy. Microscop. 85, 63-72.
2. Roberts, R.M. and Loewus, F. (1966) Inositol Metabolism in Plants III.
Conversion of Myo-inositol-2-3H to Cell Wall Polysaccharides in Sycamore
(Acer pseudoplatanus L.) Cell Culture. Plant Physiol. 41, 1489-1498.
3. Roberts, R.M. (1967) Incorporation of 14C-labeled D-glucuronate and
D-galactose into Segments of the Root Tip of Corn. Phytochemistry 6,
525-533.
4. Roberts, R.M., Shah, R., and Loewus, F. (1967) Conversion of Myo-inositol-
2-14C to Labeled 4-0-methyl-glucuronic Acid in the Cell Wall of Maize Root
Tips. Arch. Biochem. Biophys. 119, 590-593.
5. Roberts, R.M., Shah, R.H. and Loewus, F. (1967) Inositol Metabolism in Plants.
IV. Biosynthesis of Apiose in Lemna and Petroselinum. Plant Physiol. 42,
659-666.
6. Roberts, R.M. and Butt, V.S. (1967) Rates of Incorporation of Label from
D-[14C] Glucose into Different Tissues of the Root-tip of Corn. New Phytologist
66, 563-567.
7. Roberts, R.M. and Butt, V.S. (1967) Patterns of Cellulose Synthesis in Maize
Root-tips. Exper. Cell Res. 46, 495-510.
8. Roberts, R.M., Shah, R., Golebiewski, A. and Loewus, F. (1967) Incorporation
of Methanol into Pectic Substances. Plant Physiol. 42, 1737-1742.
9. Roberts, R.M. and Butt, V.S. (1968) Patterns of Incorporation of Pentose
and Uronic Acid into Cell Walls of Maize Root-tips. Exp. Cell Res. 51,
519-530.
10. Roberts, R.M., Deshusses, J. and Loewus, F. (1968) Inositol Metabolism
in Plants. V. Conversion of Myo-inositol to Uronic Acid and Pentose Units of
Acidic Polysaccharides in Root-tips of Zea mays. Plant Physiol. 43, 979-989.
11. Roberts, R.M., Deshusses, J. and Loewus, F. (1968) Inositol Metabolism in
Plants. Metabolism of Myo-inositol Metabolism in Plants. VI. Conversion
of Myo-inositol to Phytic Acid in Wolffiella floridana. Plant Physiol. 43,
1710-1716.
12. Roberts, R.M. (1968) The Metabolism of L-fucose by Sycamore (Acer
pseudoplatanus L.) Cell Cultures. Arch. Biochem. Biophys. 128, 818-820.
13. Roberts, R.M. and Butt, V.S. (1969) Patterns of Incorporation of D-galactose
into Cell Wall Polysaccharide of Growing Maize Roots. Planta 84, 250-262.
14. Roberts, R.M. and Tovey, K.C. (1969) Trehalase Activity in Selaginella
martensii. Arch. Biochem. Biophys. 133, 408-412.
15. Roberts, R.M. (1970) The Incorporation of D-glucosamine-14C into Root
Tissues of Higher Plants. Plant Physiol. 45, 263-267.
804 Wolf Prize in Agriculture
16. Roberts, R.M. and Tovey, K.C. (1970) A Critical Study of the DEAE paper
Disc Technique Used in Radiochemical Enzyme Assays. Anal. Biochem. 34,
582-590.
17. Tovey, K.C. and Roberts, R.M. (1970) An Artifact in the Chromatography of
Sugar Nucleotides Using Solvents Containing Ammonium Acetate. J. Chromatog.
47, 287-290.
18. Tovey, K.C. and Roberts, R.M. (1970) Studies on the Pyrophosphorylases of
Wheat. Plant Physiol. 46, 406-411.
19. Roberts, R.M., Butt, V.S. (1970) Incorporation of 14C-L-arabinose into
Polysaccharides of Maize Root-tips. Planta (Berl.) 94, 175-183.
20. Roberts, R.M., Heishman, A. and Wicklin, C. (1971) Growth Inhibition and
Metabolite Pool Levels in Plant Tissues Fed D-glucosamine and D-galactose.
Plant Physiol. 48, 36-42.
21. Roberts, R.M. (1971) The Metabolism of D-mannose-14C to Polysaccharide
in Corn Roots. Specific Labeling of L-galactose, D-mannose and L-fucose.
Arch. Biochem. Biophys. 145, 685-692.
22. Roberts, R.M. (1971) The Formation of UDP-glucuronic Acid in Plants. UDP-
glucuronic Acid Pyrophosphorylase from Barley Seedling. J. Biol. Chem. 246,
4995-5002.
23. Kirby, E.G. and Roberts, R.M. (1971) The Localized Incorporation of
3H-L-fucose into Cell Wall Polysaccharides of Cap and Epidermis of Corn
31. Roberts, R.M. and Yuan, B. O-C. (1974) Chemical Modification of the Plasma
Membrane Polypeptides of Cultured Mammalian Cells as an Aid to Studying
Protein Turnover. Biochemistry 13, 4846-4855.
32. Schlosnagle, D.C., Bazer, F.W., Tsibris, J.C.M. and Roberts, R.M. (1974) An
Iron-containing Phosphatase Induced by Progesterone in the Uterine Fluids
of Pigs. J. Biol. Chem. 249, 7574-7579.
33. Roberts, R.M. and Pollard, W.E. (1975) The Incorporation of D-glucosamine
into Glycolipids and Glycoproteins of Membrane Preparation from Phaseolus
aureus hypocotyls. Plant Physiol. 55, 431-436.
34. Baig, M.M., Cetorelli, J.J. and Roberts, R.M. (1975) Plasma Membrane Proteins
and Glycoproteins from Human Fibroblasts Derived from Normal Individuals
and Patients with Cystic Fibrosis. J. Pediatrics 86, 72-76.
35. Welch, D.W. and Roberts, R.M. (1975) Complex Saccharide Metabolism in
Cystic Fibrosis Fibroblasts. Pediatrics Res. 9, 698-702.
36. Chen, T.T., Bazer, F.W., Gebhardt, G. and Roberts, R.M. (1975) Uterine
Secretions in Mammals: Synthesis and Placental Transport of a Purple Acid
Phosphatase in Pigs. Biol. Reprod. 13, 304-313.
37. Bazer, F.W., Chen, T.T., Knight, J.S., Schlosnagle, D.C. and Roberts, R.M. (1975)
Presence of a Progesterone-Induced, Uterine Specific Acid Phosphatase in
Allantoic Fluid of Gilts. J. Anim. Sci. 41, 1112-1119.
38. Roberts, R.M. and Yuan, B. O-C (1975) Radiolabeling of Mammalian Cells
in Tissue Culture Using Acetic Anhydride. Potential Value for Studying the
Dynamics of Protein Turnover in Living Cells. Arch. Biochem. Biophys. 171,
226-233.
39. Roberts, R.M. and Yuan, B.O-C. (1975) Turnover of Plasma Membrane
Polypeptides in Non-Proliferating Cultures of CHO Cells and Human Skin
Fibroblasts. Arch. Biochem. Biophys. 171, 234-244.
40. Stein, G.S., Roberts, R.M., Davis, J.L., Stein, J.L., Thrall, C.L., VanVeen, J. and
Welch, D.W. (1975) Are Glycoproteins and Glycosaminoglycans Components
of the Eukaryotic Genome? Nature 258, 639-641.
41. Hall, J.L. and Roberts, R.M. (1975) Biochemical Characteristics of Membrane
Fractions Isolated from Maize (Zea mays) Roots. Ann. Bot. 39, 983-993.
42. Roberts, R.M. and Bazer, F.W. (1976) Phosphoprotein Phosphatase Activity
of the Progesterone-Induced Purple Glycoprotein of the Porcine Uterus.
Biochem. Biophys. Res. Commun. 68, 450-455.
43. VanVeen, J., Roberts, R.M. and Noonan, K.D. (1976) An Analysis of Lectin-
initiated Cell Agglutinability in a Series of CHO Sub-clones which Respond
Morphologically to growth in Dibutyryl Cyclic AMP. J. Cell Biol. 70, 204-216.
44. Schlosnagle, D.C., Sander, E.G., Bazer, F.W. and Roberts, R.M. (1976) The
Requirement of an Essential Thiol Group and Ferric Iron for the Activity of
the Progesterone-Induced, Porcine Uterine, Purple Phosphatase. J. Biol. Chem.
251, 4680-4685.
806 Wolf Prize in Agriculture
45. VanVeen, J., Noonan, K.D. and Roberts, R.M. (1976) A Correlation between
Membrane Glycopeptide Composition and Changes in Agglutinability Induced
by Dibutyryl 3′:5′ Cyclic AMP in Chinese Hamster Ovary Cell. Exptl. Cell
Res. 130, 405-413.
46. Roberts, R.M., Bazer, F.W., Baldwin, N. and Pollard, W.E. (1976) Progesterone
Induction of Lysozyme and Peptidase Activities in the Porcine Uterus. Arch.
Biochem. Biophys. 177, 499-507.
47. Basha, S.M.M. and Roberts, R.M. (1977) A Simple Colorimetric Method for
the Determination of Tryptophan. Anal. Biochem. 77, 378-386.
48. Basha, S.M.M., Horst, M.N., Bazer, F.W. and Roberts, R.M. (1978) Purification
and Properties of Aminopeptidases from Reproductive Fluids of Pigs. Arch.
Biochem. Biophys. 185, 174-184.
49. Basha, S.M.M., Bazer, F.W. and Roberts, R.M. (1979) The Secretion of a
Uterine Specific, Purple Phosphatase by Cultured Explants of Porcine
Endometrium Dependency upon the State of Pregnancy of the Donor Animal.
Biol. Reprod. 20, 431-441.
50. Horst, M.N. and Roberts, R.M. (1979) Analysis of Polypeptide Turnover Rates
in Chinese Hamster Ovary Cell Plasma Membranes Using Two-Dimensional
Electrophoresis. J. Biol. Chem. 254, 5000-5007.
51. Horst, M.N., Baumbach, G. and Roberts, R.M. (1979) Two-Dimensional
Electrophoretic Analysis of Concanvalin-A Binding Components in the Plasma
Membranes of Chinese Hamster Fibroblasts. FEBS Lett. 100, 385-388.
52. Gaber, B.P., Sheridan, J.P., Bazer, F.W. and Roberts, R.M. (1979) Resonance
Raman Scattering from Uteroferrin, the Purple Glycoprotein of the Porcine
Uterus. J. Biol. Chem. 254, 8340-8342.
53. Bazer, F.W., Roberts, R.M., Basha, S.M.M., Zavy, M.T., Caton, D. and
Barron, D. (1980) Method for Obtaining Ovine Uterine Secretions from
Unilaterally Pregnant Ewes. J. Anim. Sci. 49, 1522-1527.
54. Basha, S.M.M., Bazer, F.W. and Roberts, R.M. (1980) Effect of Conceptus on
Quantitative and Qualitative Aspects of Porcine Uterine Secretions. J. Reprod.
Fert. 60, 41-48.
55. Basha, S.M.M., Bazer, F.W., Geisert, R.D. and Roberts, R.M. (1980) Progesterone-
induced Uterine Secretions in Pigs. Recovery from Pseudopregnant and
Unilaterally Pregnant Gilts. J. Anim. Sci. 50, 113-123.
56. Horst, M.N., Basha, S.M.M. Baumbach, G.A., Mansfield, E. and Roberts, R.M.
(1980) Alkaline Urea Solubilization, Two-dimensional Electrophoresis and
Lectin Staining of Mammalian Cell Plasma Membrane and Plant Seed Proteins.
Anal. Biochem. 102, 399-408.
57. Horst, M.N., Baumbach, G.A. and Roberts, R.M. (1980) Isolation of a Domain
of the Plasma Membrane in CHO Cells which Contains Iodinatable Acidic
Glycoproteins of High Molecular Weight. Biochim. Biophys. Acta 600, 48-61.
R. Michael Roberts 807
58. Mullins, D.E., Bazer, F.W. and Roberts, R.M. (1980) Secretion of a
Progesterone-induced Inhibitor of Plasminogen Activator by the Porcine
Uterus. Cell 20, 865-872.
59. Mullins, D.E., Horst, M.N., Bazer, F.W. and Roberts, R.M. (1980) Isolation
and Characterization of a Plasma Membrane Fraction Derived from the
Luminal Surface of the Pig Uterus During the Estrous Cycle and Early
Pregnancy. Biol. Reprod. 22, 1181-1192.
60. Antanaitis, B.C., Aisen, P., Lillenthal, H.R., Roberts, R.M. and Bazer, F.W.
(1980) the Novel “g′=1.74” EPR Spectrum of Pink and Purple Uteroferrin.
J. Biol. Chem. 255, 11204-11209.
61. Basha, S.M.M. and Roberts, R.M. (1981) The Glycoproteins of Plant Seeds.
Analysis by Two-Dimensional Polyacrylamide Gel Electrophoresis and by
their Lectin Binding Properties. Plant Physiol 67, 936-942.
62. Adams, K.L., Bazer, F.W. and Roberts, R.M. (1981) Progesterone-induced
Secretion of a Retinol-Binding Protein in the Pig Uterus. J. Reprod. Fert. 62,
39-47.
63. Mancarella, D.A., Basha, S.M.M., Mullins, D.E., Mansfield, E., Bazer, F.W. and
Roberts, R.M. (1981) Properties of a Membrane-associated L-Leucine-
naphthylamidase (Leucine Aminopeptidase) from the Porcine Uterus. Biol.
Reprod. 24, 879-888.
64. Bartol, F.F., Thatcher, W.W., Bazer, F.W., Kimball, F.A., Chenault, J.R.,
Wilcox, C.J. and Roberts, R.M. (1981) Effects of the Estrous Cycle and Early
Pregnancy on Bovine Uterine, Luteal and Follicular Responses. Biol. Reprod.
25, 759-776.
65. Godkin, J.D., Bazer, F.W., Moffatt, J., Sessions, F. and Roberts, R.M. (1982)
Purification and Properties of a Major, Low Molecular Weight Protein Released
by the Trophoblast of Sheep Blastocysts at Day 13-21. J. Reprod. Fert. 65,
141-150.
66. Fitzgerald, L.A., Baumbach, G.A., Horst, M.N., Noonan, K.D. and
Roberts, R.M. (1982) The Purification and Immunocytochemical Localization
of the Major, Iodinatable, Cell Surface Glycoproteins of CHO Cells. J. Cell
Sci. 52, 405-424.
67. Zavy, M.T., Sharp, D.C., Bazer, F.W., Fazleabas, A., Sessions, F. and Roberts,
R.M. (1982) Identification of Stage-Specific and Hormonally-Induced
Polypeptides in the Uterine Protein Secretions of the Mare During the Oestrous
Cycle and Pregnancy. J. Reprod. Fert. 64, 199-207.
68. Buhi, W.C., Gray, W.J., Mansfield, E.A., Chun, P.W., Ducsay, C.A., Bazer, F.W.
and Roberts, R.M. (1981) Iron Content and Molecular Weight of Uteroferrin
and a Comparison of Its Iron and Copper Forms. Biochim. Biophys. Acta
601, 32-38.
808 Wolf Prize in Agriculture
69. Denny, J.B. and Roberts, R.M. (1982) A New Immunoreactive Probe for the
Isolation and Analysis of Plasma Membrane Polypeptides. Synthesis and
Properties of Isethionyl 3-(N-2,4-Dinitrophenyl)-aminopropioimidate. J. Biol.
Chem. 257, 2460-2468.
70. Buhi, W.C., Ducsay, C.A., Bazer, F.W. and Roberts, R.M. (1982) Iron Transfer
Between the Purple Phosphatase Uteroferrin and Transferrin and Its Possible
Role in Iron Metabolism of the Fetal Pig. J. Biol. Chem. 257, 1712-1721.
71. Fazleabas, A.T., Bazer, F.W. and Roberts, R.M. (1982) Purification and
Properties of a Progesterone-Induced, Plasmin/trypsin Inhibitor from the
Uterine Secretions of Pigs and Its Immunocytochemical Localization in the
Pregnant Uterus. J. Biol. Chem. 257, 6886-6897.
72. Ducsay, C.A., Buhi, W.C., Bazer, F.W. and Roberts, R.M. (1982) Role of
Uteroferrin in Iron Transport and Macromolecular Uptake by Allantoic
Epithelium of the Porcine Conceptus. Biol. Reprod. 26, 729-743.
73. McDowell, K., Sharp, D.C., Zavy, M.T., Fazleabas, A., Roberts, R.M. (1982)
and Bazer, F.W., Partial Characterization of the Equine Uteroferrin-like Protein.
J. Reprod. Fert. Suppl. 32, 329-334.
74. Masters, R.A., Roberts, R.M., Lewis, G.S., Thatcher, W.W., Bazer, F.W. and
Godkin, J.D. (1982) High Molecular Weight Glycoproteins Released by
Expanding Pre-attachment Sheep, Pig and Cow Blastocysts in Culture.
J. Reprod. Fert. 66, 571-583.
75. Godkin, J.D., Bazer, F.W., Geisert, R.D., Lewis, G.S. and Roberts, R.M. (1982)
Synthesis and Release of Polypeptides by Pig Conceptuses During the Period
of Blastocyst Elongation and Attachment. Biol. Reprod. 27, 977-987.
76. Geisert, R.D., Renegar, R.H., Thatcher, W.W., Roberts, R.M. and Bazer, F.W.
(1982) Establishment of Pregnancy in the Pig: I. Interrelationships Between
Preimplantation Development of the Pig Blastocyst and Uterine Endometrial
Secretions. Biol. Reprod. 27, 925-939.
77. Geisert, R.D., Brookbank, J.W., Roberts, R.M. and Bazer, F.W. (1982)
Establishment of Pregnancy in the Pig: II. Cellular Remodeling of the
Porcine Blastocyst During Elongation on Day 12 of Pregnancy. Biol. Reprod.
27, 941-955.
78. Geisert, R.D., Thatcher, W.W., Roberts, R.M. and Bazer, F.W. (1982)
Establishment of Pregnancy in the Pig: III. Endometrial Secretory Response to
Estradiol Valerate Administered on Day 11 of the Estrous Cycle. Biol. Reprod.
27, 957-965.
79. Zavy, M.T., Clark, W.R., Sharp, D.C., Roberts, R.M. and Bazer, F.W. (1982)
Comparison of Glucose, Fructose, Ascorbic Acid and Glucose phosphate
Isomerase Enzymatic Activity in Uterine Flushings from Nonpregnant and
Pregnant Gilts and Pony Mares. Biol. Reprod. 27, 1147-1158.
80. Renegar, R.H., Bazer, F.W. and Roberts, R.M. (1982) Placental Transport and
Distribution of Uteroferrin in the Fetal Pig. Biol. Reprod. 27, 1247-1260.
R. Michael Roberts 809
81. Fazleabas, A.T., Geisert, R.D., Bazer, F.W. and Roberts, R.M. (1983) The
Relationship Between Release of Plasminogen Activator and Estrogen by
Blastocysts and Secretion of Plasmin Inhibitor by Uterine Endometrium in
the Pregnant Pig. Biol. Reprod. 29, 225-238.
82. Buhi, W.C., Ducsay, C.A., Bartol, F.F., Bazer, F.W. and Roberts, R.M. (1983) A
Function of the Allantoic Sac in the Metabolism of Uteroferrin and Maternal
Iron by the Fetal Pig. Placenta 4, 455-470.
83. Godkin, J.D., Bazer, F.W. and Roberts, R.M. (1984) Ovine Trophoblast Protein
1 (oTP-1), an Early Secreted Blastocyst Protein, Binds Specifically to Uterine
Endometrium and Affects Protein Synthesis. Endocrinol. 114, 120-130.
84. Fazleabas, A.T., Mead, R.A., Rourke, R.A. and Roberts, R.M. (1984) Presence
of an Inhibitor of Plasminogen Activator in Uterine Fluid of the Western
Spotted Skunk During Delayed Implantation. Biol. Reprod. 30, 311-322.
85. Ducsay, C.A., Buhi, W.C., Bazer, F.W., Roberts, R.M. and Combs, G.E. (1984)
Role of Uteroferrin in Placental Iron Transport. II. Effect of Maternal Iron
Treatment on Fetal Iron, Uteroferrin Content and Neonatal Hemoglobin.
J. Anim. Sci. 59, 1303-1308.
86. Fitzgerald, L.A., Denny, J.B., Baumbach, G., Ketcham, C.M. and Roberts, R.M.
(1984) The Effect of Altered Oligosaccharide Structure on the Cell Surface
Number, Distribution and Turnover of the High Molecular Weight Acidic
Glycoprotein of CHO Cells. J. Cell Sci. 67, 1-23.
87. Godkin, J.D., Bazer, F.W., Thatcher, W.W. and Roberts, R.M. (1984) Proteins
Released by Cultured Day 15-16 Conceptuses Prolong Luteal Maintenance
When Introduced into the Lumen of Cyclic Ewes. J. Reprod. Fert. 71, 57-64.
88. Baumbach, G.A., Saunders, P.T.K., Bazer, F.W. and Roberts, R.M. (1984)
Uteroferrin has N-asparagine linked, High Mannose-type Oligosaccharides
Which Contain Mannose-6-phosphate. Proc. Natl. Acad. Sci. USA. 81,
2985-2989.
89. Segerson, E.C., Moffatt, R.J., Bazer, F.W. and Roberts, R.M. (1984) Suppression
of Phytohemagglutinin- stimulated Lymphocyet Blastogenesis by Ovine Uterine
Milk Protein. Biol. Reprod. 30, 1175-1186.
90. Zavy, M.T., Roberts, R.M. and Bazer, F.W. (1984) Acid Phosphatase and
Leucine Aminopeptidase Activity in the Uterine Flushings of Nonpregnant
and Pregnant Gilts. J. Reprod. Fert. 72, 503-507.
91. Lang, L., Reitman, M., Tang, J., Roberts, R.M. and Kornfeld, S.
(1984) Recognition of a Protein-dependent Determinant allows Specific
Phosphorylation of Oligosaccharides present on Lysosomal Enzymes. J. Biol.
Chem. 259, 14663-14671.
92. Fazleabas, A.T., Bazer, F.W., Hansen, P.J., Geisert, R.D. and Roberts, R.M.
(1985) Differential Patterns of Secretory Protein Synthesis within the
Pig Uterine Endometrium in Response to Progesterone. Endocrinol. 116,
240-245.
810 Wolf Prize in Agriculture
93. Ketcham, C.M., Baumbach, G.A., Bazer, F.W. and Roberts, R.M. (1985) The
type 5, Acid Phosphatase from Spleen of Humans with Hairy Cell Leukemia.
J. Biol. Chem. 260, 5768-5776.
94. Saunders, P.T.K., Renegar, R.H., Raub, T.J., Baumbach, G.A., Atkinson, P.H.,
Bazer, F.W. and Roberts, R.M. (1985) The Carbohydrate Structure of Porcine
Uteroferrin and the Role of the High Mannose Chains in Promoting Uptake
by the Reticuloendothelial Cells of the Fetal Liver. J. Biol. Chem. 260,
3658-3665.
95. Raub, T.J., Bazer, F.W. and Roberts, R.M. (1985) Localization of the Iron
Transport Glycoprotein, Uteroferrin, in the Porcine Endometrium and Placenta
by Using Immunocolloidal Gold. Anat. Embryol. 171, 253-258.
96. Bartol, F.F., Roberts, R.M., Godkin, J.D., Bazer, F.W, Lewis, G.S. and
Thatcher, W.W. (1985) Characterization of Proteins produced in vitro by
Preattachment Bovine Conceptuses. Biol. Reprod. 32, 681-693.
97. Hansen, P.J., Bazer, F.W. and Roberts, R.M. (1985) Appearance of
β-Hexosaminidase and other Lysosomal-like Enzymes in the Uterine Lumen
of Gilts, Ewes and Mares in response to Progesterone. J. Reprod. Fert. 73,
411-424.
98. Godkin, J.D., Bazer, F.W. and Roberts, R.M. (1985) Protein Production by
Cultures Established from Day 14-16 Sheep and Pig Conceptuses. J. Reprod.
Fert. 74, 377-382.
99. Hansen, P.J., Anthony, R.V., Bazer, F.W., Baumbach, G.A. and Roberts, R.M.
(1985) In Vitro synthesis and secretion of ovine trophoblast protein-1
during the period of maternal recognition of pregnancy. Endocrinol. 117,
1424-1430.
100. Bartol, F., Roberts, R.M., Bazer, F.W. and Thatcher, W.W. (1985)
Characterization of proteins produced in vitro by bovine endometrial explants.
Biol. Reprod. 33, 745-759.
101. Ducsay, C.A., Buhi, W.C., Bazer, F.W., and Roberts, R.M. (1986) Role of
uteroferrin in placental iron transport in swine: relationship between
uteroferrin levels and iron deposition in the conceptus during gestation.
J. Anim. Sci. 62, 706-716.
102. Raub, T.J., Denny, J.B., and Roberts, R.M. (1986) Cell surface glycoproteins
of CHO cells. I. internalization and rapid recycling. Exptl Cell Res. 165,
73-91.
103. Raub, T.J., and Roberts, R.M. (1986) Cell surface glycoproteins of CHO cells.
II. surface distribution and pathway of internalization. Exptl. Cell Res. 165,
92-107.
104. Baumbach, G.A., Ketcham, C.M., Richardson, D.E. Bazer, F.W. and Roberts
R.M. (1986) Isolation and characterization of a high molecular weight stable
pink form of uteroferrin from uterine secretions and allantoic fluid of pigs.
J. Biol. Chem. 261, 12869-12878.
R. Michael Roberts 811
105. Fincher, K.B., Bazer, F.W., Hansen, P.J., Thatcher, W.W., and Roberts, R.M.
(1986) Proteins secreted by the sheep conceptus suppress induction of uterine
prostaglandin F2α release by oestradiol and oxytocin. J. Reprod. Fert. 76,
425-433.
106. Knickerbocker, J.J., Thatcher, W.W., Bazer, F.W., Drost, M., Barron, D.H.,
Fincher, K.B. and Roberts, R.M. (1986) Proteins secreted by day 16 to day 18
bovine conceptuses extend corpus luteum functions in cows. J. Reprod. Fert.
77, 381-391.
107. Hansen, P.J., Ing, N.H., Moffatt, R.J., Baumbach, G.A., Saunders, P.T.K.,
Bazer, F.W. and Roberts, R.M. (1987) Biochemical characterization and
biosynthesis of the uterine milk proteins of the pregnant sheep uterus. Biol.
Reprod. 36, 405-418.
108. Moffatt, R.J., Bazer, F.W., Hansen, P.J., Chun, P.W., and Roberts, R.M. (1987)
Purification, secretion and immunocytochemical localization of the uterine
milk proteins, major progesterone-induced proteins in uterine secretions of
the sheep. Biol. Reprod. 36, 419-430.
109. Couso, R., Lang, L., Roberts, R.M. and Kornfeld, S. (1986) Phosphorylation of
the oligosaccharide of uteroferrin by UDP-GlcNAc:glycoprotein N-acetyl-
glucosamine-1-phosphotransferases from rat liver. Acanthamoeba castellani,
and Dictyostelium discoideum requires α1,2-linked mannose residues. J. Biol.
Chem. 261, 6326-6331.
110. Knickerbocker, J.J., Thatcher, W.W., Bazer, F.W., Barron, D.H., and
Roberts, R.M. (1986) Inhibition of uterine prostaglandin F2α production by
bovine conceptus secretory proteins. Prostaglandins 31, 777-793.
111. Bazer, F.W., Vallet, J.L. Roberts, R.M., Sharp, D.C. and Thatcher, W.W. (1986)
Role of conceptus secretory products in establishment of pregnancy. J. Reprod.
Fert. 76, 841-850.
112. Roberts, R.M., Baumbach, G.A., Saunders, P.T.K., Raub, T.J., Renegar, R.H. and
Bazer, F.W. (1986) Possible function of carbohydrate on glycoproteins secreted
into the uterus of pigs during pregnancy. Mol. Cell. Biochem. 72, 67-79.
113. Young, K.H., Bazer, F.W., Simpkins, J.W. and Roberts, R.M. (1987) Effects
of Early Pregnancy and Acute 17β-estradiol Administration on Porcine
Uterine Secretion, Cyclic Nucleotides and Catecholamines. Endocrinol. 120,
254-263.
114. Helmer, S.D., Hansen, P.J., Anthony, R.V., Thatcher, W.W., Bazer, F.W. and
Roberts, R.M. (1987) Identification of bovine trophoblast protein-1, a
secretory protein immunologically related to ovine trophoblast protein-1.
J. Reprod. Fert. 79, 83-91.
115. Murray, M.K., Segerson, E.C., Hansen, P.J., Bazer, F.W. and Roberts, R.M.
(1987) Suppression of lymphocyte activation by a high molecular weight
glycoprotein released from preimplantation ovine and porcine conceptuses.
Amer. J. Reprod. Immunol. and Microbiol. 14, 38-44.
812 Wolf Prize in Agriculture
116. Hunt, D.F., Yates, J.R., Shabanowitz, J., Zhu, N.-Z., Zirino, T., Averill, B.A.,
Daurat-Larraque, S.T., Shewale, J.G., Roberts, R.M. and Brew, K. (1987)
Sequence homology in the metalloproteins: purple acid phosphatase from
beef spleen and uteroferrin from porcine uterus. Biochem. & Biophys. Res.
Comm. 144, 1154-1160.
117. Moffatt, R.J., Bazer, F.W., Roberts, R.M. and Thatcher, W.W. (1987) Secretory
function of the ovine uterus: effects of gestation and steroid replacement
therapy. J. Anim. Sci. 65, 1400-1410.
118. Imakawa, K. Anthony, R.V., Kazemi, M., Marotti, K.R., Polites, H.G.,
and Roberts, R.M. (1987) Interferon-like sequence of ovine trophoblast
protein secreted by embryonic trophectoderm. Nature (London) 330,
377-379.
119. Vallet, J.L., Bazer, F.W. and Roberts, R.M. (1987) The effect of ovine trophoblast
protein-one on endometrial protein secretion and cyclic nucleotides. Biol.
Reprod. 37, 1307-1316.
120. Roberts, R.M., Murray M.K., Burke M.G., and Bazer F.W. (1987) Hormonal
control and function of secretory proteins. Adv. Exp. Biol. 230, 137-50.
121. A. Simmen, R.C.M., Baumbach, G.A. and Roberts, R.M. (1988) Molecular
cloning and temporal expression during pregnancy of the messenger RNA
encoding uteroferrin, a progesterone-induced secretory protein. Mol.
Endocrinol. 2, 253-262.
a. B. Lottenberg, R., Sjak-Shie, N., Fazleabas, A.T. and Roberts, R.M. (1988)
Aprotinin inhibits urokinase but not tissue type plasminogen activator.
Thrombosis Res. 49, 549-556.
122. Kazemi, M., Malathy, P.V., Keisler, D.H. and Roberts, R.M. (1988) Ovine
trophoblast protein-1 and bovine trophoblast protein-1 are present as specific
components of uterine flushings of pregnant ewes and cows. Biol. Reprod.
39, 457-463.
123. Hansen, T.R., Imakawa, K., Polites, H.G., Marotti, K.R., Anthony, R.V. and
Roberts, R.M. (1988) Interferon RNA of embryonic origin is expressed
transiently during early pregnancy in the ewe. J. Biol. Chem. 263, 12801-
12804.
124. Anthony, R.V., Helmer, S.D., Sharif, S.F., Roberts, R.M., Hansen, P.J.,
Thatcher, W.W. and Bazer, F.W. (1988) Synthesis and processing of ovine
trophoblast protein-1 and bovine trophoblast protein-1, conceptus secretory
proteins involved in the maternal recognition of pregnancy. Endocrinol. 123,
1274-1280.
125. Wettemann, R.P., Bazer, F.W., Thatcher, W.W., Caton, D. and Roberts, R.M.
(1988) Conceptus development, uterine response, blood gases and endocrine
function of gilts exposed to increased ambient temperature during early
pregnancy. Theriogenol. 30, 57-72.
R. Michael Roberts 813
126. Ketcham, C.M., Roberts, R.M., Simmen, R.C.M. and Nick, H.S. (1989)
Molecular cloning of the type 5, iron-containing, tartrate resistant acid
phosphatase from human placenta. J. Biol. Chem. 264, 557-563.
127. Imakawa, K., Hansen, T.R., Malathy, P.V., Anthony, R.V., Polites, H.G.,
Marotti, K.R. and Roberts, R.M. (1989) Molecular cloning and characterization
of cDNA’s corresponding to bovine trophoblast protein-1. A comparison
with ovine trophoblast protein-1 and bovine interferon-αII. Mol. Endocrinol.
3, 127-139.
128. Allen, S.H., Nuttleman, P.R., Ketcham, C.M. and Roberts, R.M. (1989)
Purification and characterization of human bone tartrate-resistant acid
phosphatase. J. Bone & Mineral Res. 4, 47-55.
129. Roberts, R.M., Imakawa, K., Niwano, Y., Kazemi, M., Malathy, P.V.,
Hansen, T.R., Glass, A.A. and Kronenberg, L.H. (1989) Interferon production
by the preimplantation sheep embryo. J. Interferon Res. 9, 175-187.
130. Sharif, S.F., Francis, H., Keisler, D.H. and Roberts, R.M. (1989) Correlation
between the release of ovine trophoblast protein-1 by the conceptus and the
production of polypeptides by the maternal endometrium of ewes.
J. Reprod. Fert. 85, 471-476.
131. Hansen, T.R., Kazemi, M., Keisler, D.H., Malathy, P.V., Imakawa, K. and Roberts,
R.M. (1989) Complex binding of the embryonic interferon, ovine trophoblast
protein-1, to endometrial receptors. J. Interferon Res. 9, 215-225.
132. Ing, N.H. and Roberts, R.M. (1989) The major progesterone-modulated
proteins secreted into the sheep uterus are members of the serpin family of
serine protease inhibitors. J. Biol. Chem. 264, 3372-3379.
133. Murray, M.K., Malathy, P.V., Bazer, F.W. and Roberts, R.M. (1989) Structural
relationship, biosynthesis and immunocytochemical localization of the
uteroferrin-associated basic glycoproteins. J. Biol. Chem. 264, 4143-4150.
134. Roberts, R.M. (1989) Minireview: Conceptus interferons and maternal
recognition of pregnancy. Biol. Reprod. 40, 449-452.
135. Allen, B.S., Ketcham, C.M., Roberts, R.M., Nick, H.S. and Ostrer, H. (1989)
Localization of the Human Type 5, tartrate-resistant acid phosphatase gene
by in situ hybridization. Genomics 4, 597-600.
136. Cross, J.C. and Roberts, R.M. (1989) Porcine conceptuses secrete an interferon
during the preattachment period of early pregnancy. Biol. Reprod. 40,
1109-1118.
137. Farin, C.E., Imakawa, K. and Roberts, R.M. (1989) In situ localization of
mRNA for the interferon, ovine trophoblast protein-1, during early embryonic
development of the sheep. Mol. Endocr. 3, 1099-1107.
138. Simmen, R.C.M., Srivivas, V. and Roberts, R.M. (1989) cDNA sequence, gene
organization and progesterone induction of messenger RNA for uteroferrin,
a porcine uterine iron transport protein. DNA 8, 543-554.
814 Wolf Prize in Agriculture
protein from the uterus of the sheep is associated with crystalline inclusion
bodies in uterine epithelium and embryonic trophectoderm. Biol. Reprod.
43, 80-96.
152. Cross, J.C., Farin, C.E., Sharif, S.F. and Roberts, R.M. (1990) Character-ization
of the antiviral activity constitutively produced by murine conceptuses:
absence of placental mRNAs for interferons alpha and beta. Mol. Reprod.
Dev. 26, 122-128.
153. Klemann, S.W., Li, J., Imakawa, K., Cross, J.C., Francis, H. and Roberts, R.M.
(1990) The production, purification, and bioactivity of recombinant bovine
trophoblast protein-1 (bovine trophoblast interferon). Mol. Endocr. 4,
1506-1514 .
154. Raub, T.J., Koroly, M.J. and Roberts, R.M. (1990) Endocytosis of wheat germ
agglutinin-binding sites from the cell surface into a tubular endosomal
network. J. Cell. Physiol. 143, 1-12.
155. Raub, T.J., Koroly, M.J. and Roberts, R.M. (1990) Rapid endocytosis and
recycling of wheat germ agglutinin-binding sites on CHO cells: Evidence for
two compartments in a nondegradative pathway. J. Cell. Physiol. 144, 52-61.
156. Klemann, S.W., Imakawa, K. and Roberts, R.M. (1990) Sequence variability
among ovine trophoblast interferon cDNA. Nucl. Acids Res. 18, 6724.
157. Hansen, T.R., Leaman, D.W., Cross, J.C., Mathialagan, N., Bixby, J.A. and
Roberts, R.M. (1991) The genes for the trophoblast interferons and the related
interferon-II possess distinct 5′-promoter and 3′-flanking sequences. J. Biol.
Chem. 266, 3060-3067.
158. Cross, J.C. and Roberts, R.M. (1991) Constitutive and trophoblast-specific
expression of a class of bovine interferon genes. Proc. Natl. Acad. Sci. USA.
88, 3817-3821.
159. Schalue-Francis, T.K., Farin, P.W., Cross, J.C., Keisler, D. and Roberts, R.M.
(1991) Effect of injected bovine interferon-αI1 on oestrous cycle length and
pregnancy success in sheep. J. Reprod. Fert. 91, 347-356.
160. Wooding, F.B.P., Morgan, G. and Roberts, R.M. (1991) Quantitative immunogold
ultracryomicrotome studies of the distribution of peri-implantation proteins
in the sheep. J. Cell & Tissue Res. 265, 83-93.
161. Farin, C.E., Cross, J.C., Tindle, N.A., Murphy, C.N., Farin, P.W. and
Roberts, R.M. (1991) Induction of trophoblastic interferon expression in
ovine blastocysts after treatment with double-stranded RNA. J. Interferon
Res. 11, 151-157.
162. Hansen, T.R., Cross, J.C., Farin, C.E., Imakawa, K. and Roberts, R.M. (1991)
Slowed transcription and rapid messenger RNA turnover contribute to a
decline in synthesis of ovine trophoblast protein during in vitro culture. Biol.
Reprod. 45, 94-100.
816 Wolf Prize in Agriculture
163. Baumbach, G.A., Saunders, P.T.K., Ketcham, C.M., Bazer, F.W. and
Roberts, R.M. (1991) Uteroferrin contains complex and high mannose-type
oligosaccharides when synthesized in vitro. Mol. Cell. Biochem. 105,
107-117.
164. Xie, S., Low, B.G., Nagel, R.J., Kramer, K.K., Anthony, R.V., Zoli, A.P.,
Beckers, J-F. and Roberts, R.M. (1991) Identification of the major pregnancy-
specific antigens of cattle and sheep as inactive members of the aspartic
proteinase family. Proc. Natl. Acad. Sci. USA. 88, 10247-10251.
165. Trout, W.E., McDonnell, J.J., Kramer, K.K., Baumbach, G.A. and Roberts, R.M.
(1991) The retinol-binding protein of the expanding pig blastocyst: Molecular
cloning and expression in trophectoderm and embryonic disc. Mol. Endocr.
5, 1533-1540.
166. Francis, H., Keisler, D.H. and Roberts, R.M. (1991) Induction of the synthesis
of the pregnancy-specific protein p70 in the endometrium by intramuscular
injection of recombinant bovine interferon-αI1 in nonpregnant ewes.
J. Reprod. Fert. 93, 367-374.
167. Leaman, D.W. and Roberts, R.M. (1992) Genes for the trophoblast interferons
in sheep, goat and musk ox and distribution of related genes among mammals.
J. Interferon Res. 12, 1-11.
168. Trout, W.E., Hall, J.A., Stallings-Mann M.L., Galvin, J.M., Anthony, R.V.
and Roberts, R.M. (1992) Steroid regulation of the synthesis and secretion
of retinol-binding protein by the uterus of the pig. Endocrinol. 130,
2557-2564.
169. Garverick, H.A., Moser, M.T., Keisler, D.H., Hamilton, S.A., Roberts, R.M. and
Smith, M.F. (1992) Luteal function after intrauterine infusion of recombinant
bovine interferon-αI1 into postpartum beef cows expected to have short or
normal luteal phases. J. Reprod. Fert. 94, 319-325.
170. Hernandez-Ledezma, J.J., Sikes, J.D., Murphy, C.N., Watson, A.J., Schultz, G.A.
and Roberts, R.M. (1992) Expression of bovine trophoblast interferon in
conceptuses derived by in vitro techniques. Biol. Reprod. 47, 374-380.
171. Mathialagan, N., Bixby, J.A. and Roberts, R.M. (1992) Expression of
interleukin-6 in porcine, ovine and bovine preimplantation conceptuses. Mol.
Reprod. & Dev 32, 324-330.
172. Skopets, B., Li, J., Thatcher, W.W., Roberts, R.M. and Hansen, P.J. (1992)
Inhibition of lymphocyte proliferation by bovine trophoblast protein-1
(Type I trophoblast interferon) and bovine interferon-αI1. Vet. Immunol.
Immunopath. 34, 81-96.
173. A. Leaman, D.W., Cross, J.C. and Roberts, R.M. (1992) Genes for the
trophoblast interferons and their distribution among mammals. Reprod. Fertil.
Dev. 4, 349-353.
R. Michael Roberts 817
186. Xie, S., Low, B.G., Nagel, R.J., Beckers, J-F. and Roberts, R.M. (1994) A novel
glycoprotein of the aspartic proteinase gene family expressed in bovine
placental trophectoderm. Biol. Reprod. 51, 1145-1153.
187. Szafranska, B., Xie, X., Green, J. and Roberts, R.M. (1995) Porcine pregnancy-
associated glycoproteins: new members of the aspartic proteinase gene family
expressed in trophectoderm. Biol. Reprod. 53, 21-28.
188. Kubisch, H.M., Larson, M.A., Funahashi, H., Day, B.N. and Roberts, R.M.
(1995) Pronuclear visibility, development and transgene expression in
IVM-IVF-derived porcine embryos. Theriogenol. 44, 391-401.
189. Kubisch, H.M., Hernandez-Ledezma, J.J., Larson, M.A., Sikes, J.D. and
Roberts, R.M. (1995) Expression of two transgenes in in vitro matured and
fertilized bovine zygotes after DNA microinjection. J. Reprod. Fertil. 104,
133-139.
190. Xie, S., Green, J., Beckers, J.F. and Roberts, R.M. (1995) The gene encoding
bovine pregnancy-associated glycoprotein-1, an inactive member of the
aspartic proteinase family. Gene 159, 193-197.
191. Li, J., Alexenko, A.P. and Roberts, R.M. (1995) A bifunctional vector suitable
for both site-directed mutagenesis and recombinant expression of interferon-
τ in Escherichia coli. Prot. Express. Purif. 6, 401-407.
192. Meyer, M.D., Hansen, P.J., Thatcher, W.W., Drost, M. and Roberts, R.M. (1995)
Effect of bovine interferon-τ on body temperature and plasma progesterone
concentrations in cyclic dairy cows. J. Dairy Sci. 78, 1470-1476.
193. Meyer, M.D., Hansen, P.J., Thatcher, W.W., Drost, M., Badinga, L.,
Roberts, R.M., Li, J., Ott, T.L. and Bazer, F.W. (1995) Extension of corpus
luteum lifespan and reduction of uterine secretion of prostaglandin F2α of
cows in response to recombinant interferon-τ. J. Dairy Sci. 78, 1921-1931.
194. Senda, T., Saitoh, S-I., Mitsui, Y., Li, J. and Roberts, R.M. (1995) A
three-dimensional model of interferon-τ. J. Interferon & Cytokine Res. 15,
1053-1060.
195. Smith, G.W., Gentry, P.C., Roberts, R.M. and Smith, M.F. (1996) Ontogeny
and regulation of luteinizing hormone receptor messenger ribonucleic acid
within the ovine corpus luteum. Biol. Reprod. 54, 76-83.
196. Xie, S., Nagel, R.J., Green, J., Beckers, J-F. and Roberts, R.M. (1996)
Trophoblast-specific processing and phosphorylation of pregnancy-associated
glycoprotein-1 in day 15 to 25 sheep placenta. Biol. Reprod. 54, 122-129.
197. Smith, G.W., Gentry, P.C., Long, D.K., Smith, M.F. and Roberts, R.M. (1996)
Differential gene expression within the ovine corpus luteum: identification
of secreted protein acidic and rich in cysteine as a major secretory product
of small but not large luteal cells. Endocrinol. 137, 755-762.
198. Liu, L., Leaman, D.W., Bixby, J.A. and Roberts, R.M. (1996) A type I ovine
interferon with limited similarity to IFN-α, IFN-ω and IFN-τ: gene structure,
R. Michael Roberts 819
210. Xie, Sancai. Green. J., Bixby, J.B., Szafranska, B., DeMartini, J.C., Hecht, S.,
Roberts, R.M. (1997) The diversity and evolutionary relationships of the
pregnancy-associated glycoproteins, an aspartic proteinase subfamily
consisting of many trophoblast-expressed genes. Proc. Natl. Acad. Sci. USA
94, 12809-12816.
211. Alexenko, A.P., Leaman, D.W., Li, J., and Roberts, R.M. (1997) The
antiproliferative and antiviral properties of IFN-τ variants in human cells.
J. Interfer. Cytokine Res. 17, 769-779.
212. Ealy, A. D., Green, A. G., Alexenko, A., Keisler, D., and Roberts, R.M.
(1998) Different ovine interferon-tau genes are not expressed identically
and their protein products display different activities. Biol. Reprod. 58,
566-573.
213. Kubisch, H. Michael, Larson, M. and Roberts, R.M. (1998) Relationship
between age of blastocyst formation and interferon-τ secretion by in vitro-
derived bovine embryos. Mol. Reprod. Dev. 49, 254-260.
214. Ealy, A.D., Alexenko, A.P., Keisler, D.H., and Roberts, R.M. (1998) Loss of the
signature six carboxyl amino acid tail from ovine interferon-tau does not
affect biological activity. Biol. Reprod. 58, 1463-1468.
215. Ezashi, T., Ealy, A.D., Ostrowski, M.C., and Roberts, R.M. (1998) Control
of interferon-τ gene expression by Ets-2. Proc. Natl. Acad. Sci. USA. 95,
7882-7887.
216. Roberts, R.M., Liu, L., Guo, Q., Leaman, D., Bixby, J. (1998) The Evolution of
the Type I Interferons. J. Interferon & Cytokine Res. 18, 805-816.
217. Green, J., Xie, S., Szafranska., B, Gan, X., Newman, A., McDowell, K., and
Roberts, R.M. (1999) Identification of a New Aspartic Proteinase expressed
by the outer chorionic cell layer of the equine placenta. Biol. Reprod. 60,
1069-1077.
218. Roberts, R.M., Ealy, A.D., Alexenko, A.P., Han, C.-S., and Ezashi, T. (1999)
Trophoblast Interferons. Placenta 20, 259-264.
219. Winkelman, G.L., Roberts, R.M., Peterson, A.J., Alexenko, A.P., and Ealy, A.D.
(1999) Identification of the Expressed Forms of Ovine Interferon-Tau in the
Periimplantation conceptus: Sequence Relationships and Comparative
Biological Activities. Biol. Reprod. 61, 1592-1600.
220. Alexenko, A.P., Ealy, A.D., and Roberts, R.M. (1999) The Cross-Species Antiviral
Activities of Different IFN-τ Subtypes on Bovine, Murine and Human Cells:
Contradictory Evidence for Therapeutic Potential. J. Interferon & Cytokine
Res. 19, 1335-1341.
221. Green, J.A., Xie, S., Quan, X., Bao, B., Gan, X., Mathialagan, N., Beckers,
J.-F., and Roberts, R.M. (2000) Pregnancy-Associated Bovine and Ovine
Glycoproteins Exhibit Spatially and Temporally Distinct Expression Patterns
During Pregnancy. Biol. Reprod. 62, 1624-1631.
R. Michael Roberts 821
222. Hughes, A.L., Green, J.A., Garbayo J.M., and Roberts, R.M. (2000) Adaptive
diversification within a large family of recently duplicated, placentally
expressed genes. Proc. Natl. Acad. Sci. USA. 97, 3319-3323.
223. Hughes, A.L., and Roberts R.M. (2000) Independent origin of IFN-α and
IFN-β in birds and mammals. J. Interferon Cytokine Res. 20, 737-740.
224. Garbayo, J.A., Green, J.A., Manikkam, M., Beckers, J.F., Kiesling, D.O.,
Ealy, A.D., Roberts, R.M. (2000) Caprine pregnancy-associated glycoproteins
(PAG): Their cloning, expression and evolutionary relationship to other PAG.
Mol. Reprod. Dev. 57, 311-322.
225. Alexenko, A.P., Ealy, A.D., Bixby, J.A., & Roberts, R.M. (2000) A classification
for the interferon-τ. J. Interferon & Cytokine Res. 20, 817-822.
226. Do, H.J., Kim, J.H., Abeydeera, L.R., Han, Y.M., Green, J., Roberts, R.M., Matteri,
R.L., Day, B.N., and Prather, R.S. (2001) Expression of pregnancy-associated
glycoprotein 1 and 2 genes in in vitro-, in vivo- and parthenogenetically-
derived preimplantation pig embryos. Zygote 9, 245-250. PMID 11508744
227. Kubisch, H.M., Larson, M.A., Ealy, A.D., Murphy, C.N., Roberts R.M. (2001)
Genetic and environmental determinants of interferon-τ secretion by in vivo-
and in vitro-derived bovine blastocysts. Anim. Reprod. Sci. 66, 1-13. PMID
11343838
228. Kubisch, H.M., Larson, M.A., Kiesling, D.O., Roberts, R.M. (2001) Control of
interferon-τ secretion by in vitro-derived bovine blastocysts during extended
culture and outgrowth formation. Mol. Reprod. Dev. 58, 390-397.
229. Han C. -S., Chen Y., Ezashi, T., Roberts R.M. (2001) Antiviral activities of the
soluble extracellular domains of Type 1 Interferon Receptors. Proc. Natl.
Acad. Sci. USA. 98, 6138-6143. PMID 11344274
230. Ealy, A.D., Larson S.F., Liu L., Alexenko A.P., Winkleman G.L., Kubisch H.M.,
Bixby J.A., Roberts R.M. (2001) Polymorphic forms of expressed bovine
interferon-tau genes: Relative transcript abundance during early placental
development, promoter sequences of genes and biological activity of protein
products. Endocrinol. 142, 2906-2915. PMID 11416010
231. Chen, X., Rosenfeld C.S., Roberts R.M., Green J.A. (2001) An Aspartic
Proteinase expressed in the Yolk Sac and neonatal stomach of the mouse.
Biol. Reprod. 65, 1092-1101. PMID 11566730
232. Szafranska, B., Miura R., Ghosh D., Ezashi T., Xie S., Roberts R.M., Green J.A.
(2001) The Gene for Porcine Pregnancy-associated glycoprotein 2 (poPAG2):
Its structural Organization and Analysis of its Promoter. Mol. Reprod. Dev.
60, 137-146. PMID 11553911
233. Larson, M.A., Kimura K., Kubisch H.M., Roberts R.M. (2001) Sexual
dimorphism among bovine embryos in their ability to make the transition to
expanded blastocyst and in the expression of the signaling molecule IFN-τ.
Proc. Natl. Acad. Sci. USA. 98, 9677-9682. PMID 11481449
822 Wolf Prize in Agriculture
234. Ezashi T., Ghosh D., Roberts R.M. (2001) Repression of Ets-2 induced
transactivation of the interferon-tau promoter by Oct-4. Mol. Cell. Biol. 21,
7883-7891. PMID 11689681
235. Rosenfeld, C.S., Han C.S., Alexenko A.P., Spencer T.E., Roberts R.M. (2002)
The Expression of interferon receptor subunits, IFNAR1 and IFNAR2, in the
Ovine Uterus. Biol Reprod. 6, 847-853. PMID 12193393
236. Ghosh, D., Ezashi T., Ostrowski M.C., Roberts R.M. (2003) A central role for
Ets-2 in the transcriptional regulation and cyclic AMP-responsiveness of the
human chorionic gonadotropin-beta subunit gene. Mol. Endocrinol, 17,
11-26. PMID 12511603
237. MacLean II, J.A., Chakrabarty, A., Xie, S., Bixby, J.A., Roberts, R.M. and
Green J.A. (2003) A family of Kunitz proteins from trophoblast: Expression
of the trophoblast Kunitz domain proteins (TKDP) in cattle and sheep. Mol.
Reprod. Dev. 65, 30-40. PMID 12658631
238. Roberts, R.M., Ezashi, T., Rosenfeld, C.S., Ealy, A.D., and Kubisch, H.M. (2003)
Evolution of the Interferon-τ genes and their promoters, and maternal-
trophoblast interactions in control of their expression. Reproduction, Suppl
61, 239-251.PMID 14635939
239. Rosenfeld, C.S., Grimm K., Livingston K., Brokman A., Lamberson W.E., Roberts
R.M. (2003) Striking variation in the sex ratio of pups born to mice according
to whether maternal diet is high in fat or carbohydrate. Proc. Natl. Acad. Sci.
USA 100, 4628-4632. PMID 12672968
240. Hughes, A.L., Green J.A., Piontkivska H., Roberts R.M. (2003) Aspartic
proteinase phylogeny and the origin of pregnancy-associated glycoproteins.
Mol. Biol. Evol. 20, 1940-45. PMID 12949149
241. Green, J.A, Parks, T.E., Avalle, P.A., Teluga, B.P., McLain, A.L., Peterson,
A.J., McMillan W., Mathialagan, N., Hook, R.R., Xie, S., Roberts, R.M.
(2005) The establishment of an ELISA for the detection of preganancy-
associated glycoproteins (PAG’S) in the serum of pregnant cow and heifers.
Theriogenology. 63, 1481-503. PMID 15725453
242. Kimura, K., Spate, L.D. Green, M.P.and Roberts, R.M. (2004) Effects of
Oxidative Stress and Inhibitors of the Pentose Phosphate Pathway on Sexually
Dimorphic Production of IFN-τ by Bovine Blastocysts. Mol. Reprod. Dev. 68,
88-95. PMID 15039952
243. Kimura, K., Spate, L.D., Green, M.P., Murphy, C.N., Seidel Jr , G.E.,
Roberts, R.M. (2004) Sexual Dimorphism in Interferon-tau Production
by In vivo-Derived Bovine Embryos. Mol. Reprod. Dev. 67, 193-199. PMID
14694435
244. Roberts, R.M, Ezashi, T., Das, P. (2004) Trophoblast Gene Expression:
transcription factors in the specification of early trophoblast. Reprod. Biol. &
Endocrinology. 2, 47. PMID 15236655
R. Michael Roberts 823
245. Ezashi, T., Roberts, R.M. (2004) Regulation of Interferon-t (IFN-t) Gene
Promoters by Growth Factors that
246. Target the Ets-2 Composite Enhancer: A Possible Model for Maternal Control
of IFN-t Production by the Conceptus during Early Pregnancy. Endocrinology
145, 4452-60. PMID 15217985
247. Maclean II, J.A., Roberts, R.M. and Green, J.A. (2004) Atypical Kunitz-Type
Serine Proteinase Inhibitors Produced by the Ruminant Placenta. Biology of
Reproduction 71, 455-463. PMID 15070828
248. Wang, S. and Roberts, R.M. (2004) Interaction of Stress-Activated Protein
Kinase-Interacting Protein-1 with the Interferon Receptor Subunit IFNAR2 in
Uterine Endometrium. Endocrinology 145, 5820-5831. PMID 155345682
249. Roberts RM. (2004). Impact of Maternal Diet on Reproductive Outcome:
Foreword. Biol Reprod. 71, 1045.
250. Rosenfeld, C. S. and Roberts, R.M. (2004). “Maternal Diet and Other Factors
Affecting Offspring Sex Ratio: A Review.” Biol Reprod 71, 1063-1070. PMID
15229140
251. Wang, S. and Roberts, R.M. (2005) The evolution of the Sin1 gene product, a
little known protein implicated in stress responses and type I interferon
signaling in vertebrates. BMC Evol Biol. 7;5(1):13 PMID 15698473
252. Ghosh D, Sachdev, S., Hannink M, Roberts, R.M. (2005) Coordinate regulation
of basal and cAMP-activated expression of hCG{alpha} by Ets-2 and CREB.
Mol Endocrinol. 1049-1066. PMID 15637148
253. Wooding, F.B.P., Roberts, R.M. and Green, J.A. (2005) Light and electron
microscope immunocytochemical studies of the distribution of pregnancy
associated glycoproteins (PAGs) throughout pregnancy in the cow: possible
functional implications. Placenta. Nov. 26(10):807-27. PMID 16226131
254. Ezashi T., Das P., Roberts R.M. (2005) Low O2 tensions and the prevention
of differentiation of human embryonic stem cells. PNAS 102, 4783-4788.
PMID 15772165
255. Roberts, R.M. (2005) Embryo culture conditions: What embryos like best.
Endocrinology 146, 2140-2141. PMID 15833923
256. Green, M.P., L.D. Spate, Bixby, J.A.. Ealy, A.D Roberts, R.M. (2005) A
Comparison of the Anti-Luteolytic Activities of Recombinant Ovine Interferon-
Alpha and –Tau in Sheep. Biol Reprod 73, 1087-1093 PMID 16079305
257. Kimura, K, Spate, L.D., Green, M.P., Roberts, R.M. (2005) Effects of
D-glucose concentration, D-fructose, and inhibitors of enzymes of the pentose
phosphate pathway on the development and sex ration of bovine blastocysts.
Molecular Reproduction and Development. 72:2, 201-207. PMID 15968626
258. Chen Y Green J.A. Antoniou, E., Ealy A.E. Mathialagan N.P., Walker A.M.,
Avalle M.P., Rosenfeld C.S., Hearne, L., Roberts R.M. (2006) Effect of Interferon
tau Administration on Endometrium of Nonpregnant Ewes: A Comparison
with Pregnant Ewes. Endocrinology, 147:2127-2137. PMID 16469802
824 Wolf Prize in Agriculture
259. Chakrabarty, A, Green J.A., Roberts R.M. (2006) Origin and evolution of the
TKDP gene family. GENE, 373. 35-43 PMID 16567063
260. Chakrabarty A., MacLean J, Roberts R.M. Hughes, A Green, J.A. (2006) Rapid
Evolution of the trophoblast Kunitz domain proteins (TKDPs) – A multi-gene
family expressed in the trophoblast of ruminants. J. Mol. Evol. 63: 274-282
PMID 16830095
261. Whyte, J J.,Roberts R.M., Rosenfeld, C.S. (2007) Fluorescent in situ
hybridization for sex chromosome determination before and after fertilization
in mice. Theriogenology, 67:1022-1031. PMID 17215034
262. Roberts, R.M., Yong, H.J., Smith, S. (2006) What Drives the Formation of
Trophectoderm During Early Embryonic Development? J of Reprod & Dev,
52, S87-S97. [link to article]
263. Ezashi, T., Das. P., Roberts R.M. (2006) Regulation of IFN-tau gene expression
by uterine factors: interaction between the conceptus and maternal
environment during early pregnancy in cattle and sheep. J of Reprod & Dev,
52, S99-S109. [link to article]
264. Green, JA, Roberts RM. (2006) Establishment of an ELISA for the detection
of native bovine pregnancy-associated glycoprotein secreted by trophoblast
binucleate cells. Methods Mol Med. 122: 321-30. PMID 16511990
265. Chen, Y., Antoniou, E., Liu, Z., Hearne, L., Roberts, RM (2007) A Microarray
analysis for genes regulated by interferon-tau in Ovine luminal epithelial
cells, Reproduction 134. 123-135. PMID 17641094
266. Chakrabarty, A. and Roberts R.M. (2007) Ets-2 and C/EBP-beta are important
mediators of ovine trophoblast Kunitz domain protein-1 gene expression in
trophoblast. BMC Molecular Biology. 8:14 PMID 17326832
267. Alexenko, A., Mao, J., Ellersieck, M., Davis, A., Whyte, J.J., Rosenfeld, R.S.,
Roberts R.M. (2007) The contrasting effects of ad libitum and restricted
feeding of a diet very high in saturated fats on sex ratio and metabolic
hormones in mice. Biol Reprod. 77(4): 599-604. PMID 17522073
268. Roberts, RM (2007) Interferon-Tau, a Type 1 Interferon involved in maternal
recognition of pregnancy. Cytokine and Growth Factors Review. 18 (5-6)
403-8 PMID 17662642
269. Fountain, E.D., Mao, J., Whyte, J.J., Mueller, K.E., Ellersieck, M.R., Will, M.R.,
Roberts, R.M., MacDonald, R., Rosenfeld, C.S. (2008) Effects of diets
enriched in omega-3 and omega-6 polyunsaturated fatty acids on offspring
sex-ratio and maternal behavior in mice. Biol of Reprod. 78, 211-217. PMID
17928632
270. Das P, Ezashi, T., Gupta, R, Roberts RM. (2008) Combinatorial roles of protein
kinase A, Ets2 and CBP/p300 in the transcriptional control of interferon-tau
expression in a trophoblast cell line. Mol. Endocrinol. 22:331-43 PMID
17975022
R. Michael Roberts 825
271. Schulz, L., Ezashi, T., Das, P., Westfall, SD, Livingston, KA Roberts RM. (2008)
Human embryonic stem cells as models for trophoblast differentiation.
Placenta 29 Suppl A, Trophoblast Research. 22:S10-S16 Science Direct
272. Roberts, RM, Chen, Y., Ezashi, T & Walker A. (2008) Interferons and the
Maternal-Conceptus Dialog in Mammals Seminars in Cell and Developmental
Biology Semin Cell Dev Biol. 19:170-7. PMID 198032074
273. Das, P., Ezashi, T., Schulz, LC., Westfall, SD., Livingston, KA., Roberts, RM.
(2008) Effects of FGF2 and Oxygen in BMP4-Driven differentiation of
trophoblast from human embryonic stem cells. Stem Cell Research. 1:1
61-74. Science Direct 18735061
274. Ezashi, T., Das, P., Gupta R., Walker, A., Roberts, R.M. (2008) The Role of
Homeobox Protein Distal-less3 and its interaction with Ets2 in Regulating
Interferon-Tau Gene Expression-Synergistic transcriptional activation with
Ets2. Biol Reprod. March 5, 2008 DOI:10.1095
275. Westfall, SD., Sachdev, S., Das, P., Hearne, L., Hannink, M., Roberts, RM.,
Ezashi, T. Identification of Oxygen Sensitive Transcriptional Programs in
Human Embryonic Stem Cell. Stem Cell & Development (accepted for
Publication 1/08)
276. Ghosh, D, Srivastava, G, Xu, D, Roberts, RM. A link between SIN1 (MAPKAP1)
and Poly(rC) Binding protein-2 (PCBP2) in counteracting environmental stress.
(accepted for Publication 2/08)
283. Horst, M.N. and Roberts, R.M. (1979) Characterization of Plasma Membrane
Polypeptides from Chinese Hamster Ovary Cells by Two-dimensional
Electrophoresis. In: Isofocus 78 (Haglund, Westerfield and Ball, eds.). Elsevier-
North Holland, pp. 99-118.
284. Bazer, F.W., Sharp, D., Thatcher, W.W. and Roberts, R.M. (1980) Comparative
Approach to Mechanism in the Maintenance of Early Pregnancy. In:
Reproductive Processes and Contraception. (McKerns, K.W. ed.). Plenum Press,
N.Y. pp 581-617.
285. Mullins, D.E., Bazer, F.W. and Roberts, R.M. (1980) The Pig Uterus Secretes a
Progesterone-induced Inhibitor of Plasminogen Activator. In: Cellular and
Molecular Aspects of Implantation (Glasser, S.R. and Bullock, D.W. eds.).
Academic Press, NY
286. Stein, G.S., Roberts, R.M., Stein, J.L. and Davis, J.L. (1981) Nuclear
Glycoproteins and Glycosaminoglycans. In: The Cell Nucleus, IX Nuclear
Particles, Part B (Busch, H. ed.). pp. 341-357.
287. Denny, J.B. and Roberts, R.M. (1981) Turnover of Plasma Membrane
Polypeptides in Transformed and Normal Cells Using 125I/131I Double Label
Techniques. In: Cancer Cell Organelles: Methodological Surveys in Biochemistry,
Vol. 10 (Reid, E. and Morre, D.J. eds.). Ellis-Horwood Lts. Chichester.
pp. 388-395.
288. Baumbach, G.A., Horst, M.N., Olympio, M.A., Aldrich, H.C. and Roberts,
R.M. (1981) Isolation and Characterization of Plasma Membranes from
Cultured Cells. In: Cancer Cell Organelles: Methodological Surveys in
Biochemistry, Vol. 10 (Reid, E. and Morre, D.J. eds.). Ellis-Horwood Ltd.,
Chichester, pp. 351-370.
289. Bazer, F.W., Roberts, R.M., Sharp, D.C. and Thatcher, W.W. (1981) Uterine
Proteins Synthesized During the Progestative Period and Pregnancy. In: Uterus
et Fecondite. (Boury-Heyler, C.I., Mauleon, P. and Rochet, Y., eds.). Masson
Publishing Co., Paris, pp 17-32.
290. Bazer, F.W., Geisert, R.D., Thatcher, W.W. and Roberts, R.M. (1982) The
Establishment and Maintenance of Pregnancy. In: Control of Reproduction
in Pig. (Cole, D.I.A. and Foxcroft, G.R. eds.). Butterworth Co., London,
pp. 227-252.
291. Roberts, R.M., Baumbach, G.A., Buhi, W.C., Denny, J.B., Fitzgerald, L.A., Babelyn,
S.F. and Horst, M.N. (1983) Analysis of Membrane Polypeptides by Two-
dimensional Polyacrylamide Gel Electrophoresis. In: Receptor Biochemistry
and Methodology. Vol. III. Molecular and Chemical Characterization of
Membrane Receptors. (Ventner, J.C. and Harrison, L. eds Alan Liss, Co. NY, .)
pp. 61-113.
292. Bazer, F.W. and Roberts, R.M. (1983) Biochemical Aspects of Conceptus-
Endometrial Interactions. In: New Frontiers in Mammalian Reproduction and
Development. (Stone, W.H. ed.). J. Exp. Zool. 228, 373-383.
R. Michael Roberts 827
293. Roberts, R.M., Bazer, F.W. and Thatcher, W.W. (1984) Biochemical Interactions
Between Blastocyst and Endometrium in the Large Domestic Animals.
J. Biosciences, 6 (Suppl. 2), 63-74. (Indian Academy of Sciences). Proceedings
of a Symposium on The Blastocyst held at the Indian Institute of Sciences,
Bangalore, India, Dec. 1983.
294. Roberts, R.M. and Bazer, F.W. (1984) Uteroferrin, a Protein in Search of a
Function. Bioessays. Vol. 1, pp. 8-11.
295. Thatcher, W.W., Bartol, F.F., Knickerbocker, J.J., Curl, J.S., Wolfenson, D.,
Bazer, F.W. and Roberts, R.M. (1984) Maternal recognition of pregnancy in
cattle. J. Dairy Sci. 67, 2797-2811.
296. Thatcher, W.W., Knickerbocker, J.J., Bartol, F.F., Bazer, F.W., Roberts, R.M. and
Drost, M. (1984) Maternal recognition of pregnancy in relation to the survival
of transferred embryos: Endocrine aspects. Theriogenol. 23, 129-143.
297. Roberts, R.M. Godkin, J.D., Bazer, F.W., Fincher, K.B., Thatcher, W.W.,
Knickerbocker, J.J. and Bartol, F.F. (1985) Antiluteolysins produced by
mammalian conceptuses. In: Implantation of the Human Embryo (R.G.
Edwards, J. Purdy and S. Steptoe eds.). Academic Press, NY pp. 253-282.
298. Bazer, F.W., Roberts, R.M., Thatcher, W.W. and Sharp, D.C. (1985) Mechanisms
Related to Establishment of Pregnancy. In: Early Pregnancy Factors. (Ellendorff,
F. and Koch, E. eds.) Perinatology Press. pp. 13-23.
299. Thatcher, W.W., Knickerbocker, J.J., Helmer, S.D., Hansen, P.J., Bartol, F.F.,
Roberts, R.M. and Bazer, F.W. (1985) Characteristics of Conceptus Secretory
Proteins and Their Effect on PGF2α Secretion by the Maternal Endometrium
in Cattle. In: Early Pregnancy Factors (Ellendorff, F. and Koch, E. eds.)
Perinatology Press. pp. 25-27.
300. Thatcher, W.W., Bazer, F.W., Sharp, D.C. and Roberts, R.M. (1986)
Interrelationships Between Uterus and Conceptus to Maintain Corpus Luteum
Function in Early Pregnancy: Sheep, Cattle, Pigs and Horses. In: Uterine-
Conceptus Control of Corpus Luteum, J. Anim. Sci. 62 , 25-46.
301. Roberts, R.M., Raub, T.J. and Bazer, F.W. (1986) Role of uteroferrin in
transplacental iron transport in the pig. Fed. Proc. of the Federation of
American Societies for Experimental Biology 45, 2513-2518.
302. Roberts, R.M. and Bazer, F.W. (1988) The function of uterine secretions.
J. Reprod. Fertil. 82, 875-892.
303. Roberts, R.M., Murray, M.K., Burke, M.G., Ketcham, C.M. and Bazer, F.W.
(1987) Hormonal Control and Function of Secretory Proteins. In: Cell and
Molecular Biology of the Uterus (W. W. Leavitt, ed.) Plenum Publishing
Corporation, pp.137-150.
304. Bazer, F.W., Vallet, J.L., Ashworth, C.J., Anthony, R.V. and Roberts, R.M. (1987)
The role of ovine conceptus secretory proteins in the establishment of
pregnancy. In: Cell and Molecular Biology of the Uterus (Wendell W. Leavitt,
ed.) Plenum Publishing Corporation, pp 221-234.
828 Wolf Prize in Agriculture
305. Roberts, R.M., Kronenberg, L.A., Malathy, P.V. and Imakawa, K. (1988)
Embryonic interferons and maternal recognition of pregnancy. In: The Biology
of the Interferon System 1988 (Y. Kawade and S. Kobayashi, eds.), Kodansha
Scientific Press, Ltd., Tokyo, pp 315-321.
306. Imakawa, K. and Roberts R.M. (1989) Interferons and maternal recognition
of pregnancy. In: Development of Preimplantation Embryos and Their
Environment (K. Yoshinaga and T. Mori, eds.), Alan R. Liss, Inc., New York
NY, pp 347-358.
307. Roberts, R.M., Farin, C.E. and Imakawa, K. (1989) Embryonic mediators of
maternal recognition of pregnancy. In: Blastocyst Implantation (K. Yoshinaga
ed.), Serono Symposia USA, Adams Publishing Group, Ltd., Boston, MA.
pp 25-30.
308. Roberts, R.M., Cross, J.C., Farin, C.E., Hansen, T.R., Klemann, S.W. and
Imakawa, K. (1990) Interferons at the placental interface. In: Proc. 2nd Symp.
on the Genetic Engineering of Animals (W. Hansel and B.J. Weir, eds.)
J. Reprod. Fertil. 41, 63-74.
309. Roberts, R.M., Farin, C.E., Cross, J.C. (1990) Trophoblast proteins and maternal
recognition of pregnancy. Oxford Reviews of Reproductive Biology (S.R.
Milligan, ed.), Oxford Univ. Press, Oxford. Vol. 12, pp 147-180.
310. Roberts, R.M., Cross, J.C., Farin, C.E., Kramer, K., Schalue-Francis, F. and
Hansen, T.R. (1990) Trophoblast interferons as Antiluteolysins. In: Establishing
a Successful Human Pregnancy (R.G. Edwards, ed.), Serono Symposia
Publications, Raven Press, New York. Vol. 66, pp 257-268.
311. Roberts, R.M. (1991) A role for interferons in early pregnancy. Bioessays. 13,
pp 121-126.
312. Roberts, R.M., Klemann, S.W., Leaman, D.W., Bixby, J.A., Cross, J.C.,
Farin, C.E., Imakawa, K. and Hansen, T.R. (1991) The polypeptides and
genes for ovine and bovine trophoblast protein-1. J. Reprod. Fert, Suppl. 43,
pp 3-12.
313. Roberts, R.M., Cross, J.C., Farin, C.E., Farin, P.W., Kramer, K.K., Francis, H.,
Librach, C. and Fisher, S.J. (1991) Trophoblast Interferons. In: Growth Factors
in Reproduction, David W. Schomberg (ed.), Serono Symposia, USA, Springer-
Verlag, New York, pp 235-245.
314. Roberts, R.M. (1991) Embryonic loss and conceptus interferon production.
In: J.F. Strauss III and C.R. Lyttle (Eds.), Uterine and Embryonic Factors in
Early Pregnancy, Plenum Press, New York, pp 21-31.
315. Roberts, R.M., Cross, J.C. and Leaman, D.W. (1991) Unique features of the
trophoblast interferons. Pharmac. Ther. 51, 329-345.
316. Roberts, R.M., Anthony, R.V., Thatcher, W.W., Hansen, P.J. and Bazer, F.W.
(1991) Conceptus proteins as Antiluteolysins. In: Development of Pre-
Implantation Embryos (G.S. Toteja, R.S. Sharma, B.K. Singh, S. Mokkapati, B.N.
Saxena, eds.), Indian Council of Medical Research, New Delhi, pp 17-27.
R. Michael Roberts 829
317. Roberts, R.M., Leaman, D.W. and Cross, J.C. (1992) Minireview: Role of
interferons in maternal recognition of pregnancy in ruminants. Proc. Soc.
Exp. Biol. Med. 200, 7-18.
318. Roberts, R.M., Cross, J.C. and Leaman, D.W. (1992) Interferons as Hormones
of Pregnancy. Endocrine Rev. 13, 432-452.
319. Roberts, R.M., Trout, W.E., Mathialagan, N., Stallings-Mann, M. and Ling, P.
(1993) Uterine secretory activity and embryo Development. In: Preimplantation
Embryo Development (B.D. Bavister, ed.), Serono Symposia, Springer-Verlag,
New York, Chap. 17, pp 229-243.
320. Roberts, R.M., Leaman, D.W., Hernandez-Ledezma, J.J. and Cosby, N.C.
(1993) Trophoblast interferons: expression during development and gene
organization. In: M.J. Soares, S. Handwerger, and F. Talamantes (Eds),
Trophoblast Cells: Pathways for Maternal-embryonic Communication,
Springer-Verlag, New York, NY, pp 206-221.
321. Roberts, R.M., Xie, S. and Trout, W.E. (1993) Embryo-uterine interactions in
pigs during week 2 of pregnancy. In: G.R. Foxcroft, M.G. Hunter, C. Doberska
(eds.), Control of Pig Reproduction IV, Proceedings of the Forth International
Conference on Pig Reproduction, Univ. of Missouri-Columbia, May 1993.
J. Reprod. Fertil. 48, 171-186.
322. Roberts, R.M. and Anthony, R.V. (1994) Molecular biology of trophectoderm
and placental hormones. In: J.K. Findlay (ed.), Molecular Biology of the
Female Reproductive System, Chapter 14. Academic Press, San Diego CA,
pp 395-440.
323. Roberts, R.M., Kramer, K.K., Xie, S., Duffy, J., Nagel, R.J. and Wooding, F.B.P.
(1994) Identification of secretory proteins released by preimplantation
embryos of the sheep. In: L. Mastroianni, Jr., H.J.T. Coelingh Bennink,
S. Suzuki and H.M. Vener (eds.), Gamete and Embryo Quality, Proceedings of
the Ivth Organon, Round Table Conference, Thessaloniki, Greece, 24-25
June, 1993. The Parthenon Publishing Group, Pearl River, NY, Chapter 14,
pp 209-223.
324. Mathialagan, N. and Roberts, R.M. (1994) Review Article: A role for cytokines
in early pregnancy. Ind. J. Physiol. Pharmacol. 38, 143-162.
325. {Leaman, 1992 #3257}
326. Mathialagan, N. and Roberts, R.M. (1995) Type I interferons and their role
in maternal recognition of pregnancy and fecundity in domestic ruminant
species. In: Cytokines in Animal Health and Disease (M.J. Myers and M.P.
Murtaugh, eds.), Marcel Dekker, Inc., New York, pp 215-234.
327. Roberts, R.M., Xie, S., Nagel, R.J., Low, B., Green, J. and Beckers, J-F. (1995)
Glycoproteins of the aspartyl proteinase gene family secreted by the developing
placenta. In: Aspartic Proteinases: Structure, Function, Biology and Biomedical
Implications, K. Takahashi (ed.), Plenum Press, New York. Adv. Exp. Med.
Biol. 362, 231-240.
830 Wolf Prize in Agriculture
328. Roberts, R.M., Mathialagan, N., Duffy, J.Y. and Smith, G. (1995) Regulation
and Regulatory Role of Proteinase Inhibitors. Critical Reviews in Eukaryotic
Gene Expression. 5, 385-436.
329. Roberts, R.M., Mathialagan, N., Duffy, J.Y., Stallings-Mann, M.L. and
Trout, W.E. (1995) Proteinase inhibitors at the trophoblast-uterine interface:
roles in implantation or immunomodulation. In: S.K. Dey (ed.) Molecular
and Cellular Aspects of Periimplantation Processes, Serono Symposia USA.
Springer-Verlag, New York, pp 253-267.
330. Roberts, R.M., Xie, S. and Mathialagan, N. (1996) Maternal recognition of
pregnancy. Biol. Reprod. 54, 294-302.
331. Roberts, R.M. (1996) Interferon-τ and Pregnancy. Milstein Award Lecture,
Annual Meeting of Soc. Interferon & Cytokine Res, Nov. 6, 1995, Baltimore,
MD. J. Interferon & Cytokine Res.16, 271-273.
332. Roberts, R.M. and Liu, L. (1996) Interferon-ω. In: B.B. Aggarwal and J.U.
Gutterman (eds.) Human Cytokines, Handbook for Basic and Clinical Research,
Vol. II, Chapter 9. Blackwell Sciences, Cambridge, MA, pp 168-177.
333. Roberts, R.M. (1996) Interferons as hormones of pregnancy. In: J. Sengupta,
D. Ghosh (eds.) Cellular and Molecular Signaling in Reproduction, New Age
International Publishers, New Delhi, pp 187-194.
334. Roberts, R.M., Liu, L., Alexenko, A. (1997) New and atypical families of Type
I interferons in mammals: comparative functions, structures and evolutionary
relationships. Prog. Nucleic Acid Res. & Mol. Biol. 56, 287-326.
335. Green, Jonathan A., Xie, S., and Roberts, R.M. (1998) Pepsin-related molecules
secreted by trophoblast. Reviews of Reproduction 3, 62-69.
336. Green, Jonathan, Xie, S., Gan, X., and Roberts, R.M. (1998) An aspartic
proteinase expressed in the equine placenta. Aspartic Proteinases MNG James
(ed). Plenum Press, New York. pp 162-167.
337. Roberts, R.M., Xie, S., and Green, J. (1998) Pregnancy-associated glycoproteins.
Handbook of Proteolytic Enzymes F. Woessner, N. Rawlings, A. Barrett (eds).
Academic Press, San Diego, California. pp 911-914.
338. Green, J., Xie, S., and Roberts, R.M. (1998) Horse placental aspartic proteinase.
Handbook of Proteolytic Enzymes F. Woessner, N. Rawlings, A. Barrett (eds).
Academic Press, San Diego, California. pp 914-915.
339. Liu, L., Leaman, D., and Roberts, R.M. (1999) Possible Role of the Transcription
Factor Oct-3/4 in Control of Human Chorionic Gonadotropin Expression.
Embryo Implantation: Molecular, Cellular and Clinical Aspects DD Carson
(ed). Springer-Verlag, New York, New York. pp 261-270.
340. Ezashi, T., and Roberts, R.M. (2000) Yeast One-Hybrid System: A Genetic
System to Identify DNA-Binding Proteins. Yeast Hybrid Technologies L. Zhu,
G. J. Hannon (eds). Eaton Publishing, Natick, Massachutes, pp 177-195.
341. Roberts, R.M. (2001) The Place of Farm Species in the New Genomics World
of Reproductive Biology. Biol. Reprod. 64, 409-417.
R. Michael Roberts 831
342. Roberts, R.M. (2001) Making the Case for Productivity. Agbioforum 3,
Special Issue: Understanding the Differences in US-EU Biotech Regulation.
N. Kalaitzandonakes (ed). www.agbioforum.org.
343. Rosenfeld, C.S., Roberts, R.M., Lubahn, D.B. (2001) Estrogen Receptor- and
Aromatase Deficient Mice Provide Insight into the Roles of Estrogen within
the Ovary and Uterus. Mol. Reprod. Dev. 59, 336-346.
344. Roberts, R.M. (2001) National Bovine Genomics Projects: Present Status,
Future Directions, and why they are important. Factors Affecting Calf Crop:
Biotechnology of Reproduction, (Eds: M.J. Fields, J.V. Yelich, R.S. Sand) CRC
Press, Boca Raton, FL. pp 165-175.
345. Rosenfeld, C.S., Wagner, J.S., Roberts, R.M., Lubahn, D.B. (2001) Intraovarian
actions of Oestrogen. J. Reprod. Fertil. 122, 215-226.
346. Roberts, R.M. (2004) Interferon-τ. Encyclopedia of Endocrine Diseases.
L. Martini (ed). Elsevier Academic Press. London.
347. Green, M.P. and Roberts, R.M. (2004) Pregnancy Recognition and Signaling.
Encyclopedia of Animal Science. (eds: W.G. Pond & A.W. Bell) Marcel Dekker,
N.Y.
348. Green, J.A., and Roberts R.M. (2004) Pepsin F. In Handbook of Proteolytic
Enzymes, Second Edition (A.J. Barrett, N.D. Rawlings, and J.F. Woessner, Jr.,
eds) Elsevier Academic Press, London.
349. Roberts R.M. , Xie, S., Green, J.A. (2004) Pregnancy-associated glycoproteins.
In Handbook of Proteolytic Enzymes, Second Edition (A.J. Barrett, N.D.
Rawlings, and J.F. Woessner, Jr., eds) Elsevier Academic Press, London.
350. Jonathan A. Green and R. Michael Roberts (2006). Establishment of an ELISA
for the Detection of Native Bovine Pregnancy-Associated Glycoproteins
Secreted by Trophoblast Binucleate Cells. In Soares, M., and Hunt J.S.
Placenta and Trophoblast: Methods and Protocols, Vol II. Humana Press,
Totowa, NJ.
351. Roberts R., Ezashi, T., Westfall, S., Sachev, S., Hannink, M. (2006) Epigenetics
and Embryonic Stem Cells. In Epigenetics: Principles, Protocols and Practice
(ed. JL Xue) China: SSMPF Publishing, pp 355-372
352. Roberts, RM. (2007) Genetically modified organisms for Agricultural Food
Production. In labeling genetically modified food: The philosophical and legal
debate. Edited by Paul Weirich, Oxford University Press. pp 10-16.
353. BOOKS
354. Longman’s Publishers, England, Plant Cell Structure and Metabolism. This is
under joint authorship with J. Hall and T. Flowers. (First edition, 1974)
Second edition (1982) This book has also been translated into Japanese,
Polish and Italian.
355. Yokendo Ltd Publishers, Japan, Cloned Animal and Placentation. Eds:
R. Michael Roberts, R. Yanagimachi, T. Kariya, K. Hashizume. 2000.
832 Wolf Prize in Agriculture
356. The National Academies Press, Washington DC, Animal Biotechnology Science-
Based Concerns. Coauthor, National Research Council of the National
Academies. 2002.
357. The National Academies Press, Washington, D.C., Animal Care & Management
at the National Zoo. Chair and Co-author, National Research Council of the
National Academies, 2004.
358. PATENTS
359. “Use of interferons of the alpha family to enhance fertility in mammals”
(with P.J. Hansen, W.W. Thatcher and K. Imakawa co-inventors). U.S. Patent
No Filed 3/04/88.
360. “Compositions and Methods for early pregnancy diagnosis” U.S. Patent
No. 6,869,770, March 22 2005.
361. “Early Pregnancy Diagnosis Using PAGS (pregnancy-associated glycoproteins)
Mexican Patent 233668, January 10, 2006.
420. Society for Study of Fertility, York, England 7/08/87. Functions of uterine
secretions in early embryonic development. In the Meeting Symposium: The
milieux of the egg and early embryo.
421. Blastocyst-Implantation Discussion Group, University of Illinois 7/19/87.
Ovine trophoblast protein-1, a polypeptide implicated in mediating maternal
recognition of pregnancy in sheep, is an interferon of the alpha class.
422. Department of Biochemistry and Molecular Biology, Gainesville, FL.
9/24/87. Purification, properties and molecular cloning of ovine trophoblast
protein-1, an unusual interferon of the alpha class.
423. Biochemistry-Biophysics Program, Washington State University, Pullman, WA.
11/03/87. Characterization and molecular cloning of a novel interferon
from sheep trophoblast.
424. Department of Biochemistry, University of Kansas Medical School,
Kansas City, KA. 12/04/87. Interferons and maternal recognition of
pregnancy.
425. Abbott Laboratories (Diagnostic Division), Abbott Park, IL. 12/10/87. The
characterization of acid phosphatase derived from human placenta, spleen
and bone.
426. AAAS Annual Meeting 1988, Boston, MA. Symposium on “Frontiers of
Reproductive Biology” 2/13/88. Interferons and Maternal Recognition of
Pregnancy.
427. Indo-US Symposium on “Factors influencing the establishment of pregnancy,”
All-India Institute of Medical Sciences, New Delhi, 2/17/88. Interferons
secreted by the trophoblast are involved in maternal recognition of pregnancy
in sheep and cattle.
428. Indian Institute of Immunology, New Delhi, 2/17/88. Interferons and
maternal recognition of pregnancy.
429. Vanderbilt University, Reproductive Biology Forum 4/5/88. Characterization
and molecular cloning of a novel interferon from sheep trophoblast.
430. Lexington Hormone Research Symposium, University of Kentucky 5/20/88.
The possible role of interferons in maternal recognition of pregnancy.
431. Current Trends in Reproductive Biology Research, Tufts University School of
Veterinary Sciences 7/01/88. The role of interferons in early pregnancy in
farm animals.
432. Gordon Conference, Reproductive Tract Biology, 7/08/88, Brewster Academy.
Possible function of interferons produced by the trophoblast in maternal
recognition of pregnancy.
433. Institute of Medicine/National Acad. Sciences Conference on Basic Science
Foundation of Medically Assisted Conception, 8/23/88, Newporter Resort,
Irvine CA. Uterine Receptivity, Maternal Recognition of Pregnancy and Early
Embryonic Loss.
R. Michael Roberts 837
467. Washington University, St. Louis, Missouri, September 4, 1991. Role of uterine
secretions in early development of the embryo.
468. University of Minnesota, St. Paul, Minnesota, November 20, 1991. Trophoblast
interferons: structure, distribution, control of expression and function during
pregnancy.
469. University of Illinois, Champaign-Urbana, May 11, 1992. Interferons: Novel
roles in the endocrinology of pregnancy.
470. Serono Symposium Trophoblast Cells: Pathways for Maternal-Embryonic
Communications, Las Vegas, August 9, 1992. Interferons: Functions in
pregnancy, gene distribution among mammals and trophectoderm-restricted
expression.
471. The Procter & Gamble Company, Cincinnati OH, October 6, 1992. Tartrate-
resistant Acid Phosphatases.
472. Genzyme Corporation, Framingham MA, October 26, 1992. Interferons as
Hormones of Pregnancy.
473. University of Missouri-Kansas City, December 10, 1992. Interferons as
Hormones of Pregnancy.
474. Triangle Conference on Reproductive Biology (Conception), Research Triangle
Park NC, January 9, 1992. Factors produced by the early conceptus involved
in maternal recognition of pregnancy.
475. Department of Physiology, All Indian Institute of Medical Sciences, New
Delhi, February 4, 1993. Trophoblast Interferons.
476. Workshop on Molecular Biology Techniques in Reproduction Research,
Bangalore, India, February 11, 1993. Farming of Embryos: In Vitro
Fertilization to produce large numbers of blastocysts for experimental use.
477. Discussion Meeting on Recent Trends in Molecular Aspects of Reproductive
Biology, Indian Institute of Science, Bangalore, India, February 15, 1993.
Trophoblast Interferons.
478. Department of Physiology, Colorado State University, Ft. Collins, May 11,
1993. Cloning and characterization of secretory products of pre-implantation
ovine conceptuses.
479. Fourth International Conference on Pig Reproduction, Columbia, Missouri,
May 25, 1993. Embryo-uterine interactions in the pig.
480. IVth Organon Round Table Conference, Thessaloniki, Greece, June 24-25,
1993. Identification of secretory proteins released by preimplantation embryos.
481. VIIIth World Congress on In Vitro Fertilization and Alternate Assisted
Reproduction, Kyoto Japan, September 12-15, 1993. Interferons and other
factors released by the perimplantation conceptus.
482. The 5th International Conference on Aspartic Proteinases, Gifu, Japan,
September 19-24, 1993. Glycoproteins of the aspartyl proteinase gene family
secreted by the developing placenta.
840 Wolf Prize in Agriculture
19, 2002. Sexual dimorphism among blastocysts may provide for sex ratio
adjustment in the bovine.
540. Department of Biomedical Sciences, Southwest Missouri State University,
Springfield, MO. April 11, 2002. Interferons in Disease and Pregnancy.
541. Department of Veterinary Medicine, University of Pennsylvania, Philadelphia,
PA. April 16, 2002. The Evolution of Mechanisms of Maternal Recognition
of Pregnancy.
542. Mammalian Gametogenesis & Embryogenesis Conference, Connecticut College,
New London CT. June 30-July 5, 2002. Mechanisms of Genetic Quality
Control: The in utero environment and its influence on sex ratio.
543. Sixth International Symposium on Reproduction in Domestic Ruminants,
Crieff, Scotland, UK. August 14-17, 2002. Ovary-uterine-embryo interactions.
544. Department of Physiology, University of Cambridge, England, UK. November
15, 2002. Does maternal diet influence the sex of offspring born?
545. Department Cell Biology, University of Southampton, England, UK. November
19, 2002. Does maternal diet influence the sex of offspring born?
546. Koret School of Veterinary Medicine, Tel Aviv, Israel. May 12, 2003. The
relationship between diet and sex of offspring.
547. Annual Meeting of the Israel Fertility Association, Tel Aviv, Israel. May 14,
2003. Pregnancy recognition signals and uterine biology.
548. Havemeyer Foundation Workshop, Embryonic and Fetal Nutrition, Ravello,
Italy. May 15-18, 2003. Can maternal nutrition influence the sex of offspring?
549. Larry Ewing Memorial Lecture, John Hopkins University, Baltimore MD.
October 8-9, 2003. Sexual dimorphism during early embryonic development
and the effects of diet on sex of offspring.
550. Keynote Speaker, Kansas IDeA Biomedical Focus Group Symposium, University
of Kansas Medical Center, Kansas City KS. November 1, 2003. The Culture
of Collaboration.
551. Reproductive Biology Seminar Series, University of Illinois. May 5, 2004.
How the diet of the mother affects the sex of her offspring.
552. Missouri. Agriculture Leaders Forum with USDA Undersecretary Bill Hawks,
University of Missouri-Columbia. October 12, 2004
553. Advantages of Agriculturally Important Domestic Species as Biomedical
Models Workshop, Michigan State University. October 29-31 2004. Are the
Domestic Farm Species Redundant as Models in Biomedical Research? Does
Mighty Mouse Rule Supreme?
554. International Symposium on Stem Cells: Premises and Promises for Research
and Therapeutics. September 18-21, 2005 Mumbai India. Hypoxia and
prevention of human embryonic stem cell differentiation
555. Saturday Morning Science Lecture Jan 28, 2006. University of Missouri.
How Diet Influences Sex of Offsprings.
R. Michael Roberts 845
556. Life Sciences and Society Symposium- Invited Speaker March 8, 2006
University of Missouri. Stem Cells: The Upside and Downside of Stem Cell
Science.
557. Saturday Morning Science Lecture Sept 16, 2006. Stem Cells: Technology
Promise and Concern
558. Center for Reproductive Science Workshop, UCSF. October 12, 2006.
Embryonic Stem Cells as Models for Trophoblast Differentiation under High
and Low Oxygen Conditions.
559. William Woods University Invited Speaker October 24, 2006. Stem Cells:
Technology Promise and Concern
560. University of Connecticut Invited Speaker. November 15, 2006. Lineage
Commitment to Trophectoderm.
561. 33rd Annual Conference of the Int’l Embryo Transfer Society. Kyoto, Japan.
January 10, 2007 Lineage Specification in Mouse.
562. Second Okayama Symposium in Reproductive Biology. Okayama Japan. Jan
11, 2007. IFNT Regulated Genes.
563. Keystone Symposia Reproduction: Advances and Challenges (C1-2007) Santa
Fe, New Mexico Feb 24, 2007. Maternal Diet and Reproductive Outcome.
564. 2nd SGI Endometrial Satellite Symposium Reno, Nevada. March 13, 2007
Implantation and Interferon Response.
565. International Federation of Placenta Associations Meeting “Placenta-Platform
for Life. Kingston, Ontario Canada. August 17-21, 2007. Human Embryonic
Stem Cells as Models for Trophoblast Differentiation.
566. Seminar presentation (Invited Speaker). November 27, 2007. UC Santa
Barbara. “Human Embryonic Stem Cells as Models of Placental Cell
Development.”
567. Seminar presentation (Invited Speaker). March 7, 2008. University of Florida-
Gainesville. “Can a mother’s diet influence the sex of her offspring?”
568. Seminar presentation (Invited Speaker) April 15, 2008. University of Santa
Barbara “Interferon-tau: function, control of expression and evolutionary
origin”.
GRADUATE STUDENTS
1. Jacques Van Veen (Ph.D.): 1972-1975. “Cell Surface Properties in Relation to
Growth and Form in Chinese Hamster Ovary Cells in Culture.”
2. Daniel W. Welch (Ph.D./M.D.): 1971-1976. “Complex Carbohydrate
Metabolism in Skin Fibroblast Cultures from Patients with Cystic Fibrosis of
the Pancreas.”
3. William Buhi (Ph.D.): 1975-1981. The Role of Uteroferrin Iron Transport of
the Pig Conceptus.
846 Wolf Prize in Agriculture
Michael D. Olympio (1978-1979) was awarded 2nd Prize for Research (Watson
Clinic Award) in the Medical Student Research Competition in 1979.
John Denny won the 1st prize in the Graduate Research Competition in the College
of Medicine in 1981.
Rosalind Masters won the 1st prize for Research (Watson Clinic Award) in the
Medical Student Research Competition in 1982. Rosalind Masters won the 1st
prize for a written paper in the AOA competition for Medical Student Research
in 1982.
George Baumbach won 2nd prize in the Graduate Research Competition in the
College of Medicine, University of Florida, 1983.
Catherine Ketcham won 1st prize in the Graduate Research Competition, College of
Medicine, University of Florida, 1987.
Jay Cross won second place in the “Young Investigator” competition at the SSR
Meeting in Seattle, 1988.
848 Wolf Prize in Agriculture
Kyle Kramer won the first place prize in the biological sciences interactions portion
of the Ninth Annual Research and Creative Activities Forum at the University
of Missouri in 1992.
Doug Leaman won 1st Place in the “Young Investigator” competition at the SSR
Meeting in Raleigh NC, 1992.
George Smith won the Superior Graduate Achievement Award for 1993-94 from
the University of Missouri Graduate Association and the Graduate School.
Limin Liu won 1st Place in the “Young Investigator” competition at the SSR Meeting
in Ann Arbor, MI, 1994.
Shushong (Phillip) Wang won 1st Place in the “Young Investigator” poster
competition at the SSR Meeting in Cincinnati 2004.
Steven D. Tanksley
Cornell University, Ithaca, New York, USA
k
CURRICULUM VITAE
EDUCATION
Colorado State University B.S. 1976 Agronomy
University of California, Davis Research Assistant 1976-79 Genetics
University of California, Davis Ph.D. 1979 Genetics
University of California, Davis Postdoctoral Fellow 1979-81 Genetics
PROFESSIONAL EXPERIENCE
1994-Present Liberty Hyde Bailey Professor of Plant Breeding, Professor of Plant
Biology, Cornell University
1991-1994 Professor, Department of Plant Breeding & Biometry, Cornell
University
1985-1991 Associate Professor, Department of Plant Breeding & Biometry, Cornell
University
1981-1985 Assistant Professor, Department of Horticulture and Plant Genetic
Engineering Laboratory, New Mexico State University, Las Cruces
849
850 Wolf Prize in Agriculture
HONORS/ACTIVITIES
Editor, Genetics 1988-92
Editor, Molecular Breeding 1996-97
Appointed to Liberty Hyde Bailey Professorship, 1994
Elected to the National Academy of Sciences, USA, 1995
Award for Outstanding Contributions to Biology, University of California, Davis,
1989
Outstanding Undergraduate Mentor Award, Cornell University, 1989
Outstanding Research Award (Gamma Sigma Delta), 1996
Chair, Cornell Genomics Initiative, 1997-2005
Alexander von Humboldt Foundation Award, 1998
Martin Gibbs Medal, American Society of Plant Physiology, 1999
Wolf Foundation Prize in Agriculture, 2004
Kumho Award in Plant Molecular Biology and Biotechnology Korea, 2005
Appointed as an Einstein Professor of the Chinese Academy of Sciences, 2006
Appointed as Honorary Professor Wuhan Botanical Garden (China), 2006
Rank Prize Award, 2008
SCIENTIFIC ACHIEVEMENTS
Steve Tanksley’s research has resulted in major contributions in our understanding
of plant genome organization and in concepts and infrastructure that improve
dramatically the efficiency of plant breeding programs. He published the first
molecular map of tomato in 1992. This proof-of-concept map was followed up
with work providing some of the first clear experimental evidence of how molecular
maps and markers could be applied to agriculture. Tanksley’s early work was
extended to molecular mapping of other major crop species, including the
Solanaceous crops, potato and pepper, and cereal crops. The markers developed in
Tanksley’s lab were distributed widely to researchers throughout the world and
facilitated the flow of information and scientific collaboration. His work to produce
the molecular map of rice became a cornerstone of the Rockefeller Foundation’s
International Program on Rice Biotechnology and was pivotal in the development
of the USDA’s National Research Initiative in Plant Genomes.
In 1988, Tanksley demonstrated for the first time that quantitatively inherited
traits spanning an entire genome could be dissected into their corresponding
Mendelian factors (quantitative trait loci, QTLs) using a comprehensive RFLP linkage
map. This led to a cascade of experiments by other researchers aimed at detecting
and mapping QTLs in a wide array of other organisms. Within Tanksley’s own
group, QTL analysis in rice led to the discovery of the genetic basis of heterosis in
this key food staple. More recently, Tanksley has used the power of QTL analysis to
explore ways of efficiently utilizing wild and unadapted germplasm resources in
Steven D. Tanksley 851
plant improvement. Working in rice and tomato, he and his colleagues are applying
the molecular maps and markers they developed to identify rate-limiting genes
associated with crop performance. Recently they discovered genes from a wild
ancestor of rice that can boost the yields of domesticated rice by 15-18%. This
approach provides an effective mechanism for expanding the gene pool of cultivated
crop varieties.
The ability to clone genes of agronomic importance and to determine their
function is a major objective of biology today. Tanksley’s group used the tomato
molecular map to accomplish the first map-based cloning of a resistance gene in
any crop species in 1993. They isolated the Pto gene for resistance to bacterial
speck and demonstrated that a protein kinase played a critical role in this plant-
microbe interaction. This work has become one of the classic studies in disease
resistance gene isolation and contributed major new insights leading to an
understanding of the kinds of signal transduction pathways that are active in host-
pathogen recognition.
Tanksley’s group successfully extended map-based cloning to QTLs of agronomic
significance where nothing is known of the target gene other than the phenotype
it confers. The QTL Fw2.2 is responsible for 30% of the difference in tomato fruit
size between the domesticated tomato and their wild relatives. Fw2.2 was cloned
using a multi-tiered strategy including high resolution mapping with RFLPs and
RAPDs on near isogenic lines, physical mapping using yeast artificial chromosomes
and cosmic clones, and finally complementation tests to confirm the identity of the
gene. This seminal research set the stage for a better understanding of both evolution
under natural selection and crop improvement under human selection. Furthermore,
this outstanding achievement in resolving the genetic basis of complex phenotypes
has opened the way for similar studies in other organisms.
Another major contribution was the development of comparative maps that
clarified evolutionary relationships among genera and provided an essential
framework for exploring questions of gene homoeology in plants. Tanksley showed
that the same RFLP probes could be used to map different members of plant
families. Tanksley’s first comparative map (tomato-potato) was published in 1988.
At the level of macro-synteny, these species appeared to be differentiated by only 5
major translocations. Following the investigation of the Solanaceae family, Tanksley’s
group demonstrated the extent of macro-synteny in the distantly related sub-
families of the grasses, notably rice and maize. Today synteny is accepted as the
rule throughout crop plant genetics and the concept pervades plant genetics
research.
Tanksley’s current work demonstrates a new marker-assisted breeding strategy
that is having a direct impact on plant improvement. His group uses maps and
markers to identify new yield-enhancing genes/QTLs from wild species and transfer
them to elite cultivars of tomato and rice. Based on an “Advanced Backcross”
852 Wolf Prize in Agriculture
LIST OF PUBLICATIONS
1. Chen K, Cong B, Wing R, Vrebalov J, Tanksley S (2007) Regulatory Mutation
in a Cell Elongation Transcription Factor Responsible for the Evolution of
Autogamy in Cultivated Tomatoes. Science 318, 643.
Steven D. Tanksley 853
2. McCarthy, A., Biget, L., Lin, C., Petiard, V., Tanksley, S., and McCarthy, J.
Cloning (2007) Expression, crystallization and preliminary X-ray analysis of
the XMT and DXMT N-methyltransferases from Coffea canephora (robusta).
Acta Crystallographica F63, 304-307.
3. Tanksley S, Fulton T (2007) Dissecting quantitative trait variation—examples
from the tomato. Euphytica 145: 365-370.
4. Hobolth A, R. Nielsen, Y. Wang, F. Wu, and S. D. Tanksley. (2006) CpG +
CpNpG analysis of protein-coding sequences from tomato. Mol. Biol. Evol.
2006 23:1318-1323.
5. Hinniger C, Caillet V, Michoux F, Amor M, Tanksley S, Lin C, McCarthy J
(2006) Isolation and Characterization of cDNA Encoding Three Dehydrins
Expressed During Coffea canephora (Robusta) Grain Development. Annals
of Botany 97:755-765.
6. Carlson J, Leebens-Mack J, Wall P, Zahn L, Mueller L, Landherr L, Hu Y,
Ilut D, Arrington J, Choirean S, Becker A, Field D, Tanksley S, Ma H,
dePamphilis C (2006) EST database for early flower development in California
poppy (Eschscholzia californica Cham., Papaveraceae) tags over 6000 genes
from a basal eudicot. Plant Molecular Biology 62 351-369.
7. Wang Y, R. S. van der Hoeven, R. Nielsen, L. A. Mueller and S. D. Tanksley
(2006) Characteristics of the tomato nuclear genome as determined by
sequencing undermethylated EcoRI digested fragments. Theor Appl Genet
112: 72-84.
8. Wang Y, Tang X,Cheng Z, Mueller M, Giovannoni J, Tanksley S (2006)
Euchromatin and Pericentromeric Heterochromatin: Comparative Composition
in the Tomato GenomeGenetics 2006 172: 2529-2540.
9. Wu F, Mueller L, Crouzillat D, Petiard V(2006) Bioinformatics and
Phylogenetics to Identify Large Sets of Single Copy, Orthologous Genes (COSII)
for Comparative, Evolutionary and Systematic Studies: A Test Case in the
Euasterid Plant Clade Genetics; 174: 1407-142.
10. Cong B, Tanksley SD (2006) FW2.2 and cell cycle control in developing
tomato fruit: a possible example of gene co-option in the evolution of a
novel organ. Plant Mole Biol 62:867-880.
11. Barrero L.S., B. Cong, F. Wu, and S.D. Tanksley 2006 Developmental
characterization of the fasciated locus and mapping of candidate genes
involved in the control of floral meristem size and carpel number in tomato.
Genome 49:991-1006.
12. Frary A, Xu Y, Liu J, Mitchell Sh, Tedeschi E, Tanksley SD (2005) Development
of a set of PCR-based anchor markers encompassing the tomato genome and
evaluation of their usefulness for genetics and breeding experiments. Theor
Appl Genet 111:291.
854 Wolf Prize in Agriculture
13. Albert, V.A., D.E. Soltis, J. Carlson, W.G. Farmerie, K. Wall, D.C. Ilut,
T. M. Solow, L.A. Mueller, L.L. Landherr, Y. Hu, M. Buzgo, S. Kim, M-J. Yoo,
M.W. Frohlich, R. Perl-Treves, S. Schlarbaum, B. Bliss, X. Zhang, S. Tanksley,
D.G. Oppenheimer, P.S. Soltis, H. Ma, C.W. dePamphilis, and J.H. Leebens-
Mack. (2005) Floral gene resources from basal angiosperms for comparative
genomics research. BMC Plant Biology 5:5.
14. Yogeeswaran K, Frary A, York T, Amenta A, Lesser A, Nasrallah J, Tanksley S,
Nasrallah M (2005)Comparative genome analyses of Arabidopsis spp.:
Inferring chromosomal rearrangement events in the evolutionary history of
A. thaliana. Genome Res., Apr 2005; 15: 505-515.
15. York T, Durrett R, Tanksley S, Nielsen R (2005) Genetical Research. 85
Bayesian and maximum likelihood estimation of genetic maps 85:159-168.
16. Lin,-Chenwei, Mueller L, McCarthy J, Crouzillat D, Petiard V, Tanksley S.
2005 Coffee and tomato share common gene repertoires as revealed by deep
sequencing of seed and cherry transcripts. Theoretical-and-Applied-Genetics.
112(1): 114-130.
17. Barrero L.S., and S. D. Tanksley 2004 Evaluating the Genetic Basis of Multiple-
locule Fruit in a Broad Cross Section of Tomato Cultivars. Theor Appl Genet
109: 669-679.
18. Frary, Amy, Fritz, Lisa A., and S. D. Tanksley 2004 A Comparative Study of
the Genetic Bases of Natural Variation in Morphology of Tomato Leaves,
Sepals, and Petals. Theor Appl Genet 109: 523-533.
19. van der Knaap, E., Sanyal, A. Jackson, S.A. S.D. Tanksley 2004 High-Resolution
Fine-Mapping and FISH analysis of sun a Locus Controlling Tomato Fruit
Shape, Reveals a Region of the Tomato Genome Prone to DNA Rearrangements.
Genetics 168(4):2127-40.
20. Chen, Kai-Yi, Tanksley, S. D. High Resolution Mapping and Functional
Analysis of se2.1: a Major Stigma Exsertion QTL Associated with the Evolution
from Allogamy to Autogamy in the Genus Lycopersicon. Genetics 168,
1563-1573.
21. Yates H, Frary A, Doganlar S, Frampton A, Eannetta N, Uhlig J, Tanksley S
(2004) Comparative fine mapping of fruit quality QTLs on chromosome 4
introgressions derived from two wild tomato species Euphytica 135:283-296.
22. Frary, A., T. M. Fulton, D. Zamir, S. D. Tanksley 2004 Advanced Backcross QTL
Analysis of a Lycopersicon Esculentum x L. Pennellii Cross and Identification
of Possible Orthologs in the Solanaceae. Theor Appl Genet 108, no. 3: 485.
23. Tanksley, S. D. (2004) The Genetic, Developmental and Molecular Bases
of Fruit Size and Shape Variation in Tomato. Plant Cell Special Issue 16:
S181-S189.
24. Ronning et al 2003 Comparative analyses of potato expressed sequence Tag
libraries. Plant Phys 131:419-429.
Steven D. Tanksley 855
25. Van der Knapp E, Tanksley S D. 2003 The Making of a Bell Pepper-Shaped
Tomato Fruit: Identification of Loci Controlling Fruit Morphology In Yellow
Stuffer Tomato. Theor Appl Genet 107:139-147.
26. Cong B, Liu J, Tanksley SD. 2002. Natural Alleles at a Tomato Fruit Size
Quantitative Trait locus Differ by Heterochronic Regulatory Mutations. Proc
Natl Acad Sci 99:13606-13611.
27. Fulton, T. M., R. van der Hoeven, N.T. Eannetta, S.D. Tanksley. 2002.
Identification, Analysis and Utilization of Conserved Ortholog Set (COS)
Markers for Comparative Genomics in Higher Plants. Plant Cell 14:1457-
1467.
28. Fulton, T. M., P. Bucheli, E. Voirol, J. Lopez, V. Petiard, and S. D. Tanksley.
2002. Quantitative Trait Loci (QTL) Affecting Sugars, Organic Acids and
Other Biochemical Properties Possibly Contributing to Flavor, Identified in
Four Advanced Backcross Populations of tomato. Euphytica 127:163-177.
29. Doganlar, S., A. Frary, M-C. Daunay, R. N. Lester, and S. D. Tanksley. 2002. A
Comparative Genetic Linkage Map of Eggplant (Solanum melongena) and
its Implications for Genome Evolution in the Solanaceae Genetics 161:
1697-1711.
30. Doganlar, S., A. Frary, M-C. Daunay, R.N. Lester, and S. D. Tanksley.
2002. Conservation of Gene Function in the Solanaceae as Revealed by
Comparative Mapping of Domestication Traits in Eggplant Genetics 161:
1713-1726.
31. Durrett R T, Chen K-Y, Tanksley S 2002. A Simple Formula Useful for
Positional Cloning. Genetics 160: 353-355.
32. Giovannoni J et al. Genetic Control of Fruit Quality and Prospects for Nutrient
Modification. HortScience. 37. P. 453-456.
33. Giovannoni J. 2002. Molecular Biology of Fruit Maturation and Ripening.
Annual. Review Plant Physiol. Plant. Mol. Biol. Vol. 52: 725-749.
34. Liu, Jiping, J. Van Eck, B. Cong, and S. D. Tanksley . 2002. A New Class of
Regulatiry Genes Underlying the Cause of Pear-Shaped Tomato Fruit PNAS.
Vol. 99: 13302-13306.
35. Payton P et al. Creation of a baseline gene expression profile for tomato fruit
ripening: characterization of ripening, ethylene signaling, and light signaling
genes. 53, No. 377. 2023-2030.
36. Van der Hoeven, R., C. Ronning, J. Giovannoni, G. Martin, S.D. Tanksley.
2002. Deductions about the number, organization and evolution of genes in
the tomato genome based on analysis of a large EST collection and selective
genomic sequencing. Plant Cell 14(7):1441-56.
37. Ku H-M, Liu J, Doganlar S, Tanksley S 2001. Exploitation of Arabidopsis-
Tomato Synteny to Construct a High-Resolution Map of the ovate Locus in
Tomato. Genome 44:470-475.
856 Wolf Prize in Agriculture
74. Tanksley, Steven D. and Susan R. McCouch. 1997. Seed banks and
molecular maps: unlocking genetic potential from the wild. Science
277:1063-1066.
75. Xiao, J., J. Li, S. Grandillo, S.N. Ahn, L.P. Yuan, S.D. Tanksley and S.R. McCouch.
Identificatoin of trait-improving QTL alleles from a wild rice relative,
O. rufipogon Genetics 150:899-909.
76. Alpert, Kevin B. and Steven D. Tanksley. 1996. High-resolution mapping
and isolation of a yeast artificial chromosome contig containing fw 2.2: A
major fruit weight quantitative locus in tomato. Proc. Natl. Acad. Sci.
93:15503-15507.
77. Autrique, Enrique, Miloudi M. Nachit, Philippe Monneveux, Steven D.
Tanksley, and Mark E. Sorrels. 1996. Genetic diversity in durum wheat based
on RFLPs, morphophysiological traits, and coefficient of parentage. Crop Sci.
36:735-742.
78. Broun, Pierre and Steven D. Tanksley. 1996. Characterization and genetic
mapping of simple repeat sequences in the tomato genome. Mol. Gen. Genet
150:39-49.
79. Frary, Amy, Gernot G. Presting and Steven D. Tanksley. 1996. Molecular
mapping of the centromeres of tomato chromosomes 7 and 9. Mol. Gen.
Genet. 250:295-304.
80. Ganal, M.W. and S.D. Tanksley. 1996. Recombination around the Tm2a and
Mi resistance genes in different crosses of Lycopersicon peruvianum. Theor.
Appl. Genet. 92:101-108.
81. Grandillo, S. and S.D. Tanksley. 1996. Genetic analysis of RFLPs, GATA
microsatellites and RAPDs in a cross between L. esculentum and
L. Pimpinellifolium. Theor. Appl. Genet. 92:957-965.
82. Grandillo, S. and S.D. Tanksley. 1996. QTL analysis of horticultural traits
differentiating the cultivated tomato from the closely related species
Lycopersicon pimpinellifolium. Theor. Appl. Genet. 92:935-951.
83. Grandillo, Silvana, Hsin-Mei Ku and Steven D. Tanksley. 1996.
Characterization of fs8.1, a major QTL influencing fruit shape in tomato.
Molecular Breeding 2:251-160.
84. Hamilton, Carol M., Ane Frary, Candice Lewis, and Steven D. Tanksley. 1996.
Stable transfer of intact high molecular weight DNA into plant chromosomes.
Proc. Natl. Acad. Sci. 93:9975-9979.
85. Pillen, K., M.W. Ganal, and S.D. Tanksley. 1996. Construction of a high-
resolution genetic map and YAC-contigs int he tomato Tm-2a region. Theor.
Appl. Genet. 93:228-233.
86. Pillen, Klaus, Kevin B. Alpert, James J. Giovannoni, Martin W. Ganal,
and Steve D. Tanksley. 1996. Rapid and reliable screening of a tomato
YAC library exclusively based on PCR. Plant Mol. Biology Reporter
14:58-67.
860 Wolf Prize in Agriculture
87. Tanksley, S.D. and J.C. Nelson. 1996. Advanced backcross QTL analysis: a
metod for the simultaneous discovery and transfer of valuable QTLs
from unadapted germplasm into elite breeding lines. Theor. Appl. Genet.
92:191-203.
88. Tanksley, S.D., S. Grandillo, T.M. Fulton, D. Zmir, Y. Eshed, V. Petiard,
J. Lopez, and T. Beck-Bunn. 1996. Advanced backcross QTL analysis in a
cross between an elite processing line of tomato and its wild relative
L. pimpinellifolium. Theor. Appl. Genet. 92:213-224.
89. Xiao, Jinhua, Jiming Li, Longping Yuan and Steve D. Tanksley. 1996.
Dominance is the major genetic basis of heterosis in rice as revealed by QTL
analysis using molecular markers. Genetics 140:745-754.
90. Xiao, Jinhua, Silvana Grandillo, Sang Nag Aho, Susan R. McCouch, Steven D.
Tanksley, Jiming Li, and Longping Yuan. 1996. Genes from wild rice improve
yield. Scientific Correspondence - Nature 384:223-224.
91. Xiao, J., J. Li, L.P. Yuan, S.R. McCouch and S.D. Tanksley. 1996. Genetic
diversity and its relationship to hybrid performance and heterosis in rice as
revealed by PCR-based markers. Theor. Appl. Genet. 92:637-643.
92. Yencho, G. Craig, Meredith W. Bonierbale, Ward M. Tingey, Robert L. Plaisted,
and Steven D. Tanksley. 1996. Molecular markers locate genes for resistance
to the Colorado potato beetle, Leptinotarsa decemlineata, in hybrid Solanum
tuberosum x S. berthaultii potato progenies. Entomologia Experimentalis et
Applicata 81: 141-154.
93. Yu, Z. H., D. J. Mackill, J. M. Bonman, S. R. McCouch, E. Guiderdoni,
J. L. Notteghem, and S. D. Tanksley. 1996. Molecular mapping of genes
for resistance to rice blast (Pyricularia grisea Sacc.). Theor Appl Gene
93:859-863.
94. Azanza, F., D. Kim, S.D. Tanksley, J.A. Juvik. 1995. Genes from Lycopersicon
chmielewskii affecting tomato quality during fruit ripening. Theor. App. Genet.
91:495-504.
95. Dixon, M.S., D.A. Jones, K. Hatzixanthis, M.W. Ganal, S.D. Tanksley, and
J.D.G. Jones. 1995. High-resolution mapping of the physical location of the
tomato Cf-2 gene.
96. Ganal, Martin W., R. Simon, S. Brommonschenkel, M. Arndt, M.S. Phillips,
S. D. Tanksley, and A. Kumar. 1995. Genetic mapping of a wide spectrum
nematode resistance gene (Hero) against Globodera rostochiensis in tomato.
Molecular Plant-Microbe Interactions 6:886-891.
97. Giovannoni, James J., Erick N. Noensie, Diane M. Ruezinsky, Xianghuai Lu,
Samantha L. Tracy, Martin W. Ganal, Gregory B. Martin, Klaus Pillen, Kevin
Alpert, Steven D. Tanksley. 1995. Molecular genetic analysis of the ripening-
inhibitor and non-ripening loci of tomato: a first step in genetic map-based
cloning of fruit ripening genes. Mol Gen Genet 248:195-206.
Steven D. Tanksley 861
98. O’Donoughue, L.S., S.F. Kianian, P.J. Rayapati, G.A. Penner, M.E. Sorrells,
S.D. Tanksley, R.L. Phillips, H.W. Rines, M. Lee, G. Fedak, S.J. Molnar,
D. Hoffman, C.A. Salas, B. Wu, E. Autrique, and A. Van Deynze. 1995. A
molecular linkage map of cultivated oat. Geneome 38:368-380.
99. Roder, Marion S., Mark E. Sorrells, and Steven D. Tanklsey. 1995. Pulsed-
field gel analysis of 5S and satellite DNA in barley. Genome 38:153-157.
100. Roder, Marion, Jens Plaschke, Susanne U. Konig, Andreas Borner, Mark E.
Sorrells, Steven D. Tanksley, and Martin W. Ganal. 1995. Abundance,
variability and chromosomal location of microsatellites in wheat. Mol. Gen.
Genet. 246:327-333.
101. Tanksley, Steven D., Martin W. Ganal and Gregory B. Martin. 1995.
Chromosome landing: a paradigm for map-based gene cloning in plants
with large genomes. Trends in Genetics 11:63-68.
102. Yen, Hsiao-Ching, Sanghyeob Lee, Steven D. Tanksley, Michael B. Lanahan,
Harry J. Klee, and James J. Giovannoni. 1995. The tomato never-ripe locus
regulates ethylene-inducible gene expression and is linked to a homolog of
the arabidopsis ETR1 Gene. Plant Physiol. 107:1343-1353.
103. Yu Z.H., S.R. McCouch, T. Kinoshita, S. Sato, and S.D. Tanksley 1995.
Association of Morphological and RFLP Markers in Rice (Oryza sativa L.).
Genome 38: 38:566-574.
104. Arumuganathan, K., Gregory B. Martin, Hakan Telenius, Steven D. Tanksley,
and Elizabeth D. Earle. 1994. Chromosome 2-specific DNA clones from flow-
sorted chromosomes of tomato. Mol. Gen. Genet. 242:551-558.
105. Autrique, Enrique, Ravi P. Singh, Steven D. Tanksley, and Mark E. Sorrells.
1994. Molecular markers for four leaf rust resistance genes introgressed
into wheat from wild relatives. Crop Science Genome 38:75-83.
106. Azanza, F., T.E. Young, D. Kim, S.D. Tanksley, and J.A. Juvik. 1994.
Characterization of the effects of introgressed segments of chromosome 7
and 10 from Lycopersicon chmielewskill on tomato soluble solids, pH, and
yield. Theor. Appl. Genet. 87:965-972.
107. Bonierbale, M.W., R.L. Plaisted, O. Pineda, and S.D. Tanksley. 1994. QTL
analysis of trichome-mediated insect resistance in potato. Theor. Appl. Genet.
87:973-987.
108. Causse, M.A., T.M. Fulton, Y.G. Cho, S.N. Ahn, J. Chunwongse, K. Wu,
J. Xiao, Z. Yu, P.C. Ronald, S.E. Harrington, G. Second, S.R. McCouch, and
S.D. Tanksley. 1994. Saturated molecular map of the rice genome based on
an interspecific backcross population. Genetics 138:1251-1274.
109. Chunwongse, J., T.B. Bunn, C. Crossman, J. Jiang, and S.D. Tanksley.
1994. Chromosomal localization and molecular marker tagging of the
powdery mildew resistance gene (Lv) in tomato. Theor. Appl. Genet.
89:76-79.
862 Wolf Prize in Agriculture
110. Ma, Z.Q., M.E. Sorrells, and S.D. Tanksley. 1994. RFLP markers linked to
powdery mildew resistance genes Pm1, Pm2, Pm3 and Pm4 in wheat. Genome.
37:871-875.
111. Martin, Gregory B., Anne Frary, Tiyun Wu, Sergio Brommonschenkel, Julapark
Chunwongse, Elizabeth D. Earle, and Steven D. Tanksley. 1994. A member of
the tomato Pto gene family confers sensitivity to fenthion resulting in rapid
cell death. The Plant Cell 6:1543-1552.
112. O’Donoughue, L.S., E. Souza, S.D. Tanksley, and M.E. Sorrells. 1994.
Relationships among North American oat cultivars based on restriction
fragment length polymorphism. Crop Sci. 34:1251-1258.
113. Ori, N., Paran, I., Aviv, D., Eshed, Y., Tanksley, S., Zamir, D., and Fluhr, R.
1994. A genomic serarch for the gene conferring resistance to Fusarium wilt
in tomato. Euphytica 79:201-204.
114. Paul, E., M. Goto, and S.D. Tanksley. 1994. SolGenes - a Solanaceae database.
Euphytica 79:181-186.
115. Wing, Rod A., Hong-Bin Zhang, and Steven D. Tanksley. 1994. Map-based
cloning in crop plants. Tomato as a model system: I. Genetic and phsyical
mapping of jointless. Mol. Gen. Genet. 242:681-688.
116. Wing, Rod A., Vipin K. Rastogi, Hong-bin Zhang, Andrew H. Paterson, and
Steven D. Tanksley. 1994. An improved method of plant megabase DNA
isolation in agarose microbeads suitable for physical mapping and YAC cloning.
The Plant Journal. 4:893-898.
117. Ahn, S. and S.D. Tanksley. 1993. Comparative linkage maps of rice and
maize genomes. Proc. Natl. Acad. Sci. USA 90:7980-7984.
118. Ahn, S., J.A. Anderson, M.E. Sorrells, and S.D. Tanksley. 1993. Homoelogous
relationships of rice, wheat and maize chromosomes. Mol. Gen. Genet.
241:483-490.
119. Anderson, J.A., G.A. Churchill, J.E. Autrique, S.D. Tanksley, and M.E. Sorrells.
1993. Optimizing parental selection for genetic linkage maps. Genome
31:181-186.
120. Anderson, J.A., M.E. Sorrells, and S.D. Tanksley. 1993. Detection of
QTLs affecting pre-harvest sprouting resistance in wheat by RFLPs. Crop Sci.
33:453-459.
121. Anderson, James A., Mark E. Sorrells, and Steven D. Tanksley. 1993. RFLP
analysis of genomic regions associated with resistance to preharvest sprouting
in wheat. Crop Sci. 33:453-459.
122. Chunwongse, J., G.B. Martin, and S.D. Tanksley. 1993. Pre-germination
genotypic screening using PCR amplification of half-seeds. Theor. Appl. Genet.
86:694-698.
123. Da Silva, Jorge A.G., Mark E. Sorrells, William L. Burnquist, and Steven D.
Tanksley. 1993. RFLP linkage map and genome analysis of Saccharum
spontaneum. Genome 36:782-791.
Steven D. Tanksley 863
124. deVicente, M.C. and S.D. Tanksley. 1993. QTL analysis of transgressive
segregation in an interspecific tomato cross. Genetics 134:585-596.
125. Mackill, D.J., M.A. Salam, Z.Y. Wang, and S.D. Tanksley. 1993. A major
photoperiod sensitivity gene tagged with RFLP and isozyme markers in rice.
Theor. Appl. Genet. 85:536-540.
126. Martin, Gregory B., Sergio H. Brommonschenkel, Julapark Chunwongse, Anne
Frary, Martin W. Ganal, Rody Spivey, Tiyun Wu, Elizabeth D. Earle, and
Steven D. Tanksley. 1993. Map-based cloning of a protein kinase gene
conferring disease resistance in tomato. Science 262:1432-1436.
127. Pineda, O., M.W. Bonierbale, R.L. Plaisted, B.B. Brodie, and S.D. Tanksley. 1993.
Identification of RFLP markers linked to the H1 gene conferring resistance to
the potato cyst nematode Globodera rostochiensis. Genome 36:152-156.
128. Roder, Marion S., Nora L.V. Lapitan, Mark E. Sorrells, and Steven D. Tanksley.
1993. Genetic and physical mapping of barley telomeres. Mol. Gen. Genet.
238-303.
129. Tanksley, Steven D. 1993. Mapping polygenes. Annu. Rev. Genet. 27:205-233.
130. Tanksley, S.D., T.M. Fulton, and S.R. McCouch. 1993. Linkage map of rice
(Oryza sativa) (2N=24) in O’Brien, S.J. (ed.). Genetic Maps 6.61-6.79.
131. Wu, Kun-Sheng and Steven D. Tanksley. 1993. Genetic and physical
mapping of telomeres and macrosatellites of rice. Plant Molecular Biology
22:861-872.
132. Ahn, S. N., C. N. Bollich, and S.D. Tanksley. 1992. RFLP tagging of a gene for
aroma in rice. Theor. Appl. Genet. 84:825-828.
133. Anderson, J.A., Y. Ogihara, M.E. Sorrells, and S.D. Tanksley. 1992. Developmnt
of a chromosomal arm map for wheat based on RFLP markers. Theor. Appl.
Genet. 83:1035-1043.
134. Broun, Pierre, Martin W. Ganal, and Steven D. Tanksley. 1992. Telomeric
arrays display high levels of heritable polymorphism among closely related
plant varieties. Proc. Natl. Acad. Sci. USA 89:1354-1357.
135. Goffreda, J.C., W.B. Burnquist, S.C. Beer,S.D. Tanksley, and M.E. Sorrells.
1992. Application of molecular markers to assess genetic relationships among
accessions of wild oat, Avena sterilis. Theor. Appl. Genet. 5:146-151.
136. Ma, Z.-Q., B.S. Gill, M.E. Sorrells, and S.D. Tanksley. 1992. RFLP markers
linked to two Hessian fly-resistant genes in wheat (Triticum tauschii (coss.)
Schmal. Theor. Appl. Genet. 85:750-754.
137. Martin, Gregory B., de Vicente, M. Carmen, and Steven D. Tanksley. 1992.
High resolution linkage analysis and physical characterization of the Pto
bacterial resistance locus in tomato. MPMI.
138. Martin, Gregory B., Martin W. Ganal, and Steven D. Tanksley. 1992.
Construction of a yeast artificial chromosome library of tomato and
identification of cloned segments linked to two disease resistance loci. Mol.
Gen. Genet. 233:25-32.
864 Wolf Prize in Agriculture
151. Ganal, Martin W., Nora L. V. Lapitan and Steven D. Tanksley. 1991.
Macrostructure of the tomato telomeres. The Plant Cell 3:87-94.
152. Ganal, Martin W., Meredith W. Bonierbale, Marion S. Roeder, William D.
Park and Steven D. Tanksley. 1991. Genetic and physical mapping of the
patatin genes in potato and tomato. Mol. Gen. Genet. 225:501-509.
153. Giovannoni, James J., Rod A. Wing, Martin W. Ganal, and Steven D. Tanksley.
1991. Isolation of molecular markers from specific chromosomal intervals
using DNA pools from existing mapping populations. Nucleic Acids Research
19:6553-6558.
154. Heun, M., A.E. Kennedy, J.A. Anderson, N.L.V. Lapitan, M.E. Sorrells, and
S.D. Tanksley. 1991. Construction of a restriction fragment length
polymorphism map for barley (Hordeum vulgare). Genome 34:437-447.
155. Lapitan, Nora L.V., Martin W. Ganal, and Steven D. Tanksley. 1991.
Organization of the 5S ribosomal RNA genes in the genome of tomato.
Genome 34:509-514.
156. Martin, Gregory, John G. K. Williams and Steven D. Tanksley. 1991. Rapid
identification of markers linked to a Pseudomonas resistance gene in tomato
by using random primers and near-isogenic lines. Proc. Natl. Acad. Sci. USA
88:2336-2340.
157. McCouch, S.R., M.L. Abenes, R. Angeles, G.S. Khush, and S.D. Tanksley. 1991.
Molecular tagging of a recessive gene, Xa-5, for resistance to bacterial blight
of rice. Rice Genet. Newsl. 8:143-145.
158. Messeguer, R. M. Ganal, M.C. de Vicente, N.D. Young, H. Bolkan, and
S.D. Tanksley. 1991. High resolution RFLP map around the root knot nematode
resistance gene (Mi) in tomato. Theor. Appl. Genet. 82:529-536.
159. Messeguer, Ramon, Martin W. Ganal, John C. Steffens and Steven D. Tanksley.
1991. Characterization of the level, target sites and inheritance of cytosine
methylation in tomato nuclear DNA. Plant Molecular Biology 16:753-770.
160. Paterson, A.H., S.D. Tanksley, and M.E. Sorrells. 1991. DNA markers in plant
improvement. Adv. Agron. 46:40-90.
161. Paterson, Andrew H., Susan Damon, John D. Hewitt, Daniel Zamir, Haim D.
Rabinowitch, Stephen E. Lincoln, Eric S. Lander, and Steven D. Tanksley.
1991. Mendelian factors underlying quantitative traits in tomato: comparison
across species, generations, and environments. Genetics 127:181-197.
162. Yu, Z. H., D. J. Mackill, J. M. Bonman and S. D. Tanksley. 1991. Tagging
genes for blast resistance in rice via linkage to RFLP markers. Theor. Appl.
Genet. 81:471-476.
163. Ganal, M. W., G. B. Martin, R. Messeguer and S. D. Tanksley. 1990. Application
of RFLPs, physical mapping and large DNA technologies to the cloning
of important genes from crop plants. AgBiotech News and Information
2:835-840.
866 Wolf Prize in Agriculture
178. Bonierbale, Meredith, Robert L. Plaisted, and Steven D. Tanksley. 1988. RFLP
maps of potato and tomato based on a common set of clones reveal modes of
chromosomal evolution. Genetics 120:1095-1103.
179. Ganal, Martin W., Nora L. V. Lapitan, and Steven D. Tanksley. 1988. A
molecular and cytogenetic survey of major repeated DNA sequences in tomato
(Lycopersicon esculentum). Mole. Gen. Genet. 213:262-268.
180. Lapitan, Nora L. V., Martin W. Ganal, and Steven D. Tanksley. 1988. Generation
of a tomato somatic chromosome karyotype based on in situ hybridization of
the TGR1 satellite repeat.
181. Loaiza-Figueroa, Fernando and Steven D. Tanksley. 1988. Genetics of a second
locus determining pungency in chili peppers (Capsicum). J. Hered.
182. McCouch, S. R., G. Kochert, Z. Yu, Z. Wang, G. S. Khush, W. R. Coffman, and
S. D. Tanksley. 1988. Molecular mapping of rice chromosomes. Theor. Appl.
Genet. 76:815-829.
183. Paterson, Andrew H., Eric S. Lander, John D. Hewitt, Susan Peterson,Stephen
E. Lincoln, and Steven D.Tanksley. 1988. Resolution of quantitative traits
into Mendelian factors by using a complete RFLP linkage map. Nature
335:721-726.
184. Tanksley, S. D. and J. Hewitt. 1988. Use of molecular markers in breeding for
soluble solids content in tomato — a re-examination. Theor. Appl. Genet.
75:811-823.
185. Tanksley, Steven D., Joyce Miller, Andrew Paterson, and Robert Bernatzky.
1988. Molecular mapping of plant chromosomes. In: Chromosome Structure
and Function ( J. Gustafson and R. Appels, eds.). Plenum. pp. 157-172.
186. Tanksley, Steven D., Robert Bernatzky, Nora L. Lapitan, and James P. Prince.
1988. Conservation of gene repertoire but not gene order in pepper and
tomato. Proc. Natl. Acad. Sci. 85:6419-6423.
187. Young, Nevin D., Daniel Zamir, Martin W. Ganal, and Steven D. Tanksley.
1988. Use of isogenic lines and simultaneous probing to identify DNA markers
tightly linked to the Tm-2a gene in tomato. Genetics 120:579-585.
188. Tanksley, S. D. and E. Pichersky. 1988. Organization and evolution of
sequences in the plant nuclear genome. In: L. D. Gottlieb and S. K. Jain (eds.).
Plant Evolutionary Biology. Chapman & Hall.
189. Zamir, D. and S. D. Tanksley. 1988. Tomato genome is comprised
largely of fast-evolving low copy-number sequences. Mole. Gen. Genet.
213:254-261.
190. Pichersky, E., N. E. Hoffman, R. Bernatzky, B. Piechulla, S. D. Tanksley, and A.
R. Cashmore. 1987. Molecular characterization and genetic mapping of DNA
sequences encoding the Type l chlorophyla/b-binding polypeptide of
photosystem 1 in Lycopersicon esculentum (tomato). Plant Molecular Biology
9:205-216.
868 Wolf Prize in Agriculture
220. Tanksley, S.D. and R. A. Jones. 1981. Effect of 02 stress on tomato ADH’s:
description of a second ADH coding gene. Biochem. Genet. 19:397.
221. Tanksley, S.D., H. Medina Filho, and C. M. Rick. 1981. The effect of isozyme
selection on an interspecific backcross of tomato. Theor. Appl. Genetics
60:291-296.
222. Zamir, D., S. D. Tanksley, and R. A. Jones. 1981. Low temperature effect on
selective fertilization in pollen mixtures of wild and cultivated tomato species.
Theor. Appl. Genetics 59:235-238.
223. Zamir, D., S. D. Tanksley, and R. A. Jones. 1981. The genetic origin of plantlets
derived from male sterile tomato anthers. Plant Science Letters 21:223-227.
224. Tanksley S, Rick C (1980) Isozymic gene linkage map of the tomato:
Applications in genetics and breeding 58:161.
225. Tanksley S, Rick C (1980) Genetics of esterases in species of Lycopersicon.
Theor Appl Genet 56:209.
226. Tanksley S (1979) Linkage, chromosomal association, and expression of Adh-
I and Pgm-2 in tomato. Biochemical Genetics 17:1159.
227. Rick, C. M., Tanksley, S. D., and Fobes, J. F. (1979). A pseudoduplication in
Lycopersicon pimpinellifolium. Proc. Natl. Acad. Sci. 763435–3439.
228. Rick C, Fobes J, Tanksley (1979) Evolution of mating systems in Lycopersicon
hirsutum as deduced from genetic variation in electrophoretic and
morphological characters. Plant Systematics and Evolution 132:279.
CURRICULUM VITAE
Marital Status: Married
Date & place of birth: Sept. 7, 1930, Peking
Nationality: Chinese
Position/Title: Director General, China National Hybrid Rice Research &
Development Center
Honorary President of Hunan Academy of Agriculture Science
Degrees: Honorary Ph. D, Chinese University of Hong Kong, 2001
B.Sc. Agronomy, South-west Agricultural University, in 1953
Scientific Field: Hybrid Rice Breeding
Address (Institution): China National Hybrid Rice Research & Development Center,
Changsha, Hunan 410125, P.R. China
871
872 Wolf Prize in Agriculture
BIOGRAPHY
For his breakthrough achievement in developing the genetic materials and
technologies essential for breeding high-yielding hybrid rice varieties, Prof. Yuan
Longping Yuan 873
Longping won the World Food Prize in 2004 – the United Nations’ Food and
Agricultural Organization’s International Year of Rice.
Born in Peking in 1930, Yuan Longping graduated from Southwest Agricultural
College in China in 1953, and then taught crop genetics and breeding at Hunan
Anjiang Agricultural School. He began research there in hybrid rice development
in 1964 and was transferred to the Hunan Academy of Agricultural Sciences in
1971 to serve as a research professor.
Professor Yuan is widely acknowledged for his discovery of the genetic basis of
heterosis in rice – a phenomenon in which the progeny of two distinctly different
parents grow faster, yield more, and resist stress better than either parent. In
developing his three-line system of hybrid rice, Professor Yuan and his team soon
produced a commercial hybrid rice variety called Nan-you No. 2, which was
released in 1974. With yields 20 percent higher than previous varieties, Professor
Yuan’s new crop immediately began to improve food availability in China.
In the subsequent three decades, planting of this new crop has spread so
widely that now about 60 percent of China’s rice production area is planted in
hybrid rice with a 20 percent higher yield over conventional rice varieties. This
translates into food to nourish approximately 70 million more people per year in
China alone.
Beyond this exceptional accomplishment, Professor Yuan has built an additional
legacy of combating food shortages and hunger. He has developed a new technique
for increasing hybrid seed yields through out-pollination, an improved two-line
system of hybrid rice, and other strategies to further improve hybrid rice. His
colleagues have carried on his work with his collaboration and supervision to
develop new strains of “super hybrid rice” that produce almost 10 tons per hectare.
With higher yields, farmers have increased rice production while simultaneously
shifting millions of hectares out of rice and into alternatives such as fish, vegetables,
fruits, and other food and fiber crops, giving more balanced diets and a higher
standard of living to rural Chinese families. Additionally, based on the new
production technique Prof. Yuan developed in 1975 to obtain higher amounts of
Nan-you No. 2 seed, China was able to establish its own hybrid seed industry,
which today provides additional revenue and training opportunities to thousands
of farmers.
The impact of Prof. Yuan’s ingenuity has been felt beyond China’s rice industry.
Researchers and producers of other crops in China have successfully used the
two-line system for rice to explore similar systems for hybrid sorghum and rapeseed
with increased yields. He has also played a key role in developing hybrid rice
throughout Asia and to Africa and the Americas. Since 1980, he has trained
thousands of scientists and researchers from over 25 countries and has served as a
chief consultant to the FAO. Farmers in more than ten other countries besides
China, including the United States, have thus benefited from his work, gaining
access to a technology they may otherwise never have enjoyed.
874 Wolf Prize in Agriculture
In addition to the 2004 World Food Prize, Prof. Yuan’s honors and awards
include China’s State Supreme Science and Technology Award, the 2001 Magsaysay
Award, the UN FAO Medal of Honor for Food Security, and the 2004 Wolf Prize in
Agriculture. Prof. Yuan currently directs China’s National Hybrid Rice Research
and Development Center and is a member or leader of several national committees,
conferences, and foundations that support agriculture, science, and technology in
China.
Professor Yuan Longping’s pioneering research has helped transform China
from food deficiency to food security within three decades. His accomplishments
and clear vision helped create a more abundant food supply and, through food
security, a more stable world. Professor Yuan’s distinguished life’s work has caused
many to call him the “Father of Hybrid Rice,” while his continuing research offers
even more promise for world food security and adequate nutrition for the world’s
poor.
LIST OF PUBLICATIONS
1. A preliminary report on male sterility in rice, Oriza sativa L. 1966, Scientia
Sinica Bulletin (in Chinese with English abstract).
2. Hybrid rice breeding—practice and theory, 1977, Scientia Agricultura Sinica
(in Chinese).
3. Key techniques for hybrid rice seed production, 1977, Genetics and Breeding
(in Chinese).
4. Hybrid rice breeding in China, 1980, Selected papers from the 1979
international Rice research Conference, IRRI.
5. A Concise Course in Hybrid Rice, 1985, published by Hunan Science and
Technology Publishing House (A Chinese-English bilingual textbook).
6. Current Status of Hybrid Rice Research and Development. International
Symposium on Hybrid Rice, Changsha, Hunan, China, 6-10 Oct., 1986.
7. Organization of a Hybrid Rice Breeding Program, Hybrid Rice Proceedings of
International Symposium, 1986, Changsha, China.
8. The strategies for hybrid rice breeding, 1987, Hybrid Rice (in Chinese).
9. The Potential Vse of Apomixis in Crop Improvement, Hybrid Rice, 1988(4):1-3.
10. Breeding for Wide Compatibility Rice Line, Hybrid Rice, 1989(2):35-38.
11. Rice Line 84-15 as an Apomictic Material Needs Scientific Evidence. Hybrid
Rice, 1989(4):3-4.
12. New Progresses on Rice Apomixis Research, Hybrid Rice, 1990(1):29-32.
13. Progress of two-line system hybrid rice breeding, 1990, Scientia Agricultura
Sinica.
14. Technical Strategy of Breeding for TGMS Lines. Hybrid Rice, 1992(1):1-4.
15. Purfication of T(P)GMS Lines of Rice and Their Production of Foundation
Seeds. Hybrid Rice. 1994(6):1-3.
Longping Yuan 875
16. Technology of hybrid rice production, a manual published by FAO, 1995 (in
English).
17. Studies on Two-Line System Hybrid Rice. Hunan Agr. Sci., 1995(6):4-5.
18. Progress of Two-line System Hybrid Rice Breeding Sci. Agr. Sinica, 1990(3):1-6.
19. Prospects of Rice Yield Potential from the View Point of Plant Breeding, Hybrid
Rice, 1996(4):1-2.
20. Hybrid Rice Breeding in China. Advances in Hybrid Rice Technology:
Proceedings of the 3rd International Symposium on Hybrid Rice, 1996, India.
21. Breeding Strategies for Development of Intersubspecific Hybrid Rice. Hybrid
Rice, 1996(2):1-3.
22. Genes from Wild Rice Improve Yield. Nature. 1996, Nov. 21, 384(6606):
223-224.
23. A Strategy for Developing Wide Spectrum Compatibility Rice Line. Sci. Agr.
Sinica, 1997, 30(4):1-8.
24. Explaiting Crop Heterosis by Two-line System Hybrid: Current, Status and
Future Prospects. Proc. Int. Symp. On Two-line System Heterosis Breed in
Crops. 1997, Changsha, Hunan, China.
25. Current Status, Objectives and Prospects of Research and Development of
Two-Line Hybrid Rice. Research of Agricultural Modernization, 1997(1):
1-3 Preliminary Experiences on SRI for Growing Super Hybrid Rice.
26. Hybrid Rice Breeding for Super High Yield. Hybrid Rice. 1997(6):1-6.
27. Hybrid Rice Development and Use: Innovative Approach and Challenges. The
19th Session of International Rice Commission, 1998, Cairo, Egypt.
28. Hybrid Rice Breeding for Super High Yield XVIIIth Int. Congr. Genet., 1998,
Beijing, China.
29. Retrospect, Current Status and Outlook of Hybrid Rice Breeding. China Rice,
1999(4)3-6.
30. Hybrid Rice — Genetics, Breeding and Seed Production. Plant Breeding Review,
Vol. 17. 1999.
31. The System of Rice Intensification (SRI). Hybrid Rice, 2001(4):1-3.
32. Preliminary Experiences on SRI for Growing Super Hybrid Rice. SRI
International Conference, 2002, Sanya, China.
33. Hybrid Rice Technology for Food Security in the world. The 4th International
Hybrid Rice Symposium, 2002, Hanoi, Vietnam.
34. The Second Generation of Hybrid Rice in China, The Commission’s 20th Session,
Bangkok, Thailand, July 23-26, 2002.
35. Recent Progress in Breeding Super Hybrid Rice in China. International Rice
Congress 2002, Beijing, China.
36. L.P. Yuan. Hybridriceology, Published by China Agriculture Press, 2002, Beijing.
37. J. Yu, S. Hu, J. Wang, G. K. S. Wong, S. Li, B. Liu, Y. Deng, J. Sun, J. Tang,
Y. Chen, X. Huang, W. Lin, C. Ye, W. Tong, L. Cong, J. Geng, Y. Han, L. Li,
W. Li, G. Hu, X. Huang, W. Li, J. Li, Z. Liu, L. Li, J. Liu, Q. Qi, J. Liu, L. Li,
876 Wolf Prize in Agriculture
that hybrid rice outyields the modern improved inbred varieties by around 20%
for many years.
2. Development of hybrid rice seed production technology. Previously, it seemed
impossible to obtain high seed yield because of the thought that the self-
pollinating characteristic in rice would restrict its outcrossing rate. Prof. Yuan
and his assistants, however, developed the hybrid seed production technique in
1975. On average the hybrid seed yield is around 2.7 tons per ha and the ratio
of field areas between hybrid seed production and commercial production of
hybrid rice is 1 to 120-150 nowadays.
3. Development of strategies for further improvement of hybrid rice.
Prof. Yuan is not content with what he has achieved. He has always been
pursuing to evolve new approaches to enhance the heterosis level and to simplify
the methodology in hybrid rice technology. He proposed an overall strategy for
further improvement of hybrid rice, i.e. the “three developmental phases of
hybrid rice breeding”:
To enhance the heterosis level: intervarietal heterosis (phase I) →
intersubspecific heterosis (phase II) → distant heterosis (phase III).
To simplify the methodology: three line or CMS system (phase I) → two line
or P(T)GMS system (phase II) → one line or apomixis system (phase III).
Each of the phases would mark a new breakthrough in rice breeding and
result in a great increase in yield if it is attained.
4. Development of practicable two-line system hybrid rice. Two-line system has
two advantages over the three-line system, namely, simpler procedure of seed
production and higher chance of developing better heterotic hybrids. Under
Prof. Yuan’s wise leadership and technical guidance, his research team succeeded
in developing a whole package of two-line system hybrid rice technology in
1995. The planting area under two-line hybrid rice reached 3.5 million ha
recently in China and the newly released two-line hybrids outyielded the three-
line check hybrids by about 10%.
5. Developing super-high yielding hybrid rice and exploring new approaches to
further increase yield potential of rice. Prof. Yuan is an imaginative scientist.
He realized early on that there is always a yield stagnation after a breakthrough
in rice breeding and dedicated to further improve the yield potential of hybrid
rice. Recently, he proposed a unique morphological model and a strategy of
utilizing inter-subspecific (indica/japonica) heterosis for developing super-high
yielding hybrid rice, i.e. super hybrid rice. Several inter-subspecific hybrids
developed according to his model yielded 12 t/ha at multi-location yield trials.
The average yield of the super hybrid rice is 8.5 t/ha on very large scale
commercial production (2 million ha) in recent years.
878 Wolf Prize in Agriculture
present president Arroyo and the former president Estrada who witnessed the
remarkable yield advantage in farmers’ fields strongly encourage the nation’s
farmers to plant hybrid rice for increasing rice yield and farmers’ income and
ensuring the food security of the country. Because of the success of developing
hybrid rice outside China in recent years, the important role of hybrid rice
technology in enhancing grain yield was emphasized at the 19th session of the
International Rice Commission (held in Cairo, 7-9 September 1998). Also at
the session, it was suggested that FAO should take active measures to speed up
the development of hybrid rice in the world.
3. Offering a feasible approach to enhance rice yield in the world this century.
The successful development of two-line hybrid rice and the ongoing research
on developing super hybrid rice offer a bright opportunity to achieve food
sufficiency in the new century in China and other developing countries as well.
In 2010, the population in China will reach 1.36 billion and 50 million ton
more food is needed. The situation may be even worse in some other Asian
countries. Because it is impossible to expand the arable land, to raise the yield
per unit area is the only solution. Developing two-line system hybrid rice,
especially super hybrid rice may play a more important role in raising rice yield
to fill the food gap in this century. If 50% of the world paddy field were
covered by rice hybrids and 2 tons grains per ha were increased, it means
400-500 million more people can be fed. Therefore, the government as well as
non-governmental organizations in and outside China pay great attention to
the super hybrid rice program.
5. Directing the hybrid rice research for the future. His “three developmental
phases of hybrid rice breeding” strategies offer the future direction of hybrid
rice research and are actually followed by most hybrid rice breeders. Especially,
the newly proposed model of plant type and approaches for developing super
hybrid rice are highly appreciated and followed by many scientists in and
outside China.
880 Wolf Prize in Agriculture
Through panicle to panicle inspection at heading stage, six male-sterile rice plants
were found in the paddy fields where several varieties were grown from 1964 to
1965 in Hunan Province. The frequency of occurrence of such male-sterile rice
plants under natural conditions was estimated to he 0.13 per cent. According to
the characteristics of their anthers and pollen grains, these male-sterile plants can
he distinguished into three different types:
(1) Pollen-free type. Two plants of this type were found to be completely male-
sterile. Their anthers are smaller and thinner than those of the normal plant
as shown in Fig. 1. They are colourless, remain unbroken after flowering, and
contain no pollen grains.
(2) Pollen abortive type. Two plants of this kind were found to be completely
male-sterile. Their pale-yellow anthers, smaller than normal ones, fail to dehisce
after flowering, but contain poorly developed pollen grains. These pollen grains
are not only smaller in size, but also irregular in form. They show no blue
colour reaction when treated with KI solution.
(3) Partially male-sterile type. Of this type two plants were found. On one plant
the anthers in most of the florets fail to dehisce after flowering, but all the
anthers contain normal pollen grains. On the other plant the anthers in most
of the florets contain poorly developed pollen grains, and remain unbroken
after flowering.
The exploitation of hybrid rice has played a key role in increasing grain yield in
China. However, the heterosis employed now is inter-vatietal and the method
being used belongs to the three line system. From a strategic point of view, the
development of hybrid rice breeding can be divided into three phases according to
the heterosis level and breeding methodology.
1. In terms of methodology: three line or CMS (cytoplasmic male sterile)
system two line or TGMS (thermo sensitive genic male sterile) and PGMS
(photoperiod-sensitive genic male sterile) system → one line or apomixis system.
2. In terms of heterosis level: inter-varietal heterosis inter-subspecific heterosis →
distant heterosis.
882 Wolf Prize in Agriculture
Introduction
The success in the development of hybrid rice in China is a great breakthrough in
rice breeding and a technological innovation in rice production, being an effective
approach to markedly increasing rice yield on a large scale.
China is the first country in the world to exploit heterosis in rice commercially.
Research on breeding hybrid rice was initiated in 1964. The genetic tools (viz.,
male sterile, maintainer and restorer line) essential to develop F1 hybrid were
successfully developed in 1973. The hybrid combinations with good heterosis were
selected in 1974, and seed production techniques were fundamentally completed
in 1975. In 1976, hybrid rice was released to farmers. Since then, the acreage of
hybrid rice has increased rapidly each year. At present, there are about 29 million
hectares under rice in China of which nearly 60% is by hybrid rice.
It has been proven for many years that hybrid rice varieties have more than 20
percent yield advantage over improved inbred varieties. In recent years the
nationwide average yield of hybrid rice is 7.1 t/ha, about 1.4 t/ha higher than
that of improved inbred varieties which is 5.7 t/ha. (The worldwide average yield
is 3.9 t/ha, the yield in Japan is 6.3 t/ha, and in India is 3 t/ha.) These figures
indicate that hybrid rice has been playing a critical role in solving the food problem
of China.
Rice is the main crop food feeding about 60% of China’s population and still
has great yield potential especially in hybrid rice to be tapped. In order to meet
food requirement for all Chinese people in the 21st century, a super rice breeding
program was set up by China Ministry of Agriculture in 1996. The yield targets
for hybrid rice are listed below (Table 1).
Hybrid ricea
Phase % increase
First Second Single
cropping cropping cropping
Fig. 1. P64S/9311 was released for commercial Fig. 2. P64S/E32 created a record yield of 17.1t
production in 2001. ha-1 in an experimental plot in 1999.
Recently the efforts are focused on breeding Phase II super hybrid rice and
good progress has been made. Some promising combinations with 13 t ha-1 of
yield potential have been developed. Among them, the best one is P88S/0293,
which yielded over 12 t ha-1 at four 6.7-ha or 67-ha locations in 2003 and twelve
6.7-ha or 67-ha locations in 2004. It means the yield target of Phase II is attained
one year ahead of the plan.
Technical approaches
Crop improvement practices have indicated, up to now, that there are only
two effective ways to increase the yield potential of crops through plant
breeding, i.e., morphological improvement and the use of heterosis. However, the
potential is very limited when using morphological improvement alone and heterosis
breeding will produce undesirable results if it is not combined with morphological
improvement. Any other breeding approaches and methods, including high
technology such as genetic engineering, must be incorporated into good
morphological characters and strong heterosis; otherwise, there will be no actual
contributions to a yield increase. On the other hand, the further development of
plant breeding for a super yield target must rely on progress in biotechnology.
884 Wolf Prize in Agriculture
Morphological improvement
A good plant type is the foundation for super high yield. Since Dr. Donald proposed
the concept of ideotype in 1968, many rice breeders pay great attention to this
idea and propose various models for super high-yielding rice. Among them, the
well-known one is the “new plant type” proposed by Dr. Khush at IRRI. Its main
features are: (1) large panicles, with 250 spikelets per panicle; (2) fewer tillers,
3-4 productive tillers per plant; and (3) a short and sturdy culm. Whether these
models can realize super high yield or not has yet to be proved.
Based on our studies, especially inspired by the striking characteristics
of the high-yielding combination P64S/E32, which has made a record yield of
17.1 t ha-1, we have found that the super high-yielding rice variety has the following
morphological features:
Fig. 3. The upper three leaf blades should be long, erect, narrow, V-shape and thick.
Number of
Plant spikelets/ Number of Seed setting Actual yield
Combination height (cm) panicle spikelets/plant rate % (kg/ha)
By means of biotechnology
This is another important approach for developing super hybrid rice. So far, three
encouraging results have been obtained in this research area.
The use of favorable genes from wild rice: Based on molecular analysis and field
experiments, two yield-enhancing QTLs from wild rice (Oryza rufipogon L.) were
identified. Each of the QTLs contributed to a yield advantage of 18% over the high-
yielding check hybrid Weiyou 64 (one of the most elite hybrids). By means of
molecular marker-assisted backcrosses and field selection, an excellent R line (Q611)
carrying one of these QTLs was developed. Its hybrid, J23A/Q611, outyielded the
check hybrid by 35% in a replicated trial for the second cropping in 2001. Its
yield in farmers’ fields planted as the second cropping is around 10 t ha-1, about
2 t ha-1 higher than that of the CK.
Using genomic DNA from barnyard grass (Echinochloa crus-galli) to create a
new resource of rice: The total DNA of barnyard grass was introduced into a
restoring line (R207) by the spike-stalk injection method and variants occurred in
the D1. From these variants, new elite stable R lines containing some DNA fragments
of barnyard grass have been developed (Figs. 7 and 8). The most outstanding one
is RB207, and its agronomic characters such as number of spikelets per panicle
and grain weight are much better than those of the original R207. Particularly, its
hybrid, GD S/RB207, has a good plant type and very strong heterosis (Fig. 9). The
estimated yield was more than 15 t ha-1 in our experimental plot in 2002. Its yield
potential at different ecologic locations is now being evaluated. Preliminary results
indicated that this hybrid performed very well in higher elevation regions. At a
mountainous location (600m above sea level), its average yield was 13 t ha-1 in a
large scale (6.5ha) demonstration (Fig. 10).
Longping Yuan 887
Fig. 9. Strong heterosis of GDS/RB207. Fig. 10. GDS/RB207 on a large scale demonstration.
C4 genes from maize have been cloned and successfully transferred into rice
plant. Preliminary test showed that the photosynthetic efficiency of the leaves of
some rice plants containing C4 genes was 15-30% higher than that of the CK. By
using this transgenic plant as donor to introduce C4 genes into super hybrid rice is
underway. It is expected that the yield potential of C4 super hybrid rice can be
further increased by a big margin theoretically.
Based on the above progress, the Phase III super hybrid rice program is proposed,
in which the yield target is 13.5 t ha-1 and could be fulfilled by 2010 (Fig. 11).
Prospects
The yield standard of Phase II super hybrid rice (12 t ha-1) had been achieved by
2004. These super hybrids were released in 2006. Large scale commercial
production showed the yield of the second generation super hybrids was
10-10.5 t ha-1. Based on this achievement, a “planting three to produce four”
project is initiated, i.e. planting three ha of super hybrids to produce as much as
growing four ha of the existing varieties. It is planned that the planting area under
such super hybrids would be extended to 4 million ha in five years, in which an
equivalent of 5.3 million ha of existing rice varieties would be produced. Therefore,
implementation of this project is not only very important for food security but also
can save 1.3 million ha of land for other more profitable uses, thus to help farmers
alleviate poverty and become rich.
By reaching the target of Phase III super hybrid rice program, we can increase
a yield of 3 t ha-1, and will produce 30 million t of more rice yearly when it is
commercialized up to 10 million ha.
Conclusion
Chinese people can not only meet their food demand by themselves, but also help
other developing countries to solve their food shortage problems.
Longping Yuan 889
Source: From Science Vol. 296, No. 5565, pp. 79-92 (2002). Reprinted with permission from AAAS.
890 Wolf Prize in Agriculture
Fig. 2. Distributions for genomic GC content in Fig. 3. GC content distribution for exons and introns in A. thaliana, O. sativa, and H. sapiens. All
冘 冘
A. thaliana, O. sativa, and H. sapiens, computed exon and intron sequences were derived from cDNA-to-genomic alignments. Mean GC content is
over a bin size of 500 bp. Note that for bins/ computed on a length-weighted basis as ⬍GC⬎ ⫽ L i 䡠GCi/ L i , where GCi and L i are the GC content
10 ⫽ 100, the number of bins with that GC i i
content is 1000. and length for the ith segment (exon or intron).
Table 2. Simple repeats. Shown are tandem repeats with periods 1 to 4 (mono-, otide of n ⫽ 2. We compute mean GC contents of the observed repeats in each
di-, tri-, and tetranucleotide) and the totality of repeats with all periods. The category. Repeat content is then given as a percentage by length, normalized
index n is the number of periodic units. For example, AGT TAGT T is a tetranucle- with respect to the data set (assembled contigs, fully masked reads, or cDNAs).
% of % of % of % of % of % of
% GC % GC % GC % GC % GC % GC
data set data set data set data set data set data set
Mononucleotides 7.63 1.7847 27.65 0.0680 20.34 0.6953 21.02 0.0154 24.08 0.6303 1.31 0.7125
Dinucleotides 35.77 0.0904 13.08 0.0847 41.86 0.0553 4.38 0.0294 46.85 0.0573 31.11 0.0394
Trinucleotides 71.79 0.0454 10.08 0.0106 67.20 0.0098 20.81 0.0012 83.05 0.1335 66.67 0.0043
Tetranucleotides 28.77 0.0072 24.90 0.0032 37.35 0.0020 31.90 0.0010 50.00 0.0018 0.00 0.0000
All periods 1.9277 0.1665 0.7624 0.0469 0.8229 0.7561
Table 3. Complex repeats. Transposons identified by RepeatMasker are total and mean size. Repeat content for each family is given as a
assigned to three classes. Each class has a number of families (e.g., percentage by length, normalized with respect to the data set (assembled
tourist-like MITEs), and each family has a number of different sub- contigs, fully masked reads, or cDNAs) or with respect to all identified
families. The number of subfamilies is listed, as well as their transposons.
Class I LINEs 5 18,997 3,799 1.1905 7.43 0.1318 0.22 0.0257 2.51
SINEs 7 1,254 179 0.0888 0.55 0.0047 0.01 0.0268 2.61
gypsy-like 19 105,614 5,559 3.7285 23.28 41.6894 70.35 0.1238 12.07
copia-like 5 35,151 7,030 1.7175 10.72 15.8506 26.75 0.0869 8.47
Subtotal 6.7254 41.99 57.6766 97.33 0.2631 25.65
Class II Ac/Ds TEs 3 1,567 522 0.1099 0.69 0.0145 0.02 0.0000 0.00
En/Spm TEs 3 5,558 1,853 0.2590 1.62 0.2770 0.47 0.0000 0.00
MULEs 22 25,800 1,173 2.4500 15.30 0.6378 1.08 0.1807 17.62
Subtotal 2.8190 17.60 0.9293 1.57 0.1807 17.62
Class III Stowaway-like 70 16,112 230 2.2370 13.97 0.1247 0.21 0.1910 18.62
tourist-like 77 19,933 259 3.7405 23.35 0.3228 0.54 0.3451 33.65
Unknown MITEs 2 1,341 671 0.4950 3.09 0.2080 0.35 0.0458 4.46
Subtotal 6.4725 40.41 0.6556 1.11 0.5818 56.73
Grand Total 213 231,327 1,086 16.0169 100.00 59.2615 100.00 1.0255 100.00
A Draft Sequence of the Rice have been sequenced. The International Rice
Genome Sequencing Project (IRGSP) was
organized to achieve ⬎99.99% accurate se-
Genome (Oryza sativa L. ssp. quence using a mapped clone sequencing
strategy (8). In addition, expressed gene se-
japonica) quencing has been actively pursued. More
CURRICULUM VITAE
Date of birth: July 18th, 1959
Nationality: Belgian
Father of three lovely children (Mathieu, Anouk, Delphine)
EDUCATION:
Doctor in Veterinary Medicine, University of Liège, 1983.
Master of Science, Molecular Biology, Free University of Brussels, 1985.
Agrégation de l’Enseignement Supérieur, University of Liège, 1991.
PROFESSIONAL EXPERIENCE:
903
904 Wolf Prize in Agriculture
Figure 1: Schematic representation of the bovine myostatin gene with position and definition of
the identified DNA sequence polymorphisms. The gray boxes correspond to the untranslated leader
and trailer sequences (large diameter), and the intronic sequences (small diameter) respectively. The
blue, pink and green boxes correspond to the sequences coding for the leader peptide, N-terminal
latency-associated peptide, and bioactive carboxyterminal domain of the protein respectively. Small
green, red, and pink arrows point towards the positions of the primers used for intron amplification,
exon amplification and sequencing, and exon sequencing respectively. The position of the identified
DNA sequence polymorphisms are shown as green, blue, or red lines in the myostatin gene for silent,
conservativen, and disrupting mutations respectively. Each mutation is connected via an arrow with a
box reporting the details of the corresponding DNA sequence and eventually encoded peptide sequence.
In each box, the variant sequence is compared with the control Holstein-Friesian sequence, and
differences are highlighted in color. (from Mammalian Genome 9: 210-213, 1998)
The first research topic that was assigned to Michel Georges was the identification
of the gene(s) underlying the distinct racial feature of Belgian Blue cattle:
double-muscling. This generalized muscular hypertrophy had been hypothesized
by Dr. Hanset to be under the influence of a major gene referred to as “mh”.
This conjecture was confirmed when the predicted gene was mapped by linkage
analysis to bovine chromosome 2 (Mammalian Genome 6: 788-792, 1995). The
gene was subsequently fine-mapped to a subcentimorgan region. This chromosome
segment was shown to harbour the myostatin gene, a member of the TGFβ family
of growth and differentiation factors, which was shown to carry a series of
loss-of-function mutations in double-muscled breeds including Belgian Blue
(Nature Genetics 17: 71-74, 1997; Mammalian Genome 9: 210-213, 1998)
(Fig. 1).
906 Wolf Prize in Agriculture
Figure 2: The IGF2 quantitative trait locus in pigs. Insulin-like growth factor 2 (IGF2) was first
identified as a paternally expressed quantitative trait locus (QTL) in INTERCROSSES between the
European wild boar and Large White domestic pigs; and between Large White and Piétrain pigs. In
the wild-boar intercross, the QTL allele from the domestic pig was associated with high muscularity,
less backfat and a larger heart. Sequence analysis showed that the IGF2 haplotypes in the Swedish
Large White and Piétrain pigs were identical by descent, whereas the Belgian Large White and wild-
boar haplotypes were similar, which indicated the presence of two alleles that are denoted Q and q
for high and low muscle growth, respectively. Another intriguing observation was the large sequence
divergence (~1%) between the two haplotypes. This led to the suspicion that the two haplotypes
might have an Asian and European origin, in line with previous finding that some European breeds,
including the Large White, are hybrids of Asian and European domestic pigs. Sequence analysis of
IGF2 haplotypes segregating in an intercross between Chinese Meishan and Large White pigs confirmed
this hypothesis. The Meishan allele, which was functionally equivalent to IGF2*q, was nearly identical
to IGF2*Q at the sequence level. These data provided conclusive evidence that the causative mutation
for IGF2*Q was a G-to-A substitution at nucleotide 3,072 in intron 3. IGF2 was identified as a
positional candidate gene, but the quantitative trait nucleotide (QTN) was identified by pure genetics:
linkage analysis to deduce QTL genotypes combined with an analysis of the minimum shared haplotype.
Functional studies showed a plausible mechanism for the QTL effect (see figure). First, the mutation
does not affect the imprinting or METHYLATION STATUS of the QTN region and the region is
undermethylated in skeletal muscle. Second, the wild-type sequence binds a nuclear factor and this
interaction is abrogated by the mutation and by methylation. Third, transfection analysis indicated
that the wild-type sequence functions as a silencer element, whereas the mutant sequence is a
significantly weaker silencer. Finally, expression analysis showed an approximately threefold
upregulation of IGF2 expression in postnatal skeletal and cardiac muscle but not in prenatal muscle
or in liver. The result is consistent with phenotypic data showing that IGF2*Q are associated with high
muscle growth and a larger heart, but has no effect on birth weight or the size of the liver. The IGF2
QTL is truly adaptive from a pig production point of view as it does not affect birth weight but
supports muscle growth after birth. The photographs of the wild boar, Meishan, Piétrain and Large
White pigs were provided by B. Kristiansson, Quality Genetics AB, J.-M. Beduin and the Roslin
Institute, respectively. (from Nature Reviews Genetics 5: 202-212, 2004)
908 Wolf Prize in Agriculture
POLAR OVERDOMINANCE AT THE OVINE CALLIPYGE LOCUS AND OTHER IMPRINTED QTL
In recent years, it has become apparent that a growing number of genes do not
operate according to Mendel’s laws. It has been shown in particular that some
genes are undergoing parental imprinting, i.e. paternal and maternal alleles are
functionally non-equivalent, carrying parent-of-origin specific epigenetic marks. It
is also increasingly realized that perturbations of such imprinted genes underlie a
series of inherited diseases characterized by so-called “parent-of-origin” effects
in man.
While studying the genetic determinism of a muscular hypertrophy in sheep
(referred to as the callipyge (CLPG) phenotype) using molecular markers, the team
of Michel Georges in collaboration with Dr. N. Cockett demonstrated that this
phenotype is transmitted in a unique, non-Mendelian way which was referred to
as polar overdominance: only heterozygous individuals having received the CLPG
mutation form their sire exhibit the phenotype (Proc. Natl. Acad. Sci. USA 91:
3019-3023, 1994; Science 273: 236-238, 1996). The gene has been mapped to a
400 Kb chromosome interval (e.g. Mammalian Genome 12: 141-149, 2001;
Mammalian Genome 12: 183-185, 2001). Complete sequencing, annotation and
functional analysis of 400 kilobases within this interval lead to the identification
of six novel genes which were shown to undergo parental imprinting and be
preferentially expressed in skeletal muscle (Genome Research 11: 850-862, 2001;
submitted). It was subsequently shown that the CLPG mutation enhances the
expression of four of these genes without altering their imprinting status (Nature
Genetics 27: 367-369, 2001). The CLPG mutation was shown by others and by
Michel Georges’s team to be a single base pair substitution in a highly conserved
dodecamer motif thought to act as a skeletal muscle specific silencer element
(Genome Research 12: 1496-1506, 2002; Genetics 163: 453-456, 2003). The
fact that only the padumnal heterozygotes express the phenotype (“polar
overdominance”) was then shown to result from the trans-inhibition of the genes
expressed from the paternal allele by the genes expressed from the maternal
allele, and the consequent ectopic expression of DLK1 exclusively in skeletal
muscle of callipyge animals (Trends in Genetics 19: 248-252, 2003; Current
Biology 14: 1858-1862, 2004). Michel Georges’ team has recently shown
that this trans-inhibition involves RNA interference and micro-RNA genes expressed
exclusively from the maternal allele (Current Biology 15: 743-749, 2005)
(Fig. 3).
Polar overdominance at the ovine callipyge locus is becoming a classical topic
in the emerging epigenetic field, as testified by the fact that Michel Georges was
invited to give a plenary lecture at last years 69th Cold Spring Harbor Symposium
on Quantitative Biology devoted to “Epigenetics”.
Michel A. J. Georges 909
Figure 3: Working model for polar overdominance, involving (i) a paternally expressed growth
promoter (DLK1), (ii) a maternally expressed trans-acting (red arrow) growth repressor thought to
correspond to one or more non-coding RNA genes (ncRNA), (iii) a cis-acting (green arrow) LRCE
affected by the CLPG mutation. The crosses correspond to the parental imprinting effects. The functional
status in postnatal skeletal muscle is illustrated for the four CLPG genotypes: active elements are
shown in red, inactive elements in white. Only the +Mat/C Pat individuals express the callipyge phenotype
(boxed in red) due to ectopic expression of the DLK1 protein. (from Current Biology 14: 1858-1862,
2004)
Using the same positional cloning approach, Michel Georges’ team has recently
identified a QTL with major effect on muscular development in the sheep. The QTL
was fine-mapped to a region shown to harbour the myostatin gene known from
previous work to cause double-muscling in cattle. Detailed analysis of the ovine
myostatin gene identified a A to G transition in the 3’UTR revealing an illegitimate
target site for miRNAs that are highly expressed in skeletal muscle. miRNAs are a
recently discovered class of small non-coding RNA genes that play a key “epigenetic”
role during developmental by causing either the degradation or the translational
inhibition of target mRNAs. A very large proportion of genes is now thought to be
under the control of batteries of miRNAs. It was demonstrated that the identified A
to G transition indeed leads to the translational inhibition of the myostatin transcripts
and thus to muscular hypertrophy (Fig. 4) (Nature Genetics 38: 813-818, 2006).
Most importantly, however, subsequent bioinformatic analyses of SNPs databases
for human and mice, demonstrated that mutations creating or destroying putative
miRNA target sites are abundant and might thus be important effectors of phenotypic
variation. This information has been compiled in a publicly accessible database
(www.patrocles.org). This discovery opens a new field of investigation not only for
Figure 4: Schematic representation of the effect of the g+6723G-A mutation in the 3’UTR of the
ovine GDF8 gene. In wild-type animals (such as the Romanov sheep shown in the left picture) the
GDF8 transcripts (in blue) in the sarcoplasm don’t interact with microRNAs miR-1 and miR-206
despite their high level of expression in skeletal muscle. Hence GDF8 protein is produced at regular
levels, controlling muscle growth by retro-inhibition. In mutant animals (such as the Texel sheep
shown in the right picture), GDF8 transcripts (in red) become targets for miR-1 and miR-206 as a
result of the g+6723G-A mutation. This leads to translational inhibition of the GDF8 transcripts,
reduced GDF8 protein levels, relaxation of the retro-inhibition and hence enhanced muscle growth.
(from 71st CSH Symposium on Quantitative Biology Vol. 69, pp 477-483, 2006)
Michel A. J. Georges 911
Michel Georges’ team but for the scientific community in general (Current Opinion
in Genetics & Development 17: 1-11, 2007).
Figure 5: Results of the whole genome association for Crohn Disease. P-values (-log(p)) for the
10,000 best SNPs out of 311,882 are shown (gray circles). The position of previously described
susceptibility loci are marked by red arrows. The p-values obtained in our cohorts with the reportedly
associated SNPs/mutations are shown by the red dots, and the corresponding odds ratios (OR)
indicated. The p-values obtained with SNPs included in the Illumina panel at ≤ 50 Kb from these
SNPs/mutations are marked by black circles. Two novel susceptibility loci identified in this study are
marked by green dots and correspond respectively to the IL23R gene on chromosome 1 and the
PTGER4 gene on chromosome 5. (from PLoS Genetics e58, 2007)
912 Wolf Prize in Agriculture
These result “have placed Belgium on the map” in the extremely competitive
“GWA” field. The laboratory of Michel Georges is presently very actively
involved in a large scale meta-analysis in collaboration with the NIDDK and
Broad.
Institute in the US, and the Welcome Trust Case Control Consortium in the UK.
The results of this unique study, involving ~10,000 CD patients, will be presented
at the 2008 American Society of Human Genetics meeting and should result in a
major publication in the near future.
Michel Georges’ team continues to study the molecular mechanisms involving
PTGER4-dependent CD susceptibility as well as the contribution of Copy Number
Variation to IBD susceptibility.
More recently they have expanded their activities in rodent models to study
the genetic determinism of resistance to infectious agents in collaboration with
Dr. Desmecht. Results of genome-wide QTL scans for resistance to Mouse Pneumonia
Virus (a mouse model of RSV dependent bronchiolitis in children) and Sendai
Virus are being prepared for publication.
TEAM WORK
The previously described scientific achievements are obviously the result of intense
team work (Fig. 6). Michel Georges is presently heading a group of approximately
30, comprising 1/3 senior scientists and post-doctoral fellows, 1/3 PhD students
and 1/3 laboratory technicians. The team comprises people originating from at
least six countries with backgrounds ranging from computer engineering to
veterinarians via biologists, chemists, physicists, and agronomists. Special credit is
expressed towards the group leaders, especially Carole Charlier and Wouter
Coppieters.
The team benefits from an annual research budget of the order of 2.5 million
Euros per year of which 65% is provided by Belgian and European public funding
organizations and 35% by private industry from Belgium, the Netherlands, New
Zealand and Canada.
Figure 6: Unit of Animal Genomics 2002. (i) Dimitri Pirottin, Cécile Libioulle, Latifa Karim, Anne
Cornet, Fabienne Marcq, Luc Grobet, Alain Empain, Patricia Simon, Bernard Grisart, (ii) Carole
Charlier, Fabrice Moreau, Valérie Marot, Karin Segers, Xavier Hubin, Fabienne Davin, Catherine
Collette, Erica Davis, Wouter Coppiteres, Maria Smit, (iii) Michel Georges, Minh Nguyen, Benoît
Brouwers, Laurence Moreau, Myriam Mni, Francesca Baraldi, Nadine Cambisano, (iv) Bernadette
Marcq, Jong-Jo Kim, Paulette Berzi.
914 Wolf Prize in Agriculture
INTERNATIONAL RECOGNITION
INVITED LECTURES
Over the last 15 years, Michel Georges has been invited on average ~5 times per
year to speak at international conferences, including the meetings of the
International Society of Animal Genetics (1992, 1994, 2004), the World Congress
on Genetics Applied to Livestock Production (1990, 1994, 1998, 2002, 2006), the
Plant and Animal Genome Conference (2002, 2003, 2004, 2006, 2007), the
International Congress of Genetics (1993, 2003, 2006, 2008), HUGO’s Human
Genome Meetings (1999, 2001, 2007), Gordon Conferences (1993, 1999, 2003),
Keystone Meetings (2004, 2007, 2x2008) and Banbury Conferences (1990, 1995).
The invited lectures that he is most proud of, however, are the three plenary
lectures that he was invited to give respectively at the 68th, 69th and 71st Cold
Spring Harbour Symposia on Quantitative Biology devoted respectively to “The
Genome of Homo Sapiens” (2003), “Epigenetics” (2004), and “Regulatory RNAs”
Michel A. J. Georges 915
(2006) as well as for the Cold Spring Harbour Meeting on “The Biology of Genomes”
in 2007. In these, his contributions were respectively on the positional identification
of QTN in livestock in 2003, on the molecular dissection of callipyge in 2004, on
polymorphic miRNA-target interactions in 2006, and on the identification of a
novel risk locus for Crohn disease in 2007.
Michel Georges has been invited on several occasions to give courses including
at the University of Guelph (Canada), at the University of Leon (Spain), at
“Internordic Courses” in Uppsala (Sweden) and Lillehammer (Norway), at the
University of Helsinki (Finland), by INRA, at a Wellcome Trust Advanced Course
and at the Summer Institute in Statistical Genetics (NCSU and University of
Washington, USA).
MISCELLANEOUS
Michel Georges is or has been a member (i) of the Steering Committee for the ESF
Scientific Program on Integrated Approaches for Functional Genomics, (ii) of the
NFWO commission on “Medical cell biology and genetics”, (iii) of the Scientific
Advisory Board of the Centre National de Génotypage (CNG) – Evry, France, (iv) of
the Scientific Advisory Board of the Institut National de la Recherche Agronomique
(INRA)- France, (v) of the Scientific Advisory Board of the Consortium National de
Recherche en Génomique (CNRG) – France, and (vi) of the RIKEN SNP Research
Center Advisory Council (SRAC) – Japan.
His laboratory is part of two European Networks of Excellence: (i) The
Epigenome, devoted to the study of epigenetic phenomena (http://www. epigenome-
noe.net/) and (ii) Eadgene, devoted to the study of host-pathogen interactions
in livestock (http://lotus5.vitamib.com/hnb/eadgene/eadgene.nsf/web/frame?
openform).
916 Wolf Prize in Agriculture
MAJOR PUBLICATIONS
ARTICLES IN INTERNATIONAL JOURNALS WITH PEER REVIEW:
8. FRIES, R.; BECKMANN, J.S.; GEORGES, M.; SOLLER, M.; WOMACK, J. (1989).
The bovine gene map. Animal Genetics 20:3-29. [1.4]
9. HILBERT, P.; MARCOTTE, A.; SCHWERS, A.; HANSET, R.; VASSART, G.;
GEORGES, M. (1989). Analysis of genetic variation in the Belgian Blue Cattle
Breed using DNA sequence polymorphism at the growth hormone, low density
lipoprotein receptor, -subunit of glycoprotein hormones and thyroglobulin
loci. Animal Genetics 20:383-394. [1.4]
10. GEORGES, M.; LATHROP, M.; HILBERT, P.; MARCOTTE, A.; SCHWERS, A.;
SWILLENS, S.; VASSART, G.; HANSET, R. (1990). On the use of DNA
fingerprints for linkage studies in cattle. Genomics 6: 461-474. [3.5]
11. PERRET, J.; SHIA, Y.; FRIES, R.; VASSART, G.; GEORGES, M. (1990). A
polymorphic satellite sequence maps to the pericentric region of the bovine
Y chromosome. Genomics 6:482-490. [3.5]
12. GEORGES, M.; LATHROP, M.; BOUQUET, Y.; HILBERT, P.; MARCOTTE, A.;
SCHWERS, A.; ROUPAIN, J.; VASSART, G.; HANSET, R. (1990) Linkage
relationships among 19 genetic markers in cattle. Evidence for linkage between
two pairs of blood group systems: B-Z and S-F/V respectively. Animal Genetics
21:95-105. [1.4]
13. STREYDIO, C.; SWILLENS, S.; GEORGES, M.; SZPIRER, C.; VASSART G. (1990).
Structure, evolution and chromosomal localisation of the human pregnancy-
specific ß1 glycoprotein gene family. Genomics 6:579-592. [3.5]
14. JULIER, C.; DE GOUYON, B.; GEORGES, M.; GUENET, J.L.; AVNER, P.; LATHROP,
G.M. (1990). Minisatellite linkage maps in the mouse by cross-hybridization
with human VNTR probes. Proc. Natl. Acad. Sci. USA 87: 4585-4589.
[10.3]
15. GEORGES, M.; MASSEY, J.M. (1991) Velogenetics or the synergistic use of
Marker Assisted Selection and Germ-Line Manipulation. Theriogenology 35(1):
151-156. [1.8]
16. MOORE, S.S.; SARGEANT, L.; KING, T.J.; MATTICK, J.S.; GEORGES, M.;
HETZEL, D.J.S. (1991). The conservation of dinucleotide microsatellites among
mammalian genomes allows the use of heterologous PCR primer pairs in
closely related species. Genomics 10: 654-660. [3.5]
17. STEELE, M.; GEORGES, M. (1991) Generation of bovine multisite haplotypes
using random cosmids. Genomics 10: 889-904. [3.5]
18. GEORGES, M.; GUNAWARDANA, A.; THREADGILL, D.; LATHROP, M.;
OLSAKER, I.; MISHRA, A.; SARGEANT, L.; STEELE, M.; TERRY, Ch.; ZHAO. X.;
HOLM, T.; FRIES, R.; WOMACK, J. (1991). Characterization of a set of variable
number of tandem repeat markers conserved in Bovidae. Genomics 11:
24-32. [3.5]
19. HILBERT, P.; LINDPAINTER, K.; SERIKAWA, T.; SOUBRIER, F.; CARTWRIGHT, P.;
DUBAY, C.; JULIER, C.; TAKAHASI, S.; VINCENT, M.; BECKMANN, J.;
GANTEN, D.; GEORGES, M. & LATHROP, M. (1991). Chromosomal mapping
918 Wolf Prize in Agriculture
of two genetic loci associated with hereditary hypertension in the rat. Nature
353:521-529. [31]
20. DIETZ, A.B.; GEORGES, M.; THREADGILL, D.W.; WOMACK, J.E.;
SCHULER, L.A. (1992). Somatic cell mapping, polymorphism and linkage
analysis of bovine prolactin related proteins and placental lactogen. Genomics
14: 137-143. [3.5]
21. MACKINNON, M.J.; GEORGES, M. (1992). The effects of selection on linkage
analysis for quantitative traits. Genetics 132: 1177-1185. [4.9]
22. DELHAISE, F.; ZHAO, X.; DESSY, F.; GEORGES, M. (1993). Quantitative
estimation of chimerism in mice using microsatellite markers. Molecular
Reproduction and Development 34: 127-132. [2.5]
23. GEORGES, M.; LATHROP, M.; DIETZ, A.B.; LEFORT, A.; LIBERT, F.;
MISHRA, A.; NIELSEN, D.; SARGEANT, L.S.; STEELE, M.R.; ZHAO, X.;
LEIPOLD, H.; WOMACK, J.E. (1993). Microsatellite mapping of the gene
causing weaver disease in cattle will allow the study of an associated QTL.
Proc. Natl. Acad. Sci. USA 90: 1058-1062. [10.3]
24. GEORGES, M.; DRINKWATER, R.; LEFORT, A.; LIBERT, F.; KING, T.;
MISHRA, A.; NIELSEN, D.; SARGEANT, L.S.; SORENSEN, A.; STEELE, M.R.;
ZHAO, X.; WOMACK, J.E.; HETZEL, J. (1993). Microsatellite mapping of a
gene affecting horn development in Bos taurus. Nature Genetics 4: 206-210.
[38.8]
25. LIBERT, F.; LEFORT, A.; OKIMOTO, R.; GEORGES, M. (1993). Construction of
a bovine genomic library of large yeast artificial chromosome clones. Genomics
18:270-276. [3.5]
26. NEIBERGS, H.L.; GALLAGHER, D.S.; GEORGES, M.; WOMACK, J.E. (1993).
Physical mapping of Inhibin beta-A in domestic cattle. Mammalian Genome
4: 328-332. [2.7]
27. BARENDSE, W.; ARMITAGE, S.M.; KIRKPATRICK, B.W.; MOORE, S.S.; GEORGES,
M.; WOMACK, J.E.; HETZEL, J. (1993). A genetic map of index markers on
bovine chromosome 21. Genomics 18: 598-601. [3.5]
28. BARENDSE, W.; ARMITAGE, S.M.; RYAN, A.M.; MOORE, S.S.; CLAYTON, D.;
GEORGES, M.; WOMACK, J.E.; HETZEL, J. (1993). A genetic map of index
markers on bovine chromosome 1. Genomics 18: 602-608. [3.5]
29. BARENDSE, W.; ARMITAGE, S.M.; KOSSAREK, L.M.; KIRKPATRICK, B.W.;
RYAN, A.M.; SHALOM, A.; CLAYTON, D.; LI, L.; NEIBERGS, H.L.; ZHANG, N.;
GROSSE, W.M.; WEISS, J.; CREIGHTON, P.; MCCARTHY, F.; RON, M.;
TEALE, A.J.; FRIES, R.; MCGRAW, R.A.; MOORE, S.S.; SOLLER, M.;
GEORGES, M.; WOMACK, J.E.; HETZEL, D.J.S. (1994). A genetic map of the
bovine genome shows unexpected chromosomal rearrangements between
humans and cattle. Nature Genetics 6:227-235. [38.8]
Michel A. J. Georges 919
30. COCKETT, N.E.; JACKSON, S.P.; SHAY, T.D.; NIELSEN, D.; GREEN, R.D.;
GEORGES, M. (1994). Chromosomal localization of the callipyge gene in
sheep (Ovis aries) using bovine DNA markers. Proc. Natl. Acad. Sci. USA 91:
3019-3023. [10.3]
31. GEORGES, M.; NIELSEN, D.; MACKINNON, M.; MISHRA, A.; OKIMOTO, R.;
PASQUINO, A.T.; SARGEANT, L.S.; SORENSEN, A.; STEELE, M.R.; ZHAO, X.;
WOMACK, J.E.; HOESCHELE, I. (1995). Mapping quantitative trait loci
controlling milk production by exploiting progeny testing. Genetics 139:
907-920. [4.9]
32. CHARLIER, C.; COPPIETERS, W.; FARNIR, F.; GROBET, L.; LEROY, P.;
MICHAUX, C.; MNI, M.; SCHWERS, A.; VANMANSHOVEN, P.; HANSET, R. &
GEORGES, M. (1995) The mh gene causing double-muscling in cattle maps
to bovine chromosome 2. Mammalian Genome 6: 788-792. [2.7]
33. CHARLIER, C.; DENYS, B.; BELANCHE, J.I.; COPPIETERS, W.; GROBET, L.;
MNI, M.; WOMACK, J.; HANSET, R. & GEORGES, M. (1996). Microsatellite
mapping of a major determinant of White Heifer Disease: the bovine roan
locus. Mammalian Genome 7:138-142. [2.7]
34. CHARLIER, C.; FARNIR, F.; BERZI, P.; VANMANSHOVEN, P.; BROUWERS, B.;
GEORGES, M. (1996). IBD mapping of recessive traits in livestock: application
to map the bovine syndactyly locus to chromosome 15. Genome Research
6:580-589. [9.6]
35. GEORGES, M. & ANDERSSON, L. (1996). Livestock genomics comes of age.
Genome Research 6:907-921. [9.6]
36. COCKETT, N.; JACKSON, S.; SHAW, T.; FARNIR, F.; SNOWDER, G.; NIELSEN, D.;
GEORGES, M. (1996). Polar overdominance at the ovine callipyge locus.
Science 273:236-238. [24.7]
37. SAMSON, M.; LIBERT, F.; DORANZ, B.J.; RUCKER, J.; LIESNARD, C.; FARBER,
C-M.; SARAGOSTI, S.; LAPOUREMOULIE, C.; COGNAUX, J.; FORCEILLE, C.;
MUYLDERMANS, G.; VERHOFSTEDE, C.; BURTONBOY, G.; GEORGES, M.;
IMAI, T.; RANA, S.; YI, Y.; SMYTH, R.J.; COLLMAN, R.G.; DOMS, R.W.;
VASSART, G.; PARMENTIER, M. (1996). Resistance to HIV-1 infection in
caucasian individuals bearing mutant alleles of the CCR-5 chemokine receptor
gene. Nature 382: 722-725. [31]
38. CAI, L.; SCHALKWIJK L.C.; STEHLI, A.S.; ZEE, R.Y.L.; HAAF, T.; GEORGES, M.;
LEHRACH, H.; LINDPAINTER, K. (1996). Construction and characterization
of a 10-genome equivalent yeast artificial chromosome library for the
laboratory rat, Rattus norvegicus. Genomics 38: 385-392. [3.5]
39. MILINKOVITCH, M.C.; LEDUC, R.G.; ADACHI, J.; FARNIR, F.; GEORGES, M.;
HASEGAWA, M. (1996). Effects of character weighting and species sampling
on phylogeny reconstruction: a case study based on DNA sequence data in
cetaceans. Genetics 144:1817-1833. [4.9]
920 Wolf Prize in Agriculture
40. BARENDSE, W.; VAIMAN, D.; KEMP, S.J.; SUGIMOTO, Y.; ARMITAGE, S.M.;
WILLIAMS, J.L.; SUN, H.S.; EGGEN, A.; AGABA, M.; ALEYASIN, S.A.; BAND, M.;
BISHOP, M.D.; BUITKAMP, J.; BYRNE, K.; COLLINS, F.; COOPER, L.;
COPPETTIERS, W.; DENYS, B.; DRINKWATER, R.D.; EASTERDAY, K.;
ELDUQUE, C.; ENNIS, S.; ERHARDT, G.; FERRETTI, L.; FLAVIN, N.; GAO, Q.;
GEORGES, M.; GURUNG, R.; HARLIZIUS, B.; HAWKINS, G.; HETZEL, J.;
HIRANO, T.; HULME, D.; JORGENSEN, C.; KESSLER, M.; KIRKPATRICK, B.W.;
KONFORTOV, B.; KOSTIA, S.; KUHN, C.; LENSTRA, J.A.; LEVEZIEL, H.;
LEWIN, H.A.; LEYHE, B.; LIL, L.; BURRIEL, I.M.; MCGRAW, R.A.; MILLER, J.R.;
MOODY, D.E.; MOORE, S.S.; NAKANE, S.; NIJMAN, I.J.; OLSAKER, I.;
POMP, D.; RANDO, A.; RON, M.; SHALOM, A.; TEALE, A.J.; THIEVEN, U.;
URQUHART, B.G.D.; VAGE, D.I.; VANDEWEGHE, A.; VARVIO, S.; VELMALA, R.;
VIKKI, J.; WEIKARD, R.; WOODSIDE, C.; WOMACK, J.E.; ZANOTTI, M.;
ZARAGOZA, P. (1997) A medium-density genetic linkage map of the bovine
genome. Mammalian Genome 8: 21-28. [2.7]
41. DUNNER, S.; CHARLIER, C.; FARNIR, F.; BROUWERS, B.; CANON, J.; GEORGES,
M. Towards interbreed IBD fine mapping of the mh locus: double-muscling
in the Asturiana de los Valles breed involves the same locus as in the Belgian
Blue cattle breed.(1997) Mammalian Genome 8: 430-435. [2.7]
42. GEORGES, M. (1997). From QTL mapping to QTL cloning: mice at the rescue.
Genome Research 7: 665-666. [9.6]
43. GROBET, L.; ROYO MARTIN, L.J.; PONCELET, D.; PIROTTIN, D.; BROUWERS, B.;
RIQUET, J.; SCHOEBERLEIN, A.; DUNNER, S.; MENISSIER, F.; MASSABANDA, J.;
FRIES, R.; HANSET, R.; GEORGES, M. (1997) A deletion in the myostatin gene
causes double-muscling in cattle. Nature Genetics 17:71-74. [38.8]
44. GROBET, L.; PONCELET, D.; ROYO MARTIN, L.J.; BROUWERS, B.; PIROTTIN, D.;
MICHAUX, CH.; MENISSI ER , F.; ZANOTTI, M.; DUNNER, S.; GEORGES, M.
(1998). Molecular definition of an allelic series of mutations disrupting the
myostatin function and causing double-muscling in cattle. Mammalian
Genome 9: 210-213. [2.7]
45. COPPIETERS, W.; MACKINNON, M.; GEORGES, M. (1998). A note on the
effect of selection on linkage analysis for quantitative traits. Journal of
Heredity, 89:193-195. [1.7]
46. MACKINNON, M.; GEORGES, M. (1998) A bottom-up approach
towards marker assisted selection. Livestock Production Science 54:229-250.
[1]
47. COPPIETERS, W.; RIQUET, J.; ARRANZ, J.-J.; BERZI, P.; CAMBISANO, N.;
GRISART, B.; KARIM, L.; MARCQ, F.; SIMON, P.; VANMANSHOVEN, P.;
WAGENAAR, D.; GEORGES, M. (1998) A QTL with major effect on milk yield
and composition maps to bovine chromosome 14. Mammalian Genome 9:
540-544. [2.7]
Michel A. J. Georges 921
48. COPPIETERS, W.; KVASZ, A.; ARRANZ, J.J.; GRISART, B.; FARNIR, F.;
MACKINNON, M. & GEORGES, M. (1998). A rank-based non parametric
method to map QTL in outbred half-sib pedigrees: application to milk
production in a grand-daughter design. Genetics 149:1547-1555. [4.9]
49. ARRANZ, J.-J.; COPPIETERS, W.; BERZI, P.; CAMBISANO, N.; GRISART, B.;
KARIM, L.; MARCQ, F.; RIQUET, J.; SIMON, P.; VANMANSHOVEN, P.;
WAGENAAR, D.; GEORGES, M. (1998) A QTL affecting milk yield and
composition maps to bovine chromosome 20: a confirmation. Animal Genetics
29: 107-115. [1.4]
50. PIROTTIN, D.; PONCELET , D.; GROBET, L.; ROYO, L.J.; BROUWERS, B.;
MASABANDA, J.; TAKEDA, H.; FRIES, R.; SUGIMOTO, K.; WOMACK, J.;
DUNNER, S.; GEORGES, M. (1999). High resolution human-bovine
comparative mapping based on a closed YAC contig spanning the bovine mh
locus. Mammalian Genome 10: 289-293. [2.7]
51. NEZER, C.; MOREAU, L.; BROUWERS, B.; COPPIETERS, W.; DETILLEUX, J.;
HANSET, R.; KARIM, L.; KVASZ, A.; LEROY, P.; GEORGES, M. (1999) An
imprinted QTL with major effect on muscle mass and fat deposition maps to
the IgfII locus in pigs. Nature Genetics 21: 155-156. [38.8]
52. COCKETT, N.E.; SHAY, T.; BEEVER, J.E.; NIELSEN, D.M.; ALBRETSEN, J.;
GEORGES, M.; PETERSON, K.; STEPHENS, A.; VERNON, W.; TIMOFEEVSKAIA,
O.; SOUTH, S.; MORK, J.; MACIULIS, A.; HOLYOAK, G.R.; BUNCH, T.D. (1999).
Localization of the locus causing Spider Lamb Syndrome to the distal end of
ovine chromosome 6. Mammalian Genome 10: 35-38. [2.7]
53. LIEN, S.; COCKETT, N.E.; KLUNGLAND, H.; ARNHEIM, N.; GEORGES, M.;
GOMEZ-RAYA, L. (1999). High-resolution gametic map of the sheep callipyge
region: linkage heterogeneity among rams detected by sperm typing. Animal
Genetics 30: 42-46. [1.4]
54. RIQUET, J.; COPPIETERS, W.; CAMBISANO, N.; ARRANZ, J.-J.; BERZI, P.;
DAVIS, S.; GRISART, B.; FARNIR, F.; KARIM, L.; MNI, M.; SIMON, P.; TAYLOR, J.;
VANMANSHOVEN, P.; WAGENAAR, D.; WOMACK, J.E.; GEORGES, M. (1999).
Identity-by-descent fine-mapping of QTL in outbred populations: application
to milk production in dairy cattle. Proc. Natl. Acad. Sci. USA 96: 9252-9257.
[9.04]
55. COPPIETERS, W.; KVASZ, A.; ARRANZ, J.-J.; GRISART, B.; RIQUET, J.; FARNIR, F.;
GEORGES, M. (1999). The great-grand-daughter design: a simple strategy
to increase the power of a grand-daughter design. Genetical Research 74:
189-199. [2.5]
56. SPELMAN, R.J.; HUISMAN, A.E.; SINGIREDDY, S.R.; COPPIETERS, W.; ARRANZ,
J.J.; GEORGES, M.; GARRICK, D.J. (1999). Quantitative Trait Loci analysis on
17 nonproduction traits in the New Zealand dairy population. J Dairy Sci
82: 2514-2516. [2.1]
922 Wolf Prize in Agriculture
57. FARNIR, F.; COPPIETERS, W.; ARRANZ, J.-J.; BERZI, P.; CAMBISANO, N.;
GRISART, B.; KARIM, L.; MARCQ, F.; MOREAU, L.; MNI, M.; NEZER, C.;
SIMON, P.; VANMANSHOVEN, P.; WAGENAAR, D.; GEORGES, M. (2000).
Extensive genome-wide linkage disequilibrium in cattle. Genome Research
10: 220-227. [9.6]
58. IKONEN, T.; BOVENHUIS, H.; OJALA, M.; UOTTINEN, O.; GEORGES M.
(2001). Associations between casein haplotypes and first lactation milk
production traits in Finnish Ayrshire cows. J Dairy Sci. 84(2):507-14.
[2.1]
59. KARIM, L.; COPPIETERS, W.; GROBET, L.; VALENTINI, A.; GEORGES, M.
(2001). Convenient genotyping of six myostatin mutations causing double-
muscling in cattle using a multiplex oligonucleotide ligation assay. Animal
Genetics 31: 396-399. [1.4]
60. SEGERS, K.; VAIMAN, D.; BERGHMANS, S.; SHAY, T.; BEEVER, J.; COCKETT, N.;
GEORGES, M. & CHARLIER, C. (2001). Construction and characterization
of an ovine BAC contig spanning the callipyge locus. Animal Genetics 31:
352-359. [1.4] |corresponding author|
61. SHAY, T.; BERGHMANS, S.; SEGERS, K.; MEYERS, S.; WOMACK., J.; BEEVER, J.;
GEORGES, M.; CHARLIER, C. & COCKETT, N. E. (2001). Fine-mapping
and construction of a bovine contig spanning a 4.6 centimorgan interval
containing the CLPG locus. Mammalian Genome 12: 141-149. [2.7]
|corresponding author|
62. BERGHMANS, S.; SEGERS, K.; SHAY, T.; GEORGES, M.; COCKETT, N. E. &
CHARLIER, C. (2001). Breakpoint mapping positions the callipyge gene within
a 400 kilobase chromosome segment containing the Dlk1 and Gtl-2 genes.
Mammalian Genome 12: 183-185. [2.7] |corresponding author|
63. CHARLIER, C.; SEGERS, K.; KARIM, L.; SHAY, T.; GYAPAY, G.; COCKETT, N.;
GEORGES, M. (2001). The callipyge (CLPG) mutation enhances the expression
of the coregulated DLK1, GTL2, PEG11 and MEG8 genes in cis without
affecting their imprinting status. Nature Genetics 27:367-369. [26.5]
64. CHARLIER, C.; SEGERS, K.; WAGENAAR, D.; KARIM, L.; BERGHMANS, S.;
JAILLON, O.; SHAY, T.; WEISSENBACH, J.; COCKETT, N.; GYAPAY, G.;
GEORGES, M. (2001). Human - ovine comparative sequencing of a 250
kilobase imprinted domain encompassing the callipyge (clpg) gene and
identification of six imprinted transcripts: DLK1, DAT, GTL2, PEG11,
antiPEG11 and MEG8. Genome Research 11:850-862. [9.6]
65. PAILHOUX, E.; VIGIER, B.; VAIMAN, D.; SCHIBLER, L.; VAIMAN, A.; CRIBIU, E.;
NEZER, C.; GEORGES, M.; SUNDSTROM, J.; PELLINIEMI, L.J.; FELLOUS, M.;
COTINOT, C. (2001) Contribution of domestic animals to the identification
of new genes involved in sex determination. J. Exp. Zoology 290: 700-708.
[1.8]
Michel A. J. Georges 923
66. PAULSEN, M.; TAKADA, S.; YOUNGSON, N.A.; BENCHAIB, M.; CHARLIER, C.;
SEGERS, K.; GEORGES, M.; FERGUSON-SMITH, A. (2001) Detailed sequence
analysis of the imprinted Dlk1-Gtl2 locus in three mammalian species
identifies highly conserved genomic elements and a domain structure different
from the Igf2-H19 region. Genome Research 11: 2085-2094. [9.6]
67. BIDWELL, C.A.; SHAY, T.L.; GEORGES, M.; BEEVER, J.E.; BERGHMANS, S.;
COCKETT, N.E. (2001) Differential expression of the GTL2 gene within the
callipyge region of ovinechromosome 18. Animal Genetics 32:248-256. [1.4]
68. GRISART, B.; COPPIETERS, W.; FARNIR, F.; KARIM, L.; FORD, C.; CAMBISANO,
N.; MNI, M.; REID, S.; SPELMAN, R.; GEORGES, M. & SNELL, R. (2002).
Positional candidate cloning of a QTL in dairy cattle: Identification of a
missense mutation in the bovine DGAT gene with major effect on milk yield
and composition. Genome Research 12: 222-231. [9.6] |corresponding
author|
69. FARNIR, F.; GRISART, B.; COPPIETERS, W.; RIQUET, J.; BERZI, P.; CAMBISANO,
N.; KARIM, L.; MNI, M.; MOISIO, S.; SIMON, P.; WAGENAAR, D.; VILKKI, J.;
GEORGES, M. (2002). Simultaneous mining of linkage and linkage
disequilibrium to fine-map QTL in outbred half-sib pedigrees: revisiting the
location of a QTL with major effect on milk production on bovine chromosome
14. Genetics 161: 275-287. [4.9]
70. NEZER, C.; MOREAU, L.; WAGENAAR; D.; GEORGES, M. (2002). Results of a
whole genome scan targeting QTL for growth and carcass characteristics
in a Piétrain X Large White intercross. Genetics, Selection, Evolution 34:
371-388. [1.3]
71. AMARGER, V.; NGUYEN, M.; VAN LARE, A.S.; NEZER, C.; GEORGES, M.;
ANDERSSON, L. (2002). Comparative sequence analysis of the INS-IGF2-
H19 gene cluster in pigs. Mammalian Genome 13: 388-398. [2.7]
72. KIM, J.J.; GEORGES, M. (2002). Evaluation of a new fine-mapping method
exploiting linkage disequilibrium: a case study analysing a QTL with major
effect on milk composition on bovine chromosome 14. Asian-Aust. J. Anim.
Sci. 15: 1250-1256. [0.4]
73. SMIT, M.; SEGERS, K.; SHAY, T.; BARALDI, F.; GYAPAY, G.; SNOWDER, G.;
GEORGES, M.; COCKETT, N.; CHARLIER, C. (2003). Mosaicism of Solid Gold
supports the causality of a non-coding A to G transition in the determinism
of the callipyge phenotype. Genetics 163: 453-456. [4.9] |corresponding
author|
74. BLOTT, S.; KIM, J.-J.; MOISIO, S.; SCHMIDT-KÜNTZEL, A.; CORNET, A.;
BERZI, P.; CAMBISANO, N.; FORD, C.; GRISART, B.; JOHNSON, D.; KARIM, L.;
SIMON, P.; SNELL, R.; SPELMAN, R.; WONG, J.; VILKKI, J.; GEORGES, M.;
FARNIR, F.; COPPIETERS, W. (2003). Molecular dissection of a QTL: a
phenylalanine to tyrosine substitution in the transmembrane domain of the
924 Wolf Prize in Agriculture
84. DAVIS, E.; HARKEN JENSEN, C.; SCHRODER, H.D.; SHAY, T.; KLIEM, A.;
COCKETT, N.; GEORGES, M.; CHARLIER, C. (2004). Ectopic expression of
DLK1 protein in skeletal muscle of padumnal heterozygotes causes the
callipyge phenotype. Current Biology 14: 1858-1862. [11.9] |corresponding
author|
85. PIROTTIN, D.; GROBET, L.; ADAMANTIDIS, A.; FARNIR, F.; HERENS, C;
SCHRODER, H.D.; GEORGES, M. (2005). Transgenic engineering of a male-
specific muscular hypertrophy. Proc. Natl. Acad. Sci. USA 102: 6413-6418
[10.3]
86. DAVIS, E.; CAIMENT, F.; TORDOIR, X.; CAVAILLÉ, J.; FERGUSON-SMITH, A.;
COCKETT, N.; GEORGES, M.; CHARLIER, C. (2005). RNAi-mediated allelic
trans-interaction at the imprinted callipyge locus. Current Biology 15:
743-749 |corresponding author| [11.9]
87. SMIT, M.A., TORDOIR, X.; GYAPAY, G.; COCKETT, N.; GEORGES, M.;
CHARLIER, C. (2005). BEGAIN: a novel imprinted gene that generates
paternally expressed transcripts in a tissue- and promotor-specific manner.
Mammalian Genome 16: 801-814. |corresponding author| [2.4]
88. QUAN, X.; LAES, J.-F.; STIEBER, D.; RIVIÈRE, M.; RUSSO, J.; WEDEKIND, D.;
COPPIETERS, W.; FARNIR, F.; GEORGES, M.; SZPIRER, J.; SZPIRER, C. (2006)
Genetic identification of distinct loci controlling mammary tumor multiplicity,
latency and aggressiveness in the rat. Mammalian Genome 17: 310-321.
[2.4]
89. SANDOR, C.; FARNIR, F.; MEUWISSEN, T.; COPPIETERS, W.; GEORGES, M.
(2006). Linkage d.isequilibrium on the bovine X chromosome: characterization
-and use in QTL mapping. Genetics 173: 1777-1786. [4.3]
90. HARMEGNIES, N.; FARNIR, F.; DAVIN, F.; GEERTS, I.; BUYS, N.; GEORGES, M.;
COPPIETERS, W. (2006). Measuring the extent of linkage disequilibrium in
commercial pig populations. Animal Genetics 37: 225-231. |corresponding
author| [2.4]
91. TAKEDA, H.; CAIMENT, F.; SMIT, M.; HIARD, S.; TORDOIR, X.; COCKETT, N.;
GEORGES, M. & CHARLIER,5 C. (2006). The callipyge mutation enhances
bidirectional long-range DLK1-GTL2 intergenic transcription in cis. Proc.
Natl. Acad. Sci. USA 103: 8119-8124. |corresponding author| [10.2]
92. VIITALA, S., SZYDA, J., BLOTT, S., SCHULMAN, N., LIDAUER, M., MAKI-
TANILA, A., GEORGES, M., VILKKI, J. (2006) The role of the bovine growth
hormone receptor and prolactin receptor genes in milk, fat and protein
production in finnish ayrshire dairy cattle. Genetics 173:2151-2164. [4.3]
93. MORREEL, K., GOEMINNE, G., STORME, V., STERCK, L., RALPH, J.,
COPPIETERS, W., BREYNE, P., STEENACKERS, M., GEORGES, M., MESSENS,
E., BOERJAN, W. (2006) Genetical metabolomics of flavonoid biosynthesis in
Populus: a case study. Plant J. 47:224-37. [7.0]
926 Wolf Prize in Agriculture
94. CLOP, A.; MARCQ, F.; TAKEDA, H.; PIROTTIN, D.; TORDOIR, X.; BIBÉ, B.;
BOUIX, J.; CAIMENT, F.; ELSEN, J.M.; EYCHENNE, F.; LARZUL, C.; LAVILLE, E.;
MEISH, F.; MILENKOVIC, D.; TOBIN, J.; CHARLIER, C.; GEORGES, M.
(2006). A mutation creating a potential illegitimate miRNA target site in the
myostatin gene affects muscularity in sheep. Nature Genetics 38:813-818.
[25.8]
95. HARMEGNIES, N.; DAVIN, F.; DE SMET, S.; BUYS, N.; GEORGES, M.;
COPPIETERS, W. (2006). Results of a whole genome QTL scan for growth,
carcass composition, and meat quality in a porcine four-way cross. Animal
Genetics 37: 543-553 |corresponding author| [2.4]
96. WUNDERLICH, K.R.; ABBEY, C.A.; CLAYTON, D.R.; SONG, Y.; SCHEIN, J.E.;
GEORGES, M.; COPPIETERS, W.; ADELSON, D.L.; TAYLOR, J.F.; DAVIS, S.L.;
GILL, C.A. (2006) A 2.5-Mb contig constructed from Angus, Longhorn and
horned Hereford DNA spanning the polled interval on bovine chromosome
1. Anim Genet. 37:592-594. [2.4]
97. HAMELIN, M.; SAYD, T.; CHAMBON, C.; BOUIX, J.; BIBE, B.; MILENKOVIC, D.;
LEVEZIEL, H.; GEORGES, M.; CLOP, A.; MARINOVA, P.; LAVILLE, E. (2006)
Proteomic analysis of ovine muscle hypertrophy. J Anim Sci. 84:3266-3276.
[1.6]
98. LIBIOULLE, C.; LOUIS, E.; HANSOUL, S.; SANDOR, C.; FARNIR, F.;
FRANCHIMONT, D.; DE WIT, O.; DE VOS, M.; VERMEIRE, S.; DEMARCHE, B.;
GUT, I.; HEATH, S.; MNI, M.. ZELENIKA, D.; BELAICHE, J.; RUTGEERTS, P.;
LATHROP, M.; GEORGES, M. (2007) A novel susceptibility locus for Crohn’s
disease identified by whole genome association maps to a gene desert on
chromosome 5p13.1 and modulates the level of expression of the
prostaglandin receptor EP4. PloS Genetics 4:e58 [7.7]
99. GEORGES, M.; COPPIETERS, W.; CHARLIER, C. (2007) Polymorphic miRNA-
mediated gene regulation: contribution to phenotypic variation and disease.
Current Opinion in Genetics & Development 17: 1-11 [10]
100. GEORGES, M. (2007) Mapping, fine-mapping and cloning QTL in domestic
animals. Annual Review of Genomics and Human Genetics 8: 131-162.
[10.8]
101. CHARLIER, C.; COPPIETERS, W.; AGERHOLM, J.S.; CAMBISANO, N.; CARTA, E.;
DESMECHT, D.; DIVE,M.; FASQUELLE, C.; FRENNET, J.C.; HANSET, R.;
HUBIN, X.; JORGENSEN, C.; KARIM, L.; KENT, M.; HARVEY, K.; PEARCE, B.R.;
ROLLIN, F.; SIMON, P.; TAMA, N.; NIE, H.; VANDEPUTTE, S.; LIEN, S.; LONGERI,
M.; FREDHOLM, M.; HARVEY, R.J.; GEORGES, M. (2007) Highly effective
SNP-based association mapping and management of recessive defects in
livestock. Nature Genetics, in the press. [25.8]
Michel A. J. Georges 927
BOOK CHAPTERS
10. GEORGES, M.; CLOP, A.; MARCQ, F.; TAKEDA, H.; PIROTTIN, D.; HIARD, S.;
TORDOIR, X.; CAIMENT, F.; MEISH, F.; BIBE, B.; BOUIX, J.; ELSEN, J.M.;
EYCHENNE, F.; LAVILLE, E.; LARZUL, C.; MILENKOVIC, D.; TOBIN, J.;
CHARLIER, C. (2006) Polymorphic Polymorphic MicroRNA-Target Interactions:
A Novel Source of Phenotypic Variation. Cold Spring Harb Symp Quant Biol.
71:343-350.
CITATION ANALYSIS
Total impact factor: 819
Average impact factor per publication: 8.4
Total number of citations: 7,457
Average number of citations per publication: 51.08
h-index: 41 (41 publications with more than 40 citations)
16 publications with more than 100 citations.
SELECTED PUBLICATIONS
COCKETT, N.; JACKSON, S.; SHAW, T.; FARNIR, F.; SNOWDER, G.; NIELSEN, D.;
GEORGES, M. (1996). Polar overdominance at the ovine callipyge locus. Science
273:236-238.
GROBET, L.; ROYO MARTIN, L.J.; PONCELET, D.; PIROTTIN, D.; BROUWERS, B.;
RIQUET, J.; SCHOEBERLEIN, A.; DUNNER, S.; MENISSIER, F.; MASSABANDA, J.;
FRIES, R.; HANSET, R.; GEORGES, M. (1997) A deletion in the myostatin gene
causes double-muscling in cattle. Nature Genetics 17:71-74.
CHARLIER, C.; SEGERS, K.; KARIM, L.; SHAY, T.; GYAPAY, G.; COCKETT, N.;
GEORGES, M. (2001). The callipyge (CLPG) mutation enhances the expression
of the coregulated DLK1, GTL2, PEG11 and MEG8 genes in cis without affecting
their imprinting status. Nature Genetics 27:367-369.
CLOP, A..; MARCQ, F.; TAKEDA, H.; PIROTTIN, D.; TORDOIR, X.; BIBÉ, B.; BOUIX, J.;
CAIMENT, F.; ELSEN, J.M.; EYCHENNE, F.; LARZUL, C.; LAVILLE, E.; MEISH, F.;
MILENKOVIC, D.; TOBIN, J.; CHARLIER, C.; GEORGES, M. (2006). A mutation
creating a potential illegitimate miRNA target site in the myostatin gene affects
muscularity in sheep. Nature Genetics 38:813-818.
LIBIOULLE, C.; LOUIS, E.; HANSOUL, S.; SANDOR, C.; FARNIR, F.; FRANCHIMONT, D.;
DE WIT, O.; DE VOS, M.; VERMEIRE, S.; DEMARCHE, B.; GUT, I.; HEATH, S.;
MNI, M.; ZELENIKA, D.; BELAICHE, J.; RUTGEERTS, P.; LATHROP, M.;
GEORGES, M. (2007) A novel susceptibility locus for Crohn’s disease identified
by whole genome association maps to a gene desert on chromosome 5p13.1
and modulates the level of expression of the prostaglandin receptor EP4. PloS
Genetics 4:e58
CHARLIER, C.; COPPIETERS, W.; AGERHOLM, J.S.; CAMBISANO, N.; CARTA, E.;
DESMECHT, D.; DIVE, M.; FASQUELLE, C.; FRENNET, J.C.; HANSET, R.;
HUBIN, X.; JORGENSEN, C.; KARIM, L.; KENT, M.; HARVEY, K.; PEARCE, B.R.;
ROLLIN, F.; SIMON, P.; TAMA, N.; NIE, H.; VANDEPUTTE, S.; LIEN, S.;
LONGERI, M.; FREDHOLM, M.; HARVEY, R.J.; GEORGES, M. (2007) Highly
effective SNP-based association mapping and management of recessive defects
in livestock. Nature Genetics, in the press.
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Ronald L. Phillips
University of Minnesota
St. Paul, Minnesota, USA
k
CURRICULUM VITAE
Date and place of birth: January 1, 1940, Huntington County, Indiana, USA
EDUCATIONAL HISTORY:
1961 B.S. Purdue University – Major Crop Science.
1963 M.S. Purdue University – Major Plant Breeding and Genetics.
1966 Ph.D. University of Minnesota – Major Genetics-Cytogenetics and Plant
Breeding Specialties.
1967 Postdoctorate, Cornell University – Major Genetics.
PROFESSIONAL POSITIONS:
1966-67 NIH Trainee, Cornell University.
1967-68 Research Associate, University of Minnesota.
1968-72 Assistant Professor, University of Minnesota.
1972-76 Associate Professor, University of Minnesota.
1976-93 Professor, University of Minnesota.
931
932 Wolf Prize in Agriculture
SELECTED HIGHLIGHTS
Grants Management:
• USDA Grants Manager for the Genetic Mechanisms Program (1979) – 5 months
in Washington, D.C.
• USDA Chief Scientist in charge of the National Research Initiative (1996-98) –
50% time in Washington, D.C.
• Grants panels for USDA (1978-80), NSF (1976, 2002), DOE (1979, 1982),
USAID (Chair, 2002)
• Interagency Working Group (OSTP) on Plant Genome, Chair (1997-98). The
Working Group’s Report was prepared in response to Senator Kit Bond’s request
for a plan for Congress. The report resulted in establishment of the NSF Plant
Genome Research Program.
• Advisory Committee for University of Arizona NSF Chromatin Functional
Genomics Grant (2003-2005)
• Advisory Committee for Purdue University NSF Comparative Genomics Grant
(2004-present)
• Scientific Advisory Board, Chair, USDA RiceCAP grant (2004-present)
Special Offices:
• American Association for the Advancement of Science: Section “O” Chair
(1989-1990), Program Committee (2003-2005); Task Force on Women in
Agricultural Sciences (2006-present)
• Crop Science Society of America, President (1999-2000)
• National Academy of Sciences: Section Chair (2001-2003), Class Membership
Committee (1994-95, 2007)
• University of Minnesota Plant Molecular Genetics Institute, Co-founder and
Director (1991-94); Microbial and Plant Genomics Institute, Co-founder and
Director (2000-2005)
• International Crop Science Society, Vice President (2004-present)
• World Food Prize, Committee and Youth Institute Faculty (2004-present)
• Council of Scientific Society Presidents, Chair (2006)
Student Training:
• Advised 61 M.S. and Ph.D. student theses
• Advised 20 postdoctoral scientists
• Teaching: Graduate course in Plant Cytogenetics (1970-present)
• Cold Spring Harbor summer course in Plant Molecular and Developmental
Biology, Guest Instructor (1984, 1985, 1990, 1991)
• President’s Distinguished Faculty Mentoring Program (1988-1994, 2002-2005)
• Project Ag-Grad (African graduate student program), Founder and Co-director
(1986-present)
934 Wolf Prize in Agriculture
Symposium Organization:
• National Academy of Sciences Colloquium on “Protecting our food supply: The
value of plant genome initiatives”, Irvine, CA, Co-organizer (1997)
• University of Bologna, Italy, International Conference: “From the Green
Revolution to the Gene Revolution”, Co-organizer (2002-2005)
• University of Minnesota, National Symposium: “Intellectual Property for the
Public Good”, Organizer (2003-2004)
Publications:
• 142 Refereed Journal Articles
• 72 Chapters, or edited books
• 343 Abstracts
BRIEF BIOGRAPHY
Hallmarks of Dr. Phillips’ career involve innovative research in plant biology and
genetics leading to technology used in genetic engineering of crops, genomics, trait
selection, student education, and national/international service.
Dr. Phillips is Regents Professor and McKnight Presidential Chair in Genomics,
University of Minnesota. He earned B.S. and M.S. degrees from Purdue University
and a Ph.D. from the University of Minnesota; his postdoctoral training was at
Cornell University. Dr. Phillips has advised over 60 graduate theses and taught a
course in cytogenetics as a faculty member in the Department of Agronomy and
Plant Genetics for over 40 years. Dr. Phillips was the co-recipient of the prestigious
Wolf Prize in Agriculture in Israel in 2007 for “ground breaking research in
service of mankind.” In 1991, he was elected a member of the National Academy
of Sciences. Currently, he serves on the Board of Trustees of the premier
International Rice Research Institute in the Philippines and on the Scientific Advisory
Board of the Donald Danforth Plant Science Center. Other awards include an
Ronald L. Phillips 935
chromosome in plants. The high estimated number of copies of these genes per
cell was an unexpected result at the time.
His laboratory, along with postdoctoral scientist Ed Green, were the first to
regenerate complete corn plants from cells in tissue culture,18 thus providing in
1975 a key technology needed for the development of genetically engineered cereals,
a technology directly applied for corn and indirectly for other cereals. The use of
genetically modified corn with insect resistance, herbicide tolerance and other
traits has expanded tremendously around the world. The inbred line used in the
first successful regeneration of corn plants from cells is still widely used over
thirty years later. In addition, the corn cell line most widely used today in academia
and industry for genetic engineering was developed in Dr. Phillips’ laboratory by
Armstrong in the mid 1980s.
Green and Phillips developed a laboratory selection system for identifying cells
and/or plants high in essential amino acids,17 and lines have been obtained high in
threonine and methionine. The high methionine trait discovered in the Phillips lab
has been transferred to elite lines of maize and recently released to the public.
Variation induced during the tissue culture process was extensively
documented by Dr. Phillips and his students showing the occurrence of mutations,
chromosome breakage, DNA methylation alterations, and activation of transposable
elements.44, 46, 47, 48, 49, 51, 54, 56, 62, 69, 156, 157, 163, 164, 167, 168, 186, 189, 194
Events in the corn kernel endosperm were well described in the Phillips lab
especially with Kowles showing a dramatic but normal developmental increase in
DNA levels per cell.35, 36, 42, 55, 59, 81, 89, 105 Some cells were shown to contain nearly
200X the level of DNA in the basic chromosome complement. Because the
endosperm is 85% of the kernel, the increase in DNA leading to larger cells must
play a major role in corn yields.81
A major gene influencing flowering date in corn was identified by use of
molecular genetic markers.64 After considerable follow up in collaboration with
Tubersosa, Salvi, and DuPont/Pioneer scientists, the gene was isolated by map-
based cloning and subsequently shown, unexpectedly, to be a non-coding sequence
70kb upstream of a flowering gene.139 This gene caused a 10-day difference in
flowering in the material analyzed. Genetically engineered plants with this gene
showed expected changes in flowering time.
Dr. Phillips and his students have played a central role in the production of
useful molecular genetic marker maps for oats and wild rice and identified
chromosomal regions responsible for several important developmental, agronomic,
and nutritional traits.79, 97, 109, 110, 111, 115, 117, 125, 135, 156, 176, 179, 193, 203, 213
Phillips has presented widely-discussed ideas on how to interpret progress in
plant improvement programs with a restricted genetic base, emphasizing
de novo variation. The paper co-authored with Rasmusson85 has been the topic of
discussion in several courses.
Ronald L. Phillips 937
With colleagues, he has edited several books that advance the field of
plant genetics, including one on DNA-based Markers in Plants co-edited with
Vasil36, 178, 201 and, he has served on several prestigious editorial boards, including
the Proceedings of the National Academy of Sciences and Advances in Agronomy.
Together with Rines and several students and postdoctoral scientists, the Phillips
lab developed oat lines with individual corn chromosomes added (Oat-Maize
Addition Lines, or OMAs) that allow rapid and efficient mapping of corn
sequences to chromosome.83, 84, 114 All 10 corn chromosomes have been individually
transferred to oat by standard wide-hybridization followed by embryo rescue
techniques.113, 187 In addition to these lines being used extensively for mapping,
they have been used for chromosome sorting, centromere studies, mapping of gene
families, identification of chimeric BACs (Bacterial Artificial Chromosomes),
chromosome pairing observations, evolutionary studies, and study of C4 versus C3
photosynthesis. The OMAs were subsequently irradiated with gamma rays to break
apart the corn chromosome to form “Radiation Hybrids.”120, 207, 209 These lines
(approximately 600) allow a gene sequence to be located to a sub-region of the
chromosome.129
Recent isolation of a DNA sequence associated with corn centromeres96 has
formed the basis of developing a maize artificial chromosome by Ananiev and
colleagues at DuPont/Pioneer that should be useful for introducing multiple genes
in genetically engineered corn.
Together with Jacobs and Diez-Gonzalez, transgenic lines of corn have been
produced that possess genes coding for a toxin against the pathogenic
E. coli O157:H7136 which is the cause of many recalls of hamburger, spinach, and
other foods. The concept is that this corn feed will eliminate the pathogenic E. coli
strain from the intestinal tract of the cattle thus preventing contamination of
food.130
More recently the Phillips lab has been determining the genetic basis of a
high-oil corn that has an embryo which is 31% of the kernel and contains 50% oil
to give an overall oil level of 20%.
Professor Phillips’ national influence on plant science is notable. He chaired
the 1998 report to Congress from the President’s Office of Science and Technology
Policy that established the U.S. National Plant Genome Initiative of the NSF. He has
been a major advocate for science in agriculture as Chief Scientist for the U.S.
Department of Agriculture in charge of the National Research Initiative, as a
member of the National Academy of Sciences serving as Chair of his section, on a
National Research Council transgenic plants study commission, as Director of the
Center for Microbial and Plant Genomics (now the Microbial and Plant Genomics
Institute) with 85 faculty members and a new microbial and plant genomics
building (which he played a major role in securing), as scientific advisor to many
research institutes including the Donald Danforth Plant Science Center, the Noble
938 Wolf Prize in Agriculture
during the previous decade (1990s); the talk was videotaped and distributed in
both English and Spanish. At the 50th anniversary Crop Science Society of America
meeting in 2005, he was asked to present a major plenary talk on the development
of the science of genetics within the crop science context. His paper was entitled
“Genetic Tools from Nature and the Nature of Genetic Tools.”212 In 2007, Professor
Phillips was invited to be a centennial speaker in the 100th anniversaries of the
Department of Crop Science at Oregon State University and the Department of
Agronomy at Purdue. In addition to the book on cytogenetics used in his class,
Professor Phillips has co-edited four symposia books144, 145, 146, 147 and two editions
of the book on “DNA-Based Markers in Plants.”178, 201 He also has been a sub-
editor for an encyclopedia series on agricultural science.206
Professor Phillips has assisted in the organization of several national and
international meetings. One was an international congress held at the University
of Bologna in Italy: “In the wake of the double helix; from the green revolution to
the gene revolution,” which was attended by 250 scientists from 35 countries.
Eight papers from the meeting were published in the Proceedings of the National
Academy of Sciences, including one from the Phillips lab.209 A book and CD of
review papers have been published from the meeting.210 In conjunction with that
meeting, the U.S. Department of State asked Professor Phillips to present two
seminars at different universities and a press conference on biotechnology in Italy,
a country not generally sympathetic toward genetically modified crops. He also
was invited to present a talk entitled “Are GMOs Safe?” at a meeting of AAAS
(American Association for the Advancement of Science). He recently served on the
Program Committee of AAAS, which orchestrates the conference program for 10,000
attendees (including 1,000 journalists). The 2007 home page for the AAAS Annual
Meeting featured a picture of Professor Phillips along with quotes about the meeting
and a biographical sketch.
One of the recent symposia organized and chaired by Professor Phillips was
the University of Minnesota President’s 21st Century Interdisciplinary Conference
on “Intellectual Property Rights for the Public Good: Obligations of U.S. Universities
to Developing Countries,” co-sponsored by the Center for Microbial and Plant
Genomics (now the Microbial & Plant Genomics Institute) and the Consortium on
Law and Values in Health, Environment & the Life Sciences. The first issue of a
new journal entitled Minnesota Journal of Law, Science and Technology (edited by
students and faculty) was devoted to papers from this meeting, including an
introductory paper by Professor Phillips.132 He now serves on the Editorial Advisory
Board of the Minnesota Journal of Law, Science & Technology. The suggestion for a
“humanitarian use clause” to be included in university intellectual property policies
was a major outcome of the meeting. Such a clause would facilitate the transfer of
technology to the developing world. Professor Phillips and three colleagues in the
U of M Law School wrote a summary of the symposium published in The Scientist
(April 29, 2004).
Ronald L. Phillips 941
4. Phillips, R.L. and W.F. Keim. 1968. Seed pod dehiscence in Lotus and
interspecific hybridization involving L. corniculatus L. Crop Sci. 8:18-21.
5. Phillips, R.L. 1968. Cyanogenesis in Lotus species. Crop Sci. 8:123-124.
6. Phillips, R.L. 1968. Cytogenetic location of the gene for the purple plant
color in maize. J. Hered. 59:56.
7. Phillips, R.L. 1969. Recombination in Zea mays L. I. Location of genes and
interchanges in chromsomes 5, 6 and 7. Genetics 61:107-116.
8. Phillips, R.L. 1969. Recombination in Zea mays L. II. Cytogenetic studies of
recombination in reciprocal crosses. Genetics 61:117-127.
9. Phillips, R.L., C.R. Burnham, and E.B. Patterson. 1971. Advantages of
chromosomal interchanges that generate haplo-viable deficiency-duplications.
Crop Sci. 11:525-528.
10. Phillips, R.L., R.A. Kleese, and S.S. Wang. 1971. The nucleolus organizer
region of maize (Zea mays L.): Chromosomal site of DNA complementary to
ribosomal RNA. Chromsoma (Berl.) 36:79-88.
11. Kleese, R.A. and R.L. Phillips. 1972. Electrophoretic mutants as useful markers
for chromosome aberrations. Genetics 72:537-540.
12. Burnham, C.R., J.T. Stout, W.H. Weinheimer, R.V. Kowles, and R.L. Phillips.
1972. Chromosome pairing in maize. Genetics 71:111-126.
13. Burnham, C.R., and R.L. Phillips. 1972. Linkage Groups: Plants. Part IV, Corn.
Contribution to Biology Data Book Second Edition, Vol. 1, pp. 85-88. Fed.
Amer. Socs. Exptl. Biol.
14. Stout, J.T., and R.L. Phillips. 1973. Two independently inherited electrophoretic
variants of the lysine-rich histones of maize. Proc. Nat. Acad. Sci. U.S.A.
70:3043-3047.
15. Phillips, R.L., D.F. Weber, R.A. Kleese, and S.S. Wang. 1974. The nucleolus
organizer region of maize (Zea mays L.): Tests for ribosomal gene
compensation or magnification. Genetics 77:285-297.
16. Green, C.E., R.L. Phillips, and R.A. Kleese. 1974. Tissue cultures of maize
(Zea mays L.): Initiation, maintenance, and organic growth factors. Crop Sci.
14:54-58.
17. Green, C.E., and R.L. Phillips. 1974. Potential selection system for mutants
with increased lysine, threonine, and methionine in cereal crops. Crop Sci.
14:827-831.
18. Green, C.E., and R.L. Phillips. 1975. Plant regeneration from tissue culture of
maize. Crop Sci. 15:417-421.
19. Givens, J.F., and R.L. Phillips. 1976. The nucleolus organizer region of maize
(Zea mays L.): Ribosomal RNA gene distribution and nucleolar interactions.
Chromosoma (Berl.) 57:103-117.
20. Liang, G.H., A.S. Wang, and R.L. Phillips. 1977. Control of ribosomal RNA
gene multiplicity in wheat. Can J. Genet. Cytol. 19:425-435.
946 Wolf Prize in Agriculture
21. Phillips, R.L. 1977. Genetic engineering for crop improvement. 32nd Ann.
Corn and Sorghum Res. Conf. pp. 6-20.
22. Phillips, R.L., A.S. Wang, I. Rubenstein, and W.D. Park. 1979. Hybridization
of ribosomal RNA to maize chromosomes. Maydica 24:7-21.
23. Phillips, R.L., E.D. Garber, O.L. Miller, Jr., and H.H. Kramer. 1979. C. R.
Burnham. Maydica 24:3-6.
24. Albertsen, M.C., and R.L. Phillips. 1981. Developmental cytology of 13 genetic
male sterile loci in maize. Can. J. Genet. Cytol. 23:195-208.
25. Phillips, R.L., P.R. Morris, F. Wold, and B.G. Gengenbach. 1981. Seedling
screening for lysine-plus-threonine resistant maize. Crop Sci. 21:601-607.
26. Mascia, P.H., I. Rubenstein, R.L. Phillips, A.S. Wang, and Lu Zhen Xiang.
1981. Localization of the 5S rRNA genes and evidence for diversity in the 5S
rDNA region of maize. Gene 15:7-20.
27. Burnham, C.R., R.L. Phillips, and M.C. Albertsen. 1981. Inheritance of male
sterility in flax involving nuclear-cytoplasmic interaction, including methods
of testing for cytoplas-mic restoration. Crop Sci. 21:659-663.
28. McCoy, T.J., R.L. Phillips, and H.W. Rines. 1982. Cytogenetic analysis of
plants regenerated from oat (Avena sativa) tissue cultures; high frequency of
partial chromosome loss. Can J. Genet. Cytol. 24:37-50.
29. McMullen, M.S., R.L. Phillips, and D.D. Stuthman. 1982. Meiotic irregularities
in Avena sativa L./A. sterilis L. hybrids and breeding implications. Crop Sci.
22:890-897.
30. McCoy, T.J., and R.L. Phillips. 1982. Chromosome stability in maize (Zea
mays) tissue cultures and sectoring in some regenerated plants. Can. J. Genet.
Cytol. 24:559-565.
31. Phillips, R.L., A.S. Wang, and R.V. Kowles. 1983. Molecular and developmental
cytogenetics of gene multiplicity in maize. Stadler Symp. 15:105-118.
32. Buescher, P.J., R.L. Phillips, and R. Brambl. 1984. Ribosomal RNA contents of
maize genotypes with different ribosomal gene numbers. Biochem. Genet.
22:923-930.
33. Luby, J.J., D.D. Stuthman, and R.L. Phillips. 1985. Micronuclei frequency and
character coherence in Avena sativa L./A. fatua crosses. Theor. Appl. Genet.
69:367-373.
34. Phillips, R.L., and B.A. McClure. 1985. Elevated proteinbound methionine
in seeds of a lysine-plus-threonine resistant maize line. Cereal Chem.
62:213-218.
35. Kowles, R.V., and R.L. Phillips. 1985. DNA amplification patterns in maize
endosperm nuclei during kernel development. Proc. Natl. Acad. Sci. (USA)
82:7010-7014.
36. Phillips, R.L., R.V. Kowles, M.D. McMullen, S. Enomoto, and I. Rubenstein.
1985. Developmentally-timed changes in maize endosperm DNA. UCLA Symp.
Plant Genetics Alan R. Liss, Inc., pp. 739-754.
Ronald L. Phillips 947
37. McMullen, M.D., B. Hunter, R.L. Phillips, and I. Rubenstein. 1986. The
structure of the maize ribosomal DNA spacer region. Nucl. Acids Res.
14:4953-4968.
38. Rhodes, C.A., R.L. Phillips, and C.E. Green. 1986. Cytogenetic stability of
aneuploid maize tissue cultures. Can. J. Genet. Cytol. 28:374-384.
39. Rhodes, C.A., C.E. Green, and R.L. Phillips. 1986. Factors affecting tissue
culture initiation from maize tassels. Plant Sci. 46:225-232.
40. Tuberosa, R., and R.L. Phillips. 1986. Isolation of methotrexatetolerant cell
lines of corn. Maydica 31:215-225.
41. Wang, A.S., R.L. Phillips, and C.C. Mi. 1986. Cell cycle parameters and
accumulation of metaphase cells in maize suspension cultures. Plant Sci.
46:53-61.
42. Kowles, R.V., M.D. McMullen, and R.L. Phillips. 1986. Gene expression in
developing kernels. First Ann. Penn. State Symp. in Plant Physiol., University
Park, Penn.
43. Tuberosa, R., and R.L. Phillips. 1986. Selection and partial characterization
of corn (Zea mays L.) callus lines tolerant to methotrexate. Genetica Agraria
40:479.
44. Lee, M., and R.L. Phillips. 1987. Genomic rearrangements in maize induced
by tissue culture. Genome 29:122-128.
45. Johnson, S.S., R.L. Phillips, and H.W. Rines. 1987. Meiotic behavior in pro-
geny of tissue culture regenerated oat plants (Avena sativa L.) carrying
neartelocentric chromosomes. Genome 29:431-438.
46. Johnson, S.S., R.L. Phillips, and H.W. Rines. 1987. Possible role of hetero-
chromatin in chromosome breakage induced by tissue culture in oats (Avena
sativa L.). Genome 29:439-446.
47. Peschke, V.M., R.L. Phillips, and B.G. Gengenbach. 1987. Discovery of
transposable element activity among progeny of tissue culturederived maize
plants. Science 238:804-807.
48. Lee, M. and R.L. Phillips. 1987. Genetic variants in progeny of regenerated
maize plants. Genome 29:834-838.
49. Armstrong, C.L. and R.L. Phillips. 1988. Genetic and cytogenetic variation
in plants regenerated from organogenic and friable, embryogenic tissue
cultures of maize. Crop Sci. 28:363-369.
50. Lee, M., J.L. Geadelmann, and R.L. Phillips. 1988. Agronomic evaluation
of inbred lines derived from tissue cultures of maize. Theor. Appl. Genet.
75:841-849.
51. Benzion, G., and R.L. Phillips. 1988. Cytogenetic stability of maize tissue
cultures: A cell line pedigree analysis. Genome 30:318-325.
52. Stadler, J., R. Phillips, and M. Leonard. 1989. Mitotic blocking agents for
suspension cultures of maize ‘Black Mexican Sweet’ cell lines. Genome
32:475-478.
948 Wolf Prize in Agriculture
53. Benner, M.S., R.L. Phillips, J.A. Kirihara, and J.W. Messing. 1989. Genetic
analysis of methionine-rich storage protein accumulation in maize. Theor.
Appl. Genet. 78:761-767.
54. Phillips, R.L. 1989. Somaclonal and gametoclonal variation. Genome
31:1119-1120.
55. Kowles, R.V., F. Srienc, and R.L. Phillips. 1990. Endoreduplication of nuclear
DNA in the developing maize endosperm. Devel. Genet. 11:125-132.
56. Peschke, V.M. and R.L. Phillips. 1991. Activation of the maize transposable
element Suppressor-mutator (Spm) in tissue culture. Theor. Appl. Genet.
81:90-97.
57. Peschke, V.M., R.L. Phillips, and B.G. Gengenbach. 1991. Genetic and
molecular analysis of tissue culture-derived Ac elements. Theor. Appl. Genet.
82:121-129.
58. Sullivan, T.D., L.I. Strelow, C.A. Illingworth, R.L. Phillips, and O.E. Nelson, Jr.
1991. Analysis of maize Brittle-1 alleles and a defective Suppressor-Mutator-
induced mutable allele. Plant Cell 3:1337-1348.
59. Kowles, R.V., M.D. McMullen, G. Yerk, R.L. Phillips, S. Kraemer, and
F. Srienc. 1992. Endosperm mitotic activity and endoreduplication in maize
affected by defective kernel mutations. Genome 35:68-77.
60. Phillips, R.L. 1993. Plant genetics: out with the old, in with the new? Am. J.
Clinical Nutrition 58 (Suppl.):2595-2635.
61. Phillips, R.L. 1993. Cytogenetic manipulation of polymitotic (po). Maydica
38:85-92.
62. Kaeppler, S.M., and R.L. Phillips. 1993. DNA methylation and tissue culture-
induced variation in plants. In Vitro Cell. Dev. Biol. 29P:125-130.
63. Stapleton, A.E. and R.L. Phillips. 1993. A fertile field: Maize genetics ’93.
Plant Cell 5:723-727.
64. Phillips, R.L., T.S. Kim, S.M. Kaeppler, S.N. Parentoni, D.L. Shaver, R.E. Stucker,
and S.J. Openshaw. 1993. Genetic dissection of maturity using RFLPs. Proc.
47th Annual Corn & Sorghum Industry Res. Conf. pp. 135-150.
65. Muenchrath, D.A. and R.L. Phillips. 1993. Relationship of maize seedling
response to lysine-plus-threonine medium and whole kernel amino acid
profile. Crop Sci. 33:1095-1099.
66. Kaeppler, S.M., R.L. Phillips, and T.S. Kim. 1993. Use of near-isogenic lines
derived by backcrossing or selfing to map qualitative traits. Theor. Appl.
Genet. 87:233-237.
67. Jellen, E.N., R.L. Phillips, and H.W. Rines. 1993. C-banded karyotypes and
polymorphisms in hexaploid oat accessions (Avena spp.) using Wrights’ stain.
Genome 36:1129-1137.
68. Jellen, E.N., W.L. Rooney, R.L. Phillips, and H.W. Rines. 1993. Characterization
of the hexaploid oat Avena byzantina cv. Kanota monosomic series using
C-banding and RFLPs. Genome 36:962-970.
Ronald L. Phillips 949
69. Kaeppler, S.M. and R.L. Phillips. 1993. Tissue culture-induced DNA
methylation variation in maize. Proc. Natl. Acad. Sci. USA 90:8773-8776.
70. Kim, T.S., R.L. Phillips, and M.Y. Eun. 1993. Identification of genomic regions
controlling maturity in maize (Zea mays L.). Crop Production and
Improvement Technology in Asia. pp. 379-397. Korean Soc. Crop Sci.
71. Jellen, E.N., R.L. Phillips, and H.W. Rines. 1994. Chromosomal localization and
polymorphisms of ribosomal DNA in oat (Avena spp.). Genome 37:23-32.
72. Phillips, R.L. and E.N. Jellen. 1994. Videotaped lectures in a graduate
cytogenetics course. J. Nat. Res. Life Sci. Educ. 23:16-18.
73. Phillips, R.L., S.M. Kaeppler, and P. Olhoft. 1994. Genetic instability of plant
tissue cultures: Breakdown of normal controls. Proc. Natl. Acad. Sci. USA
91:5222-5226.
74. Young, N.D. and R.L. Phillips. 1994. Cloning plant genes known only by
phenotype. Plant Cell 6:1193-1195.
75. Phillips, R.L., H.W. Rines, E.N. Jellen, W.L. Rooney, B.C. Wu, S. Kianian, and
O. Riera-Lizarazu. 1994. Oat genome analysis via molecular markers and
oat x corn crosses. U.S.-Japan Joint Seminar, Kansas State University.
76. Rooney, W.L., H.W. Rines, and R.L. Phillips. 1994. Identification of RFLP
markers linked to crown rust resistance genes Pc91 and Pc92 in oat. Crop
Sci. 34:940-944.
77. Rooney, W.L., E.N. Jellen, R.L. Phillips, H.W. Rines, and S.F. Kianian. 1994.
Identification of homoeologous chromosomes in hexaploid oat (A. byzantina
cv Kanota) using monosomics and RFLP analysis. Theor. Appl. Genet.
89:329-335.
78. Jellen, E.N., R.L. Phillips, H.W. Rines, and W.L. Rooney. 1995. Molecular
genetic identification of Avena chromosomes related to the group 1
chromosomes of the Triticeae. Genome 38:185-189.
79. O’Donoughue, L.S., S.F. Kianian, P.J. Rayapati, G.A. Penner, M.E. Sorrells, S.D.
Tanksley, R.L. Phillips, H.W. Rines, M. Lee, G. Fedak, S.J. Molnar,
D. Hoffman, C.A. Salas, B. Wu, E. Autrique, and A. Van Deynze. 1995. A
molecular linkage map of cultivated oat. Genome 38:368-380.
80. Phillips, R.L., M.A. Matzke, and K. Oono. 1995. Treasure your exceptions.
Plant Cell 7:1522-1527.
81. Schweizer, L., G.L. Yerk-Davis, R.L. Phillips, F. Srienc, and R.J. Jones. 1995.
Dynamics of maize endosperm development and DNA endoreduplication.
Proc. Natl. Acad. Sci. USA 92:7070-7074.
82. Kianian, S.F., D.C. Bittel, R.L. Phillips, and M. Gallo-Meagher. 1996.
Postodoctoral research associates as an educational resource: opportunities
and constraints. J. Nat. Resources, Life Sciences & Education 25:144-147.
83. Riera-Lizarazu, O., H.W. Rines, and R.L. Phillips. 1996. Cytological and
molecular characterization of oat x maize partial hybrids. Theor. Appl. Genet.
93:123-135.
950 Wolf Prize in Agriculture
84. Ananiev, E.V., O. Riera-Lizarazu, H.W. Rines, and R.L. Phillips. 1997. Oat-
maize chromosome addition lines: A new system for mapping the maize
genome. Proc. Natl. Acad. Sci. USA 94:3524-3529.
85. Rasmusson, D.C. and R.L. Phillips. 1997. Plant breeding progress and
genetic diversity from de novo variation and elevated epistasis. Crop Sci
37:303-310.
86. Martin, M., R. Phillips, and G. Gardner. 1997. Conducting and administering
biotechnology research at a land-grant university. J. Agric. and Human Values.
87. Phillips, R.L. and J. Suresh. 1997. Cytogenetic basis for fertile plants in a
maize nuclear male sterility system. Maydica 42:127-131.
88. Kianian, S.F., B-C. Wu, S.L. Fox, H.W. Rines, and R.L. Phillips. 1997. Aneuploid
marker assignment in hexaploid oat with the C genome as a reference for
determining remnant homoeology. Genome 40:386-396.
89. Kowles, R.V., G.L. Yerk, K.M. Haas, and R.L. Phillips. 1997. Maternal effects
influencing DNA endoreduplication in developing endosperm of Zea mays.
Genome 40:798-805.
90. Milach, S.C.K., H.W. Rines, and R.L. Phillips. 1997. Molecular genetic mapping
of dwarfing genes in oat. Theor. Appl. Genet. 95:783-790.
91. Jellen, E.N., H.W. Rines, S.L. Fox, D.W. Davis. R.L. Phillips, and B.S. Gill. 1997.
Characterization of “Sun II” oat monosomics through C-banding and
identification of eight new “Sun II” monosomics. Theor. Appl. Genet.
95:1190-1195.
92. Milach, S.C.K., H.W. Rines, R.L. Phillips, D.D. Stuthman, and J. Morikawa.
1998. Inheritance of a new dwarfing gene in oat. Crop Sci. 38:356-360.
93. Phillips, R.L., and M. Freeling, 1998. Plant genomics and our food supply: An
introduction. Proc. Natl. Acad. Sci. USA 95: 1969-1970.
94. Editor of 15 papers in 1998 Proc. Natl. Acad. Sci. USA 95: 1971-2032:
National Academy of Sciences Colloquium on “Protecting our Food Supply:
The Value of Plant Genome Initiatives.”
95. Chen, Z.J., R.L. Phillips, and H.W. Rines, 1998. Maize DNA enrichment by
representational difference analysis. Theor. Appl. Genet. 97:337-344.
96. Ananiev, E.V., R.L. Phillips, and H.W. Rines, 1998. Chromosome-specific
molecular organization of maize (Zea mays L.) centromeric regions. Proc.
Natl. Acad. Sci. USA 95:13073-13078.
97. Kianian, S.F., M.A. Egli, R.L. Phillips, H.W. Rines, D.A. Somers, B.G. Gengenbach,
F.H. Webster, S.M. Livingston, S. Groh, L.O. O’Donoughue, M.E. Sorrells,
D.M. Wesenberg, D.D. Stuthman, and R.G. Fulcher, 1999. Association of a
major groat oil content QTL and an acetyl-CoA carboxylase gene in oat.
Theor. Appl. Genet. 98:884-894.
98. Ananiev, E.V., R.L. Phillips, and H.W. Rines, 1998. Complex structure of
knob DNA on maize chromosome 9: Retrotransposon invasion into
heterochromatin. Genetics 149: 2025-2037.
Ronald L. Phillips 951
99. Ananiev, E.V., R.L. Phillips, and H.W. Rines, 1998. A knob-associated tandem
repeat in maize capable of forming fold-back DNA segments: Are chromosome
knobs megatransposons? Proc. Natl. Acad. Sci. USA 95: 10785-10790.
100. Kennard, W., R. Phillips, R. Porter, and A. Grombacher. 1999. A comparative
map of wild rice (Zigania palustris L. 2n=2x=30). Theor. Appl. Genet.
99:793-799.
101. DeKoeyer, D.L., R.L. Phillips, and D.D. Stuthman. 1999. Changes in genetic
diversity during seven cycles of recurrent selection for grain yield in oat,
Avena sativa L. Plant Breeding 118:37-43.
102. Vladutu, C., J. McLaughlin, and R.L. Phillips. 1999. Fine mapping and
characterization of linked quantitative trait loci involved in the transition of
the maize apical meristem from vegetative to generative structures. Genetics
153:993-1007.
103. Bass, H.W., O. Riera-Lizarazu, E.V. Ananiev, S.J. Bordoli, H.W. Rines, R.L.
Phillips, J.W. Sedat, D.A. Agard, and W.Z. Cande. 2000. Evidence for the
coincident initiation of homolog pairing and synapsis during the telomere-
clustering (bouquet) stage of meiotic prophase. J. Cell Sci. 113:1033-1042.
104. Riera-Lizarazu, O., M.I. Vales, E.V. Ananiev, H.W. Rines, and R.L. Phillips.
2000. Production and characterization of maize-chromosome 9 radiation
hybrids derived from an oat-maize addition line. Genetics 156:327-339.
105. Engelen-Eigles, G., R.J. Jones, and R.L. Phillips. 2000. DNA endore-duplication
in maize endosperm cells: The effect of exposure to short-term high
temperature. Plant, Cell and Environment 23:657-663.
106. Cao, X., N.M. Springer, M.G. Muszynski, R.L. Phillips, S. Kaeppler, and
S.E. Jacobsen. 2000. Conserved plant genes with similarity to mammalian de
novo DNA methyltransferases. Proc. Natl. Acad. Sci. USA 97:4979-4984.
107. Ananiev, E.V., R.L. Phillips, and H.W. Rines. 2000. Complex structure
of knobs and centromeric regions in maize chromosomes. Tsitol Genet.
34:11-15.
108. Muehlbauer, G.J., O. Riera-Lizarazu, R.G. Kynast, D. Martin, R.L. Phillips, and
H.W. Rines. 2000. A maize-chromosome 3 addition line of oat exhibits
expression of the maize homeobox gene liguleless3 and alterations of cell
fates. Genome 43:1055-1064.
109. Kennard, W., R. Phillips, R. Porter, and A. Grombacher. 2000. A comparative
map of wild rice (Zizania palustris L. 2n=2x=30). Theor. Appl. Genet.
101:677-684.
110. Kianian, S.F., R.L. Phillips, H.W. Rines, R.G. Fulcher, F.H. Webster, and
D.D. Stuthman. 2000. Quantitative trait loci influencing β glucan content in
oat (Avena sativa, 2n=6x=42). Theor. Appl. Genet. 101:1039-1048.
111. Groh, S., A. Zacharias, S.F. Kianian, G.A. Penner, J. Chong, H.W. Rines,
and R.L. Phillips. 2001. Comparative AFLP mapping in two hexaploid oat
populations. Theor. Appl. Genet. 102:876-884.
952 Wolf Prize in Agriculture
124. Chen, G., H. Rines, K. Leonard, D. Stuthman, R. Phillips, and S. Ding. 2002.
Identification of QTLs for horizontal resistance to crown rust in oat. J. Zhejiang
Univ. (Agri. & Life Sci.) 27:151-155.
125. Wright, C.P., N.A. Tinker, S.F. Kianian, M.E. Sorrells, L.S. O’Donoughue,
D.L. Hoffman, S. Groh, G.J. Scoles, C.D. Li, F.H. Webster, R.L. Phillips,
H.W. Rines, S.M. Livingston, K.C. Armstrong, G. Fedak, and S.J. Molnar. 2003.
A molecular map in Kanota x Ogle hexaploid oat (Avena sp.) enhanced by
additional markers and a robust framework. Genome 46: 28-47.
126. Hass, B.L., J.C. Pires, R. Porter, R.L. Phillips, and S.A. Jackson. 2003.
Comparative genetics at the gene and chromosome levels between rice (Oryza
sativa) and wild rice (Zizania palustris). Theor. Appl. Genet. 107:773-782.
127. Salvi, S., M. Morgante, K. Fengler, R. Meeley, E. Ananiev, S. Svitashev,
E. Bruggemann, X. Niu, B. Li, S.V. Tingey, et al. 2003. Proc. of 57th Corn and
Sorghum Conference.
128. Okagaki, R.J., and R.L. Phillips. 2004. Maize DNA-sequencing strategies and
genome organization. Genome Biol. Vol. 5, Issue 5, Article 223, pp. 1-3.
129. Kynast, R.G., R.J. Okagaki, M.W. Galatowitsch, S.R. Granath, M.S. Jacobs,
A.O. Stec, H.W. Rines, and R.L. Phillips. 2004. Dissecting the maize genome
by using chromosome addition and radiation hybrid lines. Proc. Natl. Acad.
Sci. USA 101: 9921-9926. (cover picture).
130. Schamberger, Gerry P., Ronald L. Phillips, Jennifer L. Jacobs, and Francisco
Diez-Gonzalez. 2004. Reduction of Escherichia coli 0157:H7 populations in
cattle by addition of colicin E7-producing E. coli to feed. Appl. Environ.
Microbiol. 70:6053-6060.
131. Vales, M.I., O. Riera-Lizarazu, H.W. Rines, and R.L. Phillips. 2004.
Transmission of maize chromosome 9 rearrangements in oat-maize radiation
hybrids. Genome 47:1202-1210.
132. Phillips, R.L. 2004. Intellectual property rights for the public good: Obligations
of U.S. universities to developing countries. MN J. Law, Sci. & Tech. 6:177-185.
133. Buhinicek, I., I. Pejic, J. Suresh, A. Vragolovic, and R. L. Phillips. 2004. Genetic
divergence of elite maize inbred lines comparing to Illinois high oil source.
Austrian J. Agric. Res. 55:29-35.
134. Dinu, I., R.J. Hayes, R. Kynast, R.L. Phillips and C.A. Thill. 2005. Novel inter-
series hybrids in Solanum, section Petota. Theor. Appl. Genet., 110:403-415.
135. Portyanko, V.A., G. Chen, H.W. Rines, R.L. Phillips, K.J. Leonard, G.E. Ochochi,
and D.D. Stuthman. 2005. Quantitative trait loci for partial resistance to
crown rust, Puccinia coronata, in cultivated oat, Avena sativa L. Theor. Appl.
Genet. 111:313-324.
136. Jacobs, J.L., F. Diez Gonzalez, M.D. Stern, and R.L. Phillips. 2005. Detection
of transgenic maize Cry1Ab protein subjected to ruminal digestion. J. Am.
Feed Sci. 14:655-664.
954 Wolf Prize in Agriculture
137. Odland, W., A. Baumgarten, and R. Phillips. 2006. Ancestral rice blocks
define multiple related regions in the maize genome. The Plant Genome
1:S41-S48 (Supplement to Crop Sci. 46).
138. Baumgarten, A.M., J. Suresh, G. May, and R.L. Phillips. 2007. Mapping QTLs
contributing to Ustilago maydis resistance in specific plant tissues of maize.
Theor. Appl. Genet. 114:1229-1238.
139. Salvi, S., G. Sponza, M. Morgante, D. Tomes, X. Niu, K.A. Fengler, R. Meeley,
E.V. Ananiev, S. Svitashev, E. Bruggemann, B. Li, C.F. Hainey, S. Radovic,
G. Zaina, J.-A. Rafalski, S.V. Tingey, G.H. Miso, R.L. Phillips, and R. Tuberosa.
2007. Conserved noncoding genomic sequences associated with a
flowering-time quantitative trait locus in maize. Proc. Natl. Acad. Sci. USA
104:11376-11381.
140. Okagaki, R.J., M.S. Jacobs, A.O. Stec, R.G. Kynast, E. Buescher, H.W. Rines,
M.I. Vales, O. Riera-Lizarazu, M. Schneerman, G. Doyle, K.L. Friedman,
R.W. Staub, D.F. Weber, T.L. Kamps, I.F.E. Amarillo, C.D. Chase, and
R.L. Phillips. 2008. Maize centromere mapping: A comparison of physical
and genetic strategies. J. Hered. 92:85-93.
141. Makarevitch, I., R.L. Phillips, and N.M. Springer. 2008. Profiling expression
changes caused by a segmental aneuploid in maize. BMC Genomics 9:7.
142. Phillips, R.L., N. Magor, D. Shires, H. Leung, S. McCouch, and D. McIntosh.
2008. Student Opportunity: short-term exposure to international agriculture.
Rice 1: (in press).
143. Phillips, R.L., and C.R. Burnham, Eds. Cytogenetics. In Benchmark Papers in
Genetics Series. Dowden, Hutchinson, and Ross, Stroudsburg, PA 489 pp.
(1977).
144. Rubenstein, I., R.L. Phillips, C.E. Green, and R.J. Desnick, Eds. Molecular
Genetic Modification of Eucaryotes. Academic Press. 171 pp. (1977).
145. Rubenstein, I., R.L. Phillips, C.E. Green, and B.G. Gengenbach, Eds. Molecular
Biology of Plants. Academic Press. 343 pp. (1979)
146. Rubenstein, I., R.L. Phillips, C.E. Green, and B.G. Gengenbach, Eds. The Plant
Seed: Development, Preservation, and Germination. Academic Press. 266 pp.
(1979).
147. Rubenstein, I., B.G. Gengenbach, R.L. Phillips, and C.E. Green, Eds. Genetic
Improvement of Crops: Emergent Techniques. University of Minnesota Press.
232 pp. (1980).
148. Phillips, R.L. Molecular cytogenetics of the nucleolus organizer region. In
Maize Breeding and Genetics, Chapter 43, John Wiley & Sons, NY (1978).
149. Phillips, R.L. Plant Cytogenetics. In Staining Procedures, 4th Edition, Chapter
8. Williams and Wilkins Co., Baltimore. (pp. 341-359). 1981.
Ronald L. Phillips 955
150. Phillips, R.L. Pollen and pollen tubes. In Staining Procedures, 4th Edition,
Chapter 9. Williams and Wilkins Co., Baltimore. (pp. 361-366). 1981.
151. Phillips, R.L., and A.S. Wang. 1982. In situ hybridization with maize meiotic
cells. In Maize for Biological Research. W.F. Sheridan, Ed. Plant Molec. Biol.
Assoc., Charlottesville, VA.
152. Bashaw, E.L., J. James, J.R. Laughnan, and R.L. Phillips. Chromosomal and
cytoplasmic manipulations: Discussion panel. In Plant Breeding II. K. J. Frey,
Ed. The Iowa State University Press, Ames, IA pp. 138-146.
153. Phillips, R.L. 1983. Genetic engineering of plants: some perspectives on the
conference, the present, and the future. In Genetic Engineering of Plants,
C. Meredith and T. Kosuge, Eds., Plenum Press.
154. Phillips, R.L., and A.S. Wang. 1984. Chromosome analysis. In Cell Culture
and Somatic Cell Genetics of Plants. Vol. I. Laboratory Techniques Chapter
78, pp. 712-727.
155. Wang, A.S., and R.L. Phillips. 1984. Synchronization of suspension culture
cells. In Cell Culture and Somatic Cell Genetics of Plants. Vol. I. Laboratory
Techniques, Chapter 21, pp. 175-181.
156. Rines, H.W., S.S. Johnson, and R.L. Phillips. 1986. Tissue culture induced
variation in oats. In 2nd Intl. Oats Conf., D.A. Lawes and H. Thomas (eds.)
Aberystwyth, Wales, July 15-18, 1985. Martinas Nijhoff Publishers,
Dordrecht, Netherlands, pp. 29-33.
157. Benzion, G., R.L. Phillips, and H.W. Rines. 1986. Case histories of genetic
variability in vitro: oats and maize. In Plant Regeneration and Genetic
Variability, Vol. III. Cell Culture and Somatic Cell Genetics of Plants series.
I. K. Vasil, Ed. Academic Press, New York, Chapter 22, pp. 435-448.
158. Phillips, R.L. 1987. Lectures in molecular cytogenetics. Published by The
Beijing Agricultural University, People’s Republic of China. 111 pp.
159. Phillips, R.L., M.D. McMullen, S. Enomoto, and I. Rubenstein. 1987. Ribosomal
DNA in maize. p. 201-214. In J.P. Gustafson and R. Appels (eds.) Chromosome
Structure and Function: Impact of New Concepts. 18th Stadler Genetics Symp.
Columbia, MO. Plenum Press. New York.
160. Kowles, R.V., and R.L. Phillips. 1988. Endosperm development in maize. Intl.
Rev. Cytl. 112:97-136.
161. Phillips, R.L. 1988. Maize genetic engineering update. Proc. Corn Refiners
Assoc. September 19-22, 1988. St. Louis, MO. pp. 1-8.
162. Phillips, R.L., D.A. Somers, and K.A. Hibberd. 1988. Cell/tissue culture
and in vitro manipulation. In Corn and Corn Improvement 3rd Ed.,
(Ed. G.F. Sprague and J.W. Dudley). Amer. Soc. Agron., Madison, WI.
p. 345-387.
163. Lee, M.D., and R.L. Phillips. 1988. Chromosomal basis of somaclonal variation.
Ann. Rev. Plant Physiol. Plant Mol. Biol. 39:413-437.
956 Wolf Prize in Agriculture
164. Phillips, R.L., and V.M. Peschke. 1988. Discovery of Ac activity among progeny
of tissue culture-derived maize plants. In Plant Transposable Elements,
Ed. O. Nelson, Plenum Press, New York. p. 305-315.
165. Somers, D.A., R.L. Phillips, and H.W. Rines. 1988. Corn and oat tissue cultures
and genetic variation in regenerated plants. In Cell and Tissue Culture in
Field Crop Improvement. J. Bay-Petersen and T.M. Chu (eds.). FFTC Symp.,
Tsukuba, Japan, Food and Fertilizer Tech. Center. Book Series No. 38, Taipei,
Taiwan. pp 57-66.
166. Rines, H.W., R.L. Phillips, W.P. Bullock, and E.N. Jellen. 1989. Biotechnology
applications in oat breeding in the U.S. p. 100-107. In: B. Mattsson and
R. Lyhagen (ed.) Proc. 3rd Int’l. Oat Conf., Lund, Sweden, 4-8 July 1988.
Svälof AB, Sweden.
167. Phillips, R.L. 1990. Genomic reorganization induced by plant tissue culture.
In R.S. Sangwan and B.S. Sangwan-Norreel (eds.) The Impact of Biotechnology
in Agriculture. Proc. Intl. Conf.: The meeting point between fundamental
and applied in vitro culture research, pp. 237-246. Amiens, France. Kluwer
Academic Publ., Dordrecht.
168. Phillips, R.L., S.M. Kaeppler, and V.M. Peschke. 1990. Do we understand
somaclonal variation? In: H.J.J. Nijkamp, L.H.W. Van Der Plas, and J. Van
Aartrijk (eds.). Progress in Plant Cellular and Molecular Biology. Proc. VIIth
Int. Congr. Plant Tissue and Cell Culture, pp. 131-141. Amsterdam, The
Netherlands. Kluwer Academic Publ., Dordrecht.
169. McMullen, M.D., R.L. Phillips, and I. Rubenstein. 1991. Molecular analysis
of the nucleolus organizer region in maize. In Chromosome Engineering in
Plants: Genetics, Breeding, and Evolution. Part A. Eds. P.K. Gupta and Tsuchiya,
T. Elsevier Science Publ., Amsterdam, pp. 561-576.
170. Phillips, R.L. 1991. Prospects of cell and molecular biology/breeding interface
in crop improvement. In: Proc. 1991 PORIM International Palm Oil Conf.,
9-14 Sept. 1991, Kuala Lumpur, Malaysia.
171. Phillips, R.L., D.J. Plunkett, and S.M. Kaeppler. 1992. Novel approaches to the
induction of genetic variation and plant breeding implications. In Stalker,
H.T., and J.P. Murphy (eds.) Plant Breeding in the 1990’s. North Carolina
State University Symposium, pp. 389-408. CAB Intl Press, Oxon, England.
172. Davis, D.W., O. Riera-Lizarazu, H.W. Rines, and R.L. Phillips. 1992.
Characterization of wide-cross derived oat haploids and their progeny. Proc.
4th Intl. Oat Conf., Adelaide, South Australia, pp. 109-111.
173. Rines, H.W., R.L. Phillips, E.N. Jellen, W.L. Rooney, S.F. Kianian, and B.-C. Wu.
1992. Toward an integrated chromosomal/molecular genetic map in oat.
Proc. 4th Intl. Oat Conf., Adelaide, South Australia, pp. 97-100.
174. Peschke, V.M., and R.L. Phillips. 1992. Genetic implications of somaclonal
variation in plants. Adv. Genet. 30:41-75.
Ronald L. Phillips 957
175. Kowles, R.V., G.L. Yerk, F. Srienc, and R.L. Phillips. 1992. Maize endosperm
tissue as an endoreduplication system. In: Setlow, J.K. (ed.) Genetic
Engineering, Principles and Methods. Vol. 14, pp. 65-88. Plenum Press,
New York.
176. Rines, H.W., R.L. Phillips, and D.A. Somers. 1992. Applications of tissue
culture to oat improvement. In Marshall, H.G., and M.E. Sorrells (eds.) Oat
Science and Technology. pp. 777-791. Am. Soc. Agron., Crop Sci. Soc. Am.
Madison, WI.
177. Phillips, R.L., and S.A. Eberhart. 1993. Novel methodology in plant
breeding-discussion. International Crop Science I. eds. D.R. Buxton, R. Shibles,
R.A. Forsberg, B.L. Blad, K.H. Asay, G.M. Paulsen, and R.F. Wilson. Crop Sci.
Soc. Am., Madison, pp. 647-648.
178. Phillips, R.L., and I.K. Vasil (eds). 1994. DNA-based markers in plants. Kluwer
Academic Publishers. Dordrecht, The Netherlands 384 pp.
179. O’Donoughue, L.S., J.P. Rayapati, S.F. Kianian, M.E. Sorrells, S.D. Tanksley,
M. Lee, H.W. Rines, and R.L. Phillips. 1994. Development of RFLP-based
linkage maps in diploid and hexaploid oat (Avena sp.). In: Phillips, R.L., and
I. K. Vasil (eds.) DNA-Based Markers in Plants. Kluwer Academic Publishers,
Dordrecht, The Netherlands pp. 359-374.
180. Freeling, M., V. Walbot (eds.) 1994. The Maize Handbook. Ch. III Genetics
Protocols. R. Phillips and J. Birchler (eds.), Ch V Cell Culture Protocols.
R. Phillips (ed.). Springer Verlag, New York.
181. Kowles, R.V., G.L. Yerk, L. Schweizer, F. Srienc, and R.L. Phillips. 1994.
Flow cytometry for endosperm nuclear DNA. In The Maize Handbook.
M. Freeling and V. Walbot (eds). Springer Verlag, New York pp. 400-406.
182. Phillips, R.L. 1994. Classification of pollen abortion in the field. In The
Maize Handbook. M. Freeling and V. Walbot (eds.). Springer Verlag, New
York pp. 297-298.
183. Phillips, R.L. 1994. Chromosomal translocations involving the nucleolus
organizer region or satellite of chromosome 6. In The Maize Handbook.
M. Freeling and V. Walbot (eds.). Springer Verlag, New York pp. 342-345.
184. Livingston, S.M., and R.L. Phillips. 1994. In situ hybridization of DNA and
RNA probes to maize chromosomes. In The Maize Handbook. M. Freeling
and V. Walbot (eds.). Springer Verlag, New York pp. 504-508.
185. Kowles, R.V., G.L. Yerk, F. Srienc, and R.L. Phillips. 1994. Absorption
cytophotometry of nuclear DNA. In The Maize Handbook. M. Freeling and
V. Walbot (eds.). Springer Verlag, New York pp. 396-400.
186. Phillips, R.L., and P. Olhoft. 1995. Genomic reprogramming by tissue
culture. In Modification of gene expression and non-Mendelian inheritance.
K. Oono and F. Takaiwa (eds.). Natl. Inst. Agrobiol. Resources, Tsukuba,
Japan, pp. 1-14.
958 Wolf Prize in Agriculture
187. Rines, H.W., O. Riera-Lizarazu, and R.L. Phillips. 1995. Disomic maize
chromosome-addition oat plants derived from oat x maize crosses. In
Modification of gene expression and non-Mendelian inheritance. K. Oono
and F. Takaiwa (eds.). Natl. Inst. Agrobiol. Resources, Tsukuba, Japan,
pp. 235-251.
188. Phillips, R.L., H.W. Rines, E.N. Jellen, W.L. Rooney, B.C. Wu, S. Kianian, and
O. Riera-Lizarazu. 1995. Oat genome anlaysis via molecular markers and
oat x corn crosses. In Classical and molecular cytogenetic analysis. Proc. U.S.
Japan Joint Symp. (W.J. Raupp and B.S. Gill, eds). Kansas Ag Exp Stn Rep.
95:352-D, Manhattan, pp. 49-57.
189. Olhoft, P. and R.L. Phillips. 1995. Genetic and epigenetic changes induced
by maize tissue culture. In Use of induced mutations and molecular
techniques for crop improvement. FAO/IAEA Intl. Symp., Vienna, Austria,
June 19-23.
190. Kaeppler, S.M., R.L. Phillips, and P. Olhoft. 1996. Molecular basis of heritable
tissue culture-induced variation in plants. In Somaclonal variation and induced
mutation in crop improvement. Jain, S.M., B.S. Ahloowalia, and D.S. Brar,
eds. Kluwer Academic Publishers, The Netherlands, Dordrecht.
191. Rines, H.W., O. Riera-Lizarazu, S.B. Maguieira, and R.L. Phillips. 1996. Wide
crosses for haploids. In V Intl. Oat Conf. and VII Intl. Barley Genetics Symp.
G. Scoles and B. Rossnagal, eds. pp. 207-212.
192. Rines, H.W., O. Riera-Lizarazu, V.M. Nunez, D.W. Davis, and R.L. Phillips.
1997. Oat haploids from anther culture and from wide hybridizations. In
In vitro haploid production of haploids in higher plants. S.M. Jain,
S.K. Sopory, and R.E. Veillaux (eds.). Kluwer Acad. Publ., Dordrecht. Vol.
4:205-221.
193. Kianian, S., M. Egli, R. Phillips, H. Rines, D. Somers, B. Gengenbach,
F. Webster, S. Livingston, S. Groh, L. O’Donoughue, M. Sorrells, D. Wesenberg,
D. Stuthman, and G. Fulcher. 1998. Association of acetyl-CoA carboxylase
and groat oil content in oat. In: J. Sanchez, E. Cerdo-Olmedo, and E. Martinez-
Force (eds.), Adv. Plant Lipid Res., pp. 629-632.
194. Olhoft, P.M. and R.L. Phillips. 1999. Genetic and epigenetic instability in
tissue culture and regenerated progenies. In Plant Response to Environmental
Stresses: From phytohormones to genomic reorganization. Ed. H.R. Lerner,
M. Dekker Inc., New York, pp. 111-148.
195. Phillips, R.L. 1999. Unconventional sources of genetic diversity: de novo
variation and elevated epistasis. In: Plant Breeding in the Turn of the
Millennium. Federal University of Vicosa, Brazil, March 2-3, pp. 103-131.
196. Phillips, R.L. 1999. Research needs in heterosis. In: Genetics and exploitation
of heterosis in crops. pp. 501-508. Ed. J.G. Coors and S. Pandey. Amer. Soc.
Agron., Madison, WI.
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197. Phillips, R.L. 2000. Synteny among cereal genomes and its utilization in
agriculture. In: Challenging New Frontiers of Scientific and Technical
Importance Symp. Tsukuba, Japan.
198. Phillips, R.L. 2000. Biotechnology and agriculture in today’s world: A brief
comment on their roles for improving human nutrition. In: Improving Human
Nutrition Through Agriculture. Symp. International Food Policy Research
Institute and International Rice Research Institute, Philippines.
199. Imle, P.T., R.L. Phillips, and R.A. Porter. 2000. Molecular genetics of wild
rice. In: Wild Rice Research and Management Conference Proc.
200. Olsen, M.S. and R.L. Phillips. 2001. Molecular genetic improvement of protein
quality in maize. Encyl. Life Support Syst-UNESCO.
201. Phillips, R.L., and I.K. Vasil (eds.) 2001. DNA-based Markers in Plants.
2 nd Ed., Kluwer Academic Publishers, Dordrecht, The Netherlands,
512 pp.
202. Phillips, R.L., and I.K. Vasil. 2001. Introduction: Molecular marker maps of
major crop species. In: DNA-based markers in plants. Eds. R.L. Phillips and
I.K. Vasil. Kluwer Academic Press, The Netherlands, pp. 167-168.
203. Kianian, S.F., S.L. Fox, S. Groh, N. Tinker, L.S. O’Donoughue, P.J. Rayapati,
R.P. Wise, M. Lee, M.E. Sorrells, G. Fedak, S.J. Molnar, H.W. Rines, and
R.L. Phillips. 2001. Molecular marker linkage maps in diploid and hexaploid
oat (Avena sp.). In: DNA-based markers in plants. Eds. R.L. Phillips and
I.K. Vasil. Kluwer Academic Press, The Netherlands, pp. 443-462.
204. Riera-Lizarazu, O., M.I. Vales, and R.L. Phillips. 2001. A compilation of
molecular genetic maps of cultivated plants. In: DNA-based markers in plants.
Eds. R.L. Phillips and I.K. Vasil. Kluwer Academic Press, The Netherlands,
pp. 463-497.
205. NRC Committee on Environmental Impacts associated with Commercial-
ization of Transgenic Plants. 2002. Environmental effects of transgenic plants:
the scope and adequacy of regulation. National Academy Press. Washington,
DC, 320 pp.
206. Okagaki, R.J., R.G. Kynast, H.W. Rines, and R.L. Phillips. 2004. Radiation
hybrid mapping. In: Encyclopedia of Plant & Crop Science. Ed., R.M. Goodman,
Marcel Dekker, Inc., New York, pp. 1074-1077.
207. Phillips, R.L., H.W. Rines, R.J. Okagaki, R.G., Kynast, R. Donahue, and
W.E. Odland. 2003. Genetic and physical mapping of the maize genome
through radiation hybrids. Proc. Intl. Rice Research Conference 2003. Gurdev
Khush Symposium. In: Rice Science: Innovations and Impact for Livelihood.
Eds. T.W. Mew, D.S. Brar, S. Peng, D. Dawe, and B. Hardy, pp. 77-90.
208. Tuberosa, R., and R.L. Phillips. 2003. Progress in map-based cloning of Vgt1,
a QTL controlling flowering time in maize. 57th Corn and Sorghum Seed
Research Conf., Dec., 2002.
960 Wolf Prize in Agriculture
209. Rines, H.W., R.L. Phillips, R.G. Kynast, R.J. Okagaki, W.E. Odland, A.O. Stec,
M.S. Jacobs, and S.R. Granath. 2005. Maize chromosome additions and
radiation hybrids in oat and their use in dissecting the maize genome. In: In
the wake of the double helix: from the green revolution to the gene revolution.
Eds. Tuberosa, R., R.L. Phillips, and M. Gale, Avenue Media, Bologna, Italy,
pp. 427-442.
210. Tuberosa, R., R.L. Phillips, and M. Gale. (Eds.) 2005. In the wake of the
double helix: From the green revolution to the gene revolution. Avenue Media,
Bologna, Italy. 772 pp. CD produced in 2006.
211. Rines, H.W., S.J. Molnar, N.A. Tinker, and R.L. Phillips. 2006. Oat. In: Kole, C.
(ed.). Genome Mapping and Molecular Breeding in Plants: Cereals and Millets
Vol. 1. Springer, Inc., NY, USA. pp. 211-242.
212. Phillips, R.L. 2006. Genetic tools from nature and the nature of genetic
tools. In: CSSA Golden Anniversary Symposium. Ed. C. Stuber. Crop Sci.
46:2245-2252.
213. Phillips, R.L., W.E. Odland, and A.L. Kahler. 2007. Rice as a reference genome
and more. In: 5th Intl. Rice Genetics Symp., Eds. D.S. Brar, D. Mackill, and
B. Hardy. pp. 3-15.
214. Phillips, R.L., and H.W. Rines. 2008. Genetic analyses with oat maize addition
and radiation hybrid lines. In: The Maize Handbook: Domestication, Genetics,
and Genome, Eds. J.L. Bennetzen and S.C. Hake, Springer-Life Sciences, New
York, NY (in press).
Ronald L. Phillips 961
962 Wolf Prize in Agriculture
Ronald L. Phillips 963
964 Wolf Prize in Agriculture
Ronald L. Phillips 965
966 Wolf Prize in Agriculture
Ronald L. Phillips 967
968 Wolf Prize in Agriculture
Ronald L. Phillips 969
970 Wolf Prize in Agriculture
ABSTRACT Novel plants with individual maize chromo- that will assist in the construction of physical maps for maize
somes added to a complete oat genome have been recovered via chromosomes. The approach is based on cloning genomic
embryo rescue from oat (Avena sativa L., 2n ⴝ 6x ⴝ 42) ⴛ DNA of an oat–maize chromosome-addition line in an appro-
maize (Zea mays L., 2n ⴝ 20) crosses. An oat–maize disomic priate vector and subsequent use of maize-specific dispersed
addition line possessing 21 pairs of oat chromosomes and one repetitive DNA sequences as detection probes to isolate clones
maize chromosome 9 pair was used to construct a cosmid carrying maize genomic DNA. Such an approach was success-
library. A multiprobe (mixture of labeled fragments used as a fully applied to the isolation of human-specific DNA fragments
probe) of highly repetitive maize-specific sequences was used in cosmid libraries constructed from DNA of human-rodent
to selectively isolate cosmid clones containing maize genomic hybrid cell lines carrying individual human chromosomes. In
DNA. Hybridization of individual maize cosmid clones or their those experiments the probes included Alu-repeats (21), a
subcloned fragments to maize and oat genomic DNA revealed Cot1 DNA fraction (22), or labeled total human DNA (23).
that most high, middle, or low copy number DNA sequences Plant repetitive sequences that are apparently species-
are maize-specific. Such DNA markers allow the identification specific were first isolated from rye (24, 25) and later found in
of maize genomic DNA in an oat genomic background. Chi- many other species (26). Comparative studies revealed a strong
meric cosmid clones were not found; apparently, significant correlation between the proportion of species-specific re-
exchanges of genetic material had not occurred between the peated families in a genome and phylogenetic relationships
maize-addition chromosome and the oat genome in these novel (27, 28). About 90–95% of all randomly tested genomic
plants or in the cloning process. About 95% of clones selected repeated sequences from barley were detected in wheat and
at random from a maize genomic cosmid library could be rye (27). According to DNA reassociation studies, less-related
detected by the multiprobe. The ability to selectively detect species such as maize and wheat have less than 10% nucleotide
maize sequences in an oat background enables us to consider sequences in common (28). The success of isolating maize
oat as a host for the cloning of specific maize chromosomes or DNA from oat–maize chromosome-addition lines depends on
maize chromosome segments. Introgressing maize chromo- how many maize-specific (relative to oat) high-copy-number
some segments into the oat genome via irradiation should dispersed nucleotide sequences are found in the maize ge-
allow the construction of a library of overlapping fragments nome. Genome analysis in grasses, including maize and oat,
for each maize chromosome to be used for developing a reveals that a major portion of genomic DNA consists of
physical map of the maize genome. families of repetitive sequences dispersed throughout the
genome (26, 29, 30). According to DNA–DNA renaturation
Chromosome addition lines of different plant species (1–5) data in maize (28), unique sequences of average length 2,100
have been generated to introgress valuable genes from wild or bp are interspersed with mid-repetitive sequences. Direct
cultivated relatives into host plant species. Alien chromosome analysis of sequences adjacent to several maize genes reveals
additions have been used for gene mapping (2, 6, 7) and serve that these genes are flanked by highly repetitive DNA se-
as an enriched source of markers for positional cloning and quences (31). Thus, maize large DNA inserts cloned in an
constructing physical maps of specific chromosomes. The appropriate vector will likely carry some kind of dispersed
discovery of maize-chromosome retention in oat ‘‘haploids’’ maize-specific nucleotide sequence. A cosmid vector was
after oat ⫻ maize crosses and the recovery of stable maize chosen for this project because of the high cloning efficiency
chromosome-addition oat lines (8, 9) should allow the devel- and relatively large insertion size. Because the oat genome is
opment of a system of chromosome analysis similar to that about 11,300 Mb, an added maize chromosome with median
available in mammalian hybrid-cell systems (10–12). Such a size of about 250 Mb would constitute about 2–4% of the total
system may be used for gene assignment, isolation of chromo- nuclear DNA of an oat–maize chromosome addition line.
some-specific probes (13), flow sorting (14) and microdissec- We found that the major part of the maize genome consists
tion of chromosomes (15), development of chromosome- of nucleotide sequences that do not cross-hybridize to oat
specific ‘‘paints’’ of fluorochrome-labeled DNA fragments genomic sequences under standard hybridization conditions.
(16–18), physical mapping, and selective isolation and map- This predominant nonhomology between oat and maize
ping of cDNAs of a particular chromosome (19, 20). genomic sequences means that many maize genomic sequences
In this paper we describe an approach for isolating clones can be directly used for Southern blot hybridization on DNA
containing large-fragment maize DNA of a single-chromo- from oat–maize chromosome addition lines to selectively
some origin from the oat–maize chromosome addition lines detect sequences of maize origin. A mixture of highly repetitive
dispersed DNA sequences of maize was used as a maize-
The publication costs of this article were defrayed in part by page charge specific multiprobe to screen a cosmid library of the chromo-
payment. This article must therefore be hereby marked ‘‘advertisement’’ in some addition line. A group of maize-specific cosmids with
accordance with 18 U.S.C. §1734 solely to indicate this fact.
Copyright 䉷 1997 by THE NATIONAL ACADEMY OF SCIENCES OF THE USA Abbreviation: RFLP, restriction fragment length polymorphism; Mb,
0027-8424兾97兾943524-6$2.00兾0 megabase(s).
PNAS is available online at http:兾兾www.pnas.org. ‡To whom reprint requests should be addressed.
3524
Agricultural Sciences: Ananiev et al. Proc. Natl. Acad. Sci. USA 94 (1997) 3525
3526 Agricultural Sciences: Ananiev et al. Proc. Natl. Acad. Sci. USA 94 (1997)
Agricultural Sciences: Ananiev et al. Proc. Natl. Acad. Sci. USA 94 (1997) 3527
FIG. 3. Hybridization of unique and low-copy-number DNA sequences isolated from maize-specific cosmids shown in Fig. 2 to a blot panel of
oat–maize chromosome addition lines carrying maize chromosomes 9, 7, 4, 3, and 2, respectively; m1 and m2 are maize stocks A188 and Seneca
60; o1 and o2 are oat stocks Sun II and Starter-1. (a) The 2.5-kb EcoRI fragment from cosmid 15 shows one band on the chromosome 9 addition
line. An additional polymorphic band is present in the parental stocks of maize. (b) The 1.8-kb EcoRI fragment from cosmid 28 detects two bands.
One band, polymorphic between m1 and m2, is present in the chromosome 9 addition line. An additional nonpolymorphic band is present on
chromosome 9 and in both parental stocks. (c) The 2.1-kb EcoRI fragment from cosmid 10 shows one band on chromosome 9. (d) The 1.4-kb EcoRI
fragment from cosmid 20 detects about 20 bands in parental maize stocks and several bands among the chromosome addition lines. The band pattern
is chromosome-specific. No cross-hybridization occurred to oat DNA in a–d. (e) The 2.9-kb EcoRI fragment from cosmid 6 detects several
polymorphic bands in parental maize stocks. One nonpolymorphic band is seen in maize, oat, and chromosomes 9, 7, 4, and 2 addition lines.
Additional bands are seen in other chromosome addition lines.
Most Cloned Maize Nucleotide Sequences Are Maize- (Fig. 3 a–d). This result indicates that a significant portion
Specific. Seven out of 11 unique or low-copy-number se- (⬎50%) of low-copy-number maize sequences have diverged
quences showed no cross-hybridization to oat genomic DNA enough to show no cross-hybridization to oat genomic DNA.
FIG. 4. Hybridization of medium and highly repetitive cloned maize DNA sequences to a blot panel of oat–maize chromosome addition lines
carrying maize chromosomes 9, 7, 4, 3, and 2, respectively; m1 and m2 are maize stocks A188 and Seneca 60; o1 and o2 are oat stocks Sun II and
Starter-1. (a) EtdBr-stained 0.8% agarose gel; lane M is a 1-kb molecular weight marker ladder. (b) The 0.7-kb EcoRI fragment of cosmid 10 shows
multiple bands on different maize chromosomes as well as one common band for all chromosomes. (c) Cosmid 1, with a 40-kb insertion, shows
strong hybridization signal over maize (lines m1 and m2) and over chromosome addition lines 9, 7, 4, 3, and 2; no hybridization is seen over oat
DNA (lines o1 and o2). (d) With the 185-bp knob repeat, most of the hybridization signal is located on chromosome 9. No hybridization to oat
DNA is detected.
Ronald L. Phillips 979
3528 Agricultural Sciences: Ananiev et al. Proc. Natl. Acad. Sci. USA 94 (1997)
Eleven of 14 (80%) middle repetitive sequences (class 10–100 The number and diversity of repeats differ among clones.
copies) are also maize-specific. Two among the remaining Some of the cosmids consist entirely of several different
three showed only faint cross-hybridization to oat genomic repeated sequences; others have only one copy of the tested
DNA. Highly repetitive sequences were preselected in this repeats. Regions of maize chromosomal DNA consisting only
study, and the process of selection showed that most highly of low-copy-number sequences with a composite length ex-
repetitive maize sequences are also maize-specific (Fig. 4 b–d). ceeding 40 kb, the size of the DNA insertion in most cosmids,
would not be detected in a cosmid library by screening with the
multiprobe.
DISCUSSION Because the 29 analyzed clones of maize chromosome 9 were
Maize Species-Specific Sequences. The cross between oat selected with the help of the multiprobe, they may carry more
and maize, two species from different subfamilies of the repeated DNA sequences than a random set of clones. There-
Graminae, represents the widest combination of crop species fore, a control experiment was conducted to analyze the
to date yielding stable, fertile partial hybrids (37). However, distribution of repetitive sequences in a set of 38 cosmid clones
little is known about the comparative structure and composi- randomly chosen from a cosmid library constructed from
tion of the two genomes. Highly conserved sequences such as DNA of the parental maize line Seneca 60. Only 2 of 38 maize
rRNA genes or tubulin cDNAs cross-hybridize to correspond- cosmid clones revealed cross-hybridization to oat genomic
ing genes in the maize and oat genomes. Unique maize RFLP DNA, and they were found to be composed of rDNA se-
probes also have been used to detect sequences in the oat quences. Thirty-four of the remaining 36 cosmid clones carried
genome (9). A series of cDNA probes has shown 71% con- maize-specific highly repetitive sequences detected by the
servation of linkage associations between maize and oat multiprobe (data not shown). This experiment indicates that
according to Van Deynze et al. (38); however, almost 50% of the probability of recovering maize-specific cosmid clones in a
maize PstI unique genomic probes do not hybridize to oat genomic library of maize chromosome 9 with the help of the
genomic DNA under standard conditions (39). primary multiprobe is around 95%.
We report herein that the proportion of maize and oat Oat as a Host for Cloning Large Maize Chromosomal DNA
nucleotide sequences that cross-hybridize to each other is low Fragments. Impressive achievements have been made in clon-
under standard hybridization conditions (65⬚C, 6⫻ SCC). ing large (⬎1 Mb) DNA fragments in yeast artificial chromo-
Among the set of low-copy-number sequences isolated and somes (41). However, the difficulties encountered make this
tested, more than 50% appeared to be maize specific. The method of cloning a eukaryotic genome expensive and prob-
proportion of maize-specific sequences among the middle or lematic for wide use. Radiation hybrids (derivative cell lines
highly repetitive maize elements was about 80–95% and from irradiated somatic cell hybrids) are an attractive alter-
among those that do hybridize to oat, most hybridize only native for cloning subfragments of a chromosome. Irradiation
weakly; therefore, even total genomic DNA of maize can serve to produce chromosome breakage and segregation of genetic
as an efficient probe to identify maize-specific cosmids in an material has been applied to a number of mammalian somatic
oat genomic background. But total maize DNA is less reliable cell hybrids containing individual alien chromosomes (10). A
for this use than the described multiprobe because it gives disadvantage of this approach in some cases is that a host
more false positive clones during primary screening of the genome and an alien chromosome have too many in-common
cosmid library. The oat–maize chromosome addition lines are nucleotide sequences, thus making difficult the direct analysis,
especially attractive as a source for isolation of region-specific identification, and isolation of the alien chromosome frag-
maize DNA fragments. It is possible to identify even small ments.
portions of a maize chromosome in an oat genome by probing The results of the research reported here allow us to propose
with total maize genomic DNA, a multiprobe of maize re- the use of oat as a host for cloning maize genomic DNA. The
peated nucleotide sequences, or many other types of cloned substantial differences in the repetitive DNA composition of
maize sequences. In the case of the highly conserved nucleo- oat and maize genomes and the high level of genetic stability
tide sequences, which are common in both maize and oat of the maize-addition chromosomes make it possible to apply
genomes, RFLPs may be used to differentially identify maize conventional molecular methods to the study of maize genetic
DNA in the oat genome. material directly in the oat genome. This approach would be
Efficiency of Recovering Maize-Specific Cosmid Clones. modeled after that used in mammalian systems (10). Radiation
The efficiency of identifying maize-specific cosmid clones would be used to induce translocations of maize segments into
depends on copy number and the pattern of distribution of the oat genome, from which maize DNA could subsequently be
repeated sequences chosen to comprise the multiprobe. Stud- isolated. In comparison with other possible maize-cloning
ies of genome structure in maize (28, 40), especially analysis of systems using phages, bacteria, or yeast, the advantage of this
repeated DNA sequences around several maize genes (30, 31), approach is that all maize chromosome fragments will derive
show that unique sequences with a median size of 2.1 kb are from a known chromosome. The high proportion of maize
surrounded by a long, complex array of repeated sequences sequences that may serve as maize-specific DNA probes in this
that belong to many different families, many of which appear system allows the identification of almost any maize chromo-
to be of retrotransposon origin (31). some segment in oat or any cloned maize DNA fragment in a
Our multiprobe highlighted EcoRI subfragments in each of library. The current availability of an extensive collection of
the 29 cosmid clones of chromosome 9 analyzed, detecting mapped DNA markers for all maize chromosomes (42) may
about 100 of the total of more than 200 fragments. Each clone allow us to identify the boundaries of cloned subfragments and
contained one or more of the repeated sequences present in to generate a collection of overlapping subfragments repre-
the multiprobe. The same group of EcoRI fragments were senting the whole chromosome. This approach would allow the
classified as highly repetitive nucleotide sequences because cloning in oat of subfragments of maize chromosomes of a
they showed strong hybridization to labeled total maize wide size range. Chromosomal subfragments 5–30 Mb in size
genomic DNA. In the same set of clones at least 50 additional may be considered optimal because they can be aligned relative
EcoRI fragments showed a strong hybridization signal to to the genetic map and, if cloned in bacterial artificial chro-
maize genomic DNA and may be classified as additional highly mosomes or even in cosmid vectors, may be arranged in contigs
repetitive nucleotide sequences. These additional sequences to generate detailed physical maps for each subfragment.
could be included in our collection of maize-specific repeated Thus, 10–60 subchromosomal fragments may comprise a
nucleotide sequences to increase the efficiency of the multi- collection of overlapping large DNA fragments of one partic-
probe used in searching for maize-specific cosmids. ular maize chromosome. Taking into account the high stability
980 Wolf Prize in Agriculture
Agricultural Sciences: Ananiev et al. Proc. Natl. Acad. Sci. USA 94 (1997) 3529
of maize genetic material in the oat background, oat lines with 20. Parimo, S., Patanjali, S. R., Shukla, H., Chaplin, D. D. & Weis-
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Douglas, C., Nizetic, D., Goodfellow, P. N. & Lehrach, H. (1991)
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Ronald L. Phillips 981
We have developed from crosses of oat (Avena sativa L.) and maize A complete series of oat–maize chromosome addition lines
(Zea mays L.) 50 fertile lines that are disomic additions of individual (10) has enabled markers and genes to be physically allocated to
maize chromosomes 1–9 and chromosome 10 as a short-arm maize chromosomes without a need for detectable polymor-
telosome. The whole chromosome 10 addition is available only in phisms. These unique plant materials confirmed interchromo-
haploid oat background. Most of the maize chromosome disomic somal duplicate loci on a large scale, one of the obstacles to
addition lines have regular transmission; however, chromosome 5 whole-genome sequencing. Numerous locus duplications com-
showed diminished paternal transmission, and chromosome 10 is plicate the reassembly of a set of shotgun DNA sequences.
transmitted to offspring only as a short-arm telosome. To further Oat–maize chromosome addition lines, however, physically sep-
dissect the maize genome, we irradiated monosomic additions arate these interchromosomal maize orthologs and paralogs
with ␥ rays and recovered radiation hybrid (RH) lines providing from each other and make them accessible to mapping, sequenc-
low- to medium-resolution mapping for most of the maize chro- ing, and cloning, even in cases where the duplicated loci of
mosomes. For maize chromosome 1, mapping 45 simple-sequence interest carry genes with monomorphic allelic sequences.
repeat markers delineated 10 groups of RH plants reflecting dif- A second obstacle that impedes sequencing of the complete
ferent chromosome breaks. The present chromosome 1 RH panel maize genome by today’s technology is the repetitive nature of
dissects this chromosome into eight physical segments defined by ⬇85% of the maize DNA. Thus, sequencing strategies are being
the 10 groups of RH lines. Genomic in situ hybridization revealed tested that accomplish the targeted sequencing of less repetitive
the physical size of a distal region, which is represented by six of gene-rich regions (11, 12). These strategies must involve tech-
the eight physical segments, as being ⬇20% of the length of the nologies that are capable of arranging those gene islands along
short arm, representing ⬇one-third of the genetic chromosome 1 the chromosomes and bridging long gaps between contigs.
map. The distal ⬇20% of the physical length of the long arm of Generating random breaks in the maize chromosome in an
maize chromosome 1 is represented by a single group of RH lines identified monosomic oat–maize chromosome addition line and
that spans >23% of the total genetic map. These oat–maize RH maintaining diminutive maize chromosomes or pieces translo-
lines provide valuable tools for physical mapping of the complex cated into oat can provide DNA panels of radiation hybrid (RH)
highly duplicated maize genome and for unique studies of inter- lines, which allow for a presence vs. absence test of markers
specific gene interactions. without the need for polymorphisms (13). With sufficient reso-
lution that is determined by the number and distribution of
MN; and Mike Gale, John Innes Center, Norwich, United Kingdom. The Congress web site
maize)F1 plants with one maize chromosome added to the (www.doublehelix.too.it) reports the list of sponsors and the abstracts.
haploid oat genome (n ⫽ 3x ⫹ 1 ⫽ 22) can produce F2 offspring Abbreviations: BC, backcross; GISH, genomic in situ hybridization; RH, radiation hybrid; SSR
with one homologous maize chromosome pair added to the marker, simple-sequence repeat marker.
doubled haploid (hexaploid) oat genome (2n ⫽ 6x ⫹ 2 ⫽ 44) ‡To whom correspondence should be addressed. E-mail: phill005@umn.edu.
among other euploid and aneuploid types (9). © 2004 by The National Academy of Sciences of the USA
(Zea mays L.) lines Seneca 60, A188, A619, B73, Mo17, the
(A188 ⫻ W64A)F1 hybrid, and a line carrying the allele bz1-
mum9. More than 200 putative F1-hybrid plants indicating
successful (oat ⫻ maize) hybridization were recovered and
analyzed for vigor, seed set, maize chromosome retention in
various tissues, and maize chromosome transmission to F2
offspring (5, 10). F2 plants were tested by PCR-based markers,
test crosses, genomic in situ hybridization (GISH), and chromo-
some counting for presence, stability, and transmission of maize
chromosomes added to the oat genome (15). F2 plants were Fig. 1. PCR products from DNA of the F1 (Starter ⫻ B73) plant F1-5133-1 when
propagated for production of F3 and subsequent generations. chromosome-specific SSR markers are used; electrophoresis in a 3.5% agarose
Backcross (BC1) plants monosomic for the maize chromosome gel shows the different elimination of maize chromosomes in individual tillers.
Lanes 1, young plantlet; lanes 2, tiller F1-5133-1兾a; lanes 3, tiller F1-5133-1兾b;
addition were produced by crossing disomic oat–maize chromo-
lanes 4, tiller F1-5133-1兾c; lanes 5, tiller F1-5133-1兾d; and lane M, standard
some addition plants back to their corresponding parental oat 100-bp ladder.
lines. Batches of 150–300 BC1 seeds were irradiated with ␥ rays
from a 137Cs source at different intensities (20–50 krad) to
induce as many maize chromosome breaks as possible without stained with propidium iodide. Signals were visualized and
seriously damaging the vigor of the seeds or resulting seedlings. captured by using an Axioskop microscope equipped for epi-
BC1F2 offspring with transmitted maize chromosome deficien- fluorescence (Zeiss) and a Magnafire charge-coupled device
cies or oat–maize chromosome translocations (RH plants) were camera (Optronics International, Chelmsford, MA).
selected by a PCR assay with primers specific for Grande 1 (16)
and CentA (17) and used for physically mapping molecular Results and Discussion
markers to the particular maize chromosome segments. Maize Chromosome Elimination in (Oat ⴛ Maize)F1 Hybrids. In oat ⫻
maize crosses, maize chromosomes are occasionally retained (2).
Genomic DNA Extraction. For a limited number of PCRs (75 or This situation is distinct in that the maize genome is completely
fewer) from single plants, DNA was extracted by the use of the eliminated in hybridizations between maize and wheat or barley.
REDExtract-N-Amp Plant PCR kit (Sigma). For larger numbers There is only one report of a maize chromosome being retained
of PCRs (⬎75) from single plants, DNA was extracted by using in wheat; however, the maize chromosome was not transmitted
either the DNeasy Plant Mini kit (Qiagen, Valencia, CA) or the to offspring (23). The timing of elimination differs as well. The
acetyltrimethylammonium bromide procedure (18). For labeling maize chromosome elimination process in oat sometimes ex-
and use as probe in GISH experiments, genomic maize DNA was tends over longer periods of time compared with that in F1
extracted from leaf cell nuclei and purified through a CsCl
hybrids generated from wheat ⫻ maize and barley ⫻ maize
gradient (19).
crosses (24). In 70% of (wheat ⫻ maize)F1 embryos, one or more
maize chromosomes were eliminated at the first mitosis. By the
PCR. PCRs were accomplished by the use of the REDExtract-
eight-cell stage, the embryos had lost all maize chromosomes
N-Amp Plant PCR kit, according to the vendor’s recommenda-
(24). Maize chromosome elimination from (oat ⫻ maize)F1
tions. F1 plantlets and seedlings of the consecutive generations
embryos starts at an early stage in embryogenesis as well (4).
were screened for the presence of maize sequences by using
maize-specific primers for the long terminal repeat of the highly However, as an example of the extended time of maize chro-
dispersed retrotransposon Grande 1 (16) and for the highly mosome elimination from oat, in the (oat ⫻ maize)F1 plant
centromere-specific retrotransposon-like repeat CentA (17). F1-5133-1 with maize chromosomes 4, 7, and 10 all detected at
Individual maize chromosomes or chromosome segments in F1 a young plant stage, a consecutive elimination of individual
plantlets, addition lines, and RH seedlings were identified by maize chromosomes was detected by the PCR analysis of
using maize-specific primers for simple-sequence repeat (SSR) genomic DNA extracted from tissues of flag leaves of different
markers (10) that were selected from the maize genetics and tillers from the same plant shortly after meiosis. The first tiller
genomics database (www.maizegdb.org). BC1F2 plants (putative retained only maize chromosome 4, thus eliminated chromo-
RH plants) were tested for presence vs. absence of maize somes 7 and 10. The second tiller eliminated chromosome 10,
chromosome segments by a PCR assay with 45 SSR markers thus retained chromosomes 4 and 7. The third and fourth tillers
specific for maize chromosome 1 (p-umc1354 to p-umc2244 eliminated chromosome 7, thus retained chromosome 4 and
spanning 1,120 map units, according to the IBM2 map). Markers chromosome 10 as a short-arm telosome (Fig. 1). We observed
were selected from the maize genetics and genomics database further instances where maize chromosomes were lost in later
mentioned above. growth stages, particularly from plantlets with two or more
originally retained maize chromosomes in their complements
Cytology. Root tips (1.5–2 cm) of oat, maize, and oat–maize (results not shown).
chromosome addition and RH lines were pretreated, fixed, and
stored as described in ref. 10. Root tips for chromosome counting (Oat ⴛ Maize)F1 Hybrids and F2 Offspring in Different Genetic Back-
were prepared as described in ref. 20. Meristem cells were grounds. Our initial work was conducted with the maize chro-
squashed in 2% wt兾vol Aceto-Orcein (Carolina Biological Sup- mosome donor Seneca 60 and the oat recipient Starter. In the
ply). Root tips for GISH were prepared as described in ref. 10. last 2 years we have tested several other combinations of maize
Further steps of RNase treatment, postfixation, and in situ and oat lines. A total of 201 (oat ⫻ maize)F1 plants have been
hybridization were as described earlier in ref. 21 except that total generated from various maize and oat backgrounds, of which 68
genomic maize DNA was labeled by the use of the ULYSIS F1 hybrids retained one or more maize chromosome(s) in their
Alexa Fluor 488 nucleic acid labeling kit (Molecular Probes) and complements. All 10 maize chromosomes could be recovered,
probed on slides without using unlabeled competitor DNA. with each occurring at different frequencies as single additions
Hybridization was carried out in 40% formamide in 1.5⫻ SSC and in combination with other maize chromosomes. No obvious
(225 mM NaCl兾22.5 mM trisodium citrate, pH 7.0) at 37°C. preferential combination for two or more specific maize chro-
Posthybridization stringency washes were carried out in 40% mosomes was detected in multiple additions in haploid oats. The
formamide in 1.5⫻ SSC at 42°C. Chromosomes were counter- frequency of recovery of a particular maize chromosome, the
Table 1. Fertile oat–maize chromosome addition lines Table 2. Seed set from fertile oat–maize chromosome
10S additions
Added maize No. of
chromosome Oat host Maize donor Addition lines No. of F2 seeds
1 Starter Seneca 60 Disomic 1 F1-plant panicle Total Tested Maize-positive Disomic Monosomic
2 Starter Seneca 60 Disomic 9
F1-0289-1兾a 51 10 9 3 6
2 Starter bz1-mum9 Disomic 1
F1-0289-1兾b 59 10 9 5 4
2 Sun II Seneca 60 Disomic 2
F1-0289-1兾c 48 10 0 0 0
3 Sun II Seneca 60 Disomic 1
F1-0289-1兾d 31 10 10 8 2
3 Preakness Seneca 60 Disomic 1
F1-5133-1兾a 6 6 6 3 3
4 Starter Seneca 60 Disomic 6
F1-5133-1兾b 0 — — — —
4 Starter A188 Disomic 2
F1-5133-1兾c 1 1 1 0 1*
4 Starter B73 Disomic 1
F1-5133-1兾d 2 2 2 1† 1*
5 Starter Seneca 60 Disomic 1
5 MN-hybrid Seneca 60 Disomic 1 *Monosomic for maize chromosome 4 and monosomic for maize chromosome
5 Sun II Seneca 60 Disomic 1 10S derivative.
†Disomic for maize chromosome 4.
6 Starter Seneca 60 Disomic 1
6 MN-hybrid Seneca 60 Disomic 1
6 Sun II Seneca 60 Disomic 1
7 Starter Seneca 60 Disomic 4 Irregular Maize Chromosome 5 Transmission in Oat. Disomic maize
7 GAF-Park Seneca 60 Disomic 1 chromosome 5 additions recovered earlier in two different oat
8 Starter Seneca 60 Disomic 1 backgrounds (OMAd5.09 in Starter and OMAd5.17 in MN-
8 Starter bz1-mum9 Disomic 1 hybrid) show significantly diminished paternal transmission,
8 GAF-Park Seneca 60 Monosomic 1 whereas maternal transmission of the maize chromosome 5 is
9 Starter Seneca 60 Disomic 7 only moderately reduced. Crossing a OMAd5.09 plant as male
9 GAF-Park Seneca 60 Disomic 1 back to Starter produced four monosomic maize additions
9 Sun II Seneca 60 Disomic 1 among 58 BC1 offspring, giving a paternal transmission rate of
10S Sun II Seneca 60 Ditelosomic 1 6.9% (4 of 58). In 30 F4 offspring of a disomic addition from the
1⫹9 Starter Seneca 60 Double disomic 1 line OMAd5.09, only 2 plants were disomic additions, which
4⫹6 Starter Seneca 60 Double disomic 1 corresponds to a minimal paternal transmission of 6.7% (2 of
30). Twenty-one plants were monosomic additions, which indi-
This table lists all available addition lines and is an update of an earlier list cates a probable maternal transmission of 76.7% (23 of 30),
that involved fewer lines (15).
although one or two of the monosomic additions could be from
paternal transmission. In a further experiment analyzing 25 F4
fertility of that plant, and the stability of a maize chromosome offspring of a disomic addition from the line OMAd5.17, only
appeared to be primarily dependent on the particular maize two plants were disomic additions, indicating a paternal trans-
chromosome interacting with the oat background. Furthermore, mission of 8% (2 of 25). Seventeen plants were monosomic
additions, which indicates a probable maternal transmission of
the ability to recover a particular maize chromosome in F1
76% (19 of 25). Taking data for both oat backgrounds together,
hybrids was not correlated with the ability to produce a fertile
the irregular low paternal transmission of the maize chromosome
addition line for that chromosome. For instance, 20 F1 hybrids
5 (6.7–8%) clearly accounts for the low frequency of disomic
with maize chromosome 5 were produced, making the chromo-
addition offspring from disomic chromosome 5 addition plants.
some 5 the most frequently recovered chromosome, either as a
single chromosome addition or in combination with other maize Maize Chromosome 10 Transmission. Chromosome 10 is the small-
chromosomes; chromosome 2 addition plants were the next most est chromosome, with 190 Mbp in the Seneca 60 complement.
frequent with 15 F1 plants produced. Yet, only one of the Yet, it was the most frequently eliminated chromosome in (oat ⫻
chromosome 5 plants was fertile and transmitted the chromo- maize)F1 hybrids, indicating a low tolerance for its presence in
some 5 to offspring resulting in the fertile disomic addition oat (4, 10). The first recovered monosomic addition of chromo-
OMAd5.59 in Sun II oat background. This finding can be some 10 occurred in a haploid of GAF-Park oat background
contrasted with chromosome 4, which has been recovered as an (10). Since that time, the plant has been vegetatively propagated
addition line nine times, with three of these addition plants being by tiller cloning under short-day conditions. Periodically, tiller
fertile and transmitting the chromosome 4 to offspring. With clones have been moved to a long-day regime to induce flowering
respect to transmission of maize chromosomes, fertile disomic for self-pollination or backcrossing with GAF-Park pollen. After
addition lines fall into different categories. Addition lines car- screening thousands of spikelets over a period of ⬇3 years, we
rying chromosomes 2, 3, 4, 6, and 9 exhibited little or no recently recovered one seed. This one seed, however, did not
problems with transmission (nearly 100% maternal and paternal possess maize chromatin; thus, a maize chromosome addition
transmission rate). Lines carrying chromosomes 1 and 7 initially offspring has not been produced by this plant.
had poor transmission of the maize chromosome, but after In this new series of oat ⫻ maize crosses, we recovered 11
several generations they show good transmission of the maize plantlets that retained maize chromosome 10 as single or mul-
chromosome (⬎80% transmission rate), possibly due to selective tiple chromosome additions, 9 plantlets in Starter oat back-
breeding for stable diploid offspring. Chromosomes 1, 5, and 8 ground, and 2 plantlets in Sun II background. One chromosome
additions have fertility problems even after several generations 10-positive F1 plant (F1-0289-1, Seneca 60 ⫻ Sun II) set 189 F2
of selection, and chromosome 10 additions have transmitted only seeds in its first four panicles (Table 2). The PCR assay with
PLANT BIOLOGY
short-arm derivatives to offspring. Eleven disomic additions for Grande 1 showed that 28 F2 plants originating from three
different maize chromosomes, including 2, 4, 5, 6, 7, 8, and 9 in panicles were maize-positive, and 2 F2 plants were maize-
different oat backgrounds, have been added to the previously negative. GISH analyses revealed 16 disomic and 12 monosomic
reported set (15), making a total of 50 fertile addition lines addition plants. All 10 F2 plants originating from the fourth
(Table 1). panicle were maize-negative. In all 28 maize-positive plants, only
Kynast et al. PNAS 兩 June 29, 2004 兩 vol. 101 兩 no. 26 兩 9923
984 Wolf Prize in Agriculture
chromosome 10 short-arm-specific SSR markers (p-phi041, p- Table 3. Groups of RH lines from independent chromosome
phi117, and p-umc1293) were present; none of the long-arm- mutation events that define the same breakpoints on
specific markers (p-umc1249, p-umc1196, p-umc1176, and p- chromosome 1
umc1084) tested was detected (22). We, therefore, assume that No. of BC1F2 No. of
the chromosome transmitted to offspring in every case is a Treatment, No. of BC1 plants with independent Panel
short-arm telocentric derivative of chromosome 10 (22). The krad plants similar breaks events* group
derived line disomic for the chromosome 10 short-arm telocen-
tric was labeled OMAdt10S.20 and seeds (F3 offspring) have 40 2 3 2 1
already been distributed (Table 1). 35 3 3 3 1
The F1 hybrid F1-5133-1 originated from crosses of Starter oat 40 1 1 1 2
with maize B73. This hybrid possessed the three maize chromo- 40 1 3 1 3
somes 4, 7, and 10 in addition to the haploid oat complement at 35 1 2 1 4
a young growth stage. In DNA samples from both the third and 35 1 1 1 5
fourth tillers (F1-5133-1兾c and F1-5133-1兾d), SSR markers were 40 1 1 1 6
present for chromosome 4 and chromosome 10. Although all 35 2 2 2 7
three short-arm-specific markers for chromosome 10 were 35 2 2 2 8
present in the two DNA samples, neither sample showed evi- 40 2 20 2 9
dence for long-arm-specific markers. Therefore, we assume that 35 4 5 4 9
chromosome 10, which was accompanied by maize chromosome 35 1 1 1 10
4, also was a telocentric short-arm derivative of chromosome 10.
*Each irradiated BC1 plant producing offspring with a rearranged added
PCR analysis of the three F2 offspring from the panicles of the maize chromosome either as a deletion or a translocation with an oat
third and fourth tiller of plant F1-5133-1 (Table 2) showed that chromosome represents at least one independent mutation event. Chromo-
only two F2 plants had the short-arm-specific SSR markers for somal BC1-plant mutants, however, are not necessarily based only on one
chromosome 10, and none had any of the long-arm-specific SSR single rearrangement event because of potentially different mutations in
markers for chromosome 10. All three F2 plants had the chro- different embryo cells during irradiation. Thus, the resulting chimeric nature
mosome 4-specific SSR markers. All six F2 offspring from the of the radiated BC1 plants among their different tillers can produce and
F1-5133-1兾a panicle were positive for B73 chromosome 4, three transmit more then one maize chromosome derivative to the corresponding
as monosomic and three as disomic additions. The tiller F1-5133- BC1F2-offspring plants.
1兾b did not set seed.
The generation of a fertile disomic telocentric addition for
negative and 171 BC1F2 offspring that tested positive for maize
chromosome 10 (OMAdt10S.20) is a major breakthrough in our
chromatin in their genomes, indicating successful transmission of
efforts to develop a complete series of fertile oat–maize addition
maize segments. It is notable that after the ␥ radiation treatment
lines (22). However, this observation raises the question of why
of 300 monosomic addition seeds, only 100 plants retained their
does only the short arm of an added maize chromosome 10
maize chromosomes or a diminutive maize chromosome deriv-
transmit in oat. Does the long arm possess a gene that prevents
ative. This finding indicates that a majority of the breaks
transmission in this alien background? High sterility occurs in
generated maize fragments that were eliminated from somatic
the highly stable whole chromosome 10 addition in GAF-Park
oat and the two independent events of short-arm derivatives of tissues. Earlier results showed a certain level of somatic insta-
chromosome 10 in Sun II and Starter oat, where the long-arm bility for whole chromosome 1 addition plants resulting in
telocentrics could not be established. The situation appears chromosome loss (7, 10).
similar to the difficulties of generating a disomic euplasmic A set of 45 SSR markers distributed along maize chromosome
addition line for Betzes barley chromosome 1H and for its 1 was used to determine by a presence vs. absence test for each
long-arm telosome 1HL in Chinese Spring wheat (25). The marker approximate points of maize chromosome breakage in
difficulties in wheat (26–28) appear to be caused by the inter- the 171 BC1F2 plants. All 45 SSR markers were present in 98
action of the gene Shw (sterility in hybrid with wheat) with the BC1F2 plants, which represent 50 families. These plants were
wheat background causing sterility. However, the sterility was considered as possessing either a whole maize chromosome
alleviated by the simultaneous addition of monosomic or disomic without a break or a reciprocal oat–maize translocation. These
chromosome 6H to the 1H addition (29). Perhaps we could select plants will be self-pollinated, and offspring of those with recip-
(oat ⫻ maize)F1 hybrids for the simultaneous additions of other rocal translocations will be selected for the segregating translo-
chromosomes with chromosome 10 to possibly allow fertility and cated chromosomes. Nine BC1F2 plants, forming five families,
transmission of the whole chromosome 10. In addition, it may be showed complex rearrangements, including interstitial deletions
feasible to use additional maize genotypes that possess a differ- and multiple translocations with oat. Forty-four BC1F2 plants
ent allele of the presumed gene on chromosome arm 10L constituted 21 families, each representing one likely independent
responsible for the sterility. In the corresponding wheat–barley (chromosome rearrangement) event (Table 3). These 44 RH
addition situation, chromosome 1H of the closely related wild plants were placed into 10 panel groups, with plants within a
barley (Hordeum vulgare L. subsp. spontaneum) was added to group resulting from similar maize chromosome breaks based on
wheat without causing a severe effect of sterility (30). marker analysis (Table 3 and Fig. 2). Fig. 2 illustrates the
definition of eight segments by seven breaks in selected RH lines
Oat–Maize Chromosome 1 RHs. Monosomic oat–maize chromo- for maize chromosome 1 representing the 10 groups (Table 3).
some 1 addition seeds, the foundation for the development of Plants with only one break in their maize chromosome, and thus
oat–maize chromosome 1 RH lines, were treated with ␥ rays at possessing only one deficiency or one oat–maize translocation,
two levels. These levels were 180 BC1 seeds treated with 40 krad are shown in the first panel (Fig. 2). The markers shown in the
and 120 BC1 seeds treated with 35 krad. A total of 46 maize- left column are the first and last marker present or absent and
positive plants, as indicated by the presence of the markers frame the breakpoints. The points define six segments on the
Grande 1 and兾or CentA, were recovered from the 40-krad short arm (p-umc1354 to p-umc168), one large segment spanning
treatments. The 35-krad treatment generated 54 maize-positive the centromere region (p-umc1626 to p-mmc0041), and one
plants. These 100 BC1 plants were allowed to self-pollinate. Of additional segment on the long arm (p-bnlg1720 to p-umc2244).
these, 91 panicles produced 340 BC1F2 offspring that tested The apparently single breakpoint in the long arm of chromo-
Summary
The current set of disomic oat–maize addition lines involves all
Fig. 2. Panel of the first RH lines for maize chromosome 1. Shown are the 15 maize chromosomes in different oat backgrounds with the
SSR markers that frame the seven breakpoints, hence define the RH segments exception of chromosome 10. The maize chromosome 10 addi-
between p-umc1354 (most distal on the short arm) and p-umc2244 (most distal tion progeny has only the short arm; a fertile disomic telocentric
on the long arm) markers representing a genetic distance of more than 1,120 addition line is available. The whole chromosome 10 added to
map units according to the IBM2 map. haploid GAF-Park oat does allow the availability of DNA.
Although not fertile, and therefore not capable of producing
disomic addition offspring, we continue to maintain the original
some 1 as defined by groups 9 and 10 is represented by seven
plant vegetatively by tiller cloning under short-day growing
independent breakage events. This break is marked proximally conditions. The leaves show remarkable somatic stability for the
by the SSR marker p-mmc0041 and distally by the SSR marker added maize chromosome over a period of ⬎3 years. The plant
p-bnlg1720 (Fig. 2). The portion (7 of 21) of lines that break at serves as a source for chromosome 10 genomic DNA and RNA.
the same point or very similar points indicates a preferential The complete series of DNAs made from each maize chro-
breakage and兾or transmission of the chromosome segments to mosome addition has been used as a powerful tool to allocate
offspring. On the short arm, five independent events (5 of 21) genes and markers to chromosome. Ananiev et al. (31) used the
demonstrate a common break in the range that is marked by the oat–maize chromosome 9 addition line as the DNA source to
SSRs p-umc1071 and p-umc1727 defining segment 1. All other construct a chromosome-specific cosmid library allowing the
breakpoints are defined by one or two events. Most striking is isolation of maize-specific repetitive DNA families. The low level
that we did not observe a centric break resulting in either a of cross-hybridization under standard conditions between oat
telocentric maize chromosome or a centric maize–oat translo- and maize genomic DNA makes it possible to screen libraries for
cation. This situation left intact a large area spanning the maize species-specific sequences (31).
centromere and the proximal regions on both arms, segment 7 Oat–maize addition lines are ideal for mapping gene families
(Fig. 2), and it is in strong contrast to earlier results from the and markers that have more than one copy on different chro-
production of maize chromosome 9 RHs (12) that showed that mosomes likely because of the duplicative nature of maize. For
the level of chromosome 9 breakage across the chromosome was example, Okagaki et al. (32) mapped 350 ESTs and sequence
relatively constant. tagged sites to chromosomes by a presence vs. absence test and
PLANT BIOLOGY
Fig. 3. GISH of metaphase chromosomes from root tips of three RH plants of the maize chromosome 1 panel. (A) Plant BC1F2, 1.07.3-001.3-3 (sibling of group
7), arrow points to the deficient short arm of maize chromosome 1; the chromosome lost ⬇20% of its short arm. p-umc1626 is the most distal present marker
tested (see also Fig. 2). The yellow-painted chromosome visualizes the segments 7 and 8 representing the genetic distance of 656 – 675 map units (B) Plant BC1F2
1.07.2-007.3-4 (sibling of group 9), arrow points to the translocation fragment visualizing the RH segment 8. The translocation fragment accounts for ⬇20% of
the long-arm length representing the genetic distance of 261–332 map units (C) Plant BC1F2 1.07.1-020.3-1 (sibling of group 3), arrow points to the translocation
fragment visualizing the RH segments 1 and 2 accounting for ⬇15% of the short-arm length representing the distance of 226 –257 map units.
Kynast et al. PNAS 兩 June 29, 2004 兩 vol. 101 兩 no. 26 兩 9925
986 Wolf Prize in Agriculture
demonstrated the usefulness of the complete addition line set. fragments together with marker data helps to relate physical
However, the true power of the addition lines as a tool for maize sizes to genetic distances along the chromosome arms. Besides
genomics and genetics may be that no marker polymorphism is the use of addition and radiation hybrid lines for mapping
required for large-scale mapping. The value of the plant material purposes, the extensive dissection of the maize genome pro-
for gene expression studies has already been shown in an vides powerful material for the targeted cloning of chromo-
analysis of interchromosomal interaction with respect to ex- some-specific DNA and to study chromosome-specific struc-
pression of the maize gene liguleless 3 on chromosome 3 (33)
tures and their behavior in an alien background.
or the reduced susceptibility against the fungal pathogen
Puccinia coronata f. sp. avenae in the oat–maize chromosome
5 addition lines (unpublished data). This is a joint contribution of the Minnesota Agricultural Experiment
With the development of RH lines from the oat–maize Station and the Department of Agriculture–Agricultural Research Ser-
additions, markers can be placed to chromosome regions. vice. This material is based upon work supported by the National Science
Visualization by GISH of the rearranged maize chromosome Foundation Grant 011134.
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For many years, Professor John A. Pickett has contributed significantly to the field
of chemical ecology and agriculture worldwide, and has achieved an international
reputation equaled by few. He has opened up new areas of research with novel
approaches and then applied basic research results by developing innovative control
strategies for agricultural pests and weeds. By integrating techniques from molecular
biology, electrophysiology, biochemistry and behavior, he has positioned himself at
the cutting-edge of the field. As a result, Pickett has become a world leader in the
development of sustainable, environmentally-sound methods of insect pest
management, based on use of semio-chemicals to manipulate the behavior of pests
and their natural enemies.
CURRICULUM VITAE:
Place/date of birth: Leicester, United Kingdom, 21 April 1945
Marital status: Married, two children
Professional title and affiliation:
Scientific Director, Rothamsted Centre for Sustainable Pest and Disease
Management, and Head, Department of Biological Chemistry, Rothamsted
Research
Special Professor, School of Biology, University of Nottingham
School: King Edward VII Grammar School, Coalville (1956-1963)
987
988 Wolf Prize in Agriculture
QUALIFICATIONS:
B.Sc., Honours chemistry, 2(i), 1967, University of Surrey (1963-1967)
Ph.D., Organic chemistry, 1971, University of Surrey, Compounds from dinitriles
and hydrazine, Professor J.A. Elvidge, 1967-1970
Chartered Chemist, 1975
D.Sc., University of Nottingham, 1993
Chartered Scientist, 2004
HONOURS/AWARDS:
Special Professor, University of Nottingham, 1991
The Rank Prize, Nutrition and Crop Husbandry, 1995
Fellow of the Royal Society, 1996
Member of the Deutsche Akademie der Naturforscher Leopoldina, 2001
International Society of Chemical Ecology Medal, 2002
CBE, for Services to Biological Chemistry, 2004
Honorary Life Membership, the Association of Applied Biologists, 2004
Foreign Member of the Royal Swedish Academy of Agriculture and Forestry, 2005
Wolf Foundation Prize in Agriculture, 2008
PROFESSIONAL AFFILIATIONS:
ACADEMIC POSITIONS:
External examiner, MSc Course, Pest Management, Imperial College at Silwood
Park, 1992-1995
Chairman, Advisory Committee, School of Applied Chemistry, University of North
London, 1993-1995
Honorary Member, Academic Staff, University of Reading, 1995-
External Examiner, Environmental Science, University of Sussex, 1997-2000
CAREER:
1970-1972 - Postdoctoral fellowship, University of Manchester Institute of
Science and Technology, Synthesis and photochemistry of
perfluoroalkylpyridazines, Professor R.N. Haszeldine, FRS.
1972-1976 - Senior scientist, Chemistry Department, Brewing Research
Foundation, Redhill, Surrey. Chemical studies on flavour active
components of hops and malt.
John Anthony Pickett 989
DISTINGUISHED LECTURES:
- Distinguished Lecture in Life Sciences, 1991, Boyce Thompson Institute for Plant
Research at Cornell University
- Alfred M. Boyce Lecture, University of California, Riverside, 1993
- “Insects and Chemical Signals: A Volatile Situation”, Royal Institution Discourse,
November 1996
- “Keynotes in Natural Resources” lecture, Swedish University of Agricultural
Sciences, Uppsala, 2 March 1998
- The Woolhouse Lecture, The Society for Experimental Biology Annual Meeting,
University of York, 26 March 1998
- The 1999 Barrington Memorial Lecture, University of Nottingham, 29 April
1999
- The 2000 Cameron-Gifford Lecture, University of Newcastle, 9 February 2000
- The Andersonian Chemical Society Centenary Lecture, University of Strathclyde,
5 April 2006
- 17th Annual H.R. MacCarthy Pest Management Lecture, University of British
Columbia, Canada, “Exploiting induced and constitutive plant stress signalling
in crop protection”, 10th October 2007
- The Royal Society Croonian Lecture, 3rd June 2008
POSTS OF DISTINCTION:
- Editorial Board, Journal of Chemical Ecology, 1991-
- Councillor, International Society of Chemical Ecology, 1991-2000
- Visiting Group IPO-DLO, Netherlands, 1993
- Vice-President, the International Society of Chemical Ecology, 1994
- President, International Society of Chemical Ecology, 1995
- Expert, committee for evaluation of INCO-Copernicus, European Commission
DGXII, 1996
- Scientific Adviser, International Foundation for Science, Stockholm, Sweden, 1996-
- Member of Royal Society Conference Grants Committee, 1997-1999
- Member of Royal Society Sectional Committee 9, 1997-2000
990 Wolf Prize in Agriculture
SCIENTIFIC INTERESTS:
Head of Department: The objectives of the Department are to make crop protection
involving natural and synthetic chemical safer and more effective. The Department
is responsible for important discoveries, including the synthetic pyrethroids, and
has a prominent world position in research areas such as chemical ecology and
evolutionary response of insects to insecticidal stress.
Personal research: The chemical ecology of interactions between insects, and some
other animals, and between insects and plants. This specifically involves the chemical
characterisation of molecular structures that influence the development and
behaviour (semiochemicals) of insects and other organisms. Pickett was the first to
identify aphid, mosquito and sandfly pheromones. Research extends to the
biochemistry and molecular biology of secondary plant metabolites that act as
semiochemicals and the mechanisms by which they are employed by insects. The
long term objectives are to develop pheromones and other semiochemicals for new
methods of pest control. This is exemplified particularly by his recent work in
Africa. His recent studies have turned more to devising novel ways of controlling
vectors of pathogens attacking the human population and farm animals.
LIST OF PUBLICATIONS:
1. J.A. Elvidge and J.A. Pickett (1972) Heterocyclic imines and amines. Part 13.
3,6-Dihydrazinopyridazine and the nature of the reaction between 3,6-
dimethoxypyridazine and hydrazine. Journal of the Chemical Society Perkin
I, 1483-1488.
2. J.A. Elvidge and J.A. Pickett (1972) Heterocyclic imines and amines. Part 14.
Products from 2,5-diiminopyrrolidine (succinimidine) and hydrazine. Journal
of the Chemical Society Perkin I, 2346-235l.
3. M.G. Barlow, R.N. Haszeldine and J.A. Pickett (1978) Heterocyclic polyfluoro-
compounds. Part 26. Synthesis of 3,6-bistrifluoromethyl- pyridazines and -
di-hydropyridazines. Journal of the Chemical Society Perkin I, 378-380.
4. J.A. Pickett (1973) Use of micro-bead anion-exchange resin for direct estimation
of flavour nucleotides in complex solutions. Journal of Chromatography 81,
156-159.
5. J.A. Pickett (1974) Estimation of nucleotides in beers and their effect on
flavour. Journal of the Institute of Brewing 80, 42-47.
6. D.R.J. Laws and J.A. Pickett (1974) Brewing, malting and allied processes.
Reports on the Progress of Applied Chemistry 59, 345-357.
7. J.A. Pickett, J. Coates and F.R. Sharpe (1975) Distortion of essential oil
composition during isolation by steam distillation. Chemistry and Industry,
571-572.
992 Wolf Prize in Agriculture
8. J.A. Pickett, J. Coates and F.R. Sharpe (1975) Improvement of hop aroma in
beer. Proceedings of the European Brewery Convention, 15th Congress, Nice,
123-140.
9. D.R.J. Laws and J.A. Pickett (1975) Brewing, malting and allied processes.
Reports on The Progress of Applied Chemistry 60, 451-463.
10. J.A. Pickett (1976) Studies on flavour-active sulphur components of hops
and beer. Proceedings of the Analytical Division of the Chemical Society,
215-217.
11. J.A. Pickett, J. Coates and F.R. Sharpe (1976) Procedure for non-destructive
concentration of flavour-active components of beer. Journal of the Institute
of Brewing 82, 228-233.
12. J.A. Pickett, J. Coates and F.R. Sharpe (1976) Chemical characterisation of
differences between ales and lagers. Journal of the Institute of Brewing 82,
233-238.
13. J.A. Pickett, T.L. Peppard and F.R. Sharpe (1976) Effect of ‘sulphuring’ on
hop oil composition. Journal of the Institute of Brewing 82, 288-289.
14. J.A. Pickett, F.R. Sharpe and T.L. Peppard (1976) Stability of essential oil of
hops. Journal of the Institute of Brewing 82, 330-333.
15. J.A. Pickett and F.R. Sharpe (1976) Effect of reduction in hop oil content on
rate of deterioration of alpha-acid in hops. Journal of the Institute of Brewing
82, 333.
16. D.R.J. Laws, N.A. Bath and J.A. Pickett (1977) Production of solvent-free
isomerized extracts. Journal of the American Society of Brewing Chemists
35, 187-191.
17. D.R.J. Laws, N.A. Bath, J.A. Pickett, C.S. Ennis and A.G. Wheldon (1977)
Preparation of hop extracts without using organic solvents. Journal of the
Institute of Brewing 83, 39-40.
18. J.A. Pickett, T.L. Peppard and F.R. Sharpe (1977) Recent developments in low
temperature steam distillation of hop oil. Journal of the Institute of Brewing
83, 302-304.
19. J.A. Pickett, F.R. Sharpe and T.L. Peppard (1977) Aerial oxidation of humulene.
Chemistry and Industry, 30-31.
20. D.R.J. Laws, T.L. Peppard, F.R. Sharpe and J.A. Pickett (1978) Recent
developments in imparting hop character to beer. Journal of the American
Society of Brewing Chemists 36, 69-72.
21. J.A. Pickett (1979) Behaviour controlling chemicals. Education in Chemistry
16, 44-47.
22. A.W. Ferguson, J.B. Free, J.A. Pickett and M. Winder (1979) Techniques used
for studying honeybee pheromones involved in clustering, and experiments
on the effect of Nasonov and queen pheromones. Physiological Entomology
4, 339-344.
John Anthony Pickett 993
23. D.C. Griffiths and J.A. Pickett (1980) A potential application of aphid alarm
pheromones. Entomologia Experimentalis et Applicata 27, 199-201.
24. J.A. Pickett and D.C. Griffiths (1980) Composition of aphid alarm pheromones.
Journal of Chemical Ecology 6, 349-360.
25. J.A. Pickett, I.H. Williams, A.P. Martin and M.C. Smith (1980) The Nasonov
pheromone of the honey bee, Apis mellifera L. (Hymenoptera: Apidae).
Part I. Chemical characterisation. Journal of Chemical Ecology 6, 425-434.
26. J.A. Pickett and J.W. Stephenson (1980) Plant volatiles and components
influencing behaviour of the field slug, Deroceras reticulatum (Mu”ll). Journal
of Chemical Ecology 6, 435-444.
27. J.B. Free, A.W. Ferguson and J.A. Pickett (1981) Evaluation of the various
components of the Nasonov pheromone used by clustering honeybees.
Physiological Entomology 6, 263-268.
28. I.H. Williams, J.A. Pickett and A.P. Martin (1981) The Nasonov pheromone
of the honey bee, Apis mellifera L. (Hymenoptera: Apidae). Part II.
Bioassay of the components using foragers. Journal of Chemical Ecology 7,
225-237.
29. J.A. Pickett, I.H. Williams, M.C. Smith and A.P. Martin (1981) The Nasonov
pheromone of the honey bee, Apis mellifera L. (Hymenoptera: Apidae). Part III.
Regulation of pheromone composition and production. Journal of Chemical
Ecology 7, 543-554.
30. J.B. Free, J.A. Pickett, A.W. Ferguson and M.C. Smith (1981) Synthetic
pheromones to attract honeybee (Apis mellifera) swarms. Journal of
Agricultural Science Cambridge 97, 427-431.
31. I.H. Williams, J.A. Pickett and A.P. Martin (1981) Attraction of honeybees to
flowering plants by using synthetic Nasonov pheromone. Entomologia
Experimentalis et Applicata 30, 199-201.
32. J.A. Pickett, I.H. Williams and A.P. Martin (1982) (Z)-11-Eicosen-1-ol, an
important new pheromonal component from the sting of the honey bee,
Apis mellifera L. (Hymenoptera: Apidae). Journal of Chemical Ecology 8,
163-175.
33. B.R. Laurence and J.A. Pickett (1982) Erythro-6-Acetoxy-5-hexadecanolide,
the major component of a mosquito oviposition attractant pheromone. Journal
of the Chemical Society Chemical Communications, 59-60.
34. R.W. Gibson, A.D. Rice, J.A. Pickett, M.C. Smith and R.M. Sawicki (1982) The
effects of the repellents dodecenoic acid and polygodial on the acquisition of
non-, semi- and persistent plant viruses by the aphid Myzus persicae. Annals
of Applied Biology 100, 55-59.
35. I.H. Williams, J.A. Pickett and A.P. Martin (1982) Nasonov pheromone
of the honeybee, Apis mellifera L. (Hymenoptera: Apidae). Part IV. Comparative
electroantennogram responses. Journal of Chemical Ecology 8, 567-574.
994 Wolf Prize in Agriculture
36. J.A. Pickett, G.W. Dawson, R.W. Gibson, D.C. Griffiths, A.D. Rice, R.M. Sawicki,
M.C. Smith and C.M. Woodcock (1982) Controlling aphid behaviour. Les
Colloques de l’INRA 7, 243-252.
37. J.B. Free, A.W. Ferguson, J.A. Pickett and I.H. Williams (1982) Use of
unpurified Nasonov pheromone components to attract clustering honeybees.
Journal of Apicultural Research 21, 26-29.
38. J.B. Free, I.H. Williams, J.A. Pickett, A.W. Ferguson and A.P. Martin (1982)
Attractiveness of (Z)-11-eicosen-1-ol to foraging honeybees. Journal of
Apicultural Research 21, 151-156.
39. D.C. Griffiths, J.A. Pickett and C.M. Woodcock (1982) Behaviour of alatae of
Myzus persicae (Sulzer) (Hemiptera: Aphididae) on chemically treated surfaces
after tethered flight. Bulletin of Entomological Research 72, 687-693.
40. G.W. Dawson, D.C. Griffiths, J.A. Pickett, M.C. Smith and C.M. Woodcock
(1982) Improved preparation of (E)-β-farnesene and its activity with
economically important aphids. Journal of Chemical Ecology 8, 1111-1117.
41. G.W. Dawson, R.W. Gibson, D.C. Griffiths, J.A. Pickett, A.D. Rice and
C.M. Woodcock (1982) Aphid alarm pheromone derivatives affecting
settling and the transmission of plant viruses. Journal of Chemical Ecology
8, 1377-1388.
42. G.G. Briggs, G.W. Dawson, R.W. Gibson, D.C. Griffiths, J.A. Pickett, A.D. Rice,
M.F. Stribley and C.M. Woodcock (1983) Compounds derived from the aphid
pheromone that interfere with colonization and virus transmission by aphids.
Aspects of Applied Biology 2, 1983, Pests, Diseases, Weeds and Weed Beet in
Sugar Beet, 41-43.
43. J.A. Pickett, G.W. Dawson, D.C. Griffiths, Liu X., E.D.M. Macaulay,
I.H. Williams and C.M. Woodcock (1983) Stabilizing pheromones for field
use: propheromones. Proceedings of the lOth International Congress of Plant
Protection 1, 271.
44. D.C. Griffiths, G.W. Dawson, J.A. Pickett and C.M. Woodcock (1983) Uses of
the aphid alarm pheromone and derivatives. Proceedings of the 10th
International Congress of Plant Protection 1, 272.
45. G.G. Briggs, G.W. Dawson, R.W. Gibson, D.C. Griffiths, J.A. Pickett, A.D. Rice,
M.F. Stribley and C.M. Woodcock (1983) Compounds derived from the aphid
alarm pheromone of potential use in controlling colonization and virus
transmission by aphids. Proceedings of the Fifth International Congress of
Pesticide Chemistry (IUPAC) Kyoto, 1982 2, 117-122.
46. J.B. Free, A.W. Ferguson and J.A. Pickett (1983) Effect of the components of
the Nasonov pheromone on its release by honeybees at the hive entrance.
Journal of Apicultural Research 22, 155-157.
47. R.W. Gibson, J.A. Pickett, G.W. Dawson, A.D. Rice and C. Venables (1983)
Pyrethroid insecticides and aphid repellents as means of controlling potato
John Anthony Pickett 995
61. B.R. Laurence and J.A. Pickett (1985) An oviposition attractant pheromone in
Culex quinquefasciatus Say (Diptera: Culicidae). Bulletin of Entomological
Research 75, 283-290.
62. J.A. Pickett (1985) Production of behaviour-controlling chemicals by
crop plants. Philosophical Transactions of the Royal Society of London 310,
235-239.
63. J.B. Free, J.A. Pickett, A.W. Ferguson, J.R. Simpkins and M.C. Smith (1985)
Repelling foraging honeybees with alarm pheromones. Journal of Agricultural
Science, Cambridge 105, 255-260.
64. C. Wall, J.A. Pickett, D.G. Garthwaite and N. Morris (1985) A female sex
pheromone in the pea midge, Contarinia pisi. Entomologia Experimentalis et
Applicata 39, 11-14.
65. Liu Xun, Z-n. Zhang, Kong Jie, J.A. Pickett, Pan Yongcheng, Xie Yige and
Gu Jiechen (1985) Field attractant activity of the synthetic sex pheromone of
diamondback moth, Plutella xylostella. Acta Ecologica Sinica 5, 249-256.
66. J.A. Pickett (1986) Biotechnology in the service of pest control. Proceedings
of National Agricultural Conference, 6 March 1986.
67. G.G. Briggs, G.R. Cayley, G.W. Dawson, D.C. Griffiths, E.D.M. Macaulay,
J.A. Pickett, M.M. Pile, L.J. Wadhams and C.M. Woodcock (1986) Some
fluorine-containing pheromone analogues. Pesticide Science 17, 441-448.
68. I.K. Kigatiira, J.W.L. Beament, J.B. Free and J.A. Pickett (1986) Using synthetic
pheromone lures to attract honeybee colonies in Kenya. Journal of Apicultural
Research 25, 85-86.
69. E.D.M. Macaulay, G.W. Dawson, Liu Xun and J.A. Pickett (1986) Field
performance of synthetic diamondback moth sex pheromones. Aspects of
Applied Biology 12, 105-116.
70. J.A. Pickett and D.C. Griffiths (1986) Electrostatic sprayers for behaviour-
controlling chemicals. AFRC Science Sprays & Sprayers, 18-19.
71. G.W. Dawson, D.C. Griffiths, J.A. Pickett, L.J. Wadhams and C.M. Woodcock
(1986) Plant compounds that synergise activity of the aphid alarm
pheromone. Proceedings of the British Crop Protection Conference – Pests
and Diseases, 829-834.
72. G.W. Dawson, D.C. Griffiths, A. Hassanali, J.A. Pickett, R.T. Plumb, B.J. Pye,
L.E. Smart and C.M. Woodcock (1986) Antifeedants: a new concept for control
of barley yellow dwarf virus in winter cereals. Proceedings of the British
Crop Protection Conference – Pests and Diseases, 1001-1008.
73. J.A. Pickett (1986) Honey bee pheromones: some recent developments in
controlling honey bee behaviour. The Gooding Memorial Lecture 1985,
pp. 1-11. (Published by the Central Association of Bee–Keepers).
74. G.W. Dawson, D.C. Griffiths, N.F. Janes, A. Mudd, J.A. Pickett, L.J. Wadhams
and C.M. Woodcock (1987) Identification of an aphid sex pheromone. Nature
325, 614-616.
John Anthony Pickett 997
75. J.A. Pickett, G.W. Dawson, D.C. Griffiths, A. Hassanali, L.A. Merritt,
A. Mudd, M.C. Smith, L.J. Wadhams, C.M. Woodcock and Z-n. Zhang (1987)
Development of plant-derived antifeedants for crop protection. In: Pesticide
Science and Biotechnology, pp. 125-128. Editors R. Greenhalgh and
T.R. Roberts. (Blackwell Scientific Publications).
76. D.C. Griffiths and J.A. Pickett (1987) Novel chemicals and their formulation
for aphid control. Proceedings of the 14th International Symposium on
Controlled Release of Bioactive Materials 14, 243-244.
77. B. Mauchamp and J.A. Pickett (1987) Juvenile hormone-like activity of
(E)-β-farnesene derivatives. Agronomie 7, 523-529.
78. J.A. Pickett, G.W. Dawson, B.R. Laurence, M.M. Pile, M.C. Smith and
L.J. Wadhams (1986) Development of the oviposition attractant pheromone
for control of Culex sp. mosquitoes. Abstracts 6th International Congress
Pesticide Chemistry, IUPAC, Ottawa, 1986, 2C-07.
79. J.A. Pickett, G.R. Cayley, G.W. Dawson, D.C. Griffiths, S.H. Hockland,
B. Marples, R.T. Plumb and C.M. Woodcock (1986) Use of the alarm
pheromone and derivatives against aphid-mediated damage. Abstracts 6th
International Congress Pesticide Chemistry, IUPAC, Ottawa, 1986, 2C-08.
80. 79a. S.H. Hockland, G.W. Dawson, D.C. Griffiths, B. Marples, J.A. Pickett and
C.M. Woodcock (1986) The use of aphid alarm pheromone (E)-β-farnesene
to increase effectiveness of the entomophilic fungus Verticillium lecanii in
controlling aphids on chrysanthemums under glass. In: Fundamental and
Applied Aspects of Invertebrate Pathology, p. 252. Editors R.A. Samson,
J.M. Vlak and R. Peters. (The Netherlands: Society of Invertebrate Pathology).
81. Liu Xun, Kong Jie, Z-n. Zhang, J.A. Pickett, Pan Yongcheng and Meng
Xiaoyun (1987) Field attraction of the photosensitive propheromone to the
diamondback moth, Plutella xylostella (L). Sinozoologia 5, 15-19.
82. G.W. Dawson, D.C. Griffiths, J.A. Pickett, L.J. Wadhams and C.M. Woodcock
(1987) Plant-derived synergists of alarm pheromone from turnip aphid,
Lipaphis (Hyadaphis) erysimi (Homoptera, Aphididae). Journal of Chemical
Ecology 13, 1663-1671.
83. G.W. Dawson, D.C. Griffiths, J.A. Pickett, R.T. Plumb, C.M. Woodcock and
Z-n. Zhang (1988) Structure/activity studies on aphid alarm pheromone
derivatives and their field use against transmission of barley yellow dwarf
virus. Pesticide Science 22, 17-30.
84. J.A. Pickett (1988) Integrating use of beneficial organisms with chemical
crop protection. Philosophical Transactions of the Royal Society of London B
318, 203-211.
85. J.A. Pickett (1988) Chemical pest control – the new philosophy. Chemistry in
Britain 24, 137-142.
86. J.A. Pickett (1988) The future of semiochemicals in pest control. Aspects of
Applied Biology 17, 397-406.
998 Wolf Prize in Agriculture
87. W.A. Otieno, T.O. Onyango, M.M. Pile, B.R. Laurence, G.W. Dawson,
L.J. Wadhams and J.A. Pickett (1988) A field trial of the synthetic oviposition
pheromone with Culex quinquefasciatus Say (Diptera: Culicidae) in Kenya.
Bulletin of Entomological Research 78, 463-470.
88. G.W. Dawson, D.C. Griffiths, L.A. Merritt, A. Mudd, J.A. Pickett, L.J. Wadhams
and C.M. Woodcock (1988) The sex pheromone of the greenbug, Schizaphis
graminum. Entomologia Experimentalis et Applicata 48, 91-93.
89. D.C. Griffiths, A. Hassanali, L.A. Merritt, A. Mudd, J.A. Pickett, S.J. Shah,
L.E. Smart, L.J. Wadhams and C.M. Woodcock (1988) Highly active
antifeedants against coleopteran pests. Proceedings of the Brighton Crop
Protection Conference – Pests and Diseases, 1041-1046.
90. Y. Asakawa, G.W. Dawson, D.C. Griffiths, J-Y. Lallemand, S.V. Ley, K. Mori,
A. Mudd, M. Pezechk–Leclaire, J.A. Pickett, H. Watanabe, C.M. Woodcock and
Z-n. Zhang (1988) Activity of drimane antifeedants and related compounds
against aphids, and comparative biological effects and chemical reactivity of
(–)– and (+)– polygodial. Journal of Chemical Ecology 14, 1845-1855.
91. G.W. Dawson, N.F. Janes, A. Mudd, J.A. Pickett, A.M.Z. Slawin, L.J. Wadhams
and D.J. Williams (1989) The aphid sex pheromone. Pure and Applied
Chemistry 61, 555-558.
92. Z-n. Zhang, Liu Xun and J.A. Pickett (1988) Several aphid alarm pheromone
analogues possessing biological activity. Acta Entomologica Sinica 31,
435-438.
93. I.F. Henderson, G.G. Briggs, N.P. Coward, G.W. Dawson, J.A. Pickett, J.I. Bullock
and L.F. Larkworthy (1989) A new group of molluscicidal compounds. 1989
BCPC Monograph No. 41 Slugs and Snails in World Agriculture, 289-294.
94. W.J. Airey, I.F. Henderson, J.A. Pickett, G.C. Scott, J.W. Stephenson and
C.M. Woodcock (1989) Novel chemical approaches to mollusc control. 1989
BCPC Monograph No. 41 Slugs and Snails in World Agriculture, 301-307.
95. G.W. Dawson, J.A. Pickett and L.J. Wadhams (1989) S344 Case Study 2.
Pheromones. In: Organic chemistry: a synthesis approach, pp. 1-24. The
Open University.
96. J. Polonsky, S.C. Bhatnagar, D.C. Griffiths, J.A. Pickett and C.M. Woodcock
(1989) Activity of quassinoids as antifeedants against aphids. Journal of
Chemical Ecology 15, 993-998.
97. G.W. Dawson, D.L. Hallahan, A. Mudd, M.M. Patel, J.A. Pickett, L.J. Wadhams
and R.M. Wallsgrove (1989) Secondary plant metabolites as targets for genetic
modification of crop plants for pest resistance. Pesticide Science 27,
191-201.
98. D.C. Griffiths, J.A. Pickett, L.E. Smart and C.M. Woodcock (1989) Use of
insect antifeedants against aphid vectors of plant virus disease. Pesticide
Science 27, 269-276.
John Anthony Pickett 999
99. G.W. Dawson, B.R. Laurence, J.A. Pickett, M.M. Pile and L.J. Wadhams (1989)
A note on the mosquito oviposition pheromone. Pesticide Science 27,
277-280.
100. M.M. Blight, J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1989) Antennal
responses of Ceutorhynchus assimilis and Psylliodes chrysocephala to volatiles
from oilseed rape. Aspects of Applied Biology 23, 329-334.
101. J.A. Pickett (1989) Semiochemicals for aphid control. Journal of Biological
Education 23, 180-186.
102. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1989) Chemical ecology and
pest management: some recent insights. Insect Science and its Application
10, 741-750.
103. J.A. Pickett (1989) Towards zero pesticide residues: the biomanagement of
pests and diseases of oilseed rape. AFRC Institute of Arable Crops Research
Report for 1989, pp. 79-82.
104. G.W. Dawson, A. Mudd, J.A. Pickett, M.M. Pile and L.J. Wadhams (1990)
Convenient synthesis of mosquito oviposition pheromone and a highly
fluorinated analog retaining biological activity. Journal of Chemical Ecology
16, 1779-1789.
105. P.I. Sopp, A. Palmer and J.A. Pickett (1990) The effect of a plant-derived
antifeedant on Tetranychus urticae and Phytoseiulus persimilis: “A first look”.
SROP/WPRS Bulletin XIII/5 (1990), 198-201.
106. G.W. Dawson, D.C. Griffiths, L.A. Merritt, A. Mudd, J.A. Pickett, L.J. Wadhams
and C.M. Woodcock (1990) Aphid semiochemicals - a review, and recent
advances on the sex pheromone. Journal of Chemical Ecology 16,
3019-3030.
107. J. Hardie, M. Holyoak, J. Nicholas, S.F. Nottingham, J.A. Pickett, L.J. Wadhams
and C.M. Woodcock (1990) Aphid sex pheromone components: age-
dependent release by females and species-specific male response.
Chemoecology 1, 63-68.
108. J.A. Pickett (1990) Gas chromatography-mass spectrometry in insect
pheromone identification: three extreme case histories. In: Chromatography
and Isolation of Insect Hormones and Pheromones, pp. 299-309. Editors
A.R. McCaffery and I.D. Wilson. (Plenum Press).
109. C.A.M. Campbell, G.W. Dawson, D.C. Griffiths, J. Pettersson, J.A. Pickett,
L.J. Wadhams and C.M. Woodcock (1990) Sex attractant pheromone of
damson-hop aphid Phorodon humuli (Homoptera, Aphididae). Journal of
Chemical Ecology 16, 3455-3465.
110. D.C. Griffiths, S.P. Maniar, L.A. Merritt, A. Mudd, J.A. Pickett, B.J. Pye,
L.E. Smart and L.J. Wadhams (1991) Laboratory evaluation of pest
management strategies combining antifeedants with insect growth regulator
insecticides. Crop Protection 10, 145-151.
1000 Wolf Prize in Agriculture
111. A. Cork, D.R. Hall, R.J. Hodges and J.A. Pickett (1991) Identification of major
component of male-produced aggregation pheromone of larger grain borer,
Prostephanus truncatus (Horn) (Coleoptera: Bostrichidae). Journal of
Chemical Ecology 17, 789-803.
112. S.F. Nottingham, J. Hardie, G.W. Dawson, A.J. Hick, J.A. Pickett, L.J. Wadhams
and C.M. Woodcock (1991) Behavioral and electrophysiological responses
of aphids to host and nonhost plant volatiles. Journal of Chemical Ecology
17, 1231-1242.
113. J.A. Pickett (1991) Novel approaches to crop protection. Proceedings of the
Symposium on Poverty Alleviation through Chemistry for Improved Food
Production, Colombo, Sri Lanka, 17 May, 1991, 57-62.
114. J.A. Pickett (1991) Lower terpenoids as natural insect control agents.
In: Ecological Chemistry and Biochemistry of Plant Terpenoids, pp. 297-313.
Editors J.B. Harborne and F.A. Tomas-Barberan. (Clarendon Press, Oxford).
115. J.A. Pickett (1991) Pheromones: will their promise in insect pest control ever
be achieved? Bulletin of Entomological Research 81, 229-232.
116. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1991) New approaches to
the development of semiochemicals for insect control. Proceedings of the
Conference on Insect Chemical Ecology, Tábor 1990, 333-345. (Academia
Prague and SPB Acad. Publ., The Hague).
117. J.A. Pickett (1992) Even safer insecticides? Chemistry and Industry 1,
25-26.
118. M.M. Blight, G.W. Dawson, J.A. Pickett and L.J. Wadhams (1991) The
identification and biological activity of the aggregation pheromone of Sitona
lineatus. Aspects of Applied Biology 27, 137-142.
119. J.A. Pickett, L.J. Wadhams, C.M. Woodcock and J. Hardie (1992) The chemical
ecology of aphids. Annual Review of Entomology 37, 67-90.
120. R. Garraway, L.D. Leake, M.G. Ford and J.A. Pickett (1991) The action of a
range of volatile compounds on a tentacular preparation of the field slug,
Deroceras reticulatum (Müll.). Pesticide Science 33, 240-242.
121. F. Rodriguez, D.L. Hallahan, J.A. Pickett and F. Camps (1992) Character-
ization of the ∆11-palmitoyl-CoA-desaturase from Spodoptera littoralis
(Lepidoptera: Noctuidae). Insect Biochem. Molec. Biol. 22, 143-148.
122. J.A. Pickett (1992). Pest control - novel approaches based on plant secondary
metabolism. Proceedings V Congresso Nazionale della Società Italiana di
Fitochimica, Ischia, Napoli, 30 May-2 June, 1990. 11 pp.
123. R. Garraway, L.D. Leake, M.G. Ford and J.A. Pickett (1992) The development
of a chemoreceptive neurophysiological assay for the field slug, Deroceras
reticulatum (Müll.). Pesticide Science 34, 97-98.
124. D.L. Hallahan, J.A. Pickett, L.J. Wadhams, R.M. Wallsgrove and C.M. Woodcock
(1992) Potential of secondary metabolites in genetic engineering of
crops for resistance. In: Plant Genetic Manipulation for Crop Protection,
John Anthony Pickett 1001
pp. 215-248. Editors A.M.R. Gatehouse, V.A. Hilder and D. Boulter. (C.A.B.
International, Wallingford).
125. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1992) Molecular determinants
of pheromone activity. In: Neurotox ’91. Molecular Basis of Drug and Pesticide
Action, pp. 339-348. Editor I.R. Duce. (Elsevier Applied Science, London and
New York).
126. J.A. Pickett (1992) Potential of novel chemical approaches for overcoming
insecticide resistance. In: Proceedings SCI Symposium “Resistance ’91:
Achievements and Developments in Combating Pesticide Resistance”, July
1991, Harpenden, U.K., pp. 354-365. Editors I. Denholm, A.L. Devonshire
and D.W. Hollomon. (Elsevier Applied Science, London and New York).
127. J.A. Pickett, W. Powell, L.J. Wadhams, C.M. Woodcock and A.F. Wright (1991)
Biochemical interactions between plant-herbivore-parasitoid. Proceedings 4th
European Workshop on Insect Parasitoids, April 3-5 1991, Perugia, Italy,
pp. 1-14. Editor F. Bin. (REDIA, Vol. LXXIV, n.3, Appendice).
128. J.A. Pickett and C.M. Woodcock (1992) The future of chemical pest control.
Korean Journal of Applied Entomology 31, 304-313.
129. J.A. Pickett, B.J. Pye, L.J. Wadhams, C.M. Woodcock and C.A.M. Campbell
(1992) Potential applications of semiochemicals in aphid control. 1992 BCPC
Monograph No. 51 Insect Pheromones and Other Behaviour-Modifying
Chemicals: Application and Regulation, 29-33.
130. J.A. Pickett and C.M. Woodcock (1992) Semiochemicals: molecular
determinants of activity and biosynthesis. In: Insect Molecular Science.
Proceedings 16th Symposium of the Royal Entomological Society London,
Imperial College London, September 1991, pp. 141-149. Editors J.M.
Crampton and P. Eggleston. (Academic Press, London).
131. J.A. Guldemond, A.F.G. Dixon, J.A. Pickett, L.J.Wadhams and C.M. Woodcock
(1992) The role of host-plant odour and sex pheromones in mate recognition
in the aphid Cryptomyzus. Proceedings 8th International Symposium on
Insect-Plant Relationships, Dordrecht, pp. 119-121. Editors S.B.J. Menken,
J.H. Visser and P. Harrewijn. (Kluwer Academic Publishers).
132. M.H. Pham-Delègue, M.M. Blight, M. Le Métayer, F. Marion-Poll, A.L. Picard,
J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1992) Plant chemicals involved
in honeybee-rapeseed relationships: behavioural, electrophysiological and
chemical studies. Proceedings 8th International Symposium on Insect-Plant
Relationships, Dordrecht, pp. 129-130. Editors S.B.J. Menken, J.H. Visser and
P. Harrewijn. (Kluwer Academic Publishers).
133. G. Powell, J. Hardie and J.A. Pickett (1992) Effects of the plant-derived
antifeedant polygodial on aphid host selection behaviour. Proceedings
8th International Symposium on Insect-Plant Relationships, Dordrecht,
pp. 181-182. Editors S.B.J. Menken, J.H. Visser and P. Harrewijn. (Kluwer
Academic Publishers).
1002 Wolf Prize in Agriculture
134. M.M. Blight, A.J. Hick, J.A. Pickett, L.E. Smart, L.J. Wadhams and
C.M. Woodcock (1992) Volatile plant metabolites involved in host-plant
recognition by the cabbage seed weevil, Ceutorhynchus assimilis. Proceedings
8th International Symposium on Insect-Plant Relationships, Dordrecht,
pp. 105-106. Editors S.B.J. Menken, J.H. Visser and P. Harrewijn. (Kluwer
Academic Publishers).
135. J.A. Pickett (1993) Honey bee pheromones: some recent developments in
controlling honey bee behaviour. In: Keeping Bees, pp. 163-171. Compiled
by J.B. Free. (Republication of The Gooding Memorial Lecture 1985). (The
Central Association of Bee–Keepers, Ilford).
136. J.A. Pickett and C.M. Woodcock (1993) Chemical ecology of plants and
insects: helping crops to help themselves. Interdisciplinary Science Reviews
18, 68-72.
137. A.J. Mordue (Luntz), A. Blackwell, B.S. Hansson, L.J. Wadhams and J.A. Pickett
(1992) Behavioural and electrophysiological evaluation of oviposition
attractants for Culex quinquefasciatus Say (Diptera: Culicidae). Experientia
48, 1109-1111.
138. J.M. Cottrell, I.F. Henderson, J.A. Pickett and D.J. Wright (1993) Evidence for
glycosaminoglycans as a major component of trail mucus from the terrestrial
slug, Arion ater L. Comp. Biochem. Physiol. 104B, 455-468.
139. J. Hardie, S.F. Nottingham, G.W. Dawson, R. Harrington, J.A. Pickett and
L.J. Wadhams (1992) Attraction of field-flying aphid males to synthetic sex
pheromone. Chemoecology 3, 113-117.
140. C.A.M. Campbell, J. Pettersson, J.A. Pickett, L.J. Wadhams and C.M. Woodcock
(1993) Spring migration of damson-hop aphid, Phorodon humuli
(Homoptera, Aphididae), and summer host plant-derived semiochemicals
released on feeding. Journal of Chemical Ecology 19, 1569-1576.
141. G.W. Robertson, D.W. Griffiths, J.A.T. Woodford, A.N.E. Birch, J.A. Pickett and
L.J. Wadhams (1993) A comparison of the flower volatiles from hawthorn
and four raspberry cultivars. Phytochemistry 33, 1047-1053.
142. G. Powell, J. Hardie and J.A. Pickett (1993) Effects of the antifeedant polygodial
on plant penetration by aphids, assessed by video and electrical recording.
Entomol. exp. appl. 68, 193-200.
143. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1993) New approaches for
semiochemicals in insect control. In: New Frontiers in Rice Research,
pp. 235-240. Editors K. Muralidharan and E.A. Siddiq. (Directorate of Rice
Research, Hyderabad).
144. J.A. Guldemond, A.F.G. Dixon, J.A. Pickett, L.J. Wadhams and C.M. Woodcock
(1993) Specificity of sex pheromones, the role of host plant odour in the
olfactory attraction of males, and mate recognition in the aphid Cryptomyzus.
Physiological Entomology 18, 137-143.
John Anthony Pickett 1003
145. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1993) Exploiting behaviourally
active phytochemicals in crop protection. In: Phytochemistry and Agriculture,
pp. 62-75. Editors T.A. van Beek and H. Breteler. (Clarendon Press,
Oxford).
146. G.W. Dawson, K.J. Doughty, A.J. Hick, J.A. Pickett, B.J. Pye, L.E. Smart and
L.J. Wadhams (1993) Chemical precursors for studying the effects of
glucosinolate catabolites on diseases and pests of oilseed rape (Brassica napus)
or related plants. Pesticide Science 39, 271-278.
147. G.W. Dawson, A.J. Hick, R.N. Bennett, A. Donald, J.A. Pickett and R.M.
Wallsgrove (1993) Synthesis of glucosinolate precursors and investigations
into the biosynthesis of phenylalkyl- and methylthioalkylglucosinolates.
The Journal of Biological Chemistry 268, 27154-27159.
148. A. Blackwell, A.J. Mordue (Luntz), B.S. Hansson, L.J. Wadhams and J.A. Pickett
(1993) A behavioural and electrophysiological study of oviposition cues for
Culex quinquefasciatus. Physiological Entomology 18, 343-348.
149. A.J. Mordue (Luntz), A. Blackwell, B.S. Hansson, L.J. Wadhams and J.A. Pickett
(1993) Oviposition attractants for Culex quinquefasciatus. IOBC/WPRS
Bulletin 16, 335-340.
150. J.A. Pickett (1993) Pheromones and other semiochemicals in the control of
aphids. IOBC/WPRS Bulletin 16, 81.
151. L.E. Smart, M.M. Blight, J.A. Pickett and B.J. Pye (1994) Development of field
strategies incorporating semiochemicals for the control of the pea and bean
weevil, Sitona lineatus L. Crop Protection 13, 127-135.
152. J.A. Pickett (1994) From the laboratory to the field: the major problem in
developing pheromones and other semiochemicals for pest management. 1994
BCPC Monograph no. 59: Comparing glasshouse & field pesticide performance
II, 317-318.
153. C. Höller, S.G. Micha, S. Schulz, W. Francke and J.A. Pickett (1994) Enemy-
induced dispersal in a parasitic wasp. Experientia 50, 182-185.
154. D.L. Hallahan, S-M. C. Lau, P.A. Harder, D.W.M. Smiley, G.W. Dawson,
J.A. Pickett, R.E. Christoffersen and D.P. O’Keefe (1994) Cytochrome P-450-
catalysed monoterpenoid oxidation in catmint (Nepeta racemosa) and avocado
(Persea americana); evidence for related enzymes with different activities.
Biochimica et Biophysica Acta 1201, 94-100.
155. E.C. Cocking, N.W. Blackhall, D.S. Brar, M.R. Davey, G.S. Khush, J.K. Ladha,
J.A. Pickett, J.B. Power and P.R. Shewry (1994) Biotechnological approaches
to rice genetic improvement. Proceedings, Food Security in Asia, The Royal
Society, London, 1st November, 1994, pp. 23-26.
156. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1994) Attempts to control
aphid pests by integrated use of semiochemicals. Brighton Crop Protection
Conference – Pests and Diseases -1994, 1239-1246.
1004 Wolf Prize in Agriculture
157. J. Pettersson, J.A. Pickett, B.J. Pye, A. Quiroz, L.E. Smart, L.J. Wadhams and
C.M. Woodcock (1994) Winter host component reduces colonization by
bird-cherry-oat aphid, Rhopalosiphum padi (L.) (Homoptera, Aphididae),
and other aphids in cereal fields. Journal of Chemical Ecology 20,
2565-2574.
158. J. Hardie, R. Isaacs, J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1994)
Methyl salicylate and (-)-(1R,5S)-myrtenal are plant-derived repellents for
black bean aphid, Aphis fabae Scop. (Homoptera: Aphididae). Journal of
Chemical Ecology 20, 2847-2855.
159. K.J. Doughty, G.A. Kiddle, B.J. Pye, R.M. Wallsgrove and J.A. Pickett (1995)
Selective induction of glucosinolates in oilseed rape leaves by methyl
jasmonate. Phytochemistry 38, 347-350.
160. G. Powell, J. Hardie and J.A. Pickett (1995) Responses of Myzus persicae to
the repellent polygodial in choice and no-choice video assays with young
and mature leaf tissue. Entomologia Experimentalis et Applicata 74, 91-94.
161. D.L. Hallahan, J.M. West, R.M. Wallsgrove, D.W.M. Smiley, G.W. Dawson,
J.A. Pickett and J.G.C. Hamilton (1995) Purification and characterization of
an acyclic monoterpene primary alcohol:NADP+ oxidoreductase from catmint
(Nepeta racemosa). Archives of Biochemistry and Biophysics 318, 105-112.
162. J.A. Pickett, T.M. Butt, K.J. Doughty, R.M. Wallsgrove and I.H. Williams (1995)
Minimising pesticide input in oilseed rape by exploiting natural regulatory
processes. Proceedings of the Ninth International Rapeseed Congress, Rapeseed
Today and Tomorrow, 4-7 July, 1995, Cambridge, U.K., Volume 2, F1,
pp. 565-571. (Dorset Press, Dorchester).
163. M.M. Blight, C.H. Bock, K.J. Doughty, J.K. Fieldsend and J.A. Pickett (1995)
Release of isothiocyanates from Brassica rapa seedlings during infection by
Alternaria brassicae. Proceedings of the Ninth International Rapeseed
Congress, Rapeseed Today and Tomorrow, 4-7 July, 1995, Cambridge, U.K.,
Volume 2, F13, pp. 604-606. (Dorset Press, Dorchester).
164. M.M. Blight, A.J. Hick, J.A. Pickett, L.E. Smart, L.J. Wadhams and C.M.
Woodcock (1995) Oilseed rape volatiles cueing host-plant recognition by
the seed weevil, Ceutorhynchus assimilis: chemical, electrophysiological and
behavioural studies. Proceedings of the Ninth International Rapeseed Congress,
Rapeseed Today and Tomorrow, 4-7 July, 1995, Cambridge, U.K., Volume 3,
I21, pp. 1031-1033. (Dorset Press, Dorchester).
165. M.M. Blight, J.A. Pickett, J. Ryan, L.J. Wadhams and C.M. Woodcock (1995)
Recognition of oilseed rape volatiles by pollen beetles, Meligethes spp.:
electrophysiological and chemical studies. Proceedings of the Ninth
International Rapeseed Congress, Rapeseed Today and Tomorrow, 4-7 July,
1995, Cambridge, U.K., Volume 3, I25, pp. 1043-1045. (Dorset Press,
Dorchester).
John Anthony Pickett 1005
177. C.A. Clarke, A. Cronin, W. Francke, P. Philipp, J.A. Pickett, L.J. Wadhams and
C.M. Woodcock (1996) Mating attempts between the Scarlet Tiger Moth,
Callimorpha dominula L., and the Cinnabar Moth, Tyria jacobaeae L.
(Lepidoptera: Arctiidae), involve a common sex pheromone composition.
Experientia 52, 636-638.
178. J.A. Pickett and C.M. Woodcock (1996) The role of mosquito olfaction in
oviposition site location and in the avoidance of unsuitable hosts. In: Olfaction
in Mosquito-Host Interactions. CIBA Foundation Symposium No. 200,
109-123. Editor G. Cardew. ( John Wiley & Sons Ltd., Chichester).
179. K.J. Doughty, M.M. Blight, C.H. Bock, J.K. Fieldsend and J.A. Pickett (1996)
Release of alkenyl isothiocyanates and other volatiles from Brassica rapa
seedlings during infection by Alternaria brassicae. Phytochemistry 43,
371-374.
180. J.S. Waterhouse, J. Ke, J.A. Pickett and P.J. Weldon (1996) Volatile components
in dorsal gland secretions of the collared peccary, Tayassu tajacu (Tayassuidae,
Mammalia). Journal of Chemical Ecology 22, 1307-1314.
181. C.J. Dodds, M.G. Ford, I.F. Henderson, L.D. Leake, A.P. Martin, J.A. Pickett, L.J.
Wadhams and P. Watson (1996) Slug chemical ecology: electrophysiological
and behavioural studies. In: Slug and Snail Pests in Agriculture, British Crop
Protection Council Symposium Proceedings No. 66, 73-81. BCPC, Farnham.
182. J.G.C. Hamilton, G.W. Dawson and J.A. Pickett (1996) 9-Methylgermacrene-
B; proposed structure for novel homosesquiterpene from the sex pheromone
glands of Lutzomyia longipalpis (Diptera: Psychodidae) from Lapinha, Brazil.
Journal of Chemical Ecology 22, 1477-1491.
183. J.G.C. Hamilton, G.W. Dawson and J.A. Pickett (1996) 3-Methyl-α-
himachalene; proposed structure for the novel homosesquiterpene sex
pheromone of Lutzomyia longipalpis (Diptera: Psychodidae) from Jacobina,
Brazil. Journal of Chemical Ecology 22, 2331-2340.
184. P.M. Lösel, M. Lindemann, J. Scherkenbeck, J. Maier, B. Engelhard, C.A.M.
Campbell, J. Hardie, J.A. Pickett, L.J. Wadhams, A. Elbert and G. Thielking
(1996) The potential of semiochemicals for control of Phorodon humuli
(Homoptera: Aphididae). Pesticide Science 48, 293-303.
185. P.M. Lösel, M. Lindemann, J. Scherkenbeck, C.A.M. Campbell, J. Hardie,
J.A. Pickett and L.J. Wadhams (1996) Effect of primary-host kairomones on
the attractiveness of the hop-aphid sex pheromone to Phorodon humuli
males and gynoparae. Entomologia Experimentalis et Applicata 80, 79-82.
186. L.E. Smart, J.A. Pickett and W. Powell (1997) “Push-Pull” strategies for pest
control. Grain Legumes No. 15, 14-15.
187. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1997) First steps in the use of
aphid sex pheromones. In: Insect Pheromone Research: New Directions, pp.
439-444. Editors R.T. Cardé and A.K. Minks. (Chapman and Hall, New York).
John Anthony Pickett 1007
188. J.E. Fuentes-Contreras, W. Powell, L.J. Wadhams, J.A. Pickett and H.M.
Niemeyer (1996) Influence of wheat and oat cultivars on the development
of the cereal aphid parasitoid Aphidius rhopalosiphi and the generalist
aphid parasitoid Ephedrus plagiator. Annals of Applied Biology 128,
181-187.
189. A.J. Hick, J.A. Pickett, D.W.M. Smiley, L.J. Wadhams and C.M. Woodcock
(1997) Higher plants as a clean source of semiochemicals and genes for
their biotechnological production. In: Phytochemical Diversity: A Source of
New Industrial Products, pp. 220-236. Editors S. Wrigley, M. Hayes,
R. Thomas and E. Chrystal. The Royal Society of Chemistry, Cambridge.
190. Z.R. Khan, K. Ampong-Nyarko, P. Chiliswa, A. Hassanali, S. Kimani,
W. Lwande, W.A. Overholt, J.A. Pickett, L.E. Smart, L.J. Wadhams and
C.M. Woodcock (1997) Intercropping increases parasitism of pests. Nature
388, 631-632.
191. M.M. Blight, M. Le Métayer, M-H. Pham-Delègue, J.A. Pickett, F. Marion-Poll
and L.J. Wadhams (1997) Identification of floral volatiles involved in
recognition of oilseed rape flowers, Brassica napus by honeybees, Apis
mellifera. Journal of Chemical Ecology 23, 1715-1727.
192. B.J. Gabrys, H.J. Gadomski, Z. Klukowski, J.A. Pickett, G.T. Sobota, L.J. Wadhams
and C.M. Woodcock (1997) Sex pheromone of cabbage aphid Brevicoryne
brassicae: identification and field trapping of male aphids and parasitoids.
Journal of Chemical Ecology 23, 1881-1890.
193. G. Powell, J. Hardie and J.A. Pickett (1997) Laboratory evaluation of antifeedant
compounds for inhibiting settling by cereal aphids. Entomologia Experimentalis
et Applicata 84, 189-193.
194. J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1997) Developing sustainable
pest control from chemical ecology. Agriculture, Ecosystems and Environment
64, 149-156.
195. A. Guerrero, J. Feixas, J. Pajares, L.J. Wadhams, J.A. Pickett and C.M. Woodcock
(1997) Semiochemically induced inhibition of behaviour of Tomicus destruens
(Woll.) (Coleoptera: Scolytidae). Naturwissenschaften 84, 155-157.
196. J. Hardie, L. Peace, J.A. Pickett, D.W.M. Smiley, J.R. Storer and L.J. Wadhams
(1997) Sex pheromone stereochemistry and purity affect field catches of
male aphids. Journal of Chemical Ecology 23, 2547-2554.
197. A. Quiroz, J. Pettersson, J.A. Pickett, L.J. Wadhams and H.M. Niemeyer (1997)
Semiochemicals mediating spacing behavior of bird cherry-oat aphid,
Rhopalosiphum padi feeding on cereals. Journal of Chemical Ecology 23,
2599-2607.
198. K.S. Boo, I.B. Chung, K.S. Han, J.A. Pickett and L.J. Wadhams (1998) Response
of the lacewing Chrysopa cognata to pheromones of its aphid prey. Journal
of Chemical Ecology 24, 631-643.
1008 Wolf Prize in Agriculture
199. D.L. Hallahan, J.M. West, D.W.M. Smiley and J.A. Pickett (1998) Nepetalactol
oxidoreductase in trichomes of the catmint Nepeta racemosa. Phytochemistry
48, 421-427.
200. M.C. Luszniak and J.A. Pickett (1998) Self-defence for plants. Chemistry in
Britain 34, 29-32.
201. S. Al Abassi, M.A. Birkett, J. Pettersson, J.A. Pickett and C.M. Woodcock (1998)
Ladybird beetle odour identified and found to be responsible for attraction
between adults. Cellular and Molecular Life Sciences 54, 876-879.
202. Y. Du, G.M. Poppy, W. Powell, J.A. Pickett, L.J. Wadhams and C.M. Woodcock
(1998) Identification of semiochemicals released during aphid feeding that
attract parasitoid Aphidius ervi. Journal of Chemical Ecology 24, 1355-1368.
203. M. Subchev, A. Harizanov, W. Francke, S. Franke, E. Plass, A. Reckziegel,
F. Schröder, J.A. Pickett, L.J. Wadhams and C.M. Woodcock (1998) Sex
pheromone of female vine bud moth, Theresimima ampellophaga comprises
(2R)-butyl (7Z)-tetradecenoate. Journal of Chemical Ecology 24, 1141-1151.
204. N.G. Agelopoulos and J.A. Pickett (1998) Headspace analysis in chemical
ecology: effects of different sampling methods on ratios of volatile compounds
present in headspace samples. Journal of Chemical Ecology 24, 1161-1172.
205. G. Powell, J. Hardie and J.A. Pickett (1998) The effects of antifeedant
compounds and mineral oil on stylet penetration and transmission of potato
virus Y by Myzus persicae (Sulz.) (Hom., Aphididae). Journal of Applied
Entomology 122, 331-333.
206. J.A. Pickett, L.J. Wadhams and C.M. Woodcock. (1998) Insect supersense:
mate and host location by insects as model systems for exploiting olfactory
interactions. The Biochemist, August 1998, 8-13.
207. J.A. Pickett (1998) Insects and chemical signals: a volatile situation.
Proceedings of the Royal Institution of Great Britain 69, 243-257. (Oxford
University Press).
208. J.A. Pickett (1998) Pest semiochemicals in arable crop protection. Pesticide
Science 54, 290-291.
209. J.G.C. Hamilton, A.M. Hooper, K. Mori, J.A. Pickett and S. Sano (1999)
3-Methyl-α-himachalene is confirmed, and the relative stereochemistry
defined, by synthesis as the sex pheromone of the sandfly Lutzomyia
longipalpis from Jacobina, Brazil. Chemical Communications, 355-356.
210. A.J. Hick, M.C. Luszniak and J.A. Pickett (1999) Volatile isoprenoids that
control insect behaviour and development. Natural Product Reports 16,
39-54.
211. R.T. Glinwood, D.W.M. Smiley, J. Hardie, J.A. Pickett, W. Powell, L.J. Wadhams
and C.M. Woodcock (1999) Aphid sex pheromones: manipulation of
beneficial insects for aphid population control. Pesticide Science 55,
208-209.
John Anthony Pickett 1009
212. N. Agelopoulos, M.A. Birkett, A.J. Hick, A.M. Hooper, J.A. Pickett,
E.M. Pow, L.E. Smart, D.W.M. Smiley, L.J. Wadhams and C.M. Woodcock
(1999) Exploiting semiochemicals in insect control. Pesticide Science 55,
225-235.
213. A.M. Hooper, J.A. Bennison, M.C. Luszniak, J.A. Pickett, E.M. Pow and
L.J. Wadhams (1999) Verbena x hybrida flower volatiles attractive to Western
flower thrips, Frankliniella occidentalis. Pesticide Science 55, 660-662.
214. E.M. Pow, A.M. Hooper, M.C. Luszniak, J.A. Pickett and L.J. Wadhams (1998)
Novel strategies for improving biological control of western flower thrips on
protected ornamentals - attraction of western flower thrips to verbena plants.
Proceedings British Crop Protection Conference - Pests and Diseases, 1998,
417-422. BCPC, Farnham.
215. M.A. Birkett, B.P.S. Khambay, J.A. Pickett, L.J. Wadhams and C.M. Woodcock
(1999) Strategies for developing natural products as crop protection agents
employing neurotoxicological and other neurophysiological modes of action.
In: Progress in Neuropharmacology and Neurotoxicology of Pesticides and
Drugs. SCI Special Publication No. 232, pp. 184-192. Editor D.J. Beadle
(Royal Society of Chemistry, Cambridge).
216. J.A. Pickett, K. Chamberlain, G.M. Poppy and C.M. Woodcock (1999)
Exploiting insect responses in identifying plant signals. In: Insect-Plant
Interactions and Induced Plant Defence. Novartis Foundation Symposium
223. Editor J. Goode. pp. 253-265. ( John Wiley & Sons Ltd., Chichester).
217. N.G. Agelopoulos, A.M. Hooper, S.P. Maniar, J.A. Pickett and L.J. Wadhams
(1999) A novel approach for isolation of volatile chemicals released
by individual leaves of a plant in situ. Journal of Chemical Ecology 25,
1411-1425.
218. T.O. Olagbemiro, M.A. Birkett, A.J. Mordue (Luntz) and J.A. Pickett (1999)
Production of (5R,6S)-6-acetoxy-5-hexadecanolide, the mosquito oviposition
pheromone, from the seed oil of the summer cypress plant, Kochia scoparia
(Chenopodiaceae). Journal of Agricultural and Food Chemistry 47,
3411-3415.
219. J.A. Pickett, D.W.M. Smiley and C.M. Woodcock (1999) Secondary Metabolites
in Plant-Insect Interactions: Dynamic Systems of Induced and Adaptive
Responses. In: Advances in Botanical Research 30, pp. 91-115. (Academic
Press, London).
220. A.M. Hooper, J.G.C. Hamilton, K. Mori, J.A. Pickett and S. Sano (1999)
Volatile isoprenoids that control insect behaviour. Chem. Listy, Symposia, 93
S21-S23.
221. J. Hardie, J.A. Pickett, E.M. Pow and D.W.M. Smiley (1999) Aphids. In:
Pheromones of Non-Lepidopteran Insects Associated with Agricultural Plants,
pp. 227-250. Editors J. Hardie and A.K. Minks. (CAB International).
1010 Wolf Prize in Agriculture
222. G. Powell, S.P. Maniar, J.A. Pickett and J. Hardie (1999) Aphid responses to
non-host epicuticular lipids. Entomologia Experimentalis et Applicata 91,
115-123.
223. E.M. Pow, J.A. Bennison, M.A. Birkett, M.C. Luszniak, M. Manjunatha,
J.A. Pickett, I.S. Segers, L.J. Wadhams, L.R. Wardlow and C.M. Woodcock
(1999) Behavioural responses of western flower thrips (Frankliniella
occidentalis) to host plant volatiles. Proceedings Sixth International Symposium
on Thysanoptera, Antalya, Turkey, April 27-May 1, 1998, pp. 121-128.
224. L.E.G. Mboera, K.Y. Mdira, F.M. Salum, W. Takken and J.A. Pickett (1999)
Influence of synthetic oviposition pheromone and volatiles from soakage pits
and grass infusions upon oviposition site-selection of Culex mosquitoes in
Tanzania. Journal of Chemical Ecology 25, 1855-1865.
225. M. Manjunatha, J.A. Pickett, L.J. Wadhams and F. Nazzi (1998) Response of
western flower thrips, Frankliniella occidentalis and its predator Amblyseius
cucumeris to chrysanthemum volatiles in olfactometer and greenhouse trials.
Insect Science and its Application 18, 139-144.
226. L.M.C. Rebouças, M. do S.B. Caraciolo, A.E.G. Sant’Ana, J.A. Pickett, L.J.
Wadhams and E.M. Pow (1999) Composição química da glândula abdominal
da fêmea da mariposa Castnia licus (Drury) (Lepidoptera: Castniidae):
possíveis feromônios e precursores. Química Nova 22, 645-648.
227. J.G.C. Hamilton, A.M. Hooper, H.C. Ibbotson, S. Kurosawa, K. Mori,
S. Muto and J.A. Pickett (1999) 9-Methylgermacrene-B is confirmed as the
sex pheromone of the sandfly Lutzomyia longipalpis from Lapinha, Brazil,
and the absolute stereochemistry defined as S. Chemical Communications,
2335-2336.
228. P.A. Shah, J.A. Pickett and J.D. Vandenberg (1999) Responses of Russian Wheat
Aphid (Homoptera: Aphididae) to Aphid Alarm Pheromone. Environmental
Entomology 28, 983-985.
229. K. Chamberlain, J.A. Pickett and C.M. Woodcock (2000) Plant signalling and
induced defence in insect attack. Molecular Plant Pathology 1, 67-72.
230. N.G. Agelopoulos, K. Chamberlain and J.A. Pickett (2000) Factors affecting
volatile emissions of intact potato plants, Solanum tuberosum: variability of
quantities and stability of ratios. Journal of Chemical Ecology 26, 497-511.
231. K.S. Boo, M.Y. Choi, I.B. Chung, V.F. Eastop, J.A. Pickett, L.J. Wadhams and
C.M. Woodcock (2000) Sex pheromone of the peach aphid, Tuberocephalus
momonis, and optimal blends for trapping males and females in the field.
Journal of Chemical Ecology 26, 601-609.
232. W. Francke, E. Plass, N. Zimmermann, H. Tietgen, T. Tolasch, S. Franke,
M. Subchev, T. Toshova, J.A. Pickett, L.J. Wadhams and C.M. Woodcock (2000)
Major sex pheromone component of female herald moth Scoliopteryx libatrix
is the novel branched alkene (6Z,13)-methylheneicosene. Journal of Chemical
Ecology 26, 1135-1149.
John Anthony Pickett 1011
244. C.H. Hansen, U. Wittstock, C.E. Olsen, A.J. Hick, J.A. Pickett and B.A. Halkier
(2001) Cytochrome P450 CYP79F1 from Arabidopsis catalyzes the conversion
of dihomomethionine and trihomomethionine to the corresponding aldoximes
in the biosynthesis of aliphatic glucosinolates. The Journal of Biological
Chemistry 276, 11078-11085.
245. C.M. Hartfield, C.A.M. Campbell, J. Hardie, J.A. Pickett and L.J. Wadhams
(2001) Pheromone traps for the dissemination of an entomopathogen by the
damson-hop aphid Phorodon humuli. Biocontrol Science and Technology
11, 401-410.
246. B. Donato, S.J. Brooks, J.A. Pickett and J. Hardie (2001) Peyerimhoffina gracilis
(Schneider, 1851) (Neur.: Chrysopidae): a green lacewing new to Britain.
Entomologist’s Record 113, 131-135.
247. M.A. Birkett, K. Chamberlain, A.M. Hooper and J.A. Pickett (2001) Does
allelopathy offer real promise for practical weed management and for
explaining rhizosphere interactions involving higher plants? Plant and Soil
232, 31-39.
248. K. Chamberlain, E. Guerrieri, F. Pennachio, J. Pettersson, J.A. Pickett,
G.M. Poppy, W. Powell, L.J. Wadhams and C.M. Woodcock (2001) Can aphid-
induced plant signals be transmitted aerially and through the rhizosphere?
Biochemical Systematics and Ecology 29, 1063-1074.
249. C. Costantini, M.A. Birkett, G. Gibson, J. Ziesmann, N’F. Sagnon, H.A.
Mohammed, M. Coluzzi and J.A. Pickett (2001) Electroantennogram and
behavioural responses of the malaria vector Anopheles gambiae to human-
specific sweat components. Medical and Veterinary Entomology 15,
259-266.
250. Z.R. Khan, A. Hassanali, T.M. Khamis, J.A. Pickett and L.J. Wadhams (2001)
Mechanisms of Striga hermonthica suppression by Desmodium spp.
Proceedings of the BCPC Conference – Weeds 2001, Brighton, UK, 12-15
November 2001, 895-900.
251. J.S. Waterhouse, M. Hudson, J.A. Pickett and P.J. Weldon (2001) Volatile
components in dorsal gland secretions of the white-lipped peccary, Tayassu
pecari, from Bolivia. Journal of Chemical Ecology 27, 2459-2469.
252. E.N. Barata, H. Mustaparta, J.A. Pickett, L.J. Wadhams and J. Araujo (2002)
Encoding of host and non-host plant odours by receptor neurones in the
eucalyptus woodborer, Phoracantha semipunctata (Coleoptera: Cerambycidae).
Journal of Comparative Physiology A 188, 121-133.
253. J.A. Pickett and T.O. Olagbemiro (2002) Repellents. In Encyclopedia of Pest
Management, pp. 696-697. (Marcel Dekker).
254. Z.R. Khan, J.A. Pickett, L. Wadhams and F. Muyekho (2001) Habitat
management strategies for the control of cereal stemborers and striga in
maize in Kenya. Insect Science and Its Application. 21, 375-380.
John Anthony Pickett 1013
255. A. Ingvarsdóttir, M.A. Birkett, I. Duce, R.L. Genna, W. Mordue, J.A. Pickett,
L.J. Wadhams and A.J. Mordue (Luntz) (2002) Semiochemical strategies for
sea louse control: host location cues. Pest Management Science 58, 537-545.
256. A.M. Hooper, B. Donato, C.M. Woodcock, J.H. Park, R.L. Paul, K.S. Boo,
J. Hardie and J.A. Pickett (2002) Characterization of (1R,4S,4aR,7S,7aR)-
dihydronepetalactol as a semiochemical for lacewings, including Chrysopa
spp. and Peyerimhoffina gracilis. Journal of Chemical Ecology 28, 849-864.
257. I. Prosser, A.L. Phillips, S. Gittings, M.J. Lewis, A.M. Hooper, J.A. Pickett and
M.H. Beale (2002) (+)-(10R)-Germacrene A synthase from goldenrod,
Solidago canadensis;. cDNA isolation, bacterial expression and functional
analysis. Phytochemistry 60, 691-702.
258. Z.R. Khan, A. Hassanali, W. Overholt, T.M. Khamis, A.M. Hooper, J.A. Pickett,
L.J. Wadhams and C.M. Woodcock (2002) Control of witchweed Striga
hermonthica by intercropping with Desmodium spp., and the mechanism
defined as allelopathic. Journal of Chemical Ecology 28, 1871-1885.
259. A. Ingvarsdóttir, M.A. Birkett, I. Duce, W. Mordue, J.A. Pickett, L.J. Wadhams
and A.J. Mordue (Luntz) (2002) Role of semiochemicals in mate location by
parasitic sea louse, Lepeophtheirus salmonis. Journal of Chemical Ecology
28, 2107-2117.
260. W. Powell and J.A. Pickett (2003) Manipulation of parasitoids for aphid
pest management: progress and prospects. Pest Management Science 59,
149-155.
261. J.A. Pickett, H.B. Rasmussen, C.M. Woodcock, M. Matthes and J.A. Napier
(2003) Plant stress signalling: understanding and exploiting plant-plant
interactions. Biochemical Society Transactions 31, 123-127.
262. M.A. Birkett and J.A. Pickett (2003) Aphid sex pheromones: from discovery
to commercial production. (Molecules of Interest Review). Phytochemistry
62, 651-656.
263. G. Jones, C.A.M. Campbell, J. Hardie, J.A. Pickett, B.J. Pye and L.J. Wadhams
(2003) Integrated management of two-spotted spider mite Tetranychus urticae
on hops using hop β-acids as an antifeedant together with the predatory
mite Phytoseiulus persimilis. Biocontrol Science and Technology 13,
241-252.
264. S. Chen, E. Glawischnig, K. Jørgensen, P. Naur, B. Jørgensen, C-E. Olsen,
C.H. Hansen, H. Rasmussen, J.A. Pickett and B.A. Halkier (2003) CYP79F1
and CYP79F2 have distinct functions in the biosynthesis of aliphatic
glucosinolates in Arabidopsis. The Plant Journal 33, 923-937.
265. R. Glinwood, J. Pettersson, E. Ahmed, V. Ninkovic, M. Birkett and J. Pickett
(2003) Change in acceptability of barley plants to aphids after exposure to
allelochemicals from couch-grass (Elytrigia repens). Journal of Chemical
Ecology 29, 261-274.
1014 Wolf Prize in Agriculture
266. K.S. Boo, S.S. Kang, J.H. Park, J.A. Pickett and L.J. Wadhams (2003) Field
trapping of Chrysopa cognata (Neuroptera: Chrysopidae) with aphid sex
pheromone components in Korea. Journal of Asia-Pacific Entomology 6,
29-36.
267. T.J. Bruce, J.A. Pickett and L.E. Smart (2003) Cis-jasmone switches on plant
defence against insects. Pesticide Outlook 14, 96-98.
268. M.A. Birkett, K. Chamberlain, E. Guerrieri, J.A. Pickett, L.J. Wadhams and
T. Yasuda (2003) Volatiles from whitefly-infested plants elicit a host-locating
response in the parasitoid, Encarsia formosa. Journal of Chemical Ecology
29, 1589-1600.
269. T.J.A. Bruce, J.L. Martin, J.A. Pickett, B.J. Pye, L.E. Smart and L.J. Wadhams
(2003) cis-Jasmone treatment induces resistance in wheat plants against the
grain aphid, Sitobion avenae (Fabricius) (Homoptera: Aphididae). Pest
Management Science 59, 1031-1036.
270. M.K. Tsanuo, A. Hassanali, A.M. Hooper, Z.R. Khan, F. Kaberia, J.A. Pickett
and L.J. Wadhams (2003) Isoflavanones from the allelopathic aqueous root
exudate of Desmodium uncinatum. Phytochemistry 64, 265-273.
271. M. Matthes, J.A. Napier, J.A. Pickett & C.M. Woodcock (2003) New chemical
signals in plant protection against herbivores and weeds. Proceedings BCPC
International Congress – Crop Science & Technology 2003, 10-12 November,
SECC Glasgow, 9C-4, 1227-1236.
272. H.M. Whitney, L.V. Michaelson, O. Sayanova, J.A. Pickett and J.A. Napier
(2003) Functional characterisation of two cytochrome b5-fusion desaturases
from Anemone leveillei: the unexpected identification of a fatty acid
∆6-desaturase. Planta 217, 983-992.
273. C.A.M. Campbell, F.J. Cook, J.A. Pickett, T.W. Pope, L.J. Wadhams and
C.M. Woodcock (2003) Responses of the aphids Phorodon humuli and
Rhopalosiphum padi to sex pheromone stereochemistry in the field. Journal
of Chemical Ecology 29, 2225-2234.
274. C.H. Hansen, L. Du, P. Naur, C.E. Olsen, K.B. Axelsen, A.J. Hick, J.A. Pickett
and B.A. Halkier (2001) CYP83B1 is the oxime-metabolizing enzyme in the
glucosinolate pathway in Arabidopsis. The Journal of Biological Chemistry
276, 24790-24796.
275. L.S. Gohole, W.A. Overholt, Z.R. Khan, J.A. Pickett and L.E.M. Vet (2003)
Effects of molasses grass, Melinis minutiflora volatiles on the foraging behavior
of the cereal stemborer parasitoid, Cotesia sesamiae. Journal of Chemical
Ecology 29, 731-745.
276. M. Subchev, A. Mircheva, J. Pickett, L. Wadhams, C. Woodcock, A. dos Santos,
S. Franke and W. Francke (2003) Sex pheromone of the leaf-miner
Phyllonorycter platani: (Z10)-tetradecenyl acetate. Journal of Chemical
Ecology 29, 2391-2396. (Originally published online July 18, 2003, Rapid
John Anthony Pickett 1015
298. N. Ngumbi, A.J. Ngi-Song, E.N.M. Njagi, R. Torto, L.J. Wadhams, M.A. Birkett,
J.A. Pickett, W.A. Overholt and B. Torto (2005) Responses of the stem borer
larval endoparasitoid Cotesia flavipes (Hymenoptera: Braconidae) to plant
derived synomones: laboratory and field cage experiments. Biocontrol Science
and Technology 15, 271-279.
299. M.D. Soler Cruz, M.C. Vega Robles, J.B. Jespersen, O. Kilpinen, M. Birkett,
S. Dewhirst and J. Pickett (2005) Scanning electron microscopy of foreleg
tarsal sense organs of the poultry red mite, Dermanyssus gallinae (DeGeer)
(Acari: Dermanyssidae). Micron 36, 415-421.
300. S.P. Jacobs, A.P. Liggins, J-J. Zhou, J.A. Pickett, X. Jin and L.M. Field (2005)
OS-D-like genes and their expression in aphids (Hemiptera: Aphididae).
Insect Molecular Biology 14, 423-432.
301. J. Pettersson, V. Ninkovic, R. Glinwood, M.A. Birkett and J.A Pickett (2005)
Foraging in a complex environment – semiochemicals support searching
behaviour of the seven spot ladybird. European Journal of Entomology 102,
365-370.
302. J.A. Pickett, M.A. Birkett, T.J.A. Bruce, K. Chamberlain,R. Gordon-Weeks,
M.C. Matthes, C.B. Moraes, J.A. Napier,L.E. Smart, L.J. Wadhams and
C.M. Woodcock (2005) cis-Jasmone as an allelopathic agent through plant
defence induction.pp 122-127 In Proceedings of the 4th World Congress on
Allelopathy, 21-26 August 2005, Eds JDI Harper M An, H Wu JH Kent,
Charles Sturt University, Wagga Wagga, NSW, Australia. International
Allelopathy Society.
303. J. Pettersson, V. Ninkovic, R. Glinwood, S. Al Abassi, M. Birkett, J. Pickett and
L. Wadhams (2005) Foraging behaviour of Coccinella septempunctata (L.):
volatiles and allelobiosis. Proceedings International Symposium on Biological
Control of Aphids and Coccids, Tsuruoka, Japan, September 25-29, 2005,
187-192.
304. J.A. Pickett, T.J.A. Bruce, K. Chamberlain, A. Hassanali, Z.R. Khan, M.C.
Matthes, J.A. Napier, L.E. Smart, L.J. Wadhams and C.M. Woodcock (2006)
Plant volatiles yielding new ways to exploit plant defence. In: Chemical ecology:
from gene to ecosystem, pp. 161-173. Editors M. Dicke and W. Takken.
(Springer, Netherlands).
305. R.J.E. Bailey, M.A. Birkett, A. Ingvarsdóttir, A.J. Mordue (Luntz), W. Mordue,
B. O’Shea, J.A. Pickett and L.J. Wadhams (2006) The role of semiochemicals
in host location and non-host avoidance by salmon louse (Lepeophtheirus
salmonis) copepodids. Canadian Journal of Fisheries and Aquatic Sciences
63, 448-456.
306. A. Couty, H. van Emden, J.N. Perry, J. Hardie, J.A. Pickett and L.J. Wadhams
(2006) The roles of olfaction and vision in host-plant finding by the
diamondback moth, Plutella xylostella. Physiological Entomology 31,
134-145.
1018 Wolf Prize in Agriculture
307. J-J.Zhou, Y. Kan, J. Antoniw, J.A. Pickett and L.M. Field (2006) Genome and
EST analyses and expression of a gene family with putative functions in
insect chemoreception. Chemical Senses (advance access published March
31, 2006), 1-13.
308. Z.R. Khan, A. Hassanali and J.A. Pickett (2006) Managing polycropping
to enhance soil system productivity: a case study from Africa. In Biological
Approaches to Sustainable Soil Systems, ed. N. Uphoff, CRC Press,
pp. 575-586.
309. C.A.O. Midega, Z.R. Khan, J. van den Berg, C.K.P.O. Ogol, J.A. Pickett and
L.J. Wadhams (2006) Maize stemborer predator activity under ‘push-pull’
system and Bt-maize: a potential component in managing Bt resistance.
International Journal of Pest Management 52, 1-10.
310. K. Chamberlain, Z.R. Khan, J.A. Pickett, T. Toshova and L.J. Wadhams (2006)
Diel periodicity in the production of green leaf volatiles by wild and cultivated
host plants of stemborer moths, Chilo partellus and Busseola fusca. Journal
of Chemical Ecology 32, 565-577.
311. M.H. Beale, M.A. Birkett, T.J. Bruce, K. Chamberlain, L.M. Field, A.K. Huttly,
J.L. Martin, R. Parker, A.L. Phillips, J.A. Pickett, I.M. Prosser, P.R. Shewry,
L.E. Smart, L.J. Wadhams, C.M. Woodcock and Y. Zhang (2006) Aphid alarm
pheromone produced by transgenic plants affects aphid and parasitoid
behaviour. Proceedings of the National Academy of Sciences USA 103,
10509-10513.
312. A.M. Hooper, J-B. Farcet, N.P. Mulholland and J.A. Pickett (2006) Synthesis
of 9-methylgermacrene B, racemate of the sex pheromone of Lutzomyia
longipalpis (Lapinha), from the renewable resource, Geranium macrorrhizum
essential oil. Green Chemistry 8, 513-515.
313. Z.R. Khan, J.A. Pickett, L.J. Wadhams, A. Hassanali and C.A.O. Midega
(2006) Combined control of Striga hermonthica and stemborers by maize-
Desmodium spp. intercrops. Crop Protection 25, 989-995.
314. T. Kalule, Z.R. Khan, G. Bigirwa, J. Alupo, S. Okanya, J.A. Pickett and
L.J. Wadhams (2006) Farmers’ perceptions of importance, control practices
and alternative hosts of maize stemborers in Uganda. International Journal
of Tropical Insect Science 26, 71-77.
315. I.M. Prosser, R.J. Adams, M.H. Beale, N.D. Hawkins, A.L. Phillips, J.A. Pickett
and L.M. Field (2006) Cloning and functional characterisation of a cis-
muuroladiene synthase from black peppermint (Mentha x piperita) and direct
evidence for a chemotype unable to synthesise farnesene. Phytochemistry 67,
1564-1571.
316. K. Schönrogge, M.G. Gardner, G.W. Elmes, E.K.V. Napper, D.J. Simcox,
J.C. Wardlaw, J. Breen, B. Barr, J.J. Knapp, J.A. Pickett and J.A. Thomas (2006)
Host propagation permits extreme local adaptation in a social parasite of
ants. Ecology Letters 9, 1032-1040.
John Anthony Pickett 1019
317. Birkett, M.A., Chamberlain, K., Khan, Z.R., Pickett, J.A., Toshova, T.,
Wadhams, L.J. and Woodcock, C.M. (2006) Electrophysiological responses
of the lepidopterous stemborers Chilo partellus and Busseola fusca to
volatiles from wild and cultivated host plants. Journal of Chemical Ecology
32, 2475-2487.
318. Z.R. Khan, C.A.O. Midega, A. Hassanali, J.A. Pickett, L.J. Wadhams and
A. Wanjoya (2006) Management of witchweed, Striga hermonthica, and
stemborers in sorghum, Sorghum bicolor, through intercropping with
greenleaf desmodium, Desmodium intortum. International Journal of Pest
Management 52, 297-302.
319. S.M. Cook, Z.R. Khan and J.A. Pickett (2007) The use of push-pull
strategies in integrated pest management. Annual Review of Entomology 52,
375-400.
320. J.A. Pickett, M.A. Birkett, M.C. Blassioli Moraes, T.J.A. Bruce, K. Chamberlain,
R. Gordon-Weeks, M.C. Matthes, J.A. Napier, L.E. Smart, L.J. Wadhams and
C.M. Woodcock (2007) cis-Jasmone as allelopathic agent in inducing plant
defence. Allelopathy Journal 19, 109-118.
321. A.C. Skelton, M.A. Birkett, J.A. Pickett and M.M. Cameron (2007)
Olfactory responses of medically and economically important mites (Acari:
Epidermoptidae and Acaridae) to volatile chemicals. Journal of Medical
Entomology 44, 367-371.
322. Z.R. Khan, C.A.O. Midega, A. Hassanali, J.A. Pickett and L.J. Wadhams (2007)
Assessment of different legumes for the control of Striga hermonthica in
maize and sorghum. Crop Science 47, 728-734.
323. J.D. Blande, J.A. Pickett and G.M. Poppy (2007) A comparison of
semiochemically mediated interactions involving specialist and generalist
Brassica-feeding aphids and the braconid parasitoid Diaeretiella rapae. Journal
of Chemical Ecology 33, 767-779.
324. H.M. Mohamed, Z.R. Khan, J.M. Mueke, A. Hassanali, E. Kairu and
J.A. Pickett (2007) Behaviour and biology of Chilo partellus (Swinhoe) on
Striga hermonthica (Del.) Benth. infested and uninfested maize plants. Crop
Protection 26, 998-1005.
325. Z.R. Khan, C.A.O. Midega, L.J. Wadhams, J.A. Pickett and A. Mumuni
(2007) Evaluation of Napier grass (Pennisetum purpureum) varieties for
use as trap plants for the management of African stemborer (Busseola fusca)
in a push-pull strategy. Entomologia Experimentalis et Applicata 124,
201-211.
326. E. Kazana, T.W. Pope, L. Tibbles, M. Bridges, J.A. Pickett, A.M. Bones,
G. Powell and J.T. Rossiter (2007) The cabbage aphid: a walking mustard oil
bomb. Proceedings of the Royal Society B 274, 2271-2277.
327. T.J.A. Bruce and J.A. Pickett (2007) Plant defence signalling induced by biotic
attacks. Current Opinion in Plant Biology 10, 387-392.
1020 Wolf Prize in Agriculture
328. A.M. Hooper, S. Dufour, S. Willaert, S. Pouvreau and J.A. Pickett (2007) Synthesis
of (2S,7S)-dibutyroxynonane, the sex pheromone of the orange wheat blossom
midge, Sitodiplosis mosellana (Géhin) (Diptera: Cecidomyiidae), by
diastereoselective silicon-tethered ring-closing metathesis. Tetrahedron Letters
48, 5991-5994.
329. J.A. Pickett, Z.R. Khan, A. Hassanali and A.M. Hooper (2007) Chemicals
involved in post-germination inhibition of Striga by Desmodium: opportunities
for utilizing the associated allelopathic traits. In: Integrating New Technologies
for Striga Control: Towards Ending the Witch-hunt, pp. 61-70. Editors
G. Ejeta and J. Gressel. (World Scientific, Singapore, New Jersey, London).
330. Z.R. Khan, C.A.O. Midega, A. Hassanali and J.A. Pickett (2007) Field
developments on Striga control by Desmodium intercrops in a “push-pull”
strategy. In: Integrating New Technologies for Striga Control: Towards Ending
the Witch-hunt, pp. 241-252. Editors G. Ejeta and J. Gressel. (World Scientific,
Singapore, New Jersey, London).
331. J.A. Pickett and R.T. Glinwood (2007) Chemical Ecology. In: Aphids as Crop
Pests, pp. 235-260. Editors H.F. van Emden and R.H. Harrington. (CABI,
Wallingford).
332. S.M. Guchu, A. Yenesew, M.K. Tsanuo, N.K. Gikonyo, J.A. Pickett, A.M. Hooper
and A. Hassanali (2007) C-methylated and C-prenylated isoflavonoids from
root extract of Desmodium uncinatum. Phytochemistry 68, 646-651.
333. T.J.A. Bruce, M.C. Matthes, J.A. Napier and J.A. Pickett (2007) Stressful
“memories” of plants: evidence and possible mechanisms. Plant Science 173,
603-608.
334. J.A. Pickett, M.A. Birkett, T.J.A. Bruce, K. Chamberlain, R. Gordon-Weeks,
M.C. Matthes, J.A. Napier, L.E. Smart and C.M. Woodcock (2007)
Developments in aspects of ecological phytochemistry: the role of
cis-jasmone in inducible defence systems in plants. Phytochemistry 68, 2937-
2945. (PSE 50 Special Issue).
335. A. Hassanali, H. Herren, Z.R. Khan, J.A. Pickett and C.M. Woodcock (2008)
Integrated pest management: the push-pull approach for controlling insect
pests and weeds of cereals, and its potential for other agricultural systems
including animal husbandry. Integrated Pest Management. Philosophical
Transactions of the Royal Society London B (Theme Issue: Sustainable
Agriculture I) 363, 611-621.
336. Z.R. Khan, C.A.O. Midega, D.M. Amudavi, A. Hassanali and J.A. Pickett (2008)
On-farm evaluation of the ‘push-pull’ technology for the control of stemborers
and striga weed on maize in western Kenya. Field Crops Research 106,
224-233.
337. Z.R. Khan, D.M. Amudavi, C.A.O. Midega, J.M. Wanyama and J.A. Pickett
(2008) Farmers’ perceptions of a ‘push-pull’ technology for control of cereal
stemborers and Striga weed in western Kenya. Crop Protection 27, 976-987.
John Anthony Pickett 1021
338. C.A.O. Midega, Z.R. Khan, J. van den Berg, C.K.P.O. Ogol, A.S. Dippenaar-
Schoeman, J.A. Pickett and L.J. Wadhams (2008) Response of ground-dwelling
arthropods to a ‘push-pull’ habitat management system: spiders as an indicator
group. Journal of Applied Entomology 132, 248-254.
339. K. Schönrogge, E.K.V. Napper, M.A. Birkett, C.M. Woodcock, J.A. Pickett,
L.J. Wadhams and J.A. Thomas (2008) Host recognition by the specialist
hoverfly Microdon mutabilis, a social parasite of the ant Formica lemani.
Journal of Chemical Ecology 34, 168-178. (DOI 10.1007/s10886-007-
9417-8).
340. J.G. Logan, M.A. Birkett, S.J. Clark, S. Powers, N.J. Seal, L.J. Wadhams,
A.J. Mordue (Luntz) and J.A. Pickett (2008) Identification of human-derived
volatile chemicals that interfere with attraction of Aedes aegypti mosquitoes.
Journal of Chemical Ecology 34, 308-322.
341. T.J.A. Bruce, M.C. Matthes, K. Chamberlain, C.M. Woodcock, A. Mohib,
B. Webster. L.E. Smart, M.A. Birkett, J.A. Pickett and J.A. Napier (2008) cis-
Jasmone induces Arabidopsis genes that affect the chemical ecology of
multitrophic interactions with aphids and their parasitoids. Proceedings of
the National Academy of Sciences USA 105, 4553-4558.
342. Z.R. Khan, D.G. James, C.A.O. Midega and J.A. Pickett (2008) Chemical ecology
and conservation biological control. Biological Control 45, 210-224.
(doi:10.1016/j.biocontrol.2007.11.009).
343. Z.R. Khan, C.A.O. Midega, E.M. Njuguna, D.M.Amudavi, J.W. Wanyama and
J.A. Pickett (2008) Economic performance of the ‘push-pull’ technology for
stemborer and Striga control in smallholder farming systems in western
Kenya. Crop Protection 27, 1084-1097.
(doi:10.1016/j.cropro.2008.01.005).
J.A. Pickett, D.W.M. Smiley and L.J. Wadhams. Food safety: insects to the rescue.
ACTIN MAFF LINK Newsletter. February, 1999.
J.A. Pickett. Striga control by intercropping with Desmodium species. 2000.
In: Haustorium. (Parasitic Plants Newsletter. Official Organ of the International
Parasitic Seed Plant Research Group). Number 37, August 2000.
M.J. Uddin, N.J. Holliday, P.A. MacKay, W. Powell and J.A. Pickett (2001) IPM of
insect pests in seed alfalfa. Forage Seed News 8, 37-42.
Sex pheromone of pea aphids – an integrated pest management component as it
attracts the natural enemies. Joint Annual Meeting 2000 of the Entomological
Societies of America, Canada and Quebec.
J.A. Pickett (2002) Stem borer and striga control for maize and sorghum in Africa.
Tropical Agriculture Association Newsletter, June 2002, 25-26.
J.A. Pickett et al. (2002). “Growing the Future”. Insect Pests and their Control.
Report of a workshop held at Rothamsted Research 6-8 March 2002.
1022 Wolf Prize in Agriculture
J.A. Pickett et al. (2002) Closing discussion: the way forward. Salmon Farming:
towards an Integrated pest Management strategy for sea lice. SCI meeting at
Department of Zoology, University of Aberdeen, Scotland, 18-19 June, 2001.
Pest Management Science 58, 622-629.
Z.R. Khan, F.N. Muyekho, E. Njuguna, J.A. Pickett, L.J. Wadhams, N. Dibogo,
A. Ndiege, G. Genga and C. Lusweti (2005) A primer on planting and managing
‘push-pull’ fields for stemborer and striga weed control in maize. A step-by-
step guide for farmers. Editor A. Ng’eny-Mengech. ICIPE Science Press, 48 pp.
J.A. Pickett (2006) Book review: Insect-plant biology,second edition. Schoonhoven,
L.M., van Loon, Joop J.A., Dicke, M. (eds.). Oxford University Press, pp. 421 +
xvii. ISBN: 0-19-852594-8 (paperback, £34.95)/0-19-852594-X (hardback,
£75.00). Phytochemistry 67, 1970.
The Quiet Revolution: Push-Pull Technology and the African Farmer. Gatsby
Occasional Paper. The Gatsby Charitable Foundation, April 2005. 26 pp.
J.A. Pickett (2007) Pheromones and other scents for the alleviation, worldwide, of
many pest problems in plant, animal and human health. Transactions of the
Leicester Literary & Philosophical Society 101, 29-32.
D. Amudavi, Z. Khan and J. Pickett (2007) Enhancing the Push-Pull strategy. Leisa
Magazine, 23.4 December 2007, 8-10.
PATENTS
D.R.J. Laws and J.A. Pickett (1975) Preparation of hop oil. Brit. Pat. No. 1501098
and 9 foreign equivalents.
D.R.J. Laws, N.A. Bath, C.S. Ennis, J.A. Pickett and A.G. Wheldon (1977) Preparation
of iso-α-acids. Brit. Pat. No. l576729, and ll foreign equivalents.
J.B. Free, J.A. Pickett, A.W. Ferguson and M.C. Smith (1981) Bee pheromone. Brit.
Pat. No. 8106906, 8206254.
G.W. Dawson, D.C. Griffiths and J.A. Pickett (1981) Improvements in or relating to
pheromones. Brit. Pat. No. 2106503 and various foreign equivalents.
B.R. Laurence and J.A. Pickett (1981) Improvements in or relating to pheromones.
Brit. Pat. No. 2111481 and various foreign equivalents.
E.D.M. Macaulay and J.A. Pickett (1983) Improvements in or relating to behaviour
modifying chemicals. Brit. Pat. No. 2140421 and various foreign equivalents.
G.G. Briggs, G.R. Cayley, B.R. Laurence and J.A. Pickett (1985) Improvements in or
relating to pheromones. Brit. Pat. No. 8504483, 8604222.
A.J. Mordue (Luntz), J.A. Pickett and M.A. Birkett (23 November 2005)
Semiochemicals for sea lice monitoring and control. UK patent no. GB 2388544.
James H. Tumlinson
Pennsylvania State University
University Park, Pennsylvania, USA
k
CURRICULUM VITAE
Ralph O. Mumma Professor and Director of the Center for Chemical Ecology,
Department of Entomology, The Pennsylvania State University, 111 Chemical Ecology
Laboratory, University Park, PA 16802.
Date of Birth: February 28, 1938
EDUCATION:
PROFESSIONAL EXPERIENCE:
1964-1969 Chemist, USDA-ARS, Boll Weevil Research Laboratory, State
College, MS.
1023
1024 Wolf Prize in Agriculture
FIELD OF SPECIALIZATION:
Insect chemical communication and chemical ecology: defining chemical
communication systems, including pheromones and other semiochemicals that
mediate insect-insect and plant-insect interactions; biosynthesis of pheromones
and plant chemical signals; insect behavior, including learning, mediated by
semiochemicals. Emphasis is on developing fundamental knowledge and principles
that can be applied in environmentally safe, ecologically sound, sustainable pest
management programs.
HONORS:
1975 U.S. Department of Agriculture, Superior Service Award, (Boll Weevil
Pheromone Development Group) “In recognition of the discovery of
and for pioneering the development of the pheromone of the boll weevil
as a technique for detection, survey, suppression, or elimination of the
pest.”
1983 U.S. Department of Agriculture, Superior Service Award, “For outstanding
service in the isolation, identification, and synthesis of pheromones of a
number of major pest insects and providing science and industry with
chemicals for insect research and control.”
James H. Tumlinson 1025
My research career, which began in 1964, has been devoted to the study of
chemical ecology, in particular the chemistry of insect communication and behavior.
All of my investigations have been interdisciplinary efforts. I was Research leader
for the Chemistry Research Unit at the Center for Medical, Agricultural and
Veterinary Entomology (formerly Insect Attractants, Behavior and Basic Biology
James H. Tumlinson 1027
Research Laboratory) from 1972 until 2003. With my colleagues and students I
identified and synthesized numerous pheromones of various types, kairomones,
biologically active plant constituents, and insect herbivore-produced elicitors of
plant defenses for over 40 insect species spanning 13 families and 5 orders. Most
of my investigations dealt with insects of considerable economic importance in
one or more areas of the world and resulted in several novel discoveries that were
the first of their kind and subsequently led to similar discoveries by others. In the
last 20 years my research has focused on the chemical ecology of tritrophic insect-
plant interactions. My colleagues and I have taken an active role in transferring the
technology developed through basic research into practical tools for insect pest
management and have had several successes, notably the boll weevil and Japanese
beetle pheromones and lures for the Mediterranean fruit fly, among others. I have
given over 150 lectures on my research throughout the United States and in
several foreign countries. Some examples of my research accomplishments follow.
For my Ph.D. dissertation I isolated, identified, and synthesized the four terpenoid
components of the boll weevil pheromone9. This pheromone was applied as one of
the three primary tools in an integrated program that resulted in the successful
eradication of the boll weevil from the U.S., which had significant economic and
environmental impacts. For example, in Georgia, cotton acreage increased from
115,000 in 1983 (pre-eradication) to 1.5 million in 1995, yield from 112,000
bales to 2 million, and net crop revenues from $187 to over $451 per acre. In the
same period, the average number of insecticide treatments decreased from 14.4 to
5.4 per acre.
As a Postdoctoral Associate in the laboratory of Prof. R. M. Silverstein, at the
New York State College of Forestry, I identified the sex pheromone of the Indian
meal moth12 and the trail pheromone of the leaf cutting ant, Atta texana13.
In 1970 I accepted a position as a Research Chemist at the USDA, ARS, Insect
Attractants, Behavior and Basic Biology Laboratory in Gainesville, Florida, where
my colleagues and I isolated, identified, synthesized, and developed lures and
trapping systems for monitoring of over 20 species of pest insects. One of the most
notable examples was the sex pheromone of the Japanese beetle46. This was the
first time that an enantiomer of an optically active pheromone was clearly
demonstrated to be a potent inhibitor of attraction to the pheromone. Collaboration
with colleagues and industry scientists resulted in development of a trapping system
used around airports and other vulnerable locations throughout the world to
guard against introduction of this pest, and over 1.5 million lures per year have
been sold in the U.S. for several years.
In a collaboration spanning more than 20 years, Dr. W. J. Lewis and I, with our
students and colleagues, conducted an in depth investigation of the chemical ecology
and behavioral mechanisms that enable insect parasitoids to locate their herbivorous
insect hosts. The goal of this research was to develop more effective methods for
1028 Wolf Prize in Agriculture
Another feature of this interesting and complex interplay between insects and
plants is that plants can detect and react to chemical signals from neighboring
plants. Chemical signals from a plant damaged by insect herbivores alert neighbors
to prime their defenses so as to respond more strongly to subsequent attack than if
they had not been forewarned. Thus, corn seedlings previously exposed to “green
leaf” volatiles from damaged corn seedlings emitted at least twice the quantity of
volatile terpenes when attacked by caterpillars as seedlings that had not been
exposed to the “alarm” signals256. This ability of plants to prime their defenses to
respond more strongly when attacked, but only if attacked, has since been reported
in other plant species.
In 2003, I accepted the Ralph O. Mumma endowed chair in chemical ecology,
in the Department of Entomology at The Pennsylvania State University, where my
students and I are continuing to investigate the chemical ecology of plant-insect
interactions. In 2006, I became the director of the Center for Chemical Ecology at
Penn State. This center promotes the collaboration of 24 faculty and their students
from seven departments and two colleges across the university in interdisciplinary
investigations of the chemical ecology of numerous biological systems.
LIST OF PUBLICATIONS
1. Minyard, J. P., Tumlinson, J. H., Hedin, P. A., and Thompson, A. C. Constituents
of the cotton bud. Terpene hydrocarbons. J. Agr. Food Chem. 13:599-602.
1965.
2. Minyard, J. P., Tumlinson, J. H., Thompson, A. C., and Hedin, P. A. Constituents
of the cotton bud. Sesquiterpene hydrocarbons. J. Agr. Food Chem.
14:332-336. 1966.
3. Minyard, J. P., Tumlinson, J. H., Thompson, A. C., and Hedin, P. A. Constituents
of the cotton bud. The carbonyl compounds. J. Agr. Food Chem. 15:517-524.
1967.
4. Tumlinson, J. H., Minyard, J. P., Hedin, P. A., and Thompson, A. C. Reaction
chromatography. I. Gas-liquid/thin-layer chromatographic derivatization
technique for the identification of carbonyl compounds. J. Chromatogr.
29:80-87. 1967.
5. Minyard, J. P., Tumlinson, J. H., Thompson, A. C., and Hedin, P. A. II.
Gas-liquid/thin-layer chromatographic derivatization technique for the
identification of alcohols. J. Chromatogr. 29:88-93. 1967.
6. Tumlinson, J. H., Hardee, D. D., Minyard, J. P., Thompson, A. C., Gast, R. T.,
and Hedin, P. A. Boll weevil sex attractant: Isolation studies. J. Econ. Entomol.
61:470-474. 1968.
7. Cross, W. H., Hardee, D. D., Nichols, F., Mitchell, H. C., Mitchell, E. B.,
Huddleston, P. M., and Tumlinson, J. H. Attraction of female boll weevils to
1030 Wolf Prize in Agriculture
20. Yonce, C. E., Tumlinson, J. H., Gentry, C. R., and Mitchell, E. R. Extraction
and field bioassay of the sex pheromone of the lesser peachtree borer. Environ.
Entomol. 3(3):569-570. 1974.
21. Tumlinson, J. H., Yonce, C. E., Doolittle, R. E., Heath, R. R., Gentry, C. R., and
Mitchell, E. R. Sex pheromones and reproductive isolation of the lesser
peachtree borer and the peachtree borer. Science 185:614-616. 1974.
22. Tumlinson, J. H., Heath, R. R., and Doolittle, R. E. Application of chemical
ionization mass spectrometry of epoxides to the determination of olefin
position in aliphatic chains. Anal. Chem. 46:1309-1312. 1974.
23. Hendricks, D. E. and Tumlinson, J. H. A field cage bioassay system for testing
candidate sex pheromones of the tobacco budworm. Ann. Entomol. Soc. Am.
67(4):547-552. 1974.
24. Mitchell, E. R., Tumlinson, J. H., Copeland, W. W., Hines, R. W., and Brennan,
M. M. Tobacco budworm: Production, collection, and use of natural
pheromone in field traps. Environ. Entomol. 3(4):711-713. 1974.
25. McLaughlin, J. R., Mitchell, E. R., Chambers, D. L., and Tumlinson, J. H.
Perception of Z-7-dodecen-1-ol and modification of the sex pheromone
response of male loopers. Environ. Entomol. 3(4):677-680. 1974.
26. Gueldner, R. C., Tumlinson, J. H., III, Hardee, D. D., Hedin, P. A.,
Thompson, A. C., and Minyard, J. P. Boll Weevil Sex Attractant. U. S. Patent
Office #3,813,443, May 28, 1974. (Patent)
27. Tumlinson, J. H., III, Mitchell, E. R., Browner, S. M., Mayer, M. S., Green, N.,
Hines, R. W., and Lindquist, D. A. Method for Disrupting Pheromone
Communication with Cis-7-Dodecen-1-ol. United States Patent #3,832, 461,
Aug. 27, 1974. (Patent)
28. Tumlinson, J. H., Hendricks, D. E., Mitchell, E. R., Doolittle, R. E., and
Brennan, M. M. Isolation, identification, and synthesis of the sex pheromone
of the tobacco budworm. J. Chem. Ecol. 1(2):203-214. 1975.
29. Nielsen, D. G., Purrington, F. F., Tumlinson, J. H., Doolittle, R. E., and Yonce,
C. E. Response of male clearwing moths to caged virgin females, female
extracts, and synthetic sex attractants. Environ. Entomol. 4(3):451-454. 1975.
30. Heath, R. R., Tumlinson, J. H., Doolittle, R. E., and Proveaux, A. T. Silver
nitrate-high pressure liquid chromatography of geometrical isomers. J. Chrom.
Sci. 13:380-382. 1975.
31. McLaughlin, J. R., Mitchell, E. R., and Tumlinson, J. H. Evaluation of some
formulations for dispensing insect pheromones in field and orchard crops.
Proc. Int. Controlled Release Pesticide Symposium, Sept. 8-10, Wright State
University, Dayton, Ohio, pp 209-215. 1975. (Proceedings)
32. McLaughlin, John R., Doolittle, R. E., Gentry, C. R., Mitchell, E. R., and
Tumlinson, J. H. Response to pheromone traps and disruption of pheromone
communication in the lesser peachtree borer and the peachtree borer
(Lepidoptera: Sesiidae). J. Chem. Ecol. 2(1):73-81. 1976.
1032 Wolf Prize in Agriculture
46. Tumlinson, J. H., Klein, M. G., Doolittle, R. E., Ladd, T. L., and Proveaux, A. T.
Identification of the female Japanese beetle sex pheromone: Inhibition of
male response by an enantiomer. Science 197:789-792. 1977.
47. Tumlinson, J. H. Animal Communication by Pheromones. H. H. Shorey (ed.).
Academic Press, New York, 1976. (Book Review) Pan-Pacific Entomol.
53(3):180. 1977.
48. Barry, M. W., Nielsen, D. G., Purrington, F. F., and Tumlinson, J. H. Attractivity
of pheromone blends to male peachtree borer, Synanthedon exitiosa. Environ.
Entomol. 7(1):13. 1978.
49. Nielsen, D. G., Purrington, F. F., Campbell, R. L., Wilmot, T. R., Capizzi, J.,
and Tumlinson, J. H. Sex attractants for Sequoia pitch moth and strawberry
crown moth. Environ. Entomol. 7(4):544-546. 1978.
50. Sharp, J. L., McLaughlin, J. R., James, J., and Tumlinson, J. H. Seasonal
abundance of Synanthedon pictipes and S. exitiosa in North Central Florida.
Environ. Entomol. 7(4):589-591. 1978.
51. Voerman, S., Minks, A. K., Vanwetswinkel, G., and Tumlinson, J. H.
Attractivity of 3,13-octadecadien-1-ol acetates to the male clearwing moth
Synanthedon myopaeformis (Borkhausen) (Lepidoptera, Sesiidae). Ent. Exp.
& Appl. 23:301-304. 1978.
52. Mitchell, E. R., Tumlinson, J. H., and Baumhover, A. H. Heliothis virescens:
Attraction of males to blends of (Z )-9-tetradecen-1-ol formate and (Z )-9-
tetradecenal. J. Chem. Ecol. 4(6):709-716. 1978.
53. Heath, R. R., Proveaux, A. T., and Tumlinson, J. H. A simple terminator for
high efficiency liquid chromatography columns. J. HRC & CC 1(6):317-319.
1978.
54. Sharp, J. L., McLaughlin, J. R., James, J., Eichlin, T. D., and Tumlinson, J. H.
Seasonal occurrence of male Sesiidae in North Central Florida determined
with pheromone trapping methods. Fla. Entomol. 61(4):245-250. 1978.
55. Sales, F. M., Tumlinson, J. H., McLaughlin, J. R., and Sailer, R. I.
Comportamento do parasitoide. Trissolcus basalis (Wollaston) em resposta a
queromonios produzidos pelio hospedeiro, Nezara viridula (L.). Fitossanidade,
Fortaleza 2(3):88. 1978.
56. Sales, F. M., McLaughlin, J. R., Sailer, R. I., and Tumlinson, J. H. Temporal
analysis of the ovipositional behavior of the female egg parasitoid, Trissolcus
basalis (Wollaston). Fitossanidade, Fortaleza 2(3):80-83. 1978.
57. Tumlinson, J. H. Recent discoveries in insect pheromone chemistry,
pp. 315-322. In Geissbuhler, H. (Ed.), Advances In Pesticide Science. Pergamon
Press, Oxford & NY. 1979. (Book Chapter)
58. Tumlinson, J. H. The need for biological information in developing
strategies for applying semiochemicals, pp. 301-311. In Ritter, F. J. (Ed.),
Chemical Ecology: Odour Communication in Animals. Elsevier/North-Holland
Biomedical Press, The Netherlands. 1979.
1034 Wolf Prize in Agriculture
59. Gentry, C. R., Yonce, C. E., Blythe, J. L., and Tumlinson, J. H. Lesser peachtree
borer: Recovery of marked native males in pheromone baited traps. Environ.
Entomol. 8(2):218-220. 1979.
60. Heath, R. R., McLaughlin, J. R., Tumlinson, J. H., Ashley, T. R., and Doolittle,
R. E. Identification of the white peach scale sex pheromone, an illustration
of micro techniques. J. Chem. Ecol. 5(6):941-953. 1979.
61. Heath, R. R., Jordan, J. R., Sonnet, P. E., and Tumlinson, J. H. Potential for the
separation of insect pheromones by gas chromatography on columns coated
with cholesteryl cinnamate, a liquid-crystal phase. J. HRC & CC 2:712-714.
1979.
62. Tumlinson, J. H. The chemistry of Sesiidae pheromones, pp. 1-10. In
Pheromones of the Sesiidae (formerly Aegeriidae). USDA/SEA AAR-NE-6, 83
pp. 1979. (Proceedings)
63. Tumlinson, J. H., Klein, M. G., Doolittle, R. E., and Ladd, Jr., T. L. Sex Pheromone
Produced by the Female Japanese Beetle: Specificity of Male Response of
Enantiomers. U.S. Patent #4,179,446. 10/18/79. (Patent)
64. Chow, Y. S., Mayer, M. S., and Tumlinson, J. H. Electroantennogram response
of Plodia interpunctella to its sex pheromone and wing gland extracts. Bull.
Inst. Zool. 19(1):27-31. 1980.
65. Heath, R. R., Burnsed, G. E., Tumlinson, J. H., and Doolittle, R. E. Separation
of a series of positional and geometrical isomers of olefinic aliphatic primary
alcohols and acetates by capillary gas chromatography. J. Chromatogr.
189:199-208. 1980.
66. Doolittle, R. E., Tumlinson, J. H., Proveaux, A. T., and Heath, R. R. Synthesis of
the sex pheromone of the Japanese beetle. J. Chem. Ecol. 6(2):473-485. 1980.
67. Heath, R. R., Doolittle, R. E., Sonnet, P. E., and Tumlinson, J. H. Sex pheromone
of the white peach scale: Highly stereoselective synthesis of the stereoisomers
of pentagonol propionate. J. Org. Chem. 45(14): 2910-2912. 1980.
68. Cross, J. H., Tumlinson, J. H., Heath R. R., and Burnett, D. E. Apparatus and
procedure for measuring release rates from formulations of lepidopteran
semiochemicals. J. Chem. Ecol. 6(4):759-770. 1980.
69. Cross, J. H., Mitchell, E. R., Tumlinson, J. H., and Burnett, D. E. Selection of a
polyethylene tubing formulation of (Z )-9-tetradecen-1-ol formate and its
use in disrupting pheromone communication in Heliothis zea (Boddie).
J. Chem. Ecol. 6(4):771-779. 1980.
70. Vander Meer, R. K., Glancey, B. M., Lofgren, C. S., Glover, A., Tumlinson,
J. H., and Rocca, J. The poison sac of red imported fire ant queens: Source of
a pheromone attractant. Ann. Entomol. Soc. Am. 73(5): 609-612. 1980.
71. Tumlinson, J. H., Mitchell, E. R., and Sonnet, P. E. Sex pheromone
components of the beet armyworm, Spodoptera exigua. J. Environ. Sci. Health,
A16(2):189-200. 1981.
James H. Tumlinson 1035
72. Klein, M. G., Tumlinson, J. H., Ladd, Jr., T. L., and Doolittle, R. E. Japanese
beetle (Coleoptera: Scarabaeidae): Response to synthetic sex attractant plus
phenethyl propionate: Eugenol. J. Chem. Ecol. 7(1):1-7. 1981.
73. Tumlinson, J. H. Chemistry of the queen pheromone of the red imported
fire ant. Proc. Tall Timbers Conf. on Ecological Animal Control by Habitat
Management No. 7, pp. 155-161. 1981. (Proceedings)
74. Teal, P. E. A., McLaughlin, J. R., and Tumlinson, J. H. Analysis of the
reproductive behavior of Heliothis virescens (F.) under laboratory conditions.
Ann. Entomol. Soc. Am. 74(3):324-330. 1981.
75. Teal, P. E. A., Heath, R. R., Tumlinson, J. H., and McLaughlin, J. R. Identification
of a sex pheromone of Heliothis subflexa (Gn.) (Lepidoptera: Noctuidae) and
field trapping studies using different blends of components. J. Chem. Ecol.
7(6):1011-22. 1981.
76. Tumlinson, J. H., Teal, P. E. A., and Heath, R. R. Chemical ethology: A holistic
approach to the study of insect pheromones. Proc. lst Japan/USA Symp. on
IPM, Tsukuba, Japan, Sept. 29-30, 1981, pp. 19-31. 1981. (Proceedings)
77. Ladd, T. L., Klein, M. G., and Tumlinson, J. H. Phenethyl propionate + eugenol
+ geraniol (3:7:3) and japonilure: A highly effective joint lure for Japanese
beetles. J. Econ. Entomol. 74(6):665-667. 1981.
78. Johnson, D. W., Mitchell, E. R., Tumlinson, J. H., and Allen, G. E. Velvetbean
caterpillar: Response of males to virgin females and pheromone in the
laboratory and field. Fla. Entomol. 64(4):529-533. 1981.
79. Prokopy, R. J., Averill, A. L., Bardinelli, C. M., Bowdan, E. S., Cooley, S. S.,
Crnjar, R. M., Dundulis, E. A., Roitberg, C. A., Spatcher, P. J., Tumlinson, J. H.,
and Weeks, B. L. Site of production of an oviposition-deterring pheromone
component in Rhagoletis pomonella flies. J. Insect Physiol. 28(1):1-10. 1982.
80. Solomon, J. D., Oliveria, F. L., Tumlinson, J. H., and Doolittle, R. E. Occurrence
of clearwing borers (Sesiidae) in west central Mississippi. J. Ga. Entomol.
Soc. 17(1):4-12. 1982.
81. Guss, P. L., Tumlinson, J. H., Sonnet, P. E., and Proveaux, A. T. Identification
of a female-produced sex pheromone of the western corn rootworm.
J. Chem. Ecol. 8(2):545-556. 1982. (Book Chapter)
82. Tumlinson, J. H., Heath, R. R., and Teal, P. E. A. Analysis of chemical
communications systems of Lepidoptera, pp. 1-25. In Leonhardt, B. A., and
Beroza, M. (Eds.), Insect Pheromone Technology: Chemistry and Applications.
ACS Symp. Series No. 190. 1982.
83. Lewis, W. J., Nordlund, D. A., Gueldner, R. C., Teal, P. E. A., and Tumlinson,
J. H. Kairomones and their use for management of entomophagous insects.
XIII. Kairomonal activity for Trichogramma spp. of abdominal tips, excretion,
and a synthetic sex pheromone blend of Heliothis zea (Boddie) moths.
J. Chem. Ecol. 8(10):1323-1331. 1982.
1036 Wolf Prize in Agriculture
96. Guss, P. L., Tumlinson, J. H., Sonnet, P. E., and McLaughlin, J. R. Identification
of a female-produced sex pheromone from the southern corn rootworm,
Diabrotica undecimpunctata howardi Barber. J. Chem. Ecol. 9(9):1363-1375.
1983.
97. Teal, P. E. A., Tumlinson, J. H., McLaughlin, J. R., Heath, R. R., and Rush, R. A.
(Z)-11-Hexadecen-1-ol: A behavioral modifying chemical present in the
pheromone gland of female Heliothis zea (Lepidoptera: Noctuidae). J. Can.
Ent. 116:777-779. 1984.
98. Guss, P. L., Sonnet, P. E., Carney, R. L., Branson, T. F., and Tumlinson, J. H.
Response of Diabrotica virgifera, D. v. zeae, and D. porracea to stereoisomers
of 8-methyl-2-decyl propanoate. J. Chem. Ecol. 10(7):1123-1131. 1984.
99. Heath, R. R., and Tumlinson, J. H. Techniques for purifying, analyzing, and
identifying pheromones, pp. 287-322. In Hummel, H. E. and Miller, T. A.
(Eds.) Techniques in Pheromone Research, Springer-Verlag, New York, NY.
1984. (Book Chapter)
100. Guss, Paul L., Tumlinson, James H., III, Sonnet, Philip E., and McLaughlin,
John R. Synthetic pheromone 10-methyl-2-tridecanone and its use in
controlling the southern corn rootworm and related diabroticites. P.C. 6918,
U.S. Patent No. 4,474,991, October 2, 1984. Also: P.C. N6931, U.S. Patent
No. 4,565,695, January 21, 1986. (Patent)
101. Glancey, B. M., Rocca, J., Lofgren, C. S., and Tumlinson, J. Field tests with
synthetic components of the queen recognition pheromone of the red
imported fire ant, Solenopsis invicta Buren. Sociobiology 9(1):19-30. 1984.
102. Guss, P. L., Sonnet, P. E., Carney, R. L., Tumlinson, J. H., and Wilkin, P. J.
Response of northern corn rootworm, Diabrotica barberi Smith and
Lawrence, to stereoisomers of 8-methyl-2-decyl propanoate. J. Chem. Ecol.
11(1):21-26. 1985.
103. Tumlinson, J. H. The beetles: pheromonal chemists par excellence,
pp. 367-380. In Paul A. Hedin (ed.), Bioregulators for Pest Control. ACS
Symposium Series 276. ACS, Washington, DC. 1985. (Book Chapter)
104. Teal, P. E. A., Mitchell, E. R., Tumlinson, J. H., Heath, R. R., and Sugie, H.
Identification of volatile sex pheromone components released by the southern
armyworm, Spodoptera eridania (Cramer). J. Chem. Ecol. 11(6):717-725.
1985.
105. Snow, J. Wendell, Eichlin, T. D., and Tumlinson, J. H. Seasonal captures
of clearwing moths (Sesiidae) in traps baited with various formulations
of 3,13-octadecadienyl acetate and alcohol. J. Agric. Entomol. 2(1):73-84.
1985.
106. Doolittle, R. E., Tumlinson, J. H., and Proveaux, A. T. Determination of double
bond position in conjugated dienes by chemical ionization mass spectrometry
with isobutane. Anal. Chem. 57(8):1625-1630. 1985.
1038 Wolf Prize in Agriculture
107. Mitchell, Everett R., J. H. Tumlinson, and Jeremy N. McNeil. Field evaluation
of commercial pheromone formulations and traps using a more effective
sex pheromone blend for the fall armyworm (Lepidoptera: Noctuidae).
J. Econ. Entomol. 78:1364-1369. 1985.
108. Teal, P. E. A., Tumlinson, J. H., and Heath, R. R. Chemical and behavioral
analyses of volatile sex pheromone components released by calling
Heliothis virescens (F.) females (Lepidoptera: Noctuidae). J. Chem. Ecol.
12(1):107-126. 1986.
109. Tumlinson, J. H., Teal, P. E. A., Mitchell, E. R., and Heath, R. R. Sex pheromones
of economically important North American Spodoptera species. Int. Conf. on
Integr. Plant Prot., Budapest, Hungary, July 4-9, 1983. Proc. Int. Conf. on
Integr. Plant Prot. Vol. 4:140-146. 1986. (Proceedings)
110. Teal, P. E. A., and Tumlinson, J. H. Terminal steps in pheromone bio-synthesis
by Heliothis virescens and H. zea. J. Chem. Ecol. 12(2):353-366. 1986.
111. Heath, R. R., Coffelt, J. A., Sonnet, P. E., Proshold, F. I., Dueben, B. D., and
Tumlinson, J. H. Identification of a sex pheromone produced by female
sweetpotato weevil. J. Chem. Ecol. 12(6):1489-1503. 1986.
112. Krysan, J. L., Wilkin, P. H., Tumlinson, J. H., Sonnet, P. E., Carney, R. L., and
Guss, P. L. Responses of Diabrotica lemniscata and D. longicornis (Coleoptera:
Chrysomelidae) to stereoisomers of 8-methyl-2- decanol propanoate and
studies on the pheromone of D. longicornis. Ann. Entomol. Soc. Am.
79(4):742-746. 1986.
113. Tumlinson, J. H., Mitchell, E. R., Teal, P. E. A., Heath, R. R., and Mengelkoch,
L. J. Sex pheromone of the fall armyworm, Spodoptera frugiperda
( J. E. Smith): Identification of components critical to attraction in the field.
J. Chem. Ecol. 12(9):1909-1926. 1986.
114. Schneiderman, A. M., Hildebrand, J. G., Brennan, M. M., and Tumlinson,
J. H. Trans-sexually grafted antennae alter pheromone-directed behavior in
a moth. Nature 323:801-803. 1986.
115. Heath, R. R., and Tumlinson, J. H. Correlation of retention times on a liquid
crystal capillary column with reported vapor pressures and half-lives of
compounds used in pheromone formulations. J. Chem. Ecol. 12(11):2081-
2088. 1986.
116. Heath, R. R., Teal, P. E. A., Tumlinson, J. H., and Mengelkoch, L. J. Prediction
of release ratios of multicomponent pheromones from rubber septa.
J. Chem. Ecol. 12(12):2133-2143. 1986.
117. Szöcs, G., Tóth, M., Bestmann, H. J., Vostrowsky, O., Heath, R. R., and
Tumlinson, J. H. Polyenic hydrocarbons as sex attractants for geometrids and
amatids (Lepidoptera) found by field screening in Hungary. Z. Naturforsch
42c:165-168. 1987.
James H. Tumlinson 1039
118. Chuman, T., Guss, P. L., Doolittle, R. E., McLaughlin, J. R., Krysan, J. L.,
Schalk, J. M. and Tumlinson, J. H. Identification of a female-produced sex
pheromone from banded cucumber beetle, Diabrotica balteata LeConte
(Coleoptera: Chrysomelidae). J. Chem. Ecol. 13(7):1601-1616. 1987.
119. Teal, P. E. A., and Tumlinson, J. H. Induced changes in sex pheromone
biosynthesis of Heliothis moths (Lepidoptera: Noctuidae), pp. 79-94.
In Law, J. (ed). Molecular Entomology. Alan R. Liss, Inc., New York. 1987.
(Book Chapter)
120. Teal, P. E. A. and Tumlinson, J. H. The role of alcohols in pheromone
biosynthesis by two noctuid moths that use acetate pheromone components.
Archiv. Insect Biochem. & Physiol. 4:261-269. 1987.
121. Stowe, M., Tumlinson, J. H., and Heath, R. R. Chemical Mimicry: Bolas
spiders emit components of moth prey species sex pheromone. Science.
236:964-967. 1987.
122. Chuman, T., Landolt, P. J., Heath, R. R., and Tumlinson, J. H. Isolation,
identification and synthesis of a male-produced sex pheromone of the papaya
fruit fly, Toxotrypana curvicauda Gerstaecker (Diptera: Tephritidae). J. Chem.
Ecol. 13(9):1979-1992. 1987.
123. Tumlinson, J. H. and Teal, P. E. A. Relationship of structure and function
to biochemistry in insect pheromone systems. pp. 3-26. In Prestwich,
G. D. and Blomquist, G. J. (eds), Pheromone Biochemistry, Academic Press.
1987. (Book Chapter)
124. Sonnet, P. E., Guss, P. L., Tumlinson, J. H., McGovern, T. P. and Cunningham,
R. T. Asymmetric synthesis of selected insect pheromones, pp. 388-400. In
Synthesis and Chemistry of Agrochemicals, ACS Symposium Series 355,
Washington, D. C. 1987. (Book Chapter)
125. Noldus, L. P. J. J., Lewis, W. J., Tumlinson, J. H., and van Lenteren, J. C.
Olfactometer and wind tunnel experiments on the role of sex pheromones
of noctuid moths in the foraging behaviour of Trichogramma spp. Proc. of
2nd Int. Sym. Trichogramma and Other Egg Parasites. Guangzhou, China,
Nov. 10-15, 1986. Ed. INRA, Paris, (Les Colloques de l’INRA no. 43),
pp. 223-238. 1988. (Proceedings)
126. Eller, F. J., Tumlinson, J. H. and Lewis, W. J. Beneficial arthropod behavior
mediated by airborne semiochemicals. II. Evidence for the use of volatile
attractants by the parasitoid Microplitis croceipes (Cresson) (Hymenoptera:
Braconidae). J. Chem. Ecol. 14(2):425-434. 1988.
127. Lewis, W. J. and Tumlinson, J. H. Host detection by chemically mediated
associative learning in a parasitic wasp. Nature. 331:257-259. 1988.
128. Tumlinson, J. H. Insect pheromone systems. Comments Agric. and Food
Chemistry. 1(3):115-146. 1988.
1040 Wolf Prize in Agriculture
129. Drost, Y. C., Lewis, W. J., and Tumlinson, J. H. Beneficial arthropod behavior
mediated by airborne semiochemicals. V. Influence of rearing method, host
plant, and adult experience on host-searching behavior of Microplitis
croceipes (Cresson), a larval parasitoid of Heliothis. J. Chem. Ecol. 14:1607-
1616. 1988.
130. Heath, R. R., Coffelt, J. A., Proshold, F. I., Sonnet, P. E., and Tumlinson,
J. H., III. (Z)-3-Dodecen-1-ol (E)-2-butenoate and its use in monitoring
the sweetpotato weevil. U. S. Patent No. 4,732,756, March 22, 1988.
(Patent)
131. Guss, P. L., Tumlinson, J. H., III, Sonnet, P. E., and Proveaux, A. T. Synthetic
pheromone 8-methyl-2-decanol propanoate. U. S. Patent No. 4,732,524,
March 29, 1988. (Patent)
132. Herard, F., Keller, M. A., Lewis, W. J., and Tumlinson, J. H. Beneficial arthropod
behavior mediated by airborne semiochemicals. III. Influence of age and
experience on flight chamber responses of Microplitis demolitor Wilkinson.
J. Chem. Ecol. 14:1583-1596. 1988.
133. Herard, F., Keller, M. A., Lewis, W. J., and Tumlinson, J. H. Beneficial arthropod
behavior mediated by airborne semiochemicals. IV. Influence of host diet on
host-oriented flight chamber responses of Microplitis demolitor Wilkinson.
J. Chem. Ecol. 14:1597-1606. 1988.
134. Landolt, P. J., Heath, R. R., Agee, H. R., Tumlinson, J. H., and Calkins, C. O. A
sex pheromone-based trapping system for the papaya fruit fly, Toxotrypana
curvicauda Gerstaecker (Diptera: Tephritidae). J. Econ. Entomol. 81:1163-
1169. 1988.
135. Mitchell, E. R., Heath, R. R., Tumlinson, J. H. WORT: Wind-oriented trap
for evaluating several pheromone blends simultaneously. J. Econ. Entomol.
81:966-969. 1988.
136. Eller, F. J., Tumlinson, J. H., and Lewis, W. J. Beneficial arthropod behavior
mediated by airborne semiochemicals: Source of volatiles mediating the host-
location flight behavior of Microplitis croceipes (Cresson) (Hymenoptera:
Braconidae), a Parasitoid of Heliothis zea (Boddie) (Lepidoptera: Noctuidae).
Environ. Entomol. 17:745-753. 1988.
137. Eller, F. J., Boucias, D. G. and Tumlinson, J. H. Interactions between Microplitis
croceipes (Cresson) (Hymenoptera: Braconidae) and a nuclear polyhedrosis
virus of Heliothis zea (Boddie) (Lepidoptera: Noctuidae). Environ. Entomol.
17(6):977-982. 1988.
138. Tumlinson, J. H. Contemporary frontiers in insect semiochemical research.
J. Chem. Ecol. 14(11):2109-2130. 1988.
139. Teal, P. E. A. and Tumlinson, J. H. Properties of cuticular oxidases used for
sex pheromone biosynthesis by Heliothis zea. J. Chem. Ecol. 14(11):2131-
2145. 1988.
James H. Tumlinson 1041
140. Chuman, T., Sivinski, J., Heath, R. R., Calkins, C. O., and Tumlinson, J. H.,
Battiste, M. A., Wydra, R. L., Strekowski, L., and Nation, J. L. Suspensolide, a
new macrolide component of male Caribbean fruit fly (Anastrepha suspensa
(Loew)) volatiles. Tetrahedron Lett. 29(50):6561-6564. 1988.
141. Battiste, M. A., Rocca, J. R., Wydra, R. L., Tumlinson, J. H., and Chuman, T.
Total synthesis and structure proof of (3E,8E)-Suspensolide. Tetrahedron
Lett. 29(50):6565-6568. 1988.
142. Prokopy, R. J., Powers, P. J., Heath, R. R., Dueben, B. D., and Tumlinson,
J. H. Comparative laboratory methods for assaying behavioral responses
of Rhagoletis pomonella flies to host marking pheromone. J. Appl. Ent.
106:437-443. 1988.
143. Kovalev, B. G., Tumlinson, J. H., Bolgar, T. S., Kolomoets, T. P., Kovalenko,
V. M., and Heath. R. R. Isolation and identification of a component of the sex
pheromone of Aegeria apiformis. Translated from Khimiya Prirodnykh
Soedinenii, No. 1, 125-128. 1988.
144. Teal, P. E. A. and Tumlinson, J. H. Isolation, identification and biosynthesis of
compounds produced by male hairpencil glands of Heliothis virescens (F.)
J. Chem. Ecol. 15(1):413-427. 1989.
145. Tumlinson, J. H. Insect chemical communication systems. Pure & Appl. Chem.
61(3):559-562. 1989. (Proceedings)
146. Teal, P. E. A. and Tumlinson, J. H. Neurohormonal induction of pheromone
biosynthesis by Heliothis zea (Boddie) during the photophase. Can. Entomol.
121:43-46. 1989.
147. Nordlund, D. A., Lewis, W. J., and Tumlinson, J. H. Habitat and host location
behavior of Microplitis croceipes. Southwestern Ent. Suppl. 12:39-48. 1989.
(Proceedings)
148. Teal, P. E. A., Tumlinson, J. H., and Oostendorp. A. Enzyme-catalyzed
pheromone synthesis by Heliothis moths, pp. 332-343. In Whitaker, J. R. and
Sonnet, P. E. (Eds.) Biocatalysis in Agricultural Biotechnology, Am. Chem.
Soc. 1989. (Book Chapter)
149. Teal, P. E. A., Tumlinson, J. H. and Oberlander, H. Neural regulation of sex
pheromone biosynthesis in Heliothis moths. Proc. Natl. Acad. Sci. USA.
86:2488-2492. 1989.
150. Turlings, T. C. J., Tumlinson, J. H., Lewis, W. J. and Vet, L. E. M. Beneficial
arthropod behavior mediated by airborne semiochemicals. VIII. Learning of
host related odors induced by a brief contact experience with host by-products
in Cotesia marginiventris (Cresson), a generalist larval parasitoid. J. Insect
Behav. 2:217-225. 1989.
151. Landolt, P. J., Tumlinson, J. H., and Brennan, M. M. Attraction of
Amphion floridensis (Lepidoptera: Sphingidae) to bombykal (E,Z)-10,12-
hexadecadienal. Fla. Entomol. 72(2):324-327. 1989.
1042 Wolf Prize in Agriculture
152. Tumlinson, J. H., Brennan, M. M., Doolittle, R. E., Mitchell, E. R., Brabham,
A., Mazomenos, B. E., Baumhover, A., and Jackson, D. M. Identification of a
pheromone blend attractive to Manduca sexta (L.) males in a wind tunnel.
Archiv. Insect Biochem. & Physiol. 10:255-271. 1989.
153. Kaissling, K. E., Hildebrand, J. G., and Tumlinson, J. H. Pheromone receptor
cells in the male moth Manduca sexta. Arch. Insect Biochem. & Physiol.
10:273-279. 1989.
154. Christensen, T. A., Hildebrand, J. G., Tumlinson, J. H. and Doolittle, R. E. The
sex pheromone blend of Manduca sexta: Responses of central olfactory
interneurons to antennal stimulation in male moths. Arch. Insect. Biochem.
& Physiol. 10:281-291. 1989.
155. Tumlinson, J. H. Chemical analysis and identification of insect pheromones,
pp. 73-92. In Ridgway, R. L., Silverstein, R. M., and Inscoe, M. N. (Eds.),
Behavior-Modifying Chemicals for Insect Management, Applications of
Pheromones and Other Attractants. Marcel Dekker, Inc., New York and Basel.
1990. (Book Chapter)
156. Chuman, T., Guss, P. L., Doolittle, R. E., McLaughlin, J. R., and Tumlinson,
J. H. 6,12-Dimethylpentadecan-2-one and its use in monitoring and
controlling the banded cucumber beetle. U. S. Patent # 4,871,537, October
3, 1989. (Patent)
157. Krysan, J. L., McDonald, I. C., and Tumlinson, J. H. Phenogram based on
allozymes and its relationship to classical biosystematics and pheromone
structure among eleven diabroticites (Coleoptera: Chrysomelidae). Ann.
Entomol. Soc. Am. 82(5):574-581. 1989.
158. Turlings, T. C. J., Scheepmaker, J. W. A., Vet, L. E. M., Tumlinson, J. H., and
Lewis, W. J. How contact foraging experiences affect the preferences for
host-related odors in the larval parasitoid Cotesia marginiventris (Cresson)
(Hymenoptera: Braconidae). J. Chem. Ecol. 16:1577-1589. 1990.
159. Doolittle, R. E., Brabham, A., and Tumlinson, J. H. Sex pheromone of Manduca
sexta (L.): Stereoselective synthesis of (10E,12E,14Z )-10,12,14-
hexadecatrienal and isomers. J. Chem. Ecol. 16:1131-1153. 1990.
160. Domek, J. M., Tumlinson, J. H., and Johnson, D. T. Responses of Male Green
June Beetles Continis nitida (L.) (Coleoptera:Scarabaeidae) to Female Volatiles
in a Flight Tunnel. J. Insect Behavior. 3(2):271-276. 1990.
161. Eller, F. J., Tumlinson, J.H., and Lewis, W. J. Intraspecific competition
in Microplitis croceipes (Cresson) (Hymenoptera:Braconidae), a parasitoid
of Heliothis species (Lepidoptera:Noctuidae). Ann. Entomol. Soc. Am.
83(3):504-508. 1990.
162. Heath, R. R., Chambers, D. L., Tumlinson, J. H., and Landolt, P. J. Controlled
release of Trimedlure Isomer C from a compressed disk and the evaluation
of its attractiveness to the Mediterranean Fruit Fly (Diptera:Tephritidae).
J. Econ. Entomol. 83:(3):819-822. 1990.
James H. Tumlinson 1043
163. Schalk, J. J., McLaughlin, J. R., and Tumlinson, J. H. Field response of feral male
banded cucumber beetles to the sex pheromone. 6,12-Dimethylpentadecan-
2-one. Fla. Entomol. 1990. 73(2):271-276. 1990.
164. Turlings, T. C. J., Tumlinson, J. H., and Lewis, W. J. Exploitation of
Herbivore-Induced Plant Odors by Host-Seeking Parasitic Wasps. Science.
250:1251-1253. 1990.
165. Lewis, W. J., Vet, Louise E. M., Tumlinson, J. H., van Lenteren, J. C., and
Papaj, D. R. Variations in parasitoid behavior essential element of a sound
biological control theory. Environ. Entomol. 19(5):1183-1193. 1990.
166. Teal, P. E. A., Tumlinson, J. H., and Oberlander, H. Endogenous suppression of
pheromone production in virgin female moths. Experientia. 46:1047-1050.
1990.
167. Tumlinson, J. H., Mitchell, E. R., and Yu, H. S. Analysis and field evaluation
of the volatile blend emitted by calling virgin females of the beet armyworm
moth, Spodoptera exigua (Hübner). J. Chem. Ecol. 16(12):3411-3424. 1990.
168. Tumlinson, J. H. and Teal, P.E.A. Endogenous regulation of pheromone
biosynthesis in Heliothis moths, pp. 243-255. In Hagedorn, H. H., Hildebrand,
J. G., Kidwell, M., and Law, J. H. (Eds.), Molecular Insect Science. Plenum
Press. New York. 1990. (Book Chapter)
169. Noldus, L.P.J.J., Lewis, W. J., and Tumlinson, J. H. Beneficial Arthropod
Behavior Mediated by Airborne Semiochemicals. IX. Differential Response of
Trichogramma pretiosum, an Egg Parasitoid of Heliothis zea, to Various
Olfactory Cues. J. Chem. Ecol. 16(12):3531-3544. 1990.
170. Doolittle, R. E., Tumlinson, J. H., Brabham, A., Brennan, M. M., and
Mitchell, E. R. Sex Pheromone Blend of the Tobacco Hornworm. Identification
and Stereoselective Synthesis, pp. 491-503. In Baker, D.R., Fenyes, J. G., and
Moberg, W. K. (Eds.), Synthesis and Chemistry of Agrochemicals II, ACS
Symposium Series No. 443. Washington, DC. American Chemical Society.
1991. (Book Chapter)
171. Teal, P. E. A., and Tumlinson, J. H. Sex pheromone production in moths,
endogenous regulation of initiation and inhibition, pp. 66-77. In Hedin, P.A.,
(Ed.), Naturally Occurring Pest Bioregulators. ACS Symposium Series No.
449, Washington, DC. American Chemical Society. 1991. (Book Chapter)
172. Turlings, T. C. J., Tumlinson, J. H., Eller, F., and Lewis, W. J. Larvae-damaged
Plants: Source of volatile synomones that guide the parasitoid Cotesia
marginiventris to the microhabitat of its hosts. Entomol. exp. appl. 58:75-82.
1991.
173. McLaughlin, J. R., Tumlinson, J. H., and Mori, K. Responses of male Diabrotica
balteata (Coleoptera: Chrysomelidae) to stereoisomers of the sex pheromone
6, 12-dimethylpentadecan-2-one. J. Econ. Entomol. 84(1):99-102. 1991.
174. Turlings, T. C. J. and Tumlinson, J. H. Do parasitoids use herbivore-induced
plant chemical defenses to locate hosts? Fla. Entomol. 74(1):42-50. 1991.
1044 Wolf Prize in Agriculture
175. Lewis, W. J., Tumlinson, J. H., and Krasnoff, S. Chemically Mediated Associative
Learning; an Important Function in the Foraging Behavior of Microplitis
croceipes (Cresson). J. Chem. Ecol. 17(7):1309-1325. 1991.
176. Christensen, T. A., Itagaki, H., Teal, P. E. A., Jasensky, R. D., Tumlinson, J. H.
and Hildebrand. J. G. Innervation and neural regulation of the sex pheromone
gland in female Heliothis moths. Proc. Natl. Acad. Sci. USA. 88:4971-4975.
1991.
177. Heath, R. R., Landolt, P. J., Tumlinson, J. H., Chambers, D. L., Murphy, R.,
Doolittle, R. E., Dueben, B. D., Sivinski, J. J., and Calkins, C. O. Analysis,
synthesis, and formulation of sex pheromone components of the male
Mediterranean fruit flies attractive to females. J. Chem. Ecol. 17(9):1925-
1940. 1991.
178. Turlings, T. C. J., Tumlinson, J. H., Heath, R. R., Proveaux, A. T., and
Doolittle, R. E. Isolation and identification of allelochemicals that attract the
larval parasitoid, Cotesia marginiventris (Cresson), to the micro-habitat of
one of its hosts. J. Chem. Ecol. 17:2235-2251. 1991.
179. Tumlinson, J. H. and Lewis, W. J. Chemically Mediated Foraging Behavior of
Parasitoids: New Implications for Biocontrol, pp. 161-178, In: Vinson, S. B.,
and Metcalf, R. L. (Eds.), Entomology Serving Society: Emerging Technologies
and Challenges, Entomological Society of America, Lanham, Maryland.
Symposium held Dec. 12-13, 1989, San Antonio, Texas. 1991. (Proceedings)
180. Teal, P. E. A., and Tumlinson, J. H. Lipidic pheromones. In Current Opinion
in Structural Biology. Vol. 2, pp. 475-481. 1992. (Book Chapter)
181. Turlings, T. C. J., and Tumlinson, J. H. Systemic release of chemical signals by
herbivore-injured corn. Proc. Natl. Acad. Sci. USA. 89:8399-8402. 1992.
182. Fang, N., Tumlinson, J. H., Teal, P. E. A., and Oberlander, H. Fatty acyl
pheromone precursors in the sex pheromone gland of female tobacco
hornworm moths, Manduca sexta (L.). Insect Biochem. Molec. Biol. 22(7):621-
631. 1992.
183. Eller, F. J., Tumlinson, J. H., and Lewis, W. J. Effect of host diet and preflight
experience on the flight response of Microplitis croceipes (Cresson). Physiol.
Entomol. 17:235-240. 1992.
184. Tumlinson, J. H., Turlings, T. C. J., and Lewis, W. J. The semiochemical
complexes that mediate insect parasitoid foraging. Agric. Zool. Reviews,
Vol. 5:221-252. 1992.
185. Christensen, T. A., Lehman, H. K., Teal, P. E. A., Itagaki, H., Tumlinson,
J. H., and J. G. Hildebrand. Diel changes in the presence and physiological
actions of octopamine in the female sex-pheromone glands of heliothine
moths. Insect Biochem. Molec. Biol. 22(8):841-849. 1992.
186. Turlings, T. C. J., McCall, P. J., Alborn, H. T., and Tumlinson, J. H. Systemic
releases of volatiles by herbivore-damaged plants: what possible functions?
pp. 365-366. In Menken, S. B. J., Visser, J. H., and Harrewijn, P. (Eds.),
James H. Tumlinson 1045
Insect-Plant Relationships, Proc. 8th Int. Symp. Dordrecht: Kluwer Acad. Publ.,
1992. (Proceedings)
187. Turlings, T. C. J., Wäckers, F., Vet, L. E. M., Lewis, W. J. and Tumlinson, J. H.
Learning of host-finding cues by hymenopterous parasitoids, pp. 51-78. In
Lewis, A. C. and Papaj, D. R. (Eds.), Insect Learning: Ecological and Evolutionary
Perspectives, Chapman and Hall, New York. 1993. (Book Chapter)
188. Tumlinson, J. H., Lewis, W. J., and Vet, L. E. M. How parasitic wasps find
their hosts. Scientific American. 268(3):100-106. 1993.
189. Tumlinson, J. H., Turlings, T. C. J., and Lewis, W. J. Semiochemically medicated
foraging behavior in beneficial parasitic insects. Arch. Insect Biochem. Physiol.
22(3/4):385-391. 1993.
190. Turlings, T. C. J., McCall, P. J., Alborn, H. T., and Tumlinson, J. H. An elicitor
in caterpillar oral secretions that induces corn seedlings to emit volatiles
attractive to parasitic wasps. J. Chem. Ecol. 19(3):411-425. 1993.
191. Teal, P. E. A., Oostendorp, A. and Tumlinson, J. H. Induction of pheromone
production in females of Heliothis virescens (F.) and H. subflexa (Gn.)
(Lepidoptera: Noctuidae) during the photophase. Can. Entomol. 125:
355-366. 1993.
192. McCall, P. J., Turlings, T. C. J., Lewis, W. J., and Tumlinson, J. H. Role
of plant volatiles in host location by the specialist parasitoid Microplitis
croceipes (Cresson) (Braconidae: Hymenoptera). J. Insect Beh. 6(5):625-639.
1993.
193. Mitchell, E. R., and Tumlinson, J. H. Response of Spodoptera exigua and
S. eridania (Lepidoptera: Noctuidae) males to synthetic pheromone and
S. exigua females. Fla. Entomol. 77(2):237-247. 1994.
194. Tumlinson, J. H., Mitchell, E. R., Doolittle, R. E. and Jackson, D. M. Field tests
of Synthetic Manduca sexta sex pheromone. J. Chem. Ecol. 20(3):579-591.
1994.
195. Abernathy, R. L., Teal, P.E.A., and Tumlinson, J. H. Age and crowding affects
the amount of sex pheromone and the oviposition rates of virgin and mated
females of Helicoverpa zea (Lepidoptera: Noctuidae). Ann. Entomol. Soc. Am.
87(3):350-354. 1994.
196. Loughrin, J. H., Manukian, A., Heath, R. R., Turlings, T. C. J., and Tumlinson,
J. H. Diurnal cycle of emission of induced volatile terpenoids by herbivore-
injured cotton plants. Proc. Natl. Acad. Sci. USA. 91:11836-11840. 1994.
197. McCall, P. J., Turlings, T. C. J., Loughrin, J. H., Proveaux, A. T., and Tumlinson,
J. H. Herbivore-induced volatile emissions from cotton (Gossypium hirsutum
L.) Seedlings. J. Chem. Ecol. 20(12):3039-3050. 1994.
198. Lewis, W. J., Sheehan, W., and Tumlinson, J. H. Unraveling the story of how
parasitoids find their hosts, pp. 671-680. In Rosen, D., Bennett, F. D. and J. L.
Capinera (Eds.), Pest Management in the Subtropics: Biological Control — a
Florida Perspective, Intercept Limited, Andover. 1994. (Book Chapter)
1046 Wolf Prize in Agriculture
199. Stowe, M. K., Turlings, T. C., Loughrin, J. H., Lewis, W. J., and Tumlinson,
J. H. The chemistry of eavesdropping, alarm, and deceit. Proc. Natl. Acad.
Sci. USA. 92:23-28. 1995. (Proceedings)
200. Fang, N., Teal, P. E. A., Doolittle, R. E., and Tumlinson, J. H. Biosynthesis
of conjugated olefinic systems in the sex pheromone gland of female
tobacco hornworm moths, Manduca sexta (L.). Insect Biochem. Molec. Biol.
25(1):39-48. 1995.
201. Abernathy, R. L., Nachman, R. J., Teal, P. E. A., Yamashita, O., and Tumlinson,
J. H. Pheromonotropic activity of naturally occurring pyrokinin insect
neuropeptides (FxPRLamide) in Helicoverpa zea. Peptides. 16(2):215-129.
1995
202. Fang. N., Teal, P. E. A., and Tumlinson, J. H. PBAN regulation of pheromone
biosynthesis in female Tobacco Hornworm moths, Manduca sexta (L.). Arch.
Insect Biochem. Physiol. 29:35-44. 1995.
203. Turlings, T. C. J., Loughrin, J. H., McCall, P. J., Röse, U., Lewis, W. J., and
Tumlinson, J. H. How caterpillar-damaged plants protect themselves by
attracting parasite wasps. Proc. Nat. Acad. Sci. USA. 92:4169-4174. 1995.
(Proceedings)
204. Fang, N., Teal, P. E. A., and Tumlinson, J. H. Characterization of oxidase(s)
associated with the sex pheromone gland in Manduca sexta (L.) females.
Arch. Insect Biochem. & Physiol. 29:243-257. 1995.
205. Loughrin, J. H., Manukian, A., Heath, R. R., and Tumlinson, J. H. Volatiles
emitted by different cotton varieties damaged by feeding beet armyworm
larvae. J. Chem. Ecol. 21(8):1217-1227. 1995.
206. Berg, B. G., Tumlinson, J. H., and Mustaparta, H. Chemical communication
in Heliothine moths. IV. Receptor neuron responses to pheromone compounds
and formate analogues in the male tobacco budworm moth Heliothis virescens.
J. Comp. Physiol. A. 177:527-534. 1995.
207. Fang, N., Teal, P. E. A., and Tumlinson, J. H. Correlation between glycerolipids
and pheromone aldehydes in the sex pheromone gland of female tobacco
hornworm moths, Manduca sexta (L.). Arch. Insect Biochem. & Physiol.
30:321-336. 1995.
208. Alborn, H. T., Lewis, W. J., and Tumlinson, J. H. Host-specific recognition
kairomone for the parasitoid Microplitis croceipes (CRESSON). J. Chem. Ecol.
21(11):1697-1708. 1995.
209. Teal, P. E. A., Abernathy, R. L., Nachman, R. J., Fang, N., Meredith, J. A., and
Tumlinson, J. H. Pheromone biosynthesis activating neuropeptides: Functions
and chemistry. Peptides 17(2):337-344. 1996.
210. Tumlinson, J. H., Teal, P. E. A., and Fang, N. The integral role of triacyl
glycerols in the biosynthesis of the aldehydic sex pheromones of Manduca
sexta (L.). Bioorganic and Medicinal Chemistry. 4(3):452-460. 1996.
James H. Tumlinson 1047
211. Gazit, Y., Lewis, W. J., and Tumlinson, J. H. Arrestment of Telenomous remus
(Hymenoptera: Scelionidae) by a kairomone associated with eggs of its host,
Spodoptera frugiperda (Lepidoptera: Noctuidae). Biol. Control. 6:283-290.
1996.
212. Teal, P. E. A. and Tumlinson, J. H. Effects of interspecific hybridization between
Heliothis virescens and H. zea on the sex pheromone communication system,
pp 535-547. In Carde, R. T. and Minks, A. K. (eds.) Insect Pheromone Research:
New Directions. Proceedings of the First International Symposium on Insect
Pheromones. Chapman & Hall, New York. 1997. (Proceedings) (Book chapter).
213. Paré, P. W. and Tumlinson, J. H. Plant volatile signals in response to herbivore
feeding. Fla. Entomol. 79(2):93-103. 1996.
214. Fang, N., Teal, P. E. A., and Tumlinson, J. H. Effects of decapitation and PBAN
injection on amounts of triacylglycerols in the sex pheromone gland of
Manduca sexta (L.). Arch. Insect Biochem. Physiol. 32:249-260. 1996.
215. Röse, U. S. R., Manukian, A., Heath, R. R., and Tumlinson, J. H. Volatile
semiochemicals released from undamaged cotton leaves: a systemic response
of living plants to caterpillar damage. Plant Physiology. 111:487-495. 1996.
216. Heath, R. R., Tumlinson, J. H. and Landolt, P. J. Interfacing analytical chemistry
with IPM - the Florida experience pp. 167-181. In Rosen, D., Bennett, F. D.
and Capinera, J. L. (Eds.) Pest Management in the Subtropics: Integrated Pest
Management - A Florida Perspective. Intercept, Andover, Hants., U.K. 1996.
(Book Chapter)
217. Paré, P. W. and Tumlinson, J. H. Induced synthesis of plant volatiles. Nature.
385:30-31. 1997.
218. Alborn, H. T., Turlings, T. C. J., Jones, T. H., Stenhagen, G., Loughrin, J. H.,
and Tumlinson, J. H. An elicitor of plant volatiles from beet armyworm oral
secretion. Science. 276:945-949. 1997.
219. Stapel, J. O., Cortesero, A. M., De Moraes, C. M., Tumlinson, J. H., and
Lewis, W. J. Extrafloral nectar, honeydew, and sucrose effects on searching
behavior and efficiency of Microplitis croceipes (Hymenoptera: Braconidae)
in cotton. Environ. Entomol. 26(3):617-623. 1997.
220. Cortesero, A.M., De Moraes, C. M., Stapel, J. O., Tumlinson, J. H., and
Lewis, W. J. Comparisons and contrasts in host foraging strategies of two
larval parasitoids with different degrees of host specificity. J. Chem. Ecol.
23(6):1589-1606. 1997.
221. Röse, U. S. R., Alborn, H. T., Makranczy, G., Lewis, W. J., and Tumlinson,
J. H. Host recognition by the specialist endoparasitoid Microplitis croceipes
(Hymenoptera: Braconidae): Role of host- and plant-related volatiles. J. Insect
Behav. 10(3):313-330. 1997.
222. Paré, P. W. and Tumlinson, J. H. De novo biosynthesis of volatiles induced by
insect herbivory in cotton plants. Plant Physiology. 114(1):1161-1167. 1997.
1048 Wolf Prize in Agriculture
223. Lewis, W. J., van Lenteren, J. C., Phatak, S. C. and Tumlinson, J. H. A total
system approach to sustainable pest management. Proc. Natl. Acad. Sci. USA
94:12243-12248. 1997.
224. Paré, P. W. and Tumlinson, J. H. Cotton volatiles synthesized and
released distal to the site of insect damage. Phytochemistry. 47(4):521-526.
1997.
225. Röse, U. S. R., Lewis, W. J., and Tumlinson, J. H. Specificity of systemically
released cotton volatiles as attractants for specialist and generalist parasitic
wasps. J. Chem. Ecol. 24(2):303-319. 1998.
226. De Moraes, C. M., Lewis, W. J., Paré, P. W., Alborn, H. T., and Tumlinson,
J. H. Herbivore-infested plants selectively attract parasitoids. Nature.
393:570-573. 1998.
227. Fónagy, A., Teal, P. E. A., Meredith, J. M., Kormendy, C., and Tumlinson,
J. H. The purification, isolation and partial identification of a new
pheromonotropic peptide from the moth Mamestra brassicae (Lepidoptera).
Proceedings of the 18th Congress of European Comparative Endocrinology.
Annals NY Academy Sciences. 839:488-490. 1998. (Proceedings)
228. Paré, P. W., Alborn, H. T., and Tumlinson, J. H. Concerted biosynthesis
of an insect elicitor of plant volatiles. Proc. Natl. Aacd. Sci. USA. 95:13971-
13975. 1998.
229. Paré, P. W. and Tumlinson, J. H. Plant volatiles as a defense against insect
herbivores. Plant Physiology 121: 325-331. 1999.
230. Paré, P. W., Lewis, J. W., and Tumlinson, J. H. Induced plant volatiles:
biochemistry and effects on parasitoids, pp. 167-180. In Agrawal, A.A.,
Tuzun, S. and Bent, E. (eds.) Induced Plant Defenses Against Pathogens
and Herbivores: Biochemistry, Ecology and Agriculture. American
Phytopathological Society Press, St. Paul, MN. 1999. (Book Chapter)
231. Landolt, P. J., Tumlinson, J. H., and Alborn, H. T. Attraction of Colorado
potato beetle (Coleoptera: Chrysomelidae) to damaged and chemically-induced
potato plants. Environ. Entomol. 28(6): 973-978. 1999.
232. Tumlinson, J. H., Paré, P. W., and Lewis, W. J. Plant production of volatile
semiochemicals in response to insect derived elicitors, pp. 95-109. In
Chadwick, D.J. and Goode, J. (eds.) Insect-plant interactions and induced
plant defence. Novartis Foundation Symposium No. 223. John Wiley Journal.
1999.
233. Turlings, T. C. J., Alborn, H. T., Loughrin, J. H., and Tumlinson, J. H. Volicitin,
an elicitor of maize volatiles in the oral secretion of Spodoptera exigua: its
isolation and bio-activity. J. Chem. Ecol. 26(1): 189-202. 2000.
234. Alborn, H. T., Jones, T. H., Stenhagen, G. S., and Tumlinson, J. H. Identification
and synthesis of volicitin and related components from beet armyworm oral
secretions. J. Chem. Ecol. 26(1): 203-220. 2000.
James H. Tumlinson 1049
235. Tumlinson, J. H., Paré, P. W., Alborn, H. T., and Lewis, W. J. Chemically
mediated tritrophic plant-insect interactions. In de Wit, P. J. G. M., Bisseling, T.,
and Stiekema, W. J. (Eds.) Biology of Plant-Microbe Interactions, Vol. 2. Proc.
9th Intl. Congress on Molecular Plant-Microbe Interactions, pp 378-383.
1999.
236. De Moraes, C. M., Lewis, W. J., and Tumlinson, J. H. Examining plant-
parasitoid interactions in tritrophic systems. Annals da Sociedade Entomologica
do Brasil 29(2): 189-203. 2000.
237. Tumlinson, J. H., Alborn, H. T., Loughrin, J. H., Turlings, T. C. J., and Jones,
T. H. Plant Volatile Elicitor from Insects. U. S. Patent # 6,054,483; April 25,
2000. (M-3499); U. S. Patent # 6,207,712 B1; March 27, 2001.
238. Frey, M., Stettner, C., Paré, P. W., Schmelz, E. A., Tumlinson, J. H., and
Gierl, A. An herbivore elicitor activates the gene for indole emission in maize.
PNAS 97(26): 14801-14806. 2000.
239. Mori, N., Alborn, H. T., Teal, P. E. A., and Tumlinson, J. H. Enzymatic
decomposition of elicitors of plant volatiles in Heliothis virescens and
Helicoverpa zea. J. Ins. Phys. 47: 749-757. 2001.
240. De Moraes, C. M., Mescher, M. C., and Tumlinson, J. H. Caterpillar-induced
nocturnal plant volatiles repel conspecific females. Nature. 410: 577-580.
2001.
241. Schmelz, E.A., Alborn, H. T., and Tumlinson, J. H. The influence of
intact-plant and excised-leaf bioassay designs on volicitin- and jasmonic
acid-induced sesquiterpene volatile release in Zea mays. Planta. 214:
171-179. 2001.
242. Cardoza, Y. J., Alborn, H. T., and Tumlinson, J. H. In vivo volatile emissions
from peanut plants induced by simultaneous fungal infection and insect
damage. J. Chem. Ecol. 28(1): 161-174. 2002.
243. Engelberth, J., Schmelz, E.A., Alborn, H. T., Cardoza, Y. J., Huang, J. and
Tumlinson, J. H. Simultaneous quantification of Jasmonic acid and Sclicylic
acid in plants by vapor phase extraction and gas chromatograph-chemical
ionization-mass spectrometry. Analytical Biochemistry. 312: 242-250. 2003.
244. Schmelz, E. A., Alborn, H. T., Banchio, E., and Tumlinson, J. H. Quantitative
relationships between induced jasmonic acid levels and volatile emission in
Zea mays during Spodoptera exigua herbivory. Planta. 216:665-673. 2003.
245. Schmelz, E. A., Alborn, H. T., and Tumlinson, J. H. Synergistic interactions
between volicitin, jasmonic acid and ethylene mediate insect-induced volatile
emission in Zea mays. Physiologia Plantarum. 117: 403-412. 2003.
246. Cardoza, Y.J., C.G. Lait, E.A. Schmelz, J. Huang, and J.H. Tumlinson. Fungus-
induced biochemical changes in peanut plants and their effect on development
of beet armyworm, Spodoptera exigua Hübner (Lepidoptera: Noctuidae),
larvae. Environmental Entomology 32: 220-228. 2003.
1050 Wolf Prize in Agriculture
247. Suazo, A., Torto, B., Teal, P.E.A. and Tumlinson, J. H. Response of the small
hive beetle (Aethina tumida Murray) to honey bee (Aphis Mellifera L.) and
beehive-produced volatiles. Apidologie. 34: 525-534. 2003.
248. Olson, D. M., Rains, G. C., Meiners, T., Takasu, K., Tertuliano, M., Tumlinson,
J. H., Wackers, F. L., Lewis, W. J. Parasitic Wasps Learn and Report
Diverse Chemicals with Unique Conditionable Behaviors. Chemical Senses.
28: 545-550. 2003.
249. Aldrich, J. R., Bartelt, R. J., Dickens, J. C., Knight, A. L., Light, D. M., Tumlinson,
J. H. Insect chemical ecology research in the United States Department of
Agriculture — Agricultural Research Service. Pest Management Science. 59:
777-787. 2003.
250. Lait C.G., H.T. Alborn, P.E.A. Teal and J.H. Tumlinson III. Rapid biosynthesis
of N-linolenoyl-L-glutamine, an elicitor of plant volatiles, by membrane
associated enzyme(s) in Manduca sexta. Proceedings of the National Academy
of Sciences USA 100: 7027-7032. 2003.
251. Cardoza, Y.J., Teal, P. E. A., Tumlinson, J. H. Effect of Peanut Plant Fungal
Infection on Oviposition Preference by Spodoptera exigua and on Host-
Searching Behavior by Cotesia marginiventris. Environmental Entomology.
32: 970-976. 2003.
252. Huang, J., Cardoza, Y. J., Schmelz, E. A., Raina, R., Engelberth, J., Tumlinson,
J. H. Differential volatile emissions and salicylic acid levels from tobacco
plants in response to different strains of Pseudomonas syringae. Planta. 217:
767-775. 2003.
253. Alborn, H. T., Brennan, M. M., Tumlinson, J. H. Differential Activity and
Degradation of Plant Volatile Elicitors in Regurgitant of Tobacco Hornworm
(Manduca sexta) Larvae. Journal of Chemical Ecology. 29: 1357-1372. 2003.
254. Schmelz, E. A., Engelberth, J., Alborn, H. T., O’Donnell, P., Sammons, M.,
Toshima, H. and Tumlinson, J. H. Simultaneous analysis of phytohormones,
phytotoxins, and volatile organic compounds in plants. PNAS, 100: 10552-
10557. 2003.
255. Schmelz, E. A., Alborn, H. T., Engelberth, J., Tumlinson, J. H. Nitrogen
Deficiency Increases Volicitin-Induced Volatile Emission, Jasmonic Acid
Accumulation, and Ethylene Sensitivity in Maize. Plant Physiology. 133:
295-306. 2003.
256. Engelberth, J., Alborn, H. T., Schmelz, E. A., and Tumlinson J. H. Airborne
signals prime plants against insect herbivore attack. PNAS, 101: 1781-1785.
2004.
257. Rose, U. S. R. and Tumlinson J. H. Volatiles released from cotton plants in
response to Helicoverpa zea feeding damage on cotton flower buds. Planta
218: 824-832. 2004.
James H. Tumlinson 1051
258. De Moraes, C. M., Schultz, J. C., Mescher, M. C., and Tumlinson, J. H. Induced
plant signaling and its implications for environmental sensing. Journal of
Toxicology and Environmental Health, Part A, 67:819-834, (2004).
259. Schmelz, E. A., Engelberth, J., Tumlinson, J. H., Block, A., and Alborn, H.T.
The use of vapor phase extraction in metabolic profiling of phytohormones
and other metabolites. The Plant Journal 39: 790-808 (2004).
260. Tumlinson, J. H. and Lait, C. G. Biosynthesis of fatty acid amide elicitors of
plant volatiles by insect herbivores. Arch. Insect Biochem. Physiol. 58: 54-68
(2005)
261. Gomez, S.K., Cox, M.M., Bede, J.B., Inoue, K., Alborn, H. T., Tumlinson, J.H.,
and Korth, K.L. Lepidopteran herbivory and oral factors induce transcripts
encoding novel terpene synthases in Medicago truncatula. Archives of Insect
Biochemistry and Physiology 58:114-127 (2005).
262. Huang, J., Schmelz, E.A., Alborn, H.T., Engelberth, J., and Tumlinson, J.H.
Phytohormones mediate volatile emission during the interaction of compatible
and incompatible pathogens: The role of ethylene in Pseudomonas syringae
infected tobacco, J. Chem. Ecol. 31: 439-459 (2005).
263. Rose, U. S. R. and Tumlinson J. H. Systemic induction of volatile release
in cotton: How specific is the signal to herbivory. Planta 222: 327-335
(2005).
264. Torto, B., Suazo, A., Alborn, H., Tumlinson, J. H., and Teal, P.E.A. Response of
the small hive beetle (Aethina tumida) to a blend of chemicals identified
from honeybee (Apis mellifera) volatiles. Apidologie 36: 523-532 (2005).
265. Rose, U. S. R., Lewis, W. J. and Tumlinson J. H. Extrafloral nectar from cotton
(Gossypium hirsutum) as a food source for parasitic wasps. Functional Ecology
20: 67-74 (2006).
266. Cardoza, Y. J. and Tumlinson, J. H. Compatible and Incompatible Xanthomonas
Infections Differentially Affect Herbivore-Induced Volatile Emission by Pepper
Plants. J. Chem. Ecol. 32: 1755-1768 (2006).
267. Torto, B., Boucias, D.G., Arbogast, R.T., Tumlinson, J.H., and Teal, P.E.A.
Multitrophic interaction facilitates parasite–host relationship between an
invasive beetle and the honey bee. PNAS 104: 8374-8378 (2007).
268. Engelberth, J., Seidl-Adams, I., Schultz, J.C., and Tumlinson, J.H. Insect elicitors
and exposure to green leafy volatiles differentially up-regulate major
octadecanoids and transcripts of 12-oxo phytodienoic acid reductases in Zea
mays. Molecular Plant-Microbe Interactions 20: 707-716 (2007).
269. Alborn, H.T., Hansen, T.V., Jones, T.H., Bennett, D.C., Tumlinson, J.H.,
Schmelz, E.A., and Teal, P.E.A. Disulfooxy fatty acids from the American bird
grasshopper, Schistocerca Americana, elicitors of plant volatiles. PNAS 104:
12976-12981 (2007).
1052 Wolf Prize in Agriculture
270. Yoshinaga, N., Aboshi, A., Ishikawa, C., Fukui, M., Shimoda, M., Nishida, R.,
Lait, C.G., Tumlinson, J.H., and Mori, N. Fatty acid amides, previously identified
in caterpillars, found in the cricket Telegryllus taiwanemma and fruit fly,
Drosophila melanogaster larvae. J. Chem. Ecol. 33: 1376-1381 (2007)
271. Lelito, J.P., Fraser, I., Mastro, V.C., Tumlinson, J.H., Böröczky, K., and
Baker, T.C. Visually mediated ‘paratrooper copulations’ in the mating behavior
of Agrilus planipennis (Coleoptera: Buprestidae), a highly destructive invasive
pest of North American ash trees. J. Insect Behav. 20: 537-552 (2007).
272. Torto, B., Arbogast, R. T., Alborn, H., Suazo, A., van Engelsdorp, D., Boucias,
D., Tumlinson, J. H., And Teal, P. E. A. Composition of volatiles from fermenting
pollen dough and attractiveness to the small hive beetle Aethina tumida,a
parasite of the honeybee Apis mellifera. Apidologie 38: 380-389 (2007).
273. Gao, X., Starr, J., Göbel, C., Engelberth,J., Feussner, I Tumlinson, J., and
Kolomiets, M. Maize 9-lipoxygenase ZmLOX3 controls development, root-
specific expression of defense genes, and resistance to root-knot nematodes.
Molecular Plant-Microbe Interactions 21: 98-109 (2008).
James H. Tumlinson 1053
Source: From Science Vol. 250, No. 4985, pp. 1251-1253 (1990). Reprinted with permission from
AAAS.
1054
Wolf Prize in Agriculture
REPORTS
the active site of ODCase must provide two 10. J. B. Bell, M. E. Jones, C. W. J. Carter, Proteins 9, no. 1) (1988).
143 (1991). 24. Using the difference in the pKa of acetic acid and
features: a low dielectric interior to allow for 11. Attempts to obtain the crystal structure of ODCase malonic acid to estimate the effect of a CO2⫺ group,
pKa tuning so that the catalytic proton trans- are underway (C. Carter, personal communication). we estimate the pKa of protonated orotate to be
fer to O-4 can occur, and a perfectly placed 12. J. A. Smiley and M. E. Jones, Biochemistry 31, ⬃1.5.
lysine to effect that proton transfer to O-4, 12162 (1992). 25. J. A. Dean, in Lange’s Handbook of Chemistry
13. J. A. Smiley and S. J. Benkovic, J. Am. Chem. Soc. (McGraw-Hill, San Francisco, CA, 1992), pp. 8.19 –
concerted with decarboxylation. 117, 3877 (1995). 8.71.
14. R. B. Silverman and M. P. Groziak, ibid. 104, 6434 26. W. P. Jencks, J. Am. Chem. Soc. 94, 4731 (1972).
REFERENCES AND NOTES (1982). 27. M. K. Gilson and B. H. Honig, Biopolymers 25, 2097
___________________________ 15. K. Shostak and M. E. Jones, Biochemistry 31, (1986).
1. A. Radzicka and R. Wolfenden, Science 267, 90 12155 (1992). 28. G. King, F. S. Lee, A. Warshel, J. Chem. Phys. 95,
(1995). 16. Gaussian 94, revision B.3 (M. J. Frisch et al., Gauss- 4366 (1991).
2. R. W. McClard, M. J. Black, L. R. Livingstone, M. E. ian, Inc., Pittsburgh, PA, 1995). RHF, MP2, and ba- 29. A. Warshel and J. Åqvist, Annu. Rev. Biophys. Bio-
Jones, Biochemistry 19, 4699 (1980). sis sets are described by W. J. Hehre, L. Radom, P. phys. Chem. 20, 267 (1991).
3. T. C. Bruice and S. Benkovic, in Bioorganic Mecha- v. R. Schleyer, and J. A. Pople [Ab Initio Molecular 30. T. Simonson and C. L. Brooks III, J. Am. Chem. Soc.
nisms (Benjamin, New York, 1966), vol. 2, pp. 188 – Orbital Theory ( Wiley, New York, 1986)]. Density 118, 8452 (1996).
194. functional theory (Becke3LYP) is described by W. 31. Solvation was modeled with the SCI-PCM method,
4. M. L. Bender, in Mechanisms of Homogeneous Ca- Kohn, A. D. Becke, and R. G. Parr [J. Phys. Chem. available in Gaussian 94 (16). Solvation energies
talysis from Protons to Proteins ( Wiley-Interscience, 100, 12974 (1996)]. were obtained by calculating RHF/6-31⫹G* ener-
New York, 1971), pp. 165 –175 and 586 –594. 17. Unless otherwise specified, the calculated ⌬H‡ is for gies in solvent of a given dielectric and comparing
5. P. Beak and B. Siegel, J. Am. Chem. Soc. 98, 3601 0 K. that value to the gas-phase energy. These solvation
(1976). 18. D. Lavorato et al., J. Am. Chem. Soc. 118, 11898 energies were then used to correct MP2/6-31⫹G*
6. S. A. Acheson, J. B. Bell, M. E. Jones, R. Wolfenden, (1996), and references therein. energies to obtain ⌬H‡ values in solution.
Biochemistry 29, 3198 (1990). 19. R. Kluger, Chem. Rev. 87, 863 (1987). 32. We are grateful to NSF and NIH (postdoctoral fellow-
7. J. A. Smiley, P. Paneth, M. H. O’Leary, J. B. Bell, M. 20. D. Kern et al., Science 275, 67 (1997). ship to J.K.L.) for financial support of this work and to
E. Jones, ibid. 30, 6216 (1991). 21. A. J. Arduengo III et al., J. Am. Chem. Soc. 116, the UCLA Office of Academic Computing and the
Source: From Science Vol. 276, No. 5314, pp. 945-949 (1997). Reprinted with permission from
AAAS.
1062 Wolf Prize in Agriculture
normal-phase diol column (12). The active final rpHPLC column indicated it was greater seedlings (2). The volatiles released were
component eluted from this column with than 99% pure. the same as those induced by treatment
MeOH (Me designates methyl). Rechro- Purified active compound was applied to with BAW oral secretion. Application of
matography of the active component on the artificially damaged leaves of intact corn only buffer resulted in the release of signif-
icantly smaller quantities and different pro-
portions of volatiles (Fig. 2).
The active compound was identified by
mass and infrared spectroscopy and by
chemical transformations. Fast atom bom-
bardment mass spectrometry (FAB-MS)
analysis (13) indicated the presence of
only one compound with diagnostic peaks
at mass-to-charge ratio (m/z) 423.280 (M
⫹ H)⫹ in the positive ion mode and at m/z
421.2733 (M ⫺ H)⫺ in the negative ion
mode. The addition of sodium chloride to
the FAB matrix resulted in reduced inten-
sity of the m/z 423.280 and the appearance
of m/z 445.2628 (MH ⫹ Na)⫹. Thus, the
active compound is a weak acid with a
molecular weight of 422.274 daltons for
Fig. 2 (left). Average amount (nanograms per 2 hours) (n ⫽ 4) of volatiles The next morning at 9:00 a.m. the seedlings were cut off above the root, and
collected from three intact Ioana corn seedlings that had been artificially volatiles were collected and analyzed as described (7, 8). Bars with the same
damaged and treated with (A) 15 l of BAW oral secretion per seedling on the retention time in each graph represent the following compounds: 1, hexenyl
damage sites, (B) 15 l of oral secretion equivalents of pure natural volicitin, acetate; 2, linalool; 3, (E)-4,8-dimethyl-1,3,7-nonatriene; 4, indole; 5,
(C) 15 l of buffer (8), or (D) undamaged control plants. At 9:00 p.m. a 1-cm2 ␣-trans-bergamotene; 6, (E)--farnesene; 7, (E)-nerolidol; 8, (3E,7E)-4,8,12-
area of the second leaf of three-leaf seedlings was scratched with a clean trimethyl-1,3,7,11-tridecatetraene. Fig. 3 (right). Synthesis scheme for
razor blade and the test solution immediately rubbed over the damaged site. volicitin.
REPORTS
sistent with the electron impact (EI) mass double bond, respectively. The intensity of ing the step in which the fatty acid was
spectrum of glutamine (14). Subtraction of the 3019 cm⫺1 peak indicated three cis dou- coupled to glutamine. However, the active
glutamine, linked by an ester or amide bond, ble bonds. Absorption at 1758 cm⫺1 con- natural compound consisted exclusively of
gave C18H30O3 as the elemental composition firmed the presence of a methyl ester. the L-glutamine form. Enantiomerically
of the second part of the molecule, which is Partial reduction (19) of the methyl ester pure D- and L-glutamine forms of synthetic
consistent with a hydroxy C18 acid with three of the C18 hydroxy acid resulted in a mixture volicitin were collected from a chiral col-
double bonds. of mono- and di-unsaturated products as iden- umn (24) for bioassay.
When the active sample was lyophilized tified by GC/MS. Subsequent ozonolysis (20) The concentration of natural volicitin
and treated with MeOH and acetic anhydride of this mixture and GC/MS analysis produced was estimated by HPLC detector response
(15), GC/MS analysis (16) of the product three diagnostic GC peaks with (M ⫹ 1)⫹ to be about 20 pmol per microliter of BAW
revealed two prominent peaks. Chemical ion- ions at m/z 187, 229, and 271, corresponding oral secretion. Bioactivity of the L-glu-
ization (CI)/MS analysis of the first of these to H(CO)(CH2)n(CO)OCH3 with n ⫽ 7, 10, tamine form of the synthetic volicitin was
peaks with a retention time of 21.05 min and 13, respectively. Methyl linolenate treat- equivalent to that of oral secretion (Fig. 4).
revealed a prominent (M ⫹ 1)⫹ ion at m/z ed in the same way gave identical products. The D-glutamine form was not active (Fig.
144, and EI/MS analysis revealed a molecular Thus, the olefinic bonds in the chain are 4). Racemic 17-hydroxylinolenic acid, D-
ion at m/z 143 and diagnostic ions at m/z 84 located on carbons 9, 12, and 15, and the glutamine, and L- glutamine at 300 and 900
(base peak), 56, and 41, identifying it as the alcohol group is on either the 17th or 18th pmol per excised corn seedling showed no
methyl ester of pyroglutamate, which con- carbon. activity. Thus, the biologically active com-
firmed the presence of glutamine. The CI The methyl ester was saturated by treat- pound is N-(17-hydroxylinolenoyl)-L-glu-
mass spectrum of the second peak (retention ment with PdO/H2 overnight (21). GC/MS tamine. At this time the configuration
time of 27.55 min) contained a very weak m/z (EI) analysis of the product showed an m/z about the asymmetric 17th carbon in the
known to be synthesized de novo (30). 30 min to eliminate solid material, and the superna- was concentrated to dryness by a stream of N2, then
tant was then filtered through a 0.22-m sterilizing 50 l of CH2Cl2 was added and the sample was
Volicitin accounts for the total activity of membrane (Millex GV, Millipore, Bedford, MA). An analyzed by GC/MS.
BAW oral secretion, and boiling the secre- equal amount of 50 mM pH 3.3 phosphate buffer 16. Methanolysis products were analyzed on a Finni-
tion for 30 min did not diminish its activity; was added, and the precipitated proteins were re- gan TSQ 700 mass spectrometer (Finnigan MAT,
moved by centrifugation as before. A 0.5-ml sample
thus, there is no evidence for enzymatic ac- of the supernatant was put on a 6-ml activated, oc-
San Jose, CA) interfaced to an HP 5890 gas chro-
matograph (Hewlett-Packard, Palo Alto, CA). Injec-
tivity in eliciting volatiles in this case. In tadecyl solid-phase extraction cartridge (Bakerbond, tions were made in the splitless mode at 225°C (1
contrast, a -glucosidase in the saliva of J. T. Baker, Phillipsburg, NJ) and eluted with 2-ml
l containing the equivalent of 2 l of oral secre-
Pieris brassicae caterpillars elicits the release volumes of H2O, 50% CH3CN (Low-UV HPLC grade,
tion). A polar capillary column (OV351, 25 m long,
Burdick and Jackson) in H2O, and CH3CN. All frac-
of volatile compounds from cabbage leaves 0.25-mm ID, Scandinaviska Genetec, Kungs-
tions were concentrated to near dryness under vac-
backa, Sweden) was held at 60°C for 3 min and
(32). The sequence of events between the uum (Speed Vac rotary concentrator, Savant Instru-
then increased 10°C per minute to 250°C and held
introduction of -glucosidase and the emis- ments, Farmingdale, NY ) and redissolved in 0.5 ml of
at that temperature for 18 min. The carrier gas was
50 mM pH 8 buffer, for bioassay.
sion of volatiles is unknown (5), but the 10. The active material from solid-phase extraction was
He and the reaction gas was CH4 for chemical
ionization. The ion source temperature was 200°C
simple release of a terpenoid or other volatile fractionated by HPLC [LDC 4100 pump with in both EI and CI mode.
compound from a glycoside by the direct SM5000 diode array UV detector (LDC Analytical,
Riviera Beach, FL)], monitoring wavelengths from 17. The acetate had a shorter retention time (26.06 min)
action of such an enzyme from the attacking 190 to 360 nm. A reversed-phase column ( Waters on the polar OV-351 column than the hydroxy acid
herbivore obviously occurs by a different Nova Pac C18 4 m, 4-mm ID by 150-mm long methyl ester (27.55 min), but a longer retention time
(25.54 min) than the hydroxy acid methyl ester
mechanism than the induction of de novo column, Waters, Milford, MA) was eluted (1 ml/min)
(25.38 min) on a nonpolar column (DB1, 25 m long,
with a solvent gradient of 0 to 25% CH3CN in H2O in
biosynthesis by a small molecule like volic- 15 min, followed by an increase to 100% CH3CN in 0.25-mm ID, J&W Scientific, Folsom, CA). The CI
itin. Also, it cannot account for the delayed 15 min. Eluate was collected in 2-ml fractions. All mass spectrum showed ions at m/z 351 (weak) (M ⫹
1), m/z 309 (weak) (loss of MeOH), m/z 291 (base
release of herbivore-induced volatiles (28) or activity was in the 6- to 8-ml fraction, which was
peak) (loss of acetic acid), and m/z 259 (loss of both
further fractionated on a C18 reversed-phase column
the systemic release of induced volatiles (29,
REPORTS
J. Preston, and P. A. Cruickshank [J. Am. Chem. Soc. ml/min (10). The synthetic L-form had a retention 32. L. Mattiacci, M. Dicke, M. A. Posthumus, ibid. 92,
87, 2492 (1965); W. Konig and R. Geiger, Chem. Ber. time of 5.25 min, identical to that of the natural elic- 2036 (1995).
103, 788 (1970)]. The coupling was carried out in N,N- itor, and the D-form had a retention time of 9.03 min. 33. Significant differences in relative release rates of
dimethylformamide (DMF) (Sigma) with addition of 5 to 25. E. E. Farmer and C. A. Ryan, Plant Cell 4, 129 (1992). volatiles were tested by Tukey’s studentized range
10% H2O to increase solubility of the salt and 1.2 equiv- 26. R. Kramell et al., J. Plant Growth Regul. 14, 29 test after analysis of variance with a significance level
alents of 1-hydroxybenzotriazole (Sigma) to reduce ra- (1995). of 5% (SYSTAT, Systat Inc., Evanston, IL ).
cemization of glutamine. The product was purified by 27. K. E. Ricker and R. M. Bostock, Physiol. Mol. Plant 34. This project was funded in part by a grant from the
rpHPLC (10) on the YMC column eluted with 25% Pathol. 44, 65 (1994), and references therein.
Swedish Natural Science Research Council. We
28. J. H. Loughrin, A. Manukian, R. R. Heath, T. C. J. thank H. Karlsson, A. T. Proveaux, and D. Powell for
CH3CN in 0.4 mM ammonium acetate buffer (Aldrich) at
Turlings, J. H. Tumlinson, Proc. Natl. Acad. Sci.
a flow rate of 1.2 ml/min. The retention time of the assistance with mass spectrometric analysis, J.
U.S.A. 91, 11836 (1994).
synthetic and natural elicitor was identical. Lockerman and S. Sharp for oral secretion collec-
29. U. S. R. Röse, A. Manukian, R. R. Heath, J. H. Tum- tion, and M. Brennan for technical assistance. We
24. The D- and L-glutamine forms of synthetic volicitin linson, Plant Physiol. 111, 487 (1996).
were separated on a 250-mm, 4.6-mm ID chirobiotic also thank J. G. Millar, C. A. Ryan, and G. G. Still for
30. P. W. Pare and J. H. Tumlinson, Nature 385, 30
T column (Advanced Separation Technologies, helpful comments on the manuscript.
(1997).
Whippany, NJ) eluted with 10% CH3CN in 10 mM 31. T. C. J. Turlings and J. H. Tumlinson, Proc. Natl.
ammonium acetate buffer, pH 4.5, at a flow rate of 1 Acad. Sci. U.S.A. 89, 8399 (1992). 24 January 1997; accepted 17 March 1997
ABSTRACT A variety of agricultural plant species, in- volatiles from plants fed on by different herbivores suggests
cluding corn, respond to insect herbivore damage by releasing that each insect species may produce its own signature mole-
large quantities of volatile compounds and, as a result, become cule(s) that allows plants to differentiate among herbivorous
highly attractive to parasitic wasps that attack the herbivores. attackers (11, 12). The current study was undertaken to
An elicitor of plant volatiles, N-(17-hydroxylinolenoyl)-L- determine the biogenetic origin of volicitin. By feeding beet
glutamine, named volicitin and isolated from beet armyworm armyworms corn seedlings labeled with 13CO2, we obtained
caterpillars, is a key component in plant recognition of chemical evidence that the caterpillars acquire linolenic acid
damage from insect herbivory. Chemical analysis of the oral [an essential fatty acid in the diet of Lepidoptera (13)] from
secretion from beet armyworms that have fed on 13C-labeled plants and that the insects subsequently hydroxylate and
corn seedlings established that the fatty acid portion of conjugate the fatty acid with glutamine. Thus, the modification
volicitin is plant derived whereas the 17-hydroxylation reac- by the insect of linolenic acid of plant origin provides a distinct
tion and the conjugation with glutamine are carried out by the chemical cue that allows a plant to differentiate between
caterpillar by using glutamine of insect origin. Ironically, herbivore damage and other types of wounding that trigger the
these insect-catalyzed chemical modifications to linolenic acid octadecanoid defense signaling pathway.
are critical for the biological activity that triggers the release
of plant volatiles, which in turn attract natural enemies of the
caterpillar. MATERIALS AND METHODS
Plant Growth and Labeling Conditions. Corn seeds, Zea
Several studies have shown the active role of herbivore- mays L., variety LG11 sweet corn, were given 4 days to
damaged plants in the attraction of insect predators and germinate under a moist paper towel in the dark. To reduce the
parasitoids. Volatile plant compounds released in response to amount of stored 12C-labeled carbon available for the growing
insect feeding serve as a chemical signal for herbivore natural seedling, seeds were trimmed by using a razor blade to remove
enemies (for a review, see refs. 1 and 2). Recent work suggests ⬇60–70% of the endosperm without cutting into the germi-
that this as well as other plant defense responses are triggered nated seedling. Nine excised seedlings were planted in steril-
by an active component or components associated with the ized Metromix 300 potting soil (Scotts-Sierra Horticulture,
feeding herbivore that allows the plant to differentiate be- Marysville, OH). The pot was placed in a 48- ⫻ 16-cm
tween general wounding and damage caused by chewing (diameter) glass growth chamber, and synthetic premixed air
insects. In cotton, induced volatiles that are synthesized in (Cambridge Isotope Laboratories, Cambridge, MA and Airco,
response to wounding are released in greater quantities as a Riverton, NJ), which contained 1,800 M兾liter CO2 (13C
result of caterpillar feeding than with mechanical damage 99%), 20.7% oxygen, and a balance of nitrogen was introduced
alone (3), and, in tobacco, higher concentrations of the by flushing the chamber at 500 ml min⫺1 for 10 min and then
defense-signaling molecule jasmonic acid result from herbi- reducing to a flow of 50 ml min⫺1. Synthetic air passed up over
vore damage by hornworm caterpillars than from mechanical the potted plants contained within the volatile collection
damage designed to mimic herbivory (4). At the transcriptional apparatus and out one of eight ports from the glass lid through
level, potato mRNAs involved in plant defense accumulate a plastic tube into a water bubbler. One metal halide and two
more rapidly with insect-derived elicitor(s) in contact with the 400-W high-pressure sodium lamps positioned 10 cm above the
damaged leaves than with mechanical damage alone (5). chamber provided a 16-h light兾8-h dark photoperiod; an
Thus far, two elicitors of plant volatiles have been identified air-cooled glass panel directly below the lights insulated the
and reported from chewing insects. Dicke’s group has identi- plants from the lamp heat source.
fied a -glucosidase, present in the regurgitant of Pieris Collection of Plant Volatiles. Corn seedling leaves were
brassicae caterpillars that triggers the same emissions of vola- damaged with a stainless steel wire, fed through one of the top
tiles in cabbage plants as larvae that feed on the plant (6). ports of the growth chamber to scrape the leaves against the
Because enzyme activity in the regurgitant is retained when glass chamber. Plant volatiles then were collected from 1,200–
caterpillars are fed on -glucosidase free diet, enzyme activity 1,500 h by drawing synthetic 13C-labeled air (100 ml min⫺1)
does not appear to be plant derived. Volicitin, N-(17- through Super-Q adsorbent traps (Alltech Associates). Com-
hydroxylinolenoyl)-L-glutamine, identified from the oral se- pounds were eluted from the adsorbent filters with 150 l of
cretion of beet armyworm caterpillars, induces corn seedlings dichloromethane; 1-l aliquots were analyzed by capillary GC
to synthesize and release volatile chemical signals (7) that on a 50-m ⫻ 0.25-mm (i.d.) fused silica column with a
attract parasitic wasps. (ref. 8; for review, see refs. 9 and 10). 0.25-m-thick bonded methyl silicone stationary phase (Qua-
Volicitin has not been found in plant tissues, although the drex, New Haven, CT). The column was held at 60°C for 5 min
structure of the molecule suggests that it may interact with the and then was increased 5°C per min to 225°C and was held at
octadecanoid signaling pathway in plants (7). The collection of that temperature for 30 min. Helium was used as a carrier gas
at a linear flow velocity of 18 cm sec⫺1. To determine the
The publication costs of this article were defrayed in part by page charge amount of 13C incorporated into each compound, samples
payment. This article must therefore be hereby marked ‘‘advertisement’’ in were analyzed by a Finnigan-MAT ITS40 (ion trap) mass
accordance with 18 U.S.C. §1734 solely to indicate this fact.
0027-8424兾98兾9513971-5$0.00兾0 *To whom reprint requests should be addressed. e-mail: jtumlinson@
PNAS is available online at www.pnas.org. gainesville.usda.ufl.edu.
13971
13972 Plant Biology: Paré et al. Proc. Natl. Acad. Sci. USA 95 (1998)
FIG. 1. HPLC profile of volicitin (A) and other fatty acid derivatives detected in the oral secretions of beet armyworms at a wavelength of 200
nm. Compounds and approximate retention times include: (A) N-(17-hydroxylinolenoyl)-L-glutamine, 7.5 min; (a) N-(17-hydroxylinoleoyl)-L-
glutamine, 8.0 min; (B) 17-hydroxy linolenic acid, 11.1 min; (C) N-linolenoyl-L-glutamine, 11.9 min; (b) 17-hydroxy linoleic acid, 12 min; (IS)
N-palmitoleoyl-L-glutamine (internal standard), 12.3 min; (c) N-linoleoyl-L-glutamine, 12.6 min; (D) linolenic acid, 15.2 min; (d) linoleic acid, 16.2
min; (E) unknown, 16.6 min; (d ) oleic acid, 17.1 min; and (F) impurity, 17.8 min.
spectrometer (Finnigan-MAT, San Jose, CA) in the chemical- and was redissolved in H2O (400 l), and the supernatant, after
ionization mode, with isobutane as the reagent gas. Injections centrifugation, was collected. The aqueous extract was con-
were made via a septum-equipped programmable injector held centrated to dryness and derivatized with 300 l MeOH:Ac2O
at 40°C for 0.25 min, then programmed at 170°C min⫺1 to (5:1) by heating for 10 min at 100°C (16). Identification of plant
270°C onto a 30-m ⫻ 0.25-mm (i.d.) fused silica column with fatty acids, glutamine, and glutamic acid methyl esters were
0.25-m-thick bonded 5% phenyl methyl silicone (DB-5MS; J confirmed by comparison with methylated synthetic standards
& W Scientific, Folsom, CA) held at 40°C for 5 min, then by GC-MS analysis following the same procedures as the
programmed at 5°C min⫺1 to 260°C; He carrier gas linear flow analysis of plant volatiles except that samples were injected
velocity was 19 cm sec⫺1. Source temperature was adjusted to onto a polar capillary column (OV351, 25-m ⫻ 0.25-mm i.d.,
120 ⫾ 20°C to optimize the molecular ion abundance. Selected Quadrex, New Haven, CT), which was held at 60°C for 5 min
mass ions were quantified via computer software analysis. The and then was increased 5°C per min to 230°C and held at that
fraction of each compound that incorporated 13C was com- temperature for 30 min.
puted on a molecule basis (14). Caterpillar Regurgitant Collection and Analysis. Beet ar-
Plant Chemical Analysis. Lipid and amino acid analysis was
myworms were reared on an artificial pinto bean diet following
based on procedures of Harborne (15). For lipid extraction,
the method of King and Leppla (17). Insects were transferred
frozen plant tissue (0.1 g) was homogenized with chilled
to feed on corn seedlings at least 48 h before labeling exper-
isopropanol (5 ml) and was extracted with diethyl ether (5 ml);
the supernatant fractions, after centrifugation, were combined iments. Regurgitation was induced by holding fourth instar
and concentrated to dryness. For methanolysis, 2 ml MeOH: beet armyworm caterpillars with forceps and gently pinching
HCl (3:1) was added to the extract and was heated for 30 min behind the head with a second pair (18). The oral secretion
at 100°C. Hydrolyzed lipids were diluted with H2O (5 ml), were from five larvae was collected, 5 l of an aqueous N-
extracted with CH2Cl2 (2 ⫻ 4 ml), and were rinsed with palmitoleoyl-L-glutamine solution (1 g l⫺1) was added as an
saturated NaHCO3 (2 ⫻ 4 ml). A concentrated extract was internal standard, and the mixture was centrifuged. The su-
redissolved in a minimum amount of Hex:EtOAc (5:1) and was pernatant was fractioned by HPLC [LDC 4100 pump with
applied to a 6-ml activated silica gel solid-phase extraction SM5000 diode array UV detector (LDC Analytical, Rivera
cartridge that was eluted with the same solvent. Fractions Beach, FL)] monitoring wavelength 200 nm on a reversed-
containing oleic, linoleic, and linolenic methyl esters initially phase column (C18 ODS-AQ S-5 200 Å, 250 mm long, 4.6-mm
were identified by comigration with standards on silica gel i.d., YMC, Kyoto). Components were eluted (1 ml min⫺1) with
TLC plates (Merck). Glutamine and glutamic acid were ex- a solvent gradient of 40 to 100% CH3CN, containing 0.8%
tracted from frozen tissue (0.1 g) with 65% EtOH (5 ml); the glacial acetic acid, in H2O, containing 0.4% glacial acetic acid,
supernatant, after centrifugation, was concentrated to dryness over 10 min and then were held at 100% CH3CN, still
1068 Wolf Prize in Agriculture
Plant Biology: Paré et al. Proc. Natl. Acad. Sci. USA 95 (1998) 13973
FIG. 2. Chemical ionization mass spectra of methylated linolenic F IG . 3. Chemical ionization mass spectra of methyl 17-
acid from: beet armyworms (BAW) fed on unlabeled corn seedlings hydroxylinolenate prepared by transesterification of volicitin from oral
for 48 h (A), extracted corn seedlings grown for 12 days with 13CO2 secretion of caterpillars fed 6 h on unlabeled corn seedlings (A), 6 h
enrichment (B), and beet armyworms fed on unlabeled corn plants and on 13C-labeled corn seedlings (B), and 12 h on 13C-labeled seedlings
then fed for 6 h on 13C-labeled corn seedlings (C). with unlabeled linolenic acid added to the leaves after 6 h (C). The 291
ion resulted from loss of water from the molecular ion (M ⫹ 1 ⫺ 18)⫹.
containing 0.8% acetic acid, for 10 min. Fractions were
methylated with MeOH and Ac2O as noted above (10). into which synthetic premixed air that contained 1,800 M兾
Unlabeled Linolenic Acid Applications. Linolenic acid (Sig-
liter 13CO2 flowed continuously. As a preliminary experiment
ma) in EtOH (1:9) was diluted to a 1% acid solution with H2O.
to estimate the degree of incorporation of 13C into the fatty
The solution was applied to labeled corn seedling with an
atomize sprayer to cover the corn leaves with a thin mist of acids of the plant, leaves were damaged mechanically and the
unlabeled acid. hexenals and hexenols, released as a result of lipoxygenase
activity, were collected and analyzed by GC-MS (19). These
analyses indicated that linolenic acid in 9-day-old seedlings had
RESULTS incorporated substantial levels of 13C [e.g., 84% enrichment of
To assess rate of synthesis and the source of the chemical (Z)-3-hexenol]. Seedlings grown for 12 days in an enclosed
components used by beet armyworm (Spodoptera exigua Hüb- glass chamber to ensure a high level of 13C labeling were used
ner) to assemble volicitin, caterpillars were fed on corn to feed beet armyworm larvae. Subsequent to the feeding
seedlings that were labeled uniformly with 13C. Plants were experiment (see below), the remaining portions of the leaves
labeled isotopically by growing seedlings in a closed container of each plant were extracted and analyzed to determine the
James H. Tumlinson 1069
13974 Plant Biology: Paré et al. Proc. Natl. Acad. Sci. USA 95 (1998)
(Fig. 3C) and free linolenic acid, both collected from the beet
armyworm, indicates that hydroxylation of the relatively large
amount of free unlabeled linolenic acid sprayed onto the plant
FIG. 4. Chemical ionization mass spectra of methyl pyroglutamate
(the product of glutamine methylation) from extracted corn seedlings leaves is done by the beet armyworm. The linolenic and linoleic
that were grown for 12 days with 13CO2 enrichment (A), volicitin from acid components from caterpillars that had fed on unsprayed
13C-labeled seedlings for the same total time of 12 h main-
caterpillars that fed on unlabeled corn seedlings for 48 h (B), and
volicitin from beet armyworms that fed on unlabeled corn plants and tained high 13C incorporation levels. To ensure that the
then fed 6 h on 13C-labeled corn seedlings (C). decrease in 13C incorporation with application of unlabeled
linolenic acid was a specific response for linolenic acid deriv-
percentage of incorporation of the label in linolenic acid and atives, linoleic acid was collected from caterpillars that had fed
glutamine. on sprayed seedlings and was found to sustain the high level of
Early fourth instar beet armyworms fed on an artificial diet 13C labeling. The 17-hydroxylinolenic acid methyl ester and the
were transferred to unlabeled corn plants for a minimum of 17-hydroxylinoleic acid methyl ester were not detected in
48 h before being placed on the labeled corn seedlings, to derivatized plant extracts, although these compounds were
ensure that the ratio of fatty acids in the oral secretions was identified when plant tissue was spiked with the corresponding
based on plant constituents and not our rearing diet. Oral synthetic acid before extraction and methylation.
secretion was collected by gently squeezing caterpillars after In contrast to the fatty acid portion of volicitin, which rapidly
they had fed on labeled seedlings for 6 h, causing them to incorporated 13C and showed a similar mass spectral labeling
regurgitate. The oral secretion supernatant from the caterpil- pattern as the linolenic acid from the plant (Fig. 2B), the
lars was injected, without further purification, onto a reverse glutamine incorporated little 13C relative to the glutamine
phase HPLC column and was eluted with an acetonitrile-water from the plant (Fig. 4), indicating that the plant was not
gradient. Peaks eluting from this column (Fig. 1) were col- catalyzing the coupling of glutamine to the fatty acids. It also
lected, lyophilized, and treated with methanol and acetic appears that labeled glutamine from the plant ingested by the
1070 Wolf Prize in Agriculture
Plant Biology: Paré et al. Proc. Natl. Acad. Sci. USA 95 (1998) 13975
insect was diluted with an unlabeled insect source of glutamine 1. Tumlinson, J. H., Lewis, W. J. & Vet, L. E. M. (1993) Sci. Am.
before coupling to form the volicitin molecule. This could be 268, 100–106.
caused by either a large glutamine pool present in the cater- 2. Paré, P. W. & Tumlinson, J. H. (1996) Florida Entomol. 79,
93–103.
pillar’s oral secretions or a physically separate glutamine 3. Paré, P. W. & Tumlinson, J. H (1997) Nature (London) 385,
source. Because the plants are 13C-labeled from the time that 30–31.
photosynthesis begins, the possibility that unlabeled glutamine 4. McCloud, E. S. & Baldwin, I. T. (1998) Planta 203, 430–435.
is derived from an isolated source separate from current 5. Korth, K. L & Dixon, R. A. (1997) Plant Physiol. 115, 1299–1305.
photosynthate and apart from the major pool of plant glu- 6. Mattiacci, L., Dicke, M. & Posthumus, M. A. (1995) Proc. Natl.
Acad. Sci. USA 92, 2036–2040.
tamine is unlikely. 7. Alborn, H. T., Turlings, T. C. J., Jones, T. H., Stenhagen, G.,
These biochemical data demonstrate that the plant supplies Loughrin, J. H. & Tumlinson, J. H. (1997) Science 276, 945- 949.
linolenic acid, which is required for growth and development 8. Turlings, T. C. J., Tumlinson, J. H. & Lewis, W. J. (1990) Science
of beet armyworms, and also provides this fatty acyl chain for 250, 1251–1253.
the synthesis of volicitin, the modified elicitor of plant volatiles 9. Dicke, M. (1994) J. Plant Physiol. 143, 465–472.
10. Takabayashi, J. & Dicke, M. (1996) Trends Plant Sci. 1, 109–113.
that is central to signaling between plants and natural enemies 11. De Moraes, C. M., Lewis, W. J., Paré, P. W., Alborn, H. T. &
of the caterpillars that attack them. It is not clear why the Tumlinson, J. H. (1998) Nature (London) 393, 570–572.
caterpillar (or gut symbionts in the caterpillar) adds L- 12. Takabayashi, J., Dicke, M. & Posthumus, M. A. (1991) Chemo-
glutamine and the hydroxy group to linolenic acid or whether ecology 2, 1–6.
volicitin plays a role in the metabolism of the herbivore. It is 13. Chapman, R. F. (1982) The Insects: Structure and Function
also not clear how volicitin interacts on a biochemical level to (Harvard Univ. Press, Cambridge, MA).
14. Paré, P. W. & Tumlinson, J. H (1997) Plant Physiol. 114,
induce synthesis of plant volatiles; however, we do know that 1161–1167.
volicitin provides a rapid, clear, and reliable signal for initia- 15. Harborne, J. B (1984) Phytochemical Methods (Chapman & Hall,
tion of synthesis and release of volatile compounds. Thus the London).
plant and the herbivore are inexorably linked through a 16. Mee, J. M. L. (1977) Biomed. Mass Spectrom. 4, 178–182.
signaling molecule of dual origin and effect. 17. King, E. G. & Leppla, N. C. (1984) Advances and Challenges in
Insect Rearing. (U. S. Government Printing Service, Washington,
D. C).
We are grateful for technical assistance from P. M. Brennan and 18. Turlings, T. C. J., McCall, P. J., Alborn, H. T. & Tumlinson, J. H.
A. T. Proveaux. We thank C. A. Ryan, R. B. Croteau, J. Gershenzon, (1993) J. Chem. Ecol. 19, 411–425.
F. C. Schroeder, P. E. A. Teal, and T. C. J. Turlings for their 19. Vick, B. A. & Zimmerman, D. C. (1987) Plant Physiol. 85,
constructive comments concerning the initial manuscript. 1073–1078.
James H. Tumlinson 1071
Plant Physiology, October 1999, Vol. 121, pp. 325–331, www.plantphysiol.org © 1999 American Society of Plant Physiologists
Leaves normally release small quantities of volatile 1993). In the latter case, a parasitoid female injects her eggs
chemicals, but when a plant is damaged by herbivorous when she stings, and the eggs hatch into wasp larvae inside
insects, many more volatiles are released. The chemical the caterpillar. Once the caterpillar has been stung, its
identity of the volatile compounds varies with the plant reproductive cycle is terminated and a new generation of
species and with the herbivorous insect species. These wasps is produced.
volatiles attract both parasitic and predatory insects that In all plants reported thus far, there are notable similar-
are natural enemies of the herbivores. They may also in- ities in the structure of the volatile compounds that are
duce defense responses in neighboring plants. Such chem- emitted from insect-damaged leaves and from leaves distal
icals, which function in communication between and to the site of damage. The structural uniformity in the
among species, as well as those that serve as messengers chemical emissions from different plants with insect feed-
between members of the same species, are called semio- ing suggests the activation of a common set of biosynthetic
chemicals (from the Greek “semeion,” a mark or signal) pathways shared by a wide range of plant families, and
(Law and Regnier, 1971). that the products are detectable to a broad spectrum of
Semiochemicals emitted from a diverse group of plants insect parasitoids and predators (Fig. 2). The ability of
and insects mediate key processes in the behavior of spe- host-seeking insects to recognize and respond to such
cific insects. Volatile phytochemicals can serve as airborne chemical cues and differentiate them from background
semiochemicals, promoting or deterring interactions be- odors indicates that insect-damaged plants emit volatile
tween plants and insect herbivores. For example, wheat chemicals that are clearly distinguishable from those re-
seedlings without herbivore damage attract aphids, leased in response to other types of damage or those re-
whereas odors released from wheat seedlings with a high leased from undamaged plants. The plant’s ability to dif-
density of aphids repel other aphids (Quiroz et al., 1997). For ferentiate between herbivore damage and a general wound
swallowtail butterflies, volatiles from host plants enhance response suggests the presence of elicitors associated with
the effect of contact stimulants, increasing landing rates and insect feeding that are absent from other types of leaf
oviposition relative to non-host plants (Feeny et al., 1989). damage.
In addition to the bouquet of compounds that render
leaves attractive or disagreeable to herbivores, volatile ter-
penoids and other compounds emitted from leaves in re- PLANTS RESPOND TO INSECT FEEDING DAMAGE
sponse to insect damage allow insect parasitoids (such as BY RELEASING GREATER AMOUNTS OF A VARIETY
parasitic wasps) and predators to distinguish between in- OF VOLATILES
fested and noninfested plants, and thus aid in locating
hosts or prey (Fig. 1). These phytodistress signals, which An undamaged plant maintains a baseline level of vola-
result in an active interaction between herbivore-damaged tile metabolites that are released from the surface of the leaf
plants and a third trophic level, have been described for and/or from accumulated storage sites in the leaf. These
several agro-ecosystems. Examples include lima bean and constitutive chemical reserves, which often include mono-
apple plants, which produce volatiles that attract predatory terpenes, sesquiterpenes, and aromatics, accumulate to
mites when damaged by spider mites (Takabayashi and high levels in specialized glands or trichomes (Paré and
Dicke, 1996), and corn and cotton plants, which release Tumlinson, 1997a). In addition, green-leaf odors consisting
volatiles that attract hymenopterous parasitoids that attack of a blend of saturated and unsaturated six-carbon alco-
larvae of several Lepidoptera species (Tumlinson et al., hols, aldehydes, and esters are produced by autolytic oxi-
dative breakdown of membrane lipids and are released
when leaves are mechanically damaged. This pattern of
constitutive compounds has been analyzed in the field for
1
Present address: Department of Chemistry and Biochemistry, perennials, including beech (Tollsten and Müller, 1996) and
Texas Tech University, Lubbock, TX 79409. ash (Markovic et al., 1996) trees, as well as under green-
* Corresponding author; e-mail jtumlinson@gainesville.usda.ufl.
house conditions for many herbaceous annuals, including
edu; fax 352–374 –5707.
325
1072 Wolf Prize in Agriculture
Figure 1. Schematic representation indicating an increase of volatile compounds released by plants in response to insect
feeding triggered by an interaction of elicitors such as volicitin in the oral secretions of insect herbivores with damaged plant
tissue. Volatile semiochemicals are then used by natural enemies of herbivores such as parasitoid wasps to locate their hosts.
brussels sprouts (Mattiacci et al., 1994) and cucumber (Tak- breakage of leaf glands causes stored terpenes to be re-
abayashi et al., 1994). leased in much higher levels, and the emissions of lipoxy-
Plants respond to insect feeding damage by releasing a genase pathway green-leaf volatiles are also increased.
variety of volatiles from the damaged site, and the profile While the release of these metabolites correlates closely
of the volatiles emitted is markedly different from those of with leaf damage from insect feeding (Loughrin et al.,
undamaged or mechanically damaged plants. In cotton, 1994), a subset of terpenes, the nitrogen-containing com-
James H. Tumlinson 1073
Figure 2. Biosynthetic pathways leading to the release of plant volatiles. Indole, a product of the shikimic acid pathway, is
formed from indole-3-glycerol-P either as an intermediate in Trp biosynthesis or by a Trp-independent pathway leading to
a family of nitrogen-containing defense compounds (e.g. 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one) (Frey et al.,
1997). Sesquiterpenes are synthesized via the isopentenyl pyrophosphate (IPP) intermediate following the classical meva-
lonate pathway, whereas monoterpenes and diterpenes are synthesized via an alternative IPP pathway with glyceraldehyde-
3-P and pyruvate identified as the direct precursors of IPP (Lichtenthaler et al., 1997). The mevalonate pathway is localized
in the cytosol and reactions for the non-mevalonate pathway are localized in plastids. The homoterpene (E)-4,8-dimethyl-
1,3,7-nonatriene and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene are derived from their 15 and 20 carbon precursors,
farnesyl- and geranylgeranyl-pyrophosphate, respectively, by a series of enzymatic steps with the overall loss of four carbon
units (Donath and Boland, 1994). The green-leaf volatiles are derived from linolenic acid via a 13-hydroperoxylinolenic acid
intermediate (Blee, 1998). This oxidized linolenic acid, instead of losing water and committing the molecule down the
defense signaling jasmonic acid pathway, is cleaved to form two fragments of 12 and six carbon units (Fig. 3). The variety
of green-leaf volatiles are formed from this second pathway by multiple rearrangement steps of the six-carbon (Z)-3-hexenal.
pound indole, and hexenyl acetate are also released in RELEASE OF VOLATILES FROM UNDAMAGED LEAVES
much higher levels with insect feeding, but in a diurnal cycle OF A DAMAGED PLANT INDICATES A
that is decoupled from short-term insect damage. These SYSTEMIC SIGNAL
compounds, linalool and (E)--ocimene (monoterpenes),
(E,E)-␣-farnesene and (E)--farnesene (sesquiterpenes), non- In addition to the release of volatiles at the site of herbi-
atriene and tridecatetraene (homoterpenes), and indole and vore feeding, analysis of volatile emissions from unharmed
(Z)-3-hexenyl acetate, have an emissions profile more simi- leaves of insect-damaged plants has established that there
lar to the light cycle, with low emissions at night and high is a systemic response. In both corn (Turlings and Tumlin-
levels during the periods of maximal photosynthesis. son, 1992) and cotton (Röse et al., 1996), leaves distal to the
Chemical labeling studies have established that the com- site of herbivore feeding showed an increase in the release
pounds released in much greater quantities during the day of volatiles. The chemical blend of volatiles from undam-
and specifically in response to insect damage are synthe- aged cotton leaves differs from the volatiles collected from
sized de novo and are not stored in the plant (Paré and the entire plant. The products of the lipoxygenase pathway,
Tumlinson, 1997b). These induced compounds rapidly in- including the hexenals and hexenols, which are released
corporate a high level of label when plants damaged by from freshly cut or damaged tissue, are not detected in the
feeding caterpillars are held in volatile collection chambers systemically released volatiles, with the exception of (Z)-3-
under an atmosphere containing 13C-CO2. The high incor- hexenyl acetate. One explanation is that these six-carbon
poration of 13C detected by mass spectral analysis, and the compounds can only be released from undamaged leaf
rapid turnover of this label in experiments where short tissue when they are converted to the acetate form (Paré
pulses of 13C-CO2 were used indicate that its production is and Tumlinson, 1998).
tightly coupled with photosynthesis. A consistent, several- The activation of the lipoxygenase pathway in undam-
hour delay between when insect feeding begins and emis- aged leaves suggests a mechanism analogous to that pro-
sion of the induced volatiles supports the hypothesis that a posed by Farmer and Ryan (1992), wherein a mobile signal
series of biochemical reactions, including gene expression, such as systemin can transmit information from the dam-
protein assembly, and/or enzyme induction, is required aged site to distal leaves, triggering the lipoxygenase path-
for the synthesis and release of these compounds. way and resulting in a cascade of signals activating several
1074 Wolf Prize in Agriculture
defense responses in plants. Some of the monoterpenes and production costs in terms of reproductive success can de-
sesquiterpenes, as well as indole and isomeric hexenyl pend on the level of herbivory. When herbivore levels are
butyrates and 2-methyl butyrates, are also only released low, chemically induced wild-type tobacco plants produce
from damaged leaves (Röse et al., 1996). The induced ter- fewer seeds than their noninduced counterparts. With in-
penoids that are synthesized de novo in cotton leaves in termediate herbivory, chemically induced plants experi-
response to herbivore damage are also released systemi- ence less feeding on the foliage and have a higher fitness
cally from undamaged leaves of a caterpillar-damaged level than noninduced, insect-damaged control plants
plant. Chemical labeling experiments established that the (Baldwin, 1998; Mitchell-Olds et al., 1998). It appears that
systemic volatiles are synthesized at the site of release, volatiles need to be judiciously synthesized and safely
suggesting that a mobile chemical messenger is trans- stored, as increased synthesis can be costly and potentially
ported from the damage location to distal, undamaged toxic to the plant. However, decreases in terpene accumu-
leaves to trigger synthesis and volatile release (Paré and lation may make an individual plant more vulnerable to
Tumlinson, 1998). insect pest attacks or temperature stress.
Chemical labeling experiments using herbivore- With or without insect feeding, plants usually release a
damaged plants in combination with an analysis of the variety of terpenes during periods of high temperature.
volatiles released has only been reported for cotton. How- Although the biological function of terpene production is
ever, since many of the compounds emitted from corn not fully understood, one proposed explanation for these
during the day have also been shown to be induced in emissions is that it is a strategy for responding to high
cotton, and the quantities released increase with increased temperatures (Mlot, 1995). It has been suggested that fat-
light intensity, it can be speculated that these volatiles are soluble hydrocarbons dissolve into the thylakoid mem-
also synthesized de novo in corn plants. It is interesting brane and keep the chloroplast from degrading when tem-
that similar compounds are emitted in response to insect peratures exceed the plant’s biological optimum. These
herbivore damage in several agricultural species, including hydrocarbons evaporate as the temperature rises, so that
cucumber, apple, lima bean, corn, and cotton (see Table I). terpene volatilization cools the chloroplasts. However,
Both among individual plants of the same species and since the evaporative cooling of terpenes is relatively small
between different plant species, whether the blend of vol- compared with a solvent such as water, this explanation is
atile compounds is induced through a common signaling not universally accepted.
pathway or if their emissions are triggered by different
signaling mechanisms is not yet known.
VOLATILES FROM INSECT-DAMAGED PLANTS
ATTRACT NATURAL ENEMIES OF THE HERBIVORES
THE SYNTHESIS OF VOLATILES HAS A HIGH
The task for a female parasitoid to locate lepidopteran
METABOLIC COST
caterpillar hosts would most often be unproductive if she
Terpenes are an important source of olefinic compounds were simply to rely on visual cues. Unlike insect pollina-
involved in the formation of phytotoxic products. For ex- tors seeking out well-marked flower targets, parasitoids
ample, in conifers (Buchbauer et al., 1994) and broadleaf are searching for small herbivores that are often well cam-
tree species (Monson and Fall, 1989), an array of terpene ouflaged and mostly inhabit the undersides of leaves.
hydrocarbons are released from plants during times of Therefore, the chances of parasitoids finding hosts by ran-
photosynthesis. These naturally produced isoprenoids are dom searching are remote. Both McCall et al. (1993) and
known to form photooxidants and ozone in combination Steinberg et al. (1993) have shown by wind tunnel flights
with nitrogen oxides. As a result, increased amounts of and GC analysis the weak allure and low abundance that
terpenes can act as pollutants, increasing the stress to the herbivore odors alone provide for parasitoids. In contrast,
plant. The metabolic cost of these phytochemical emissions the chemicals released from herbivore-damaged plants ap-
can also be high. In particular, terpenoids are more expen- pear to contain critical chemical information that draws
sive to manufacture per gram than most other primary and parasitoids to air streams spiked with these plant odors in
secondary metabolites due to the need for extensive chem- the laboratory and to damaged plants placed among a
ical reduction (Gershenzon, 1994). Defensive compound group of undamaged neighbors in the field.
Table I. Diverse plant species with shared volatile terpenes released in response to herbivory
(E,E)-4,8,12-
(E)-- (E)-4,8-Dimethyl- (E,E)-␣- (E)--
Plant Linalool Trimethyl-1,3,7,11- Reference
Ocimene 1,3,7-Nonatriene Farnesene Farnesene
Tridecatetraene
Cucumber ⫹ ⫹ ⫹ ⫹ Takabayashi et al. (1994)
Apple ⫹ ⫹ ⫹ ⫹ Takabayashi et al. (1991)
Lima bean ⫹ ⫹ ⫹ ⫹ ⫹ Takabayashi et al. (1994)
Cotton ⫹ ⫹ ⫹ ⫹ ⫹ ⫹ Paré and Tumlinson (1997a)
Corn ⫹ ⫹ ⫹ ⫹ ⫹ ⫹ Turlings et al. (1990)
Tobacco ⫹ ⫹ ⫹ ⫹ De Moraes et al. (1998)
Potato ⫹ ⫹ ⫹ ⫹ Bolter et al. (1997)
James H. Tumlinson 1075
To examine whether systemically released chemicals less water available for the plant, elevated levels of vola-
alone provide sufficient chemical cues to attract parasitic tiles are released from infested individuals relative to non-
wasps, herbivore-damaged leaves were removed immedi- water-stressed controls. Correlating this with insect pref-
ately before flight tests. Wind tunnel experiments showed erence showed that predatory mites selected plants that
that systemically released components were detectable at were infested and water-stressed over those that were in-
levels sufficient to direct parasitoids to their hosts (Corte- fested but not water-stressed (Takabayashi et al., 1994). The
sero et al., 1997). In cotton and tobacco field trials using addition of high levels of mineral and/or organic nitrogen
female wasps (Cardiochiles nigriceps), the ratio of landings fertilizers significantly decreased the constitutive volatiles
on host (tobacco budworm) damaged versus undamaged extracted from celery (Van Wassenhove et al., 1990). With
plants was high: approximately 95% to 5%, respectively, in volatile analysis and flight studies for plants under differ-
systemic or whole-plant volatile emissions (De Moraes et ent nutritional conditions, the role of these volatiles in
al., 1998). Interestingly, this specialist parasitic wasp, using attracting wasps to their herbivore hosts may be more
chemical cues released by the plant, can distinguish plants clearly assigned.
infested by her host Heliothis virescens from those infested
by Helicoverpa zea, a closely related, non-host herbivore
ENZYMES AND ELICITORS FROM INSECT HERBIVORES
species. In tobacco, cotton, and maize, each plant produces
TRIGGER VOLATILE RELEASE
a herbivore-specific blend of volatile components in re-
sponse to a particular herbivore species feeding on the Key to the emissions of plant signals for the foraging
leaves, and these differences are observable by GC chemi- success of parasitoids are substances in the oral secretion of
cal analyses and detectable by parasitic wasps. herbivores. Recent work suggests that volatile emissions
and other plant defense responses are potentiated by a
component or components associated with the feeding her-
PARASITIC WASPS LEARN CHEMICAL CUES
bivore that allows the plant to differentiate between gen-
ASSOCIATED WITH HOSTS
eral wounding and damage due to chewing insects. In
Although the volatile compounds released by insect her- cotton, induced volatiles that are synthesized in response
bivore damage are similar among the several plant species to wounding are released in greater quantities as a result of
studied thus far, the specific blends are quite distinct, caterpillar feeding than as a result of mechanical damage
varying in both the number of compounds and the actual alone (Paré and Tumlinson, 1997a). In tobacco, higher con-
structures produced. Thus, the task of finding a host is centrations of the defense-signaling molecule jasmonic acid
more complicated for the parasitoid when the host feeds on result from herbivore damage by hornworm caterpillars
several different plant species. The wasps have overcome than from mechanical damage designed to mimic her-
this obstacle by developing the ability to learn chemical bivory (McCloud and Baldwin, 1998). At the transcrip-
cues associated with the presence of a host (Lewis and tional level, potato mRNAs involved in plant defense ac-
Tumlinson, 1988). The chemicals to which a female wasp is cumulate more rapidly with insect-derived elicitor(s) in
exposed during interactions with her host familiarize her contact with the damaged leaves than with mechanical
with particular host location cues. A successful host expe- damage alone (Korth and Dixon, 1997).
rience increases the wasp’s responsiveness to host- Thus far, two oral secretion products from chewing in-
associated chemicals. For example, an oviposition experi- sects have been identified that augment the release of plant
ence on the plant-host complex significantly increases the volatiles. A -glucosidase present in the regurgitant of
oriented flight and landing responses of females of the Pieris brassicae caterpillars triggers the same emissions of
aphid parasitoid Aphidius ervi relative to those that aren’t volatiles in cabbage plants as induced by feeding caterpil-
allowed to sting but that are exposed to undamaged or lars (Mattiacci et al., 1995). Since enzyme activity in the
host-damaged plants (Du et al., 1997). This underscores the regurgitant is retained when caterpillars are fed a
importance of the oviposition experience in combination -glucosidase-free diet, enzyme activity does not appear to
with host-damaged plant cues. Interestingly, female wasps be plant derived. Presumably, the enzyme acts to cleave
can also learn volatile odors associated with food sources sugars coupled to organic compounds that then become
and use them to locate necessary food (Lewis and Takasu, more volatile and are released. In contrast, a low-Mr fatty
1990). acid derivative, N-(17-hydroxylinolenoyl)-l-Gln (volicitin),
has been identified from the oral secretions of beet army-
worm caterpillars and induces corn seedlings to release
ENVIRONMENTAL CONDITIONS MODULATE
volatile chemical signals (Alborn et al., 1997).
VOLATILE EMISSIONS
Analysis of volicitin from beet armyworms fed 13C-
Differences in the amount of volatiles released by indi- labeled corn seedlings demonstrated that the caterpillar
vidual plants can vary with environmental conditions that synthesizes this elicitor by adding a hydroxyl group and
influence the plant’s physiology. Several species, including Gln to linolenic acid obtained directly from the plant on
corn, cotton, and lima bean, respond to reduced light (due which the caterpillar feeds (Paré et al., 1998). Thus, al-
to either lower light intensity or shorter daylength) with a though the precursor of volicitin is obtained from plants,
decline in the release of herbivore-induced volatiles. Based the bioactive product has only been found in the caterpil-
on studies with lima bean, water stress also seems to di- lar. This strongly suggests that these molecules play an
rectly affect volatile release (Takabayashi et al., 1994). With important yet still unknown role in metabolism or some
1076 Wolf Prize in Agriculture
other process critical to the life of the herbivorous insects. ever, we don’t know the biochemical mechanisms by which
Although it is known that the plant provides linolenic acid, these elicitors trigger biosynthesis and release of plant
which is essential for most lepidopteran larvae (Stanley- volatiles. Do they interact with the octadecanoid signaling
Samuelson, 1994), it is seemingly detrimental to the insect pathway, and if so, how? Do they regulate the release of
to chemically convert this fatty acid into an elicitor that linolenic acid, the production of jasmonic acid, or the acti-
triggers plant defense. The full implications of this are not vation of the oxidative burst, all of which are associated
yet understood. with the wounding of plant tissue? Also, we have no
It has been suggested that jasmonic acid, which is pro- knowledge of the mechanism leading to the systemic re-
duced from linolenic acid by the octadecanoid signaling lease of volatiles. Does the original, herbivore-produced
pathway (see Fig. 3), is a key regulatory component in the elicitor serve as a mobile messenger, triggering whole-
transduction sequence that triggers synthesis and release of plant volatile synthesis? Or are secondary messengers em-
volatile compounds by plants (Krumm et al., 1995). Jas- ployed to transmit the signal to sites distal to the site of
monates also stimulate other physiological and defensive damage? Furthermore, why do herbivores produce com-
processes in plants (Farmer and Ryan, 1992), and the amino pounds that activate plant chemical defenses? What func-
acid conjugates of jasmonic acid are involved in physiolog- tion, if any, do these compounds serve in herbivore metab-
ical and developmental processes in many plants (Kramell olism or defense?
et al., 1995). Therefore, the structure of volicitin, an octa- The answers to these and similar questions should lead
decatrienoate conjugated to an amino acid, suggests that to the development of more effective methods for the bio-
the elicitor molecule interacts with the octadecanoid path- logical control of insect pests with natural enemies. It may
way in herbivore-damaged plants. also lead to the development of new plant varieties with
enhanced chemical defenses or to methods of “inoculating”
plants with elicitors to increase their resistance to insect
THE MECHANISMS THAT REGULATE THE SYNTHESIS
pests.
AND RELEASE OF PLANT VOLATILES ARE
POORLY UNDERSTOOD
Received May 20, 1999; accepted June 16, 1999.
There is still much to learn about the chemical interac-
tions between plants and insect herbivores that lead to the
synthesis and release of volatiles by the plants. Only one LITERATURE CITED
herbivore-specific volatile elicitor, volicitin, has been iden-
tified, but we know from preliminary investigations of the Alborn HT, Turlings TC, Jones TH, Stenhagen G, Loughrin JH,
Tumlinson JH (1997) An elicitor of plant volatiles from beet
chemistry and activity of oral secretions of other insect armyworm oral secretion. Science 276: 945–949
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1078 Wolf Prize in Agriculture
letters to nature
Acknowledgements night, and that female moths exploit these speci®c night-time
We thank D. Wake and R. Carroll for discussion; K. Monoyios for editorial assistance; signals to avoid oviposition on previously damaged plants.
M. Ellison for illustrations; and A. Davidson for preparing GMV 1606. For help in ®eld Gas chromatographic analysis of volatiles collected in two-hour
collection of specimens, we thank Z. Cheng, X. Xu, Y. Jin and Z. Chen. N.H.S. was
supported by a grant from the John Simon Guggenheim Memorial Foundation. Field
intervals continuously for seven days revealed consistent differences
work was supported by the National Geographic Society and the Frick Funds of the in the composition of volatile blends released by H. virescens-
American Museum of Natural History. infested tobacco plants (n = 6) during the light and dark phases
of the photoperiod (Fig. 1a). Visual and auditory observations
Correspondence and requests for materials should be addressed to K.-Q.G.
(e-mail: kgao@amnh.org).
con®rmed that larvae fed during both the light and dark phases.
Seven major compounds were consistently released during both
light and dark phases, but usually in lesser amounts during the dark
phase (Fig. 1a). In addition, ®ve compounds ((Z)-3-hexenyl buty-
rate, (Z)-3-hexenyl isobutyrate, (Z)-3-hexenyl acetate, (Z)-3-hex-
................................................................. enyl tiglate, and one unidenti®ed compound) were produced only
during the dark phase. OthersÐ(E)-2-hexenal and three unidenti-
Caterpillar-induced nocturnal plant ®ed compoundsÐwere produced in signi®cantly larger amounts
during the dark than the light period. Thus, the qualitative and
volatiles repel conspeci®c females quantitative composition of volatile blends emitted by tobacco
plants in response to feeding by H. virescens larvae can differ
Consuelo M. De Moraes*, Mark C. Mescher² & James H. Tumlinson* signi®cantly between night and day.
Repetition of our analysis using two other species of lepidopteran
* USDA-ARS, CMAVE, PO Box 14565, Gainesville, Florida 32604, USA larvae (n = 6 per species), Manduca sexta and Helicoverpa zea,
² Department of Entomology, University of Georgia, Athens, Georgia 30602, USA provided further evidence of induced volatile release from tobacco
.............................................................................................................................................. plants during the dark period (Fig. 1b). Although the volatile
Plants respond to insect herbivory by synthesizing and releasing
complex blends of volatile compounds, which provide important
host-location cues for insects that are natural enemies of herbi-
vores1±3. The effects of these volatile blends on herbivore behav- a
iour have been investigated to only a limited extent4,5, in part
because of the assumption that herbivore-induced volatile emis- Heliothis virescens
sions occur mainly during the light phase of the photoperiod6,7. daytime volatiles
Because many mothsÐwhose larvae are some of the most impor- IS ISIS
tant insect herbivoresÐare nocturnal, herbivore-induced plant
volatiles have not hitherto been considered to be temporally
available as host-location cues for ovipositing females. Here we 6 9 10 13 14 1516
present chemical and behavioural assays showing that tobacco Heliothis virescens
plants (Nicotiana tabacum) release herbivore-induced volatiles IS night-time volatiles
IS
during both night and day. Moreover, several volatile compounds
are released exclusively at night and are highly repellent to female
1 2 3 4567 8 9 11 12 13 14 15 16
moths (Heliothis virescens). The demonstration that tobacco
plants release temporally different volatile blends and that lepi-
dopteran herbivores use induced plant signals released during the
dark phase to choose sites for oviposition adds a new dimension to b
our understanding of the role of chemical cues in mediating Helicoverpa zea
night-time volatiles
tritrophic interactions. IS IS
Feeding by insect herbivores induces plants to release chemical
signals that serve as important foraging cues for parasitoids and 1 3 4567 8 9 11 12 13 15 16
predators, and thus enhance the plants' defence1±3,8±10. Synthesis and
release of these chemical signals is an active physiological process Manduca sexta
triggered by substances in the oral secretion of herbivores11,12. The night-time volatiles
IS
recent discovery that plant volatiles can transmit herbivore-speci®c IS
NATURE | VOL 410 | 29 MARCH 2001 | www.nature.com © 2001 Macmillan Magazines Ltd 577
James H. Tumlinson 1079
letters to nature
pro®les revealed quantitative differences in plant response to the P , 0.0001) for untreated tobacco plants over those with septa
three herbivores, all caterpillar species induced release of the same releasing the synthetic blends (Fig. 2B, b). In control trials, the
volatile compounds from tobacco during the night (Fig. 1). To response to plants with septa containing only solvent (hexane) was
determine whether plants continue nocturnal volatile production in statistically indistinguishable (F1,10 = 1.91, P . 0.05) from that to
the absence of continuous feeding by the caterpillars, H. virescens untreated plants (Fig. 2B, a). Thus, female H. virescens are able to
were removed from the plants at 15:00 after 48 hours of feeding. In identify infested plants on the basis of chemical cues consisting of
this case, plants still emitted volatiles during the dark phase volatile compounds emitted by the plants at night.
although in smaller amounts. Some of the volatiles in the nocturnal volatile pro®le were also
In behavioural trials, mated H. virescens females spent a signi®- present in the diurnal pro®le. However, others were released only at
cantly greater proportion of time (80% of the observed hour) in an night. To determine the importance of volatiles produced at night,
area with only undamaged plants than in an area with both damaged relative to those produced during the day, we repeated the behav-
and undamaged plants (Fig. 2A, a). Moths exhibited a tendency to ioural assays using the diurnal blends. Although these daytime
¯y south when ®rst released; however, if the damaged plants were volatiles produced avoidance behaviour (Fig. 2B, c; F1,10 = 55, P ,
placed to the south, the moths would often turn and ¯y in the 0.001), they were signi®cantly less repellent than the nocturnal
opposite direction. The same preference was displayed in ovi- volatiles (Fig. 2B, b), indicating that the moths are speci®cally
position. Females selected only non-infested plants for oviposition repelled by the night-time volatile blends. To further examine the
(Fig. 2A, b; F1,10 = 427, P , 0.0001) and also avoided uninfested extent to which the repellence of the night-time volatiles is due to
plants close to infested plants. the presence of compounds produced exclusively at night, the
Because the preference of H. virescens for uninfested plants might experiment was repeated using a synthetic blend containing only
conceivably be explained by visual cues associated with plant these compounds. The presence of these exclusively nocturnal
damage, we repeated the behavioural assays using synthetic volatile compounds alone was suf®cient to explain the moth repellence
blends. Synthetic blends of the major volatile compounds that were effect (Fig. 2B, d; F1,10 = 145, P , 0.0001).
produced in signi®cant amounts and emitted by the plants during It is well established that induced plant volatiles function as
the dark period were formulated on rubber septa19,20 to release signals between plants and the natural enemies of insect herbivores.
volatiles in approximately the same proportions and amounts as The discovery that the herbivores themselves exploit the informa-
released by herbivore-damaged plants. In behavioural assays, tion present in night-time volatile blends to avoid oviposition on
H. virescens demonstrated a signi®cant preference (F1,10 = 134, previously damaged plants reveals a new dimension of chemically
A
100 100
a b
80 80
Oviposition (%)
Time spent (%)
60 60
40 40
20 20
0 0
Undamaged plants Damaged + undamaged Undamaged plants Damaged + undamaged
plants plants
B
100 100
a b
80 80
Oviposition (%)
Oviposition (%)
60 60
40 40
20 20
0 0
Undamaged plants Hexane Undamaged plants Night-time blend
100 100
c d
80 80
Oviposition (%)
Oviposition (%)
60 60
40 40
20 20
0 0
Undamaged plants Daytime blend Undamaged plants Exclusive night-time
compounds
Figure 2 Response of female moths to plant volatiles. A, Nocturnal ¯ight responses of with synthetic blend. On one side of the cage only undamaged plants (6) were used, on the
three mated H. virescens females to tobacco plants. On one side of the cage, only other side two plants were treated (synthetic blends on rubber septa) placed among four
undamaged plants (6) were used. On the other side, two plants that had been infested by undamaged and untreated plants. a, Solvent (hexane) used as control. b, A nocturnal
ten third-instar H. virescens larvae were included. Larvae were removed from the plants blend compared to undamaged plants. c, A diurnal blend compared to undamaged plants.
before moths were introduced. a, Per cent of time during one hour of observation spent on d, A blend containing compounds released exclusively at night compared to undamaged
each side of the cage, b, per cent of eggs oviposited on each side of the cage. plants.
B, Oviposition preference by H. virescens females for tobacco plants that had been treated
578 © 2001 Macmillan Magazines Ltd NATURE | VOL 410 | 29 MARCH 2001 | www.nature.com
1080 Wolf Prize in Agriculture
letters to nature
mediated plant±insect interactions and raises a number of issues. 19:00 to 20:00) and the time (min.) spent by moths on each side of the cage was
recorded. Egg numbers per plant on each side of the cage were counted at 06:00 the next
The ®tness advantages to herbivores of avoiding oviposition on
morning. Similar procedures were used for assays measuring oviposition and behaviour
induced plants are obvious, as such plants are likely to host not only of female moths in response to plants with or adjacent to synthetic blends, but in this
larvae that represent potential competitors for the moth's offspring case plants on both sides of the cages were undamaged. For these treatments, pots of
but also potentially a population of natural enemies attracted by the undamaged plants with volatile-releasing rubber septa on wooden sticks (three per
volatile blend21. Moreover, the associated induction of direct plant) replaced the damaged plants. We evaluated the response of moths to three
synthetic volatile blends (nocturnal volatiles, exclusively nocturnal volatiles, and diurnal
defence mechanisms means that infested plants are likely to contain volatiles). Rubber septa without the synthetic blend were placed on the control side to
chemical toxins and to be of lower nutritional value than uninfested neutralize any visual preference. Each bioassay was conducted on three days (two
plants22,23. It is less clear whether plants bene®t signi®cantly from the repetitions each) to account for day-to-day variation. Preference between the
release of nocturnal volatiles or whether such release is merely a undamaged and damaged plants by the herbivores was subjected to analysis of variance
(an arc-sine square root transformation of the percentage data was used; SAS Institute,
physiological by-product associated with diurnal volatile produc- Cary, North Carolina).
tion. The protection enjoyed by undamaged plants that reside near
induced plants is interesting, but it is not certain whether this
Synthetic blends
phenomenon would have been signi®cant in the ancestral environ-
Synthetic blends were formulated according to a method developed to predict the release
ment or represents only an effect of high-density agricultural ratio of volatile compounds from rubber septa19,20. Calculations of predicted release ratios
cultivation. If plants do bene®t from advertising their status to are based on relative vapour pressures19 of the components and the original amounts
herbivores, it raises the question of whether herbivore-induced released in the natural blend. Blends were dissolved in hexane and 0.3 ml of a blend
signals ®rst evolved as parasitoid attractants or as herbivore repel- solution was pipetted onto a rubber septum. Volatiles released by the blend when
formulated on rubber septa were sampled and analysed. The relative amounts were
lents; alternatively, the dual functions of herbivore-induced plant adjusted to correct for slight deviations from the predicted amount. Compounds used to
signals may have evolved simultaneously. prepare synthetic blends were obtained from Sigma-Aldrich or from Chemical Samples
The recognition that plant volatile signals, long known to be and analysed by gas chromatography and mass spectrometry to determine purity. All
important in the mediation of plant±parasitoid interactions, also synthetic compounds were at least 98% pure except for ocimene where both isomers were
present (60% trans and 40% cis).
transmit information to herbivores expands our view of tritrophic Release rates of synthetic blends: daytime blend 1, (E)-2-hexenal (200 ng h-1); 2, (E)-b-
systems and is signi®cant with regard to our understanding of the ocimene (5,500 ng h-1); 3, b-caryophyllene (7,300 ng h-1); 4, a-humulene (200 ng h-1); 5,
selective pressures governing the evolution of such signals. We are (E,E)-a-farnesene (1,180 ng h-1). Night-time blend 1, (E)-2-hexenal (500 ng h-1); 2, (Z)-
currently exploring whether moths can interpret the herbivore- 3-hexen-1-ol (850 ng h-1); 3, (Z)-3-hexenyl acetate (1,000 ng h-1); 4, (E)-b-ocimene
(1,500 ng h-1); 5, (Z)-3-hexenyl isobutyrate (100 ng h-1); 6, (Z)-3-hexenyl butyrate
speci®c information that these signals convey to parasitoids13 or
(500 ng h-1); 7, (Z)-3 hexenyl tiglate (1,200 ng h-1); 8, b-caryophyllene (2,200 ng h-1);
rather use these signals only as generalized indicators of insect 9, (E,E)-a-farnesene (900 ng h-1). Exclusive night-time blend 1, (Z)-3-hexen-1-ol
feeding. If moths can interpret the higher-order information con- (850 ng h-1); 2, (Z)-3-hexenyl acetate (1,000 ng h-1); 3, (Z)-3-hexenyl isobutyrate
tent of these signals, then they may be making sophisticated choices (100 ng h-1); 4, (Z)-3-hexenyl butyrate (500 ng h-1); 5, (Z)-3-hexenyl tiglate (1,200 ng h-1).
based on the likely presence of particular larval competitors and Received 19 December 2000; accepted 31 January 2001.
perhaps even of particular predators and parasitoids. M
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screened cage (4 ´ 2 ´ 3 m) with six plants on each side of the cage (80 cm between plants). Entomol. Exp. Appl. 67, 79±85 (1993).
On one side only undamaged plants were used, on the other side two of the plants had been 19. Heath, R. R. & Tumlinson, J. H. Prediction of release ratios of multicomponent pheromones from
fed on by 10 laboratory-reared third-instar H. virescens (two larvae per leaf) for 48 h. rubber septa. J. Chem. Ecol. 12, 2081±2088 (1986).
Larvae were removed from plants before moth release. Three mated H. virescens females 20. Heath, R. R., Teal, P. E. A., Tumlinson, J. H. & Mengelkoch, L. J. Correlation of retention times on
(18 females per treatment) were released in the central sector of the cage at dusk. To liquid crystal capillary column with reported vapor pressures and half-lives of compounds used in
account for a possible directional preference by the moths, two cages were used so in one pheromone formulations. J. Chem. Ecol. 12, 2133±2143 (1986).
the damaged plants were on the north end and in the other they were on the south end of 21. Thaler, J. S. Jasmonate-inducible plant defenses cause increased parasitism of herbivores. Nature 399,
the cage. The moths' behaviour was visually observed for one hour (from approximately 686±588 (1999).
NATURE | VOL 410 | 29 MARCH 2001 | www.nature.com © 2001 Macmillan Magazines Ltd 579
James H. Tumlinson 1081
letters to nature
22. Agrawal, A. A. Induced responses to herbivory and increased plant performance. Science 279, 1201± by synaesthetes, although idiosyncratic, remain highly stable over
1202 (1998).
23. Coley, P. D., Bryant, J. P. & Chapin, F. S. Resource availability and plant antiherbivore defense. Science
time4.
230, 895±899 (1985). Colour-graphemic synaesthesia may be triggered by automatic
24. Heath, R. R. & Manukian, A. An automated-system for use in collecting volatile chemicals released co-activation of independent brain areas responsible for processing
from plants. J. Chem. Ecol. 20, 593±608 (1994).
colour and symbolic form12,13. This ®ts with synaesthetes' subjective
accounts of the involuntary nature of their experiences. We there-
Acknowledgements fore manipulated the physical colours of alphanumeric characters so
We thank G. W. G. De Moraes for discussions; H. T. Alborn, J. G. Hildebrand, P. J. Landolt, that they differed from the synaesthetic colours induced. Our
W. J. Lewis, K. G. Ross and J. R. Ruberson for comments on the manuscript; and B. Dueben
approach was based on the Stroop effect14, in which naming the
and M. Sammons for technical assistance.
print colour of an incongruent colour word (for example, RED
Correspondence and requests for materials should be addressed to J.H.T. printed in blue) takes signi®cantly longer than naming the print
(e-mail: jtumlinson@gainesville.usda.u¯.edu).
colour of a congruent colour word (for example, RED printed in
red), or a colour patch. This slowing re¯ects interference arising
from an involuntary word-reading response15.
We began by comparing synaesthetes and controls on the stan-
................................................................. dard Stroop task to check for any baseline differences in their
susceptibility to interference. Colour words were displayed in
Unconscious priming eliminates either congruent or incongruent colours; solid colour patches
were used in a baseline condition (see Methods). Participants
automatic binding of colour and named the colour of each stimulus aloud. As expected, both
groups were signi®cantly slower to name colours in the incongruent
alphanumeric form in synaesthesia than the congruent condition (F1,28 = 119.72, P , 0.001). Critically,
there was no overall difference between the groups (F1,28 = 0.66,
Jason B. Mattingley*, Anina N. Rich*, Greg Yelland² P . 0.10) and no interaction (F1,28 = 1.56, P . 0.10; Fig. 2a),
& John L. Bradshaw² indicating equivalent interference for synaesthetes and controls.
Moreover, their colour naming times in the baseline condition were
* School of Behavioural Science, University of Melbourne, Victoria 3010, Australia the same (535 ms versus 536 ms; t28 = 0.03, P . 0.10).
² Department of Psychology, Monash University, Victoria 3800, Australia If synaesthesia is an involuntary phenomenon, then having
.............................................................................................................................................. participants judge the physical colour of an alphanumeric character
Synaesthesia is an unusual perceptual phenomenon in which that elicits an incongruent synaesthetic colour should yield signi®-
events in one sensory modality induce vivid sensations in cant interference, and slow response times accordingly. We therefore
another1,2. Individuals may `taste' shapes3, `hear' colours4, or constructed unique stimulus ensembles for each synaesthete, which
`feel' sounds5. Synaesthesia was ®rst described over a century
ago6, but little is known about its underlying causes or its effects
on cognition. Most reports have been anecdotal or have focused
on isolated unusual cases3,7±9. Here we report an investigation of
15 individuals with colour-graphemic synaesthesia, each of whom
experiences idiosyncratic but highly consistent colours for letters
and digits. Using a colour±form interference paradigm, we show
that induced synaesthetic experiences cannot be consciously
suppressed even when detrimental to task performance. In con-
trast, if letters and digits are presented brie¯y and masked, so that
they are processed but unavailable for overt report, the synaesthe-
sia is eliminated. These results show that synaesthetic experiences
can be prevented despite substantial processing of the sensory
stimuli that otherwise trigger them. We conclude that automatic
binding of colour and alphanumeric form in synaesthesia arises
after initial processes of letter and digit recognition are complete.
We studied 15 individuals with colour-graphemic synaesthesia
and 15 non-synaesthetic controls. Each synaesthete reported vivid
and immediate sensations of colour for speci®c letters and digits. All
reported having had synaesthesia since childhood, and many had
biological relatives with the phenomenon, consistent with previous
reports10. Our study focused on colour-graphemic synaesthesia
because it is the most common form10, and because it has received
considerable attention11.
A test of consistency veri®ed the presence of synaesthesia in our
group4. Participants were each given a 150-item list containing
letters (A±Z), digits (0±9) and words. They described their synaes-
thetic colour for each item (or an arbitrary colour in the case of non- Category
synaesthetic controls). Three months later, without warning, the
synaesthetes were given the same list and again asked to indicate Figure 1 Mean (+1 s.e.) consistency of colour associations for 150 items (letters, arabic
their synaesthetic colour for each item. For controls, the retest was numerals and words), plotted separately for each of 11 categories tested. Results for
given just one month later, thus giving them a potential advantage. synaesthetes (®lled bars) represent performance with a 3-month retest interval; those for
The synaesthetes were highly consistent in their responses overall, non-synaesthetic controls (open bars) represent performance with a 1-month retest
signi®cantly more so than the controls (F1,28 = 162.56, P , 0.0001; interval. For every category tested synaesthetes were more consistent in their colour
Fig. 1). These ®ndings show that the unusual sensations experienced associations than non-synaesthetic controls.
580 © 2001 Macmillan Magazines Ltd NATURE | VOL 410 | 29 MARCH 2001 | www.nature.com
1082 Wolf Prize in Agriculture
Green leafy volatiles (GLV), six-carbon aldehydes, alcohols, and with (E)-2-hexenal released significantly greater quantities of
esters commonly emitted by plants in response to mechanical VOC than control plants, they released significantly less than
damage or herbivory, induced intact undamaged corn seedlings to plants treated with MeJA or damaged by insect herbivores (13).
rapidly produce jasmonic acid (JA) and emit sesquiterpenes. More Also, in other studies, treatment of plants with six-carbon
importantly, corn seedlings previously exposed to GLV from neigh- aldehydes induced less than the complete set of defense-related
boring plants produced significantly more JA and volatile sesquit- genes and resulted in a moderate plant response relative to
erpenes when mechanically damaged and induced with caterpillar MeJA at both the physiological and molecular levels (11, 12).
regurgitant than seedlings not exposed to GLV. The use of pure Two important questions arose from these findings: Which
synthetic chemicals revealed that (Z)-3-hexenal, (Z)-3-hexen-1-ol, signaling pathways are involved, and is the immediate but rather
and (Z)-3-hexenyl acetate have nearly identical priming activity. moderate activation of plant defense responses the main func-
Caterpillar-induced nocturnal volatiles, which are enriched in GLV, tion of this signaling? To answer these questions, we have started
also exhibited a strong priming effect, inducing production of a comprehensive investigation of the effects of naturally released
larger amounts of JA and release of greater quantities of volatile GLV on neighboring plants. We used GLV from wounded plant
organic compounds after caterpillar regurgitant application. In tissue, caterpillar-induced night-time volatiles enriched in GLV,
contrast, GLV priming did not affect JA production induced by and pure C6 compounds to examine their effects on intact corn
mechanical wounding alone. Thus, GLV specifically prime neigh- plants. We discovered not only that GLV stimulate transient
boring plants against impending herbivory by enhancing inducible jasmonic acid (JA) biosynthesis and VOC release in corn, but
chemical defense responses triggered during attack and may play also that exposure to GLV primed corn plant defenses to
a key role in plant–plant signaling and plant–insect interactions. respond more strongly against subsequent attack by herbivorous
insects by increasing JA biosynthesis and VOC release.
PLANT BIOLOGY
and predators, natural enemies of the actively feeding arthro-
pods (2, 3). Herbivore-induced VOC have also been shown to first and early second instar larvae were selected for the induc-
decrease oviposition rates and increase egg predation on the tion of nocturnal volatiles.
emitting plant in nature (4, 5). Although insect-induced VOC
can serve direct and indirect defense functions, neighboring (or Preparation of Crude Regurgitant Elicitor (CRE) from Larvae of BAW.
receiver) plants may also perceive and respond to these signals. BAW were transferred to feed on corn seedlings at least 48 h
Pathogen infection of plants often leads to the release of methyl before collection of regurgitant. Regurgitation was induced by
salicylate (6), which serves as a potential signal for the induction holding fourth instar BAW caterpillars with forceps and gently
of defense-related genes in neighboring plants. Also, after pinching behind the head with a second pair. The regurgitant
wounding and herbivore damage some plants emit methyl jas- from 40–50 caterpillars was collected, boiled for 5 min to
monate (MeJA), which has been shown to effectively turn on inactivate degrading enzymes (16), centrifuged to remove cell
defense genes (7). However, not all plants release MeJA, calling debris and denatured proteins, and the supernatant diluted 1:1
into question its role as a general volatile defense signal. Green in buffer (50 mM sodium phosphate, pH 8) before use (referred
leafy volatiles (GLV) consist mainly of degradation products to as CRE).
derived from C18 fatty acids (linolenic and linoleic acid), which,
after being transformed to a hydroperoxide by a lipoxygenase, Chemicals. (Z)-3-HAL (50% in triacetin), (Z)-3-HOL (98%
are cleaved into C12 and C6 components by hydroperoxide lyase pure), cis jasmone (85% pure), methyl salicylate, and MeJA were
(HPL). Depending on the C18-substrate, HPL produces either purchased from Sigma–Aldrich. (Z)-3-HAC was made from
(Z)-3-hexenal [(Z)-3-HAL)] or hexanal (8). Further processing (Z)-3-HOL by acetylation with acetyl chloride and purity esti-
by alcohol dehydrogenase, acetylation, and isomerization leads mated by GC and GC兾MS analysis (95% pure). Dihydro MeJA
to the production of the remaining C6-components, like (Z)-3- (Bedoukian Research, Danbury, CT) was converted to dihydro
hexenol [(Z)-3-HOL)], (Z)-3-hexenyl acetate [(Z)-3-HAC], and
the respective E-isomers. The C12-component is processed to
Abbreviations: JA, jasmonic acid; SA, salicylic acid; GLV, green leafy volatiles; CIV, caterpil-
traumatin, which has long been hypothesized to play an impor-
lar-induced volatiles; VOC, volatile organic compounds; (Z)-3-HAL, (Z)-3-hexenal; (Z)-3-
tant role in the wound response of plants (9). GLV are typically HOL, (Z)-3-hexenol; (Z)-3-HAC, (Z)-3-hexenyl acetate; MeJA, methyl jasmonate; CRE, crude
released locally by plants immediately after wounding or herbi- regurgitant elicitor; FW, fresh weight.
vore damage (8) but can also be induced and released system- *Present address: Department of Entomology, Pennsylvania State University, Pesticide
ically (10). Previous studies indicated that GLV induce certain Research Laboratory, University Park, PA 16802.
defense-related genes (11–13). However, although plants treated †To whom correspondence should be addressed. E-mail: jht2@psu.edu.
JA (DhJA) by alkaline hydrolysis. [2H6]salicylic acid (SA) was Table 1. Amount of GLV (in ng) released by source plants
purchased from CDN Isotopes (Pointe-Claire, QC, Canada). All upstream from receiver plants during periods of treatment
solvents used were analytical grade. CIV GLV Control
Plant Treatments. Two different sets of experiments were con- 30 min Z-3-HAL 356 ⫾ 95 4,872 ⫾ 633 ND
ducted, one to measure the direct effect of GLV on JA, SA, and Z-3-HOL 108 ⫾ 53 2,736 ⫾ 546 ND
VOC production in plants, and the second to determine the Z-3-HAC 595 ⫾ 569 3,720 ⫾ 1,052 ND
effect of GLV treatment on subsequent plant response to Overnight Z-3-HAL 2,177 ⫾ 470 653 ⫾ 339 44 ⫾ 37
wounding and treatment with caterpillar regurgitant. To mea- Z-3-HOL 1,210 ⫾ 128 3,137 ⫾ 245 44 ⫾ 30
sure the short-term production of JA and SA, intact corn plants Z-3-HAC 3,540 ⫾ 833 4,102 ⫾ 663 207 ⫾ 184
(receiver plants) were exposed to various volatiles in 6-l Plexiglas
Data are from caterpillar-infested corn plants (CIV), cut leaf material (GLV),
cylinders. After 0, 30, 60, and 180 min, the intact receiver plants and control plants during the 30-min and overnight incubation period. Data
were removed from the chamber and the leaves frozen in liquid represent mean ⫾ SD (n ⫽ 4). ND, not detected.
nitrogen for further analysis. For short-term exposure to GLV
vapors, 2 g of cut-leaf material from 2- to 3-wk-old corn plants
was placed on a small dish and added to the chamber with the plants, and air was drawn through the trap at 200 ml兾min for
receiver corn plant. Control plants were held in the chambers for various periods of time, depending on the experiment. VOC
equal periods of time in ambient air. For induction with synthetic were eluted from the trap with dichloromethane, nonyl acetate
compounds, 20 g of (Z)-3-HAL, (Z)-3-HOL, (Z)-3-HAC, or added as an internal standard, and samples were analyzed by GC
cis jasmone (17) (dissolved in dichloromethane, 1 g兾l), was and GC兾MS (3). The identity of each compound was confirmed
pipetted onto a cotton ball in the Plexiglas cylinder. Controls by comparison of retention times and mass spectra with those of
consisted of a plant in a chamber with 20 l of pure dichlo- authentic chemicals, and quantities were determined by com-
romethane or 20 g of triacetin in dichloromethane [for treat- parison of peak areas with peak area of internal standard. The
ments with (Z)-3-HAL)] on a cotton ball. For short-term average amounts of GLV released from the cut-leaf material and
exposure to caterpillar-induced volatiles (CIV) emitted from after caterpillar damage are shown in Table 1.
neighboring infested plants, 2-wk-old intact corn plants held in
200-ml glass tubes were infested with 20–25 BAW caterpillars Quantification of JA and SA. Extraction and quantification were
(late first to early second instar) in the morning. After 5 h, each performed as described (18, 19). In brief, plant tissues were
glass tube with a caterpillar-infested corn plant (source plant) frozen in liquid N2, and ⬇100 mg of each sample was transferred
was connected by Teflon tubing (i.d. 5 mm) to a 6-liter Plexiglas to 2-ml screw-cap FastPrep tubes (Qbiogene, Carlsbad, CA)
cylinder containing an intact receiver plant. A vacuum was containing 1 g of Zirmil beads (1.1 mm; SEPR Ceramic Beads
attached to each Plexiglas cylinder, and fresh charcoal-purified and Powders, Mountainside, NJ). Dihydro JA and [2H6]SA (100
air was pulled at ⬇200 ml兾min over the infested plant and then ng) were added to the 2-ml tubes before sample addition. The
through the Plexiglas cylinder to allow the entrained nocturnal samples were mixed with 300 l of 1-propanol兾H2O兾HCl
volatiles to flow over the intact plants. As a control, uninfested (2:1:0.002) and shaken for 30 s in a FastPrep FP 120 tissue
corn seedlings were used as source plants. homogenizer (Qbiogene). Dichloromethane (1 ml) was added to
A second control was performed by using CRE-induced corn each sample, reshaken for 10 s in the homogenizer, and centri-
plants as a source for VOC. Plants were induced through
fuged at 11,300 ⫻ g for 30 s. The bottom dichloromethane兾1-
application of 5 l of CRE on one wounded site on each leaf
propanol layer was then transferred to a 4-ml glass screw-cap
(three leaves total) and then transferred to the 200-ml (vol) glass
vial, with care taken to avoid transfer of the upper aqueous layer.
tubes. After 5 h, the tubes were connected to the 6-l Plexiglas
The organic phase was evaporated by a constant airstream and
cylinders with the receiver plants as described above for CIV.
100 l of diethyl ether兾methanol (9:1, vol兾vol) added. Carbox-
For overnight exposure to GLV, cut corn leaf material was
added to the 200-ml glass cylinder, and GLV was drawn over the ylic acids were converted into methyl esters by the addition of 2
receiver plants in 6-liter cylinders at 200 ml兾min, as described l of a 2.0 M solution of trimethylsilyldiazomethane in hexane.
above. In the controls, the 200-ml cylinders were empty. For The vials were then capped, vortexed, and allowed to sit at room
induction with synthetic compounds, 20 g of (Z)-3-HAL, temperature for 30 min. Excess trimethylsilyldiazomethane was
(Z)-3-HOL, or (Z)-3-HAC (dissolved in dichloromethane, 1 then destroyed by adding an equivalent molar amount of acetic
g兾l) was pipetted onto a cotton ball in the Plexiglas cylinder. acid to each sample.
For controls, 20 l of dichloromethane was added to the cotton Volatile metabolites were separated from the complex mixture
ball. Corn plants were exposed to CIV overnight (15 h) in the by vapor-phase extraction as described in ref. 19. The trapped
same manner described for short-term exposure to CIV (see volatiles were then eluted with 150 l of dichloromethane and
above). analyzed by chemical ionization-GC兾MS (19).
For induction with CRE, intact corn plants were exposed to
GLV, CIV, or the respective synthetic chemical overnight, as Analysis of Released VOC. Plants exposed to GLV, CIV, and pure
described above. After 15 h, plants were removed from Plexiglas synthetic compounds overnight (15 h) were transferred to 200-ml
cylinders. An area of ⬇2 ⫻ 10 mm on the third leaf of each plant glass cylinders and VOC collected for 1 h without further
was scratched with a razor blade and 5 l of CRE from BAW treatment. In a second experiment, intact corn plants were
immediately added to the wounded site. For controls, buffer only exposed overnight as described above. After 15 h, plants were
was added to the wounded site. Plants were harvested 0, 30, 60, removed from the Plexiglas incubation chamber and induced
and 180 min after application of CRE, and JA and SA quantified with CRE as described above. Plants were subsequently trans-
(see below). In a separate experiment, VOC were collected from ferred to the 200-ml glass cylinder, and VOC were collected for
plants exposed to GLV, CIV, and synthetic compounds over- 1-h periods beginning 30 min after induction. Between sequen-
night and then treated with CRE (see below). tial volatile collections from the same plant, there was a 30-min
To estimate the amount of each compound in the volatiles to delay, during which the plant was watered. Volatiles were
which the corn plants were exposed (Table 1), a Super Q collected by pulling purified air at 200 ml兾min over the plants
filter-trap (Alltech Associates) was connected to the down- and through Super Q filter traps. Analysis of the trapped VOC
stream side of the Plexiglas cylinders containing the treated was performed as described in ref. 3. The amounts of linalool,
PLANT BIOLOGY
plants were used as a source of volatiles in the controls. (C) Induction of VOC indicators of the induction of defense responses. After overnight
after overnight exposure to GLV, synthetic C6 compounds, CIV, or controls, as exposure to GLV, CIV, and synthetic C6 compounds, the resting
described above. The amounts of linalool, 4,8-dimethylnona-1,3,7-triene, levels of JA in these plants were the same as in the control plants
-caryophyllene, bergamotene, and -farnesene emitted by the plants during (8 ng兾g FW). Thirty minutes after induction with CRE, the level
the volatile collection period were measured and summed to obtain total of endogenous JA rose in control plants to 96 ng兾g FW, whereas
volatiles. in GLV-pretreated plants, 190 ng兾g FW were found (Fig. 2 A and
B). The level of JA in GLV-pretreated plants remained higher
over a period of 3 h but followed the same trend as JA in
4,8-dimethylnona-1,3,7-triene, -caryophyllene, bergamotene, CRE-induced control plants (Fig. 2A). The same effect was
and -farnesene emitted by the plants during the volatile col- observed for pretreatment with pure C6 compounds and CIV
lection period were measured and summed to obtain an estimate (Fig. 2B). Surprisingly, wound-induced JA was not affected by
of total volatiles. previous exposure of corn plants to GLV (Fig. 2C).
The consequences of GLV pretreatment were further dem-
Statistical Analysis. At least three replicates of all experiments onstrated by their effect on the release of VOC induced by
were conducted. Data were analyzed for significance with t test application of CRE. Corn plants exposed overnight to GLV,
(P ⬍ 0.05). Treatments were compared to appropriate controls. CIV, and pure C6 compounds released VOC without further
treatment. However, this effect was significantly enhanced by
Results
induction with CRE. The GLV-pretreated plants released ⬇4 g
GLV Induce JA Production and Volatile Emission. When we exposed of total VOC 4–6 h postinduction compared to 2.5 g from
intact hydroponically grown corn seedlings to wound-induced nonprimed control plants (Fig. 3A). The same effect was ob-
GLV by adding cut-leaf material to the incubation chamber, JA served for pretreatment with CIV (Fig. 3B) and pure C6
was induced transiently, reaching a maximum [52 ng兾g fresh compounds (Fig. 3C). GLV-pretreated plants reached the max-
weight (FW)] 30 min after exposure (Fig. 1A). This initial burst imum release rate of unprimed plants earlier and exhibited a
of JA declined rapidly and reached the baseline levels of higher absolute release rate
untreated control plants after 2–3 h (9 ng兾g FW; control 8 ng兾g
FW). The analysis of the released GLV revealed that predom- Discussion
inantly the (Z)-3-isomers were released after wounding and Our results clearly demonstrate a specific function for GLV in
caterpillar infestation. When individual seedlings in incubation priming the defenses of corn plants against herbivorous insects.
chambers were exposed to the vapors of (Z)-3-HAL, (Z)-3- Both JA production, which is important to direct and indirect
Engelberth et al. PNAS 兩 February 10, 2004 兩 vol. 101 兩 no. 6 兩 1783
James H. Tumlinson 1085
mechanism beneficial only under conditions with a high prob- Additionally, parasitization of attacking insect herbivores in-
ability of attack. However, priming by GLV provides a different creased seed production in mature corn plants (26), demonstrat-
way of responding to the threat of insect herbivory. Defense- ing clearly the fitness benefits of this defense measure. It is
related processes are turned on but incompletely compared to obvious that the effectiveness of this strategy strongly depends
actual herbivore damage (11, 12). More importantly, the plant on the timely release of a strong VOC signal. Priming corn plants
prepares, by a yet-unknown mechanism, to respond more in- with GLV results in a faster and more intense release of these
tensely when it is subsequently attacked, as demonstrated herein VOC when induced with CRE and could give them a competitive
for the induction of JA and release of VOC. In this way, the plant advantage over nonprimed plants.
avoids great biochemical investments, which would affect the Our results demonstrate that GLV induce defense responses
general physiology significantly unless actually attacked (24, 25). in neighboring plants via induction of JA followed by the release
A further, equally important aspect of priming is the specificity of low levels of typical herbivore-induced VOC. Furthermore, a
of the signaling process. As mentioned previously, GLV induce specific priming of corn plants against subsequent insect herbi-
a subset of defense-related genes (11, 12) or cause the release of vore attack is initialized, allowing them to respond more rapidly
VOC (13), all processes related to JA. However, in addition to by enhanced JA production and an increased release of VOC
the induction of defense responses, JA is involved in various (26, 27). The principle of priming plants against pathogen
developmental processes and responses to other environmental infections by chemicals that mimic endogenous defense-related
factors in plants (21, 22). This raises the question of which JA signaling compounds is well established (28). A comparable
signaling processes are induced by GLV. In corn, there are at mechanism has not been shown to date for priming against insect
least three different ways of inducing JA, all related to wounding herbivore attack.
but different in their regulation. GLV induces JA transiently; A further indication for the specificity of this signaling is the
wounding also results in JA production; application of CRE on inactivity of other VOC in this system. However, other classes of
wounded sites combines the wound response and an elicitor VOC have been demonstrated to be interplant defense signals in
response, resulting in higher amounts of induced JA. Priming other plant species (11, 29–31). The mechanism of priming may
with GLV specifically promoted only the CRE-induced JA benefit receiver plants by reducing investment in defenses until
production, whereas wound-induced JA or JA induced by a the onset of actual herbivory. Thus, the effect of GLV is
second application of GLV after 15 h (data not shown) was not far-reaching and influences both directly and indirectly the
affected. Furthermore, SA was never affected by any of the entire tritrophic complex of plants, insect herbivores, and natural
treatments with GLV, giving further proof for the specificity of enemies of the herbivores. Future research is now directed
the GLV signal in insect herbivore defense response. toward the underlying molecular mechanism of this process in
The induction and release of VOC by plants as a response to plants.
insect herbivore damage have been demonstrated to be an
effective defense strategy. Recruiting parasites and predators of We thank the two reviewers for helpful advice that significantly improved
the herbivore (2, 3) as well as repelling female moths, thereby the manuscript. This work was supported in part by a grant from the
avoiding egg deposition (4), helps the plant to reduce damage. Defense Advanced Research Project Agency.
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25. Baldwin, I. T. (1998) Proc. Natl. Acad. Sci. USA 95, 8113–8118.
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& Takabayashi, J. (2002) Plant J. 29, 87–98. 553–565.
13. Farag, M. A. & Pare, P. W. (2002) Phytochemistry 61, 545–554. 27. Fritzsche Hobalah, M. E., Tamo, C. & Turlings, T. C. J. (2002) J. Chem. Ecol.
14. Schmelz, E. A., Alborn, H. T. & Tumlinson, J. H. (2001) Planta 214, 171–179. 28, 951–968.
15. King, E. G. & Leppla, N. C. (1984) Advances and Challenges in Insect Rearing 28. Conrath, U., Pieterse, C. M. J. & Mauch-Mani, B. (2002) Trends Plant Sci. 7,
(U.S. Govt. Printing Office, Washington, DC). 210–216.
16. Mori, N., Alborn, H. T., Teal, P. E. & Tumlinson, J. H. (2001) J. Insect Physiol. 29. Arimura, G., Ozawa, R., Shimoda, T., Nishioka, T., Boland, W. & Takabayashi,
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Engelberth et al. PNAS 兩 February 10, 2004 兩 vol. 101 兩 no. 6 兩 1785
James H. Tumlinson 1087
Contributed by James H. Tumlinson, June 27, 2007 (sent for review April 23, 2007)
A previously unidentified class of compounds has been isolated cow pea releases VOC when fed on by fall armyworm larvae, but
from the regurgitant of the grasshopper species Schistocerca not when treated with fatty acid–amino acid elicitors. This
americana. These compounds (named here ‘‘caeliferins’’) are com- phenomenon led to the identification of a previously unidenti-
posed of saturated and monounsaturated sulfated ␣-hydroxy fatty fied type of peptide elicitor in fall armyworm regurgitant,
acids in which the -carbon is functionalized with either a sulfated derived by proteolysis from the chloroplastic ATP synthase of
hydroxyl or a carboxyl conjugated to glycine via an amide bond. the plant on which the herbivore feeds (26). Thus, Spodoptera
The regurgitant contains a series of these compounds with fatty larvae produce at least two different types of elicitors, both
acid chains of 15–20 carbons and in varying proportions. Of these, derived in part from the plant, that share similar activity, i.e.,
the 16-carbon analogs are predominant and are also most active in induced release of volatiles, but on different host plants of the
inducing release of volatile organic compounds when applied to herbivores. Salivary enzymes of insect herbivores also may
damaged leaves of corn seedlings. Caeliferins are nonlepidopteran influence plant defense responses (27). Therefore, complex
elicitors identified in insect herbivores. This adds a category of interactions between herbivore-produced enzymes and elicitors
insect herbivore-produced elicitors of plant responses, providing are likely responsible for the specificity in the blend of VOC
further evidence of the ability of plants to detect and respond to released by plants attacked by Lepidoptera herbivores.
a broad range of insect herbivore-produced compounds. Corn seedlings fed on by the grasshopper Schistocerca Amer-
icana or treated with its regurgitant were previously found to
American grasshopper 兩 insect herbivory 兩 regurgitant 兩 caeliferin release VOC similar to that induced by caterpillar feeding or
regurgitant treatment (28). Here we describe the isolation,
Fig. 1. HPLC separation and bioassay results of collected fractions. (A) Fig. 2. Electro-spray LC/MS total negative ion trace of assigned active
Chromatographic trace (using 200-nm UV detection and pH 4.5 solvent) and fraction 1 (AF1) (A) and AF2 (B) (in Fig. 1C). Negative ion analyses gave a strong
1-ml fraction collection of protein-precipitated and solid-phase extraction (M-1)⫺ ion at m/z 445 (MW 446 amu) for AF1 and (M-1)⫺ ion at m/z 447 (MW
(SPE)-purified S. Americana regurgitant. (B) Bioassay results (n ⫽ 20) of 448 amu) for AF2. Both compounds gave strong double-charged ions (M-2)2⫺
combined fractions showing strong activity in the 15- to 17-min region and at m/z 222 and m/z 223, respectively. Positive ion analyses gave three charac-
some activity in the 13- to 14-min fraction. (C) Bioassay result (n ⫽ 12) of the teristic ions at MZ 464, 481, and 498 for AF1 and at MZ 466, 483, and 499 for
combined fractions 15–17 min from B after repeated HPLC. Separation was AF2, indicating compounds with a MW of 446 and 448 amu containing three
achieved using a neutral (pH 7) solvent and a slow gradient. The active 13- to acidic sites forming ammonium salts by the solvent buffer.
14-min fraction was assigned active fraction 1 (AF1) and the 15- to 16-min
fraction was assigned AF2. The excised plant assay was used for all bioassays,
and the induced volatile release was normalized and analyzed statistically (P ⬍
and m/z 303 for AF2. This finding suggested methyl esters of the
0.01) as described in Methods.
same fragments as the (M-1)⫺ ions at m/z 285 and 287 in the
LC/MS analyses (and the loss of two 80-amu groups during
reverse-phase (C18) solid-phase extraction column with 50% methanolysis). An indicative fragmentation pattern in the elec-
acetonitrile in water (water fraction, 4.6 ⫾ 1.4%; 50% acetoni- tron impact (EI) spectra was the loss of ⫺59, ⫺18, and ⫺18 amu
trile, 101.8 ⫾ 20.3%, 100% acetonitrile, 15.1 ⫾ 5.4%; n ⫽ 8). at m/z 241, 223, and 205, respectively, for AF1 (Analysis 3 in SI
Further fractionation of the 50% acetonitrile solid-phase eluant Text and SI Fig. 7) and 243, 225, and 207, respectively, for AF2,
by reverse-phase C18 HPLC with an acidic mobile phase (Fig. 1A) which indicated two alcohols and a carboxylic acid methylester
resulted in partial activity in the 13- to 14-min fraction and strong with the first alcohol in the position ␣ to the acid. Cleavage ␣ to
activity in the 15- to 17-min fractions (Fig. 1B). No obvious peaks the COOCH3 gave the characteristic M-59 fragment. The strong
corresponding to the activity could be seen in the UV trace at M-30 ion for AF2 at m/z 272 (and a weaker ion at m/z 270 for
200 nm or at higher wavelengths. The main active fraction (15–17 AF1) indicated a long-range proton transfer to the carbonyl
AGRICULTURAL
min) was further separated on the same column by using neutral followed by the loss of CH2O. Consequently, the second alcohol
SCIENCES
buffer and a modified gradient. This fraction now separated into was located on the -carbon. On the basis of this information and
two active fractions (Fig. 1C), designated AF1 (13–14 min) and NMR analyses of intact AF1 and AF2, these compounds ap-
AF2 (15–16 min). Reanalyzing AF1 on HPLC showed a peak peared to be 2,16-dihydroxy C16 fatty acids with the addition of
with what appeared to be a very low response factor at 200-nm two unknown 80-amu groups. The only difference between the
or lower wavelengths, whereas AF2 produced no detectable UV two compounds was the presence of a double bond in AF1,
response at any wavelength. explaining its weak UV absorption. To test the one double-bond
The isolated compounds were analyzed using negative and hypothesis, the AF1 methyl ester was subjected to hydrogena-
positive ion electrospray LC/MS. In negative ion mode, both AF1 tion, which, as expected, gave a GC/MS peak identical to that of
and AF2 gave very strong M-1 ions at m/z 445 and 447, the AF2 methyl ester. The presence of two alcohols was con-
respectively (Fig. 2). They also gave strong double-charged ions firmed by acetylation that resulted in the expected increase in
at m/z 222 and 223 (M-2)/2. Repeated daughter ion analyses [see molecular weight of 2 ⫻ 42 amu for both compounds (Analysis
Analysis 1 in supporting information (SI) Text and Fig. 5] showed 2 in SI Text and SI Fig. 6). Finally, the presence of only one
that both compounds readily lost two 80 atomic mass units carboxylic acid was confirmed by ethanolysis and chemical
(amu)-neutral fragments to give stable ions at m/z 285 and 287, ionization (CI) GC/MS analyses that, for both compounds, gave
respectively. In positive ion mode, both compounds showed an ethyl ester with M ⫹ 1 ions 14 amu higher than for the
three prominent ions 17 mass units apart (Fig. 2), indicating corresponding methyl esters.
ammonium salts of three acidic sites and confirming molecular CI GC/MS analyses of ozonized acetylated AF1-methyl ester
weights of 446 amu and 448 amu. gave 10-acetoxydecanal with an M ⫹ 1 ion at m/z 215 and a
Chemical ionization GC/MS analyses (Analysis 2 in SI Text and compound with an M ⫹ 1 ion at m/z 203 containing a 6-carbon
SI Fig. 6) of methanolysed AF1 gave (M ⫹ 1)⫹ ions at m/z 301 chain with aldehyde and carboxyl methyl ester functions on the
Fig. 3. The structures of caeliferin A16:1, and A16:0 and caeliferin B16:1 and
B16:0. The H-NMR chemical shifts were obtained for isolated natural com-
pounds dissolved in D2O. Caeliferin A16:1 ⫽ (E)-2,16 disulfooxy-6-hexadecenoic
acid; caeliferin A16:0 ⫽ 2,16 disulfooxyhexadecanoic acid; caeliferin B16:1 ⫽ Fig. 4. Negative ion LC/MS separation and bioassay results of collected
N-[(E)-15-sulfooxy,15-carboxy-10-pentadecenoyl] glycine; and caeliferin fractions. (A) Total ion trace of regurgitant from laboratory-reared S. Amer-
B16:0 ⫽ N-(15-sulfooxy,15-carboxy pentadecanoyl) glycine. icana. The numbers indicate fractions that were collected for intact plant
bioassays. Fraction 2, caeliferin B16:1; fraction 4, caeliferin B16:0; fraction 7,
caeliferin A16:1; fraction 9, caeliferin A16:0; fraction 10, caeliferin A17:0; and
opposite ends of the chain and an acetoxy ␣ to the methyl ester. fraction 11, caeliferin A18:0. (B) Normalized result of intact seedling bioassay
Consequently, the double bond was located between carbon 6 at natural relative concentrations (0.5-l regurgitant equivalents), n ⫽ 12. (C)
and 7 in the 16-carbon chain. Bioassay of selected fractions with all of the concentrations adjusted to that
The AF1 methyl ester was also analyzed by GC/FTIR. Two of caeliferin A16:1 (100 pmol in 0.5-l regurgitant equivalents). Different
letters indicate significant differences (P ⬍ 0.01).
weak absorptions at 3,643 and 3,568 cm⫺1 for AF1 confirmed two
nonidentical alcohols. Furthermore, absorption at 959 cm⫺1
indicated the presence of a trans double bond. NMR analysis of Natural and synthetic caeliferin A16:1 gave a relative response
the original intact AF1 (not subjected to methanolysis) gave 1H of 82 ⫾ 14% versus 87 ⫾ 14%, and caeliferin A 16:0 gave 86 ⫾
at ␦ ⫽ 4.45 ppm (broad), indicative of a proton on an alcohol- 17% and 100 ⫾ 19%, respectively. However, all treatments were
substituted carbon ␣ to a carboxylic acid, and 2H at ␦ ⫽ 3.92 significantly different (P ⬍ 0.01) from the buffer control with a
ppm, suggesting two protons on an alcohol-substituted -carbon. relative response of 23 ⫾ 2%. Although it appears chirality is not
Two protons at ␦ ⫽ 5.42 and 5.36 ppm, J15 Hz, indicated a trans critical for the biological activity, this will still be addressed in an
double bond for AF1. However, comparison with spectra of improved synthesis. Preliminary data indicated that the activity
commercial ␣-hydroxy and -hydroxy palmitic acid indicated the is reduced as the number of sulfates on synthetic caeliferin A16:0
␣- and -protons did not have chemical shifts identical to the is reduced. However, this could also have been the result of a
comparable protons of AF1 and AF2, confirming that the two diminishing solubility in water.
substituents on AF1 could not be simple alcohols. Furthermore, The initial bioassays during isolation and purification indi-
the NMR analyses did not indicate the presence of any other cated that there might be related, but less-active compounds
organic structure than a di-O(H) substituted C16 fatty acid. present in the regurgitant (Fig. 1). This was confirmed by LC/MS
There was also no indication of phosphate. However, sulfate analyses of the supernatant after precipitation of protein from
esters would explain the consecutive loss of 80 amu in LC/MS crude regurgitant (Fig. 4A). Both C17 and C18 (as well as traces
analyses [loss of two SO3; molecular weight (MW), 80 amu] as of C15, C19, and C20) versions of caeliferin A16:0 were found, with
well as the presence of three acidic sites (two sulfates, one C17 predominant over C18, although there appeared to be only
carboxylic acid) and the water solubility of the natural products. the C16 version of caeliferin A16:1 at detectable amounts. There
The most likely candidates for AF1 and AF2 were therefore was also a set of related but different analogs. Of those, peak 4
2,16-disulfooxy-E6-hexadecenoic acid and 2,16-disulfooxyhexa- in Fig. 4A, with a molecular weight of 439 amu, was the most
decanoic acid shown in Fig. 3, which we named caeliferin A16:1 likely compound responsible for the weak activity in fraction
and caeliferin A16:0, respectively. 13–14 in Fig. 1B. Sequential daughter ion analyses of this peak
Both proposed (racemic forms of) dihydroxy acids were revealed only one sulfate to be present. GC/MS analyses of the
synthesized and transformed to disulfate esters (Analysis 4 in SI methyl ester gave a MW of 330 amu, whereas the ethyl ester gave
Text and SI Fig. 8). The synthetic versions of caeliferin A16:1 and a MW of 358 amu (330 ⫹ 2 ⫻ 14), indicating a diethyl ester. The
A16:0 were analyzed by HPLC, LC/MS, NMR, and, after acetylated dimethyl ester had a MW of 372 amu (330 ⫹ 42),
derivatizing, EI and CI GC/MS and shown to be identical to the showing the addition of one acetate as expected. EI spectra of
natural products. Synthetic and natural products were purified the dimethyl ester gave a clear M-59 ion at m/z 271 (M-
on HPLC and bioassayed (n ⫽ 16) at their respective regurgitant COOCH3). Thus, peak 4 appeared to be ␣-sulfooxy hexade-
concentration (4 nmol/l for A16:0 and 100 pmol/l for A16:1) canedioic acid with a calculated molecular weight of 382 amu.
by using the intact plant assay, which is a better mimic of natural However, that is 57 mass units less than the MW given by the
damage than the excised plant assay (29). It also required less LC/MS analyses and indicated the presence of nitrogen. Com-
material per bioassay [0.5-l regurgitant equivalents (MRE) per pound 4 was therefore subjected to a mild methyl ester/acetate
seedling]. The bioassay showed none of the treatments to be procedure especially suitable for compounds containing amino
significantly different from the crude regurgitant treatment. acids (20, 30). GC/MS analysis with on-column injection of the
product resulted in three peaks; the mono-acetylated dimethyl pected C16 and C18 to dominate over C17 and, if the origin was
ester of the complete compound (minus the sulfate) with a MW plant lipids, then we would expect more C18 than C16 and also
of 429, the acetylated methyl ester seen before with a MW of 372 double bonds to be cis. However, in S. americana regurgitant, the
and finally the acetylated methyl ester of glycine (MW 131), relative abundance of caeliferin A16:0, A17:0, and A18:0 is
giving the final structure of caeliferin B16:0 in Fig. 3. Also, ⬇30:5:1, and the double bond in caeliferin A16:1 is trans. We
homologs with C17 and C18 fatty acid chains (with traces of C15 have not found hydroxylation at any but the 2 and 16 positions.
and C19) were found in analogy with caeliferin A16:0. There The 2-hydroxylation might explain the unusual lengths in carbon
were also traces of an unsaturated version of caeliferin B16:0 chains as the first step in a one-carbon chain-shortening se-
(peak 1–3 in Fig. 4A), consequently named caeliferin B16:1 (Fig. quence. Furthermore, the -hydroxyl might be the result of
3). Proton NMR analysis of fraction 4 showed the same single enzymatic reduction of an amide precursor (caeliferin B). How-
proton at a substituted carbon ␣ to a carboxylic acid (␦ ⫽ 4.45 ever, that still leaves us with a unique and unknown precursor.
ppm) (broad) but, as expected, with no indication that an It appears that all caeliferins in all species investigated are totally
alcohol-substituted -carbon was present. However, the two sulfated in the pH 5 regurgitant, whereas alkaline or stronger
protons at ␦ ⫽ 2.14 ppm confirmed the amide, and finally a acid conditions result in partially or completely desulfated
singlet with 2 protons at ␦ ⫽ 3.62 ppm confirmed the presence compounds.
of glycine. The range of biological activity and function of caeliferins in
LC/MS made it possible to collect and bioassay the identified grasshoppers is not fully understood. Sulfated compounds, for
caeliferins and several of the related peaks in Fig. 4A. However, example, glycosaminoglycans, and proteins, have been found in
the LC/MS trace could not be used for quantitative estimates due insect ovaries, eggs, and fat bodies (33–35). The hydrophilic
to different response factors for caeliferin A and B. Therefore sulfate groups make caeliferins especially useful in bridging
the absolute amount of the collected compounds in each fraction lipids and water, and as such they may function in digestion.
was established by derivatization and GC analysis. In the first Additionally, they may also be an important part of chemical
intact plant bioassay, we used the same regurgitant equivalents defense, because grasshoppers regurgitate copious amounts of
(0.5 l) for each fraction tested, maintaining their natural frothy secretions as a defense against attack. Indeed, investiga-
relative concentrations (as in Fig. 1). The results (Fig. 4B) were tions (36–38) have shown that both compounds from host plants
also very comparable to the earlier results. Caeliferin A16:1 is and insect produced water-soluble components of the regurgi-
naturally only a minor component (⬇200 pmol/l) compared tant have deterrent activity (36). Although the deterrent effect
with caeliferin A16:0 (⬇4 nmol/l) and B16:0 (⬇500 pmol/l). of caeliferins remains to be tested, it is probable that the
In a second assay (Fig. 4C) we bioassayed the major compounds emulsifying properties are important in aiding the water-based
at the same concentration (100 pmol per seedling). In this regurgitant to carry lipophillic toxic compounds.
experiment caeliferin A16:1 was significantly more active than It has been well demonstrated that induced plant volatiles play
any other component, including caeliferin A16:0. The latter was a role in attracting natural enemies of Lepidopteran herbivores,
now approximately as active as its C17 and C18 analogs and but there is no behavioral evidence that we are aware of that
caeliferin B16:0. indicates this occurs for grasshoppers. Jasmonic acid signaling is
a key player in induced plant response to wounding and feeding
Discussion damage. Recently it has been shown that a sulfotransferase in
We report a new class of insect herbivore-produced plant volatile Arabidopsis thaliana inactivates 12-hydroxyjasmonate by trans-
elicitors with strikingly different chemical structures than those forming it to its corresponding sulfate (39). Caeliferins might
previously described. Yet, the basic moiety of these structures is interact with this system, but at this stage we don’t know the full
a fatty acid, such as the fatty acid–amino acid conjugates of the range of activity of caeliferins on plants and signal cascades.
Lepidopteran elicitors, although the most active, as well as the It has been reported that S. americana avoids diet with added
most abundant, compounds from S. americana have 16- rather monoterpenoids (40). We know, from electroantennogram ex-
than 18-carbon chains. Preliminary analyses of regurgitant col- periments, that S. americana detects most of the volatile com-
lected from several other species of Orthoptera indicates that the pounds released by induced corn seedlings. It is therefore likely
caeliferins may be present in most, if not all grasshoppers, that, under natural conditions, the solitary S. americana will stop
members of the suborder Caelifera, but not in crickets or feeding on and even avoid induced plants. Thus, the plants will
katydids in suborder Ensifera. This was the basis for naming directly benefit from the release of VOC in response to grass-
AGRICULTURAL
these new compounds caeliferins. Interestingly, regurgitant of at hopper feeding and caeliferins. Most of the time the American
SCIENCES
least some crickets (31) and katydids (unpublished data) contain grasshopper lives a solitary life, but occasionally they transform
fatty acid amides, although we have not been able to find even into a gregarious stage, for example, when crowded. At that
traces of these in grasshopper regurgitant. stage, plant volatiles could aid in aggregation. Caging will
In the Lepidoptera, the fatty acid moiety of the elicitors is transform wild, solitary, nymphal as well as adult S. americana
derived from the diet on which the larvae feed (25), and variation from solitary to gregarious stage, with the typical change in
in proportions of dietary fatty acids results in similar changes in coloration occurring within a few days. This has so far made it
proportions of the corresponding elicitors (32). The composition difficult to study changes in behavior, including their response to
of the regurgitant of laboratory-reared S. americana is very induced plant volatiles. This physical change occurs in parallel
consistent and does not change with diet or maturing of the with the change in caeliferin composition, mentioned earlier.
insect (data not shown). However, regurgitant from field- S. americana feeds on a broad range of plants (41). There is
collected insects can contain up to 5-fold more of caeliferin now evidence for dramatic variation in the response, or lack of
A16:1, as well as more of the glycine-containing caeliferin B16:1 response, to volicitin- and inceptin-types of elicitors from Lep-
and B16:0 than laboratory-reared insects. Furthermore, the idopteran larvae among plant species (26). It is an intriguing
regurgitant composition in wild grasshoppers will change into possibility that analogs of caeliferin A and B have different levels
laboratory-reared proportions within a week of caging in the of activity on the wide variety of host plants of S. americana or
laboratory, independent of what diet the insects are fed. even on different varieties of the same species of plants.
Whether this is diet-related or due to other factors, such as Induced release of volatiles is just one of many levels of
crowding, is not yet known. The origin, or precursors of the S. defense against phytophagous insects, and it would be surprising
americana elicitors is not known. On the basis of the natural if the known elicitors do not also trigger nonvolatile defenses. An
preference for even carbon chain lengths, we would have ex- investigation of the activity of these elicitors on several plant
species in inducing volatile emission, as well as increased levels in refs. 22 and 28, and were analyzed by gas chromatography
of jasmonic acid and other plant hormones. The discovery of (Analysis 5 in SI Text and SI Fig. 9). Volatiles were collected for
caeliferins provides new biological tools and directions to ex- 2 h for the cut-stem assays and for 1 h for the intact-plant assays.
plore the physiological ecology and interactions of insects and The response by the plant to each test sample was measured as
plants. the combined amount of four induced terpenoids: E--
caryophyllene, E- ␣ -bergamotene, ␣ -humulene, and E-  -
Materials and Methods farnesene. Although several more volatile components are
Insect Rearing. S. americana were either obtained from a colony released by Delprim cultivar in response to insect herbivory (5),
maintained at the Department of Entomology and Nematology these four components show the strongest induced response. All
at the University of Florida or field-collected in Gainesville, FL. bioassays also included positive controls (0.5 or 5 l of crude
The insects were fed lettuce, wheat, or corn leaves and kept in regurgitant, depending on bioassay) and a negative control
cages in a temperature-controlled greenhouse (25°C during the (buffer only). All samples were replicated three to four times
day/20°C during the night) with natural light, supported by a within each bioassay and each bioassay was repeated three to
mixture of high-pressure sodium and metal halide lamps (400W four times (giving a minimum total of n ⫽ 9 per treatment). To
Lucalox; GE, Piscataway, NJ) to provide 14 h of day/10 h of reduce the variation in response between bioassays, a correlation
night. To facilitate reproduction and egg laying, cups with moist factor was calculated for each bioassay to give the mean release
vermiculite were kept in the cages. of induced volatiles for the regurgitant treatments a response of
100. The correlation factor was then used for all samples within
Plants. All plants were grown in 1-gallon containers (Miracle that bioassay. Data were analyzed using a Tukey pairwise
Grow no. 92695 potting soil fertilized with 1 teaspoon of multiple comparison test.
Osmocote 14-14-14; Scotts-Sierra Horticulture, Marysville,
OH). Feed corn variety Delprim (Delley Seeds and Plants, Collection and Initial Fractionation of Regurgitant. Regurgitant was
Delley, Switzerland) was planted 10 seeds per pot and main- collected from both wild and laboratory-reared nymphs and
tained on a 12-h dark and 12-h light cycle. One week after adults as described earlier for Lepidoptera caterpillars (22). The
germination, the number of seedlings per pot were either regurgitant was diluted 50:50 with acetonitrile, centrifuged at
reduced to six of uniform size for the cut stem bioassay or 16,000 ⫻ g for 15 min to remove precipitated proteins, and then
transferred to hydroponic chambers (18). The light intensity was filtered through 0.45- and 0.22-m sterilizing membranes. The
305 mol䡠m⫺2䡠S⫺1 of photosynthetically active radiation sup- supernatant was concentrated to original volume by a gentle
plied by a mixture of high-pressure sodium and metal halide stream of N2 and stored at ⫺70°C until used. A C18 solid-phase
lamps (400W GE Lucalox) at the top of the plant canopy with extraction column (mega bond elute; Waters, Milford, MA) was
day/night temperatures at 25°C/20°C, respectively. The relative
activated by eluting with 40 ml of acetonitrile and then with 40
humidity ranged from 60% to 70%. For both bioassays, 10- to
ml of water. Two milliliters of concentrated supernatant was
12-day-old seedlings that contained three to four leaves were
added to the top of the column and separated into three fractions
used for treatments and volatile collections.
by eluting with 20 ml of water, followed by 20 ml of 50:50
water:acetonitrile and then by 20 ml of acetonitrile with gravity
Plant Bioassay. For both bioassays, plants were treated ⬇2 h into
flow. Each fraction was concentrated to the original volume
the scotophase and incubated for ⬇16–17 h on the same
day/night schedule under which they were grown but with before bioassaying.
reduced daytime light (2,000 lux) before volatile collection.
HPLC Purification. The active 50% acetonitrile-water fraction was
Excised Plant. At the time of treatment, the corn seedlings were
further fractionated by HPLC under acidic conditions (AS3500
cut off near the base of the stem with a razor blade, and the cut Autosampler, Constametric 4100 Pump, and Spectromonitor
end was immediately immersed in 600 l of 50 mM Na2HPO4 3200 variable wavelength detector; Thermo Separation Prod-
buffer (pH 8) (control) or a test solution in 600 l of the same ucts, Riviera Beach, FL), while monitoring UV absorption at 220
buffer, contained in a 1.5-ml vial. Five-microliter regurgitant nm. With the temperature maintained at 60°C, a C18 reverse-
equivalents (MRE) of the crude regurgitant or subsequent phase column (ODS-AMQ, S-5 m, 200 A, 250 ⫻ 4.6 mm i.d.;
fractions thereof were used to treat each seedling in all bioassays. YMC, Kyoto, Japan) was eluted with a solvent gradient (1.0
After incubation, the stem of each plant was recut, wrapped in ml/min) of 10–70% acetonitrile in water for 20 min followed by
wet cotton, and placed in a glass volatile collection chamber, and 70% acetonitrile for 10 min., with both solvents containing 1 mM
volatiles were collected as described below. NH4Ac:AcOH buffer, pH 4.5 [acetic acid (Aldrich, Milwaukee,
WI), acetonitrile UV (B & J Brand High Purity Solvent,
Intact Plant. At the time of treatment the two oldest leaves of Muskegon, MI), and water (Milli-Q UV Plus System; Millipore,
individual plants each received two superficial damage sites by Bedford, MA)]. Combined fractions from several repeated
scratching the surface (2 mm ⫻ 10 mm) with a razor blade separations were evaporated under vacuum to near dryness and
approximately equidistant between the base and tip of the leaf desalted on activated C18 solid-phase extraction columns (SPE
(29). Immediately after damage, 5 l of the same buffer solution C18; J.T. Baker, Phillipsburg, NJ) that were rinsed with 2 ml of
as above containing 0.5-l regurgitant equivalents (MRE) was water, followed by 2 ml of 50% acetonitrile in water. The latter
applied evenly to the four damage sites. The plants remained in fraction was reduced to dryness and redissolved in bioassay
their hydroponic solution during incubation and then were buffer (50 mM phosphate buffer, pH 8) before bioassaying.
removed from their container, their roots were gently wrapped The most active fractions from the first HPLC separation were
in soft tissue and dipped in hydroponic solution, the seedlings combined and further fractionated by a modified solvent gradi-
were placed in the volatile collection chambers, and volatiles ent and neutral conditions by using the same column and system
were collected as described below. as above with UV absorption monitored at 210 nm. The column
was eluted with a solvent gradient (1.0 ml/min) of 15–60%
Volatile Collections. Volatile chambers (30 cm long ⫻ 4 cm i.d.) acetonitrile in water in 20 min followed by 60% acetonitrile for
were placed under the same type and intensity of light as used 10 min, with both solvents containing 5 mM NH4Ac buffer, pH
in rearing and volatiles, were collected on Super Q 80/100 7.0. Two-minute fractions were collected and desalted as de-
(catalog no. 2735; Alltech Associates, Deerfield, IL) as described scribed above before bioassaying.
LC/MS Conditions. A Thermo Finnigan LCQ Deca XP Max was same amount of regurgitant equivalents of HPLC fraction, was
used with electro spray ionization (sheet gas, 30 arbitrary units; added to a vial and evaporated to dryness with N2. Two hundred
sweep gas, 5 arbitrary units; spray voltage, 5.00 kV; capillary microliters of dry methanol/HCl (10:1) or ethanol/HCl was
temperature 275°C; and capillary voltage, 3.0 V). The Thermo added, and the sealed vial was heated to 100°C for 30 min. The
Separations spectra HPLC system consisted of a quaternary mixture was allowed to cool to room temperature, 500 l of
pump P4000, autosampler AS 3000, and diode array detector methylene chloride was added, and the solution was extracted
UV6000. The tertiary solvents consisted of (i) methanol with twice with 500 l of water. The organic phase was evaporated to
0.05% formic acid, (ii) water with 10 mM ammonium formate, dryness with N2, and 500 l of fresh methylene chloride was
and (iii) 90% acetonitrile/10% water with 10 mM ammonium added for GC/MS analyses.
formate. With the column temperature maintained at 60°C and For acetylation, 20 l of acetyl chloride was added to the above
a solvent flow of 1.0 ml/min, the C18 column (ODS-AMQ, S-5 solution, which then was heated to 90°C for 30 min. The solution
m, 200 A, 250 ⫻ 4.6 mm i.d.; YMC) was eluted with a solvent was then extracted three times with water before GC/MS
composition starting with 4:90:6 (i:ii:iii) for 1 min, followed by a analyses.
gradient to 4:50:30 in 13 min and a fast ramping to 4:0:96 in 2 min HPLC fractions derivatized for GC/MS analyses (acetylated
and was then kept at that composition for 5 min. UV absorption methanolysis products) were ozonized as described in refs. 20
was monitored at 190–400 nm, and the solvent flow between the and 42 and analyzed by GC/MS by using on-column injection.
UV detector and MS electro spray interface was split 9:1 with a Double bonds were reduced by gently drying methylene
low-volume micro needle valve splitter P450 (Upchurch Scien- chloride solutions of acetylated methylesters of pure HPLC
tific, Oak Harbor, WA) making it possible to obtain spectra of fractions with N2 and redissolving in ethyl acetate. A few grains
eluted compounds and simultaneously collect 90% of the in- of Pd on carbon were added, H2 was gently bubbled through the
jected material for bioassaying. solution for 1 h, and the solution analyzed on GC/MS.
GC/MS Conditions. Derivatized samples were analyzed by GC/MS NMR Analyses. 1H NMR spectra were obtained with Varian (Palo
(6,890/5,973 GC/MS; Agilent, Palo Alto, CA) in both EI and Alto, CA) Unity 600 and Bruker (Billerica, MA) Avance 500
isobutane CI mode. Samples were injected in the splitless mode instruments by using a Wilmad (Buena, NY) 520 1A microbore
at 220°C or with cold on-column. The methyl silicone column, tube. Samples were dissolved in D2O, and 1H NMR chemical
(HP1, 30 m ⫻ 0.25 mm i.d. ⫻ 0.1-m film thickness; Agilent) was shifts are reported with respect to internal tetramethylsilane.
kept at 40°C for 1 min and was then temperature programmed
at 10°C/min to 260°C. The He carrier gas flow rate was 30 cm/sec We thank Peggy Brennan, Mary Searle, Spencer Walse (all at the Center
(constant flow), and the transfer line temperature was 250°C. for Medical, Agricultural, and Veterinary Entomology at the United
States Department of Agriculture) and Jim Rocca (University of Florida,
The ion source temperature was 220°C in EI mode and 250°C in Gainesville, FL) for technical assistance; and Paul Pare (Texas Tech
CI mode. University, Lubbock, TX), Naoki Mori (Kyoto University, Kyoto, Ja-
pan), Ted Turlings (University of Neuchatel, Neuchatel, Switzerland),
Derivatizations for GC/MS. For methanolysis and ethanolysis 10 l and John Pickett (Rothamsted Experimental Station, Harpenden, U.K.)
of regurgitant supernatant after protein precipitation, or the for helpful comments.
1. Turlings TCJ, Tumlinson JH, Lewis WJ (1990) Science 250:1251–1253. 23. Mori N, Alborn HT, Teal EA, Tumlinson JH (2001) J InsectPhysiol 47:749–757.
2. Kessler A, Baldwin IT (2001) Science 291:2141–2144. 24. Pohnert G, Jung V, Haukioja E, Lempa K, Boland W (1999) Tetrahedron
3. Loughrin JH, Manukian A, Heath RR, Tumlinson JH (1995) J Chem Ecol 55:11275–11280.
1217–1227. 25. Paré PW, Alborn HT, Tumlinson JH (1998) Proc Natl Acad Sci USA 95:13971–
4. Gouinguene S, Degen T, Turlings TCJ (2001) Chemoecol 11:9–16. 13975.
5. Paré PW, Tumlinson JH (1997) Nature 385:30–31. 26. Schmelz EA, Carroll MJ, LeClere S, Phipps SM, Meredith J, Chourey PS,
6. Rose USR, Manukian A, Heath RR, Tumlinson JH (1996) Plant Physiol Alborn HT, Teal PEA (2006) Proc Natl Acad Sci USA 103:8894–8899.
111:487–495. 27. Musser RO, Hum-Musser SM, Eichenseer H, Peiffer M, Ervin G, Murphy JB,
7. Mithofer A, Wanner G, Boland W (2005) Plant Physiol 137:1160–1168. Felton GW (2002) Nature 416:599–600.
8. Spiteller D, Pohnert G, Boland W (2001) Tetrahedron Lett 42:1483–1485. 28. Turlings TCJ, McCall PJ, Alborn HT, Tumlinson JH (1993) J Chem Ecol
9. Arimura G, Huber DPW, Bohlman J (2004) Plant J 37:603–616. 19:411–425.
10. Degenhardt DC, Lincoln DE (2006) J Chem Ecol 32:725–743. 29. Schmelz EA, Alborn HT, Banchio E, Tumlinson JH (2003b) Planta 216:665–
AGRICULTURAL
11. De Moraes CM, Mescher MC, Tumlinson JH (2001) Nature 410:577–580. 673.
SCIENCES
12. Litvak ME, Monson RK (1998) Oecologia 114:531–540.
30. Mee JML, Korth J, Halpern B (1977) Biomed Mass Spectrom 4:178–181.
13. Roese USR, Tumlinson JH (2005) Planta 222:327–335.
31. Yoshinaga N, Aboshi T, Ishikawa C, Fukui M, Shimoda M, Nishida. R., Lait
14. Rodrigues-Saona C, Crafts-Brandner SJ, Williams L, Pare PW (2002) J Chem
CG, Tumlinson JHT, Mori N (2007) J Chem Ecol 33:1376–1381.
Ecol 28:1733–1747.
32. De Moraes CM, Mescher MC (2004) Proc Natl Acad Sci USA 101:8993–8997.
15. Turlings TCJ, Bernasconi M, Bertossa R, Bigler F, Caloz G, Dorn S (1998)
33. Costa A, Werneck CC, Nasciutti LE, Masuda H, Atella GC, Silva LCF (2001)
Biolog Cont 11:122–129.
Insect Biochem Mol Biol 31:31–40.
16. Turlings TCJ, Alborn, HT. Loughrin JH, Tumlinson JH (2000) J Chem Ecol
26:189–202. 34. Giorgi F, Cecchettini A, Locci MT, Masetti M, Bradley JT (1995) Develop Biol
17. Williams L, Rodriguez-Saona C, Pare PW, Crafts-Brandner SJ (2005) Arch 167:379–387.
Biochem Phys 58:84–96. 35. Giorgi F, Falleni A, Cecchettini A, Gremigni V (1998) Biol Cell 90:183–197.
18. De Moraes CM, Lewis WJ, Pare PW, Alborn HT, Tumlinson JH (1998) Nature 36. Ortego F, Evans PH, Bowers WS (1997) J Chem Ecol 23:1941–1950.
393:570–573. 37. Sword GA (2001) Oecologia 128:416–421.
19. Du YJ, Poppy GM, Powell W, Pickett JA, Wadhams LJ, Woodcock CM (1998) 38. Calcagno MP, Avila JL, Rudman I, Otero LD, Alonso-Amelot ME (2004)
J Chem Ecol 24:1355–1368. Physiol Entomol 29:123–128.
20. Alborn HT, Turlings TCJ, Jones TH, Stenhagen G, Loughrin JH, Tumlinson 39. Gidda SK, Miersch O, Levitin J, Wasternack C, Varin L (2003) J Biol Chem
JH (1997) Science 276:945–949. 278:17895–17900.
21. Alborn HT, Jones TH, Stenhagen GS, Tumlinson JH (2000) J Chem Ecol 40. Bernays EA, Chapman RF (1977) Ecol Entomol 2:1–18.
26:203–220. 41. Capinera JH (1993) Pop Ecol 22:127–133.
22. Alborn HT, Brennan MM, Tumlinson JHT (2003) J Chem Ecol 29:1357–1372. 42. Beroza and Bierl (1966) Anal Chem 38:1976–1986.
CURRICULUM VITAE
Supervisory Research Entomologist (Super Grade), United States Department of
Agriculture, Agricultural Research Service (USDA-ARS); Crop Protection and
Management Research Laboratory; Tifton, Georgia, 31793, USA (Retired September,
2006)
EDUCATION:
Ph.D.; 1968 Mississippi State University; major, Entomology; minor, Zoology
M.S.; 1965 Mississippi State University; major, Entomology; minor, Genetics
B.S.; 1964 Mississippi State University; major, Entomology; minor, Composite;
(Outstanding Senior of The Year Award, Department of Entomology)
A.A. 1962 Copiah-Lincholn Jr. College; major, Engineering.
RESEARCH EXPERIENCE:
1093
1094 Wolf Prize in Agriculture
Association for the Advancement of Science, etc. His services have consisted of
numerous Office roles, organizing many symposia/conferences, editorial duties,
and a range of professional board activities. Specific examples include:
dedicated to the subject of biological control. The journal has been highly
successful and is established as major forum for the discipline. He served as
Editor, 1989-1998 and on Editorial Board 1998-2005.
- Nominations Committee for Nomination of Officers, Georgia Entomological
Society, 1998.
insect learning. The importance of these findings and their impact on the
science of learning in general are documented in recent books: “Insect Learning:
Ecological and Evolutionary Perspectives” (D. R. Papaj and A. C. Lewis Editors -
Chapman & Hall, 1993); “Chemical Ecology of Insects 2” (R. T. Carde and
W. J. Bell Editors - Chapman & Hall, 1995) including two and one key chapters,
respectively, by Dr. Lewis and his colleagues. Moreover, this accomplishment
specifically resulted in research on parasitoid learning and foraging behavior
being incorporated as part of a major funding initiative in the Department of
Defense’s Controlled Biological Systems program targeted toward developing
novel detection and tracking technology. (Publications: #95, #101, #122, and
#135)
F. In cooperation with Dr. J. H. Tumlinson and students discovered that salivary
secretions from caterpillar hosts of Cotesia marginiventris induce host plants
to release terpenoid volatiles (apparently as a defense against the caterpillar)
which the searching parasitoids learn to exploit as a means of locating the
host caterpillars. Subsequently, they demonstrated with the Heliothis/
Helicoverpa complex and Cardiochiles nigriceps that these plant signals were
herbivore-specific and could be used selectively by parasitoids to discern sites
infested by host versus non-host herbivore species (1990-97). Impact: Though
previous workers had shown the attraction of parasitoids to plant odors and
suggested an active interaction between the herbivore-damaged plant and the
third trophic level, no specific herbivore-induced factor used by the foraging
parasitoid had been pinpointed. These breakthroughs demonstrate the closely
interwoven evolutionary relationships among different levels of the tritrophic
system and have opened exciting new possibilities for joint use of plant breeding/
engineering and entomophages for pest management and prevention. Extensive
interdisciplinary interest, fundamentally and applied, has been demonstrated
by researchers and commercial practitioners in these and related discoveries by
Dr. Lewis and co-workers as part of a wave of international interest in plant
signaling. This interest and impact of these findings are exemplified by an
invitation to chronicle these findings in Scientific American and by similar
initiatives subsequently spawned in the United Kingdom, The Netherlands,
Switzerland, and France. (Publications: #111, #141, and #157)
G. With Post-Doctorates and colleagues elucidated the important role of extra
floral nectar as an food for foraging parasitoids, showed that plants can
synchronize the elevated provision of nectar and foraging signals with
herbivore damage, and that the parasitoids can separately learn and use
odors associated with competing host and food needs (1993-2000). Impact:
Little was previously known about the importance of adult food for parasitoids,
the role of plants in providing the food, nor how learning, hunger state and
previous feeding experiences influenced the parasitoid foraging behavior. These
1106 Wolf Prize in Agriculture
findings fill major gaps in understanding past failures and insuring future
reliability of biological control agents, and based on these reports similar research
has been initiated in the USA, Switzerland, France, and Japan. The sophisticated
foraging system of parasitoids demonstrated by Dr. Lewis and others is a central
premise for Dr. E. F. Knipling’s recent book “Principles of Insect Parasitism
Analyzed from New Perspectives: Practical Implications for Regulating Insect
Populations by Biological Means”. Furthermore, these findings in combination
with 5 & 6 above have received extensive popular press, including Business
Week, Kiplinger Agricultural Letter, Organic Gardening, New York Times, BBC
Wildlife, CNN Science and Technology, and documentaries by the discovery
channel. (Publications: #106, #140, 147, #155, and #158)
H. With Dr. J. H. Tumlinson and a team of associated cooperators, used cotton
production as a model example, to formulate and demonstrate, a fundamental
shift to a total system approach to sustainable pest management involving the
use of ecosystem management, crop attributes/multitrophic level interactions,
and minimal use of therapeutics — the basis for which is largely drawn from
the fundamental discoveries of Dr. Lewis and cooperators (1993-99). Impact:
Despite major efforts toward alternatives to conventional pesticides, pest
management science has continued by-and-large to regress to a continued array
of new therapeutic intervention tools and failed to make a mainstream shift to
understanding and redesigning cropping systems so that natural and renewable
strengths keep pests within acceptable bounds on a sustained basis. On-farm
work, was conducted directly with cotton growers, and showed a $60/acre
higher net annual profit along with major benefits of improved soil and water
health and reduced environmental contamination. These demonstrations
catalyzed rapid expansion of the acreage under sustainable practices in Georgia
and other areas of the region. For example, sustainable practices in Coffee
County, Georgia increased from 200 acres in the early 1990’s to over 30,000
acres. These concepts, approaches, and demonstrations are having major impact
on overall crop management philosophy and practices at consultant and grower
levels, research directions, and funding and policy decisions. For example, the
SARE agency has employed their seminal PNAS paper and associated work for
nationwide use as a standard in guiding constituents toward systems approaches,
major funding decisions and research directions by EPA, ARS, and other
institutions are being shaped by the work, and various institutions are using the
work as a key focus in educational/training programs. (Publications: #151,
#174, #149, #159, #165 and #175)
I. In cooperation with Dr. J. H. Tumlinson, Post-Doctorates and other colleagues,
and with major funding by the Department of Defense, developed and
demonstrated methodology for training parasitic wasps to quickly learn
any chemical of interest in association with food or hosts and to display a
W. Joe Lewis 1107
work, and various institutions are using the work as a key focus in educational/
training programs.
Also, Dr. Lewis and cooperators formulated principles, derived from natural
ecosystems, for guiding sustainable human communities, including publishing a
benchmark publication (through the Kerr Center for Sustainable Agriculture and
Rural Communities) with an analysis of how modern trends are eroding inherent
strengths in local communities and proposed fundamental redirections for achieving
ecologically-based human communities, and is working directly with local
governments, educational systems, business leaders, and advocacy organizations to
implement redirections toward sustainable communities. As Vice–Mayor, of his
hometown, Tifton, Georgia, he has led implementation of these principles in
successful “smart growth” land-use patterns, greenway development, historic
preservation, and local “self-sufficiency” in waste management and fiber optic/
digital technology. Tifton’s success is highly recognized at the State level and used
through the Georgia Municipal Association and Department of Community Affairs
as a model for wise strategies in sustainable community development (He was
presented Special Congressional recognition in May, 2000). More recently, with
the cooperation of key education organizations, he is underway with the
development of pilot programs for schools in South Georgia to connect students
with ecological values and the importance of sustainable living practices. A Radio
Netherlands documentary, Rural Renaissance, broadcast internationally, recently
highlighted his work in Tifton. He is working in cooperation with the Kerr Center,
SARE-USDA, and other organizations to implement these sustainable practices on a
National and International basis.
LIST OF PUBLICATIONS
1. Vinson, S. B. and Lewis, W.J. A method of host selection by Cardiochiles
nigriceps. J. Econ. Entomol. 58: 869-871. 1965.
2. Lewis, W.J. and Brazzel, J.R. Biological relationships between Cardiochiles
nigriceps and the Heliothis complex. J. Econ. Entomol. 59: 8320-823.
1966.
3. Lewis, W. J. and Brazzel, J. R. Incidence of parasitic insects on the Bollworm
in Mississippi. Mississippi State Univ. 727: 1-7 pp. 1966.
4. Lewis W. J. and Brazzel, J. R.and Vinson, S.B. Heliothis subflexa, a host for
Cardiochiles nigriceps. J. Econ. Entomol. 60: 615-616. 1967.
5. Lewis, W. J. Studies of the suitability of certain Heliothis spp. as a host for
Cardiochile nigriceps Vierick. Ph.D. Thesis Miss. State Univ. pp. 56. 1968.
6. Lewis, W.J. and Brazzel, J.R. A three-year study of parasites of the bollworm
and the tobacco budworm in Mississippi. J. Econ. Entomol. 61: 673-676.
1968.
1110 Wolf Prize in Agriculture
22. Barras, D. J., Kisner, R. L., Lewis, W. J., and Jones, R. L. Effects of the parasitoid,
Microplitis croceipes, on the haemolymph proteins of the corn earworm,
Heliothis zea. Comp. Biochem. Physiol. 43: 940-947. 1972.
23. Lewis, W. J., Jones, R. L., and Sparks, A. N. A host-seeking stimulant for
the egg parasite Trichogramma evanescens: its source and a demonstration
of its laboratory and field activity. Ann. Entomol. Soc. Am. 65: 1087-1089.
1972.
24. Lewis, W. J., Sparks, A. N., Jones, R. L., and Barras, D. J. Efficiency of
Cardiochiles nigriceps as a parasite of Heliothis virescens on cotton. Environ.
Entomol. 1: 468-471. 1972.
25. Lewis, W. J. and Young, J. R. Parasitism by Trichogramma evanescens of
eggs from tepa-sterilized and normal Heliothis zea. J. Econ. Entomol. 65:
705-711. 1972.
26. Snow, J. W., Sparks, A. N., and Lewis, W. J. Seasonal capture of corn earworm
adults in light traps near Tifton, Georgia, compared with captures in traps
baited with virgin females. J. Georgia Entomol. Soc. 7: 85-89. 1972.
27. Snow, J. W., Young, J. R., Lewis, W. J., and Jones, R. L. Sterilization of adult
fall armyworms by gamma irradiation and its effect on competitiveness.
J. Econ. Entomol. 65(5): 1431-1433. 1972.
28. Jones, R. L., Lewis, W. J., Beroza, M., Bierl, B. A., and Sparks, A. N. Host-
seeking stimulants (kairomones) for the egg parasite, Trichogramma
evanescens. Environ. Entomol. 2: 593-596. 1973.
29. Vinson, S. B. and Lewis, W. J. Teratocytes: growth and numbers in the hemocoel
of Heliothis virescens attacked by Microplitis croceipes. J. Invertebr. Pathol.
22: 351-355. 1973.
30. Nordlund, D. A., Lewis, W. J., Gross, H. R., Jr., and Harrell, E. A. Description
and evaluation of a method for field application of Heliothis zea eggs and
kairomones for Trichogramma. Environ. Entomol. 3: 981-984. 1974.
31. Gross, H. R., Jr., Lewis, W. J., Jones, R. L., and Nordlund, D. A. Kairomones
and their use for management of entomophagous insects: III. Stimulation of
Trichogramma achaeae, T. pretiosum, and Microplitis croceipes with host-
siiking stimuli at time of release to improve their efficiency. J. Chem. Ecol. 1:
431-438. 1975.
32. Lewis, W. J., Jones, R. L., Nordlund, D. A., and Sparks, A. N. Kairomones and
their use for management of entomophagous insects: I. Evaluation for
increasing rates of parasitization by Trichogramma spp. in the field.
J. Chem. Ecol. 1: 343-360. 1975.
33. Lewis, W. J., Jones, R. L., Nordlund, D. A., and Gross, H. R., Jr. Kairomones
and their use for management of entomophagous insects: II. Mechanism
causing increase in rate of parasitization by Trichogramma spp. J. Chem.
Ecol. 1: 349-360. 1975.
1112 Wolf Prize in Agriculture
45. Nordlund, D. A., Lewis, W. J., Todd, J. W., and Chalfant, R. B. Kairomones and
their use for management of entomophagous insects: VII. The involvement of
various stimuli in the differential response of Trichogramma pretiosum Riley
to two suitable hosts. J. Chem. Ecol. 3: 513-518. 1977.
46. Lewis, W. J., Beevers, M., Nordlund, D. A., Gross, H. R., Jr., and Hagen, K. S.
Kairomones and their use for management of entomophagous insects: IX.
Investigations of various kairomone-treatment patterns for Trichogramma
spp. J. Chem. Ecol. 5: 673-680. 1979.
47. Sauls, C. E., Nordlund, D. A., and Lewis, W. J. Kairomones and their use for
management of entomophagous insects: VIII. Effect of diet on the kairomonal
activity of frass from Heliothis zea (Boddie) larvae for Microplitis croceipes
(Cresson). J. Chem. Ecol. 5: 363-369. 1979.
48. Lewis, W. J. and Nordlund, D. A. Employment of parasitoids and predators
for fall armyworm control. Fla. Entomol. 63: 433-438. 1980.
49. Lewis, W. J., Hagen, K. S., Roelofs, W. L., and Schoonhoven, L. M. Status and
potential use of behavioural chemicals in pest management. FAO Plant Protect.
Bull. 28: 121-128. 1980.
50. Morrison, G., Lewis, W. J., and Nordlund, D. A. Spatial differences in Heliothis
zea egg density and the intensity of parasitism by Trichogramma spp.: An
experimental analysis. Environ. Entomol. 9: 79-85. 1980.
51. Altieri, M. A., Lewis, W. J., Nordlund, D. A., Gueldner, R. C., and Todd, J. W.
Chemical interactions between plants and Trichogramma wasps in Georgia
soybean fields. Protect. Ecol. 3: 259-263. 1981.
52. Beevers, M., Lewis, W. J., Gross, H. R., Jr., and Nordlund, D. A. Kairomones
and their use for management of entomophagous insects: X. Laboratory studies
on manipulation of host-finding behavior of Trichogramma pretiosum Riley
with a kairomone extracted from Heliothis zea (Boddie) moth scales. J. Chem.
Ecol. 7: 635-648. 1981.
53. Gross, H. R., Jr., Harrell, E. A., Lewis, W. J., and Nordlund, D. A. Trichogramma
spp.: concurrent ground application of parasitized eggs, supplemental Heliothis
zea host eggs, and host-seeking stimuli. J. Econ. Entomol. 74: 227-229. 1981.
54. Gross, H. R., Jr. Employment of kairomones in the management of parasitoids,
pp. 137-150. In: D. A. Nordlund, R. L. Jones, and W. J. Lewis (ed.),
Semiochemicals: Their Role in Pest Control. Wiley, NY, 306 pp. 1981.
55. Gross, H. R., Jr., Lewis, W. J., and Nordlund, D. A. Trichogramma pretiosum:
effect of prerelease parasitization experience on retention in release areas
and efficiency. Environ. Entomol. 10: 554-556. 1981.
56. Jackson, R. D. and Lewis, W. J. Summary of significance and employment
strategies for semiochemicals, pp. 283-295. In: D. A. Nordlund, R. L. Jones,
and W. J. Lewis (ed.), Semiochemicals: Their Role in Pest Control. Wiley, NY,
306 pp. 1981.
1114 Wolf Prize in Agriculture
69. Gross, H. R., Jr., Lewis, W. J., Beevers, M., and Nordlund, D. A. Trichogramma
pretiosum (Hymenoptera: Trichogrammatidae): effects of augmented densities
and distributions of Heliothis zea (Lepidoptera: Noctuidae) host eggs
and kairomones on field performance. Environ. Entomol. 13: 981-985.
1984.
70. Gueldner, R. C., Nordlund, D. A., Lewis, W. J., Thean, J. E., and Wilson, D. M.
Kairomones and their use for management of entomophagous insects. XV.
Identification of several acids in scales of Heliothis zea moths and comments
on their possible role as kairomones for Trichogramma pretiosum. J. Chem.
Ecol. 10: 245-251. 1984.
71. King, E. G., Bouse, L. F., Bull, D. L., Dickerson, W. M., Lewis, W. J., Liapis, P.,
Lopez, J. D., Morrison, R. K., and Phillips, J. R. Potential for management of
Heliothis spp in cotton by augmentative releases of Trichogramma pretiosum,
pp. 232-236. Proceedings Beltwide Cotton Prod Res Conf, Memphis, TN,
National Cotton Council of Am., 381 pp. 1984.
72. Lewis, W. J. and Nordlund, D. A. Semiochemicals influencing fall armyworm
parasitoid behavior: implications for behavioral manipulation. Fla. Entomol.
67: 343-349. 1984.
73. Morrison, G. and Lewis, W. J. Inferring parasitoid searching behaviors
from host mortality data: A note of caution. Environ. Entomol. 13: 7-14.
1984.
74. Nordlund, D. A., Chalfant, R. B., and Lewis, W. J. Arthropod populations,
yield and damage in monocultures and polycultures of corn, beans and
tomatoes. Agric. Ecosys. Environ. 11: 353-367. 1984.
75. Dmoch, J., Lewis, W. J., Martin, P. B., and Nordlund, D. A. Role of host-
produced stimuli and learning in host selection behavior of Cotesia
(=Apanteles) marginiventris (Cresson). J. Chem. Ecol. 11: 453-463. 1985.
76. Keller, M. A. and Lewis, W. J. Movements by Trichogramma pretiosum
(Hymenoptera: Trichogrammatidae) released into cotton. Southwest. Entomol.
8: 99-109. 1985.
77. Keller, M. A., Lewis, W. J., and Stinner, R. E. Biological and practical significance
of movement by Trichogramma species: A review. Southwest. Entomol. 8:
138-155. 1985.
78. Lewis, W. J., Gross, H. R., Jr., and Nordlund, D. A. Biological control of
bollworm and tobacco budworm in cotton by augmentative releases of
Trichogramma — behavioral manipulation of Trichogramma (Hymenoptera:
Trichogrammatidae). Southwest. Entomol. Suppl. No. 8: 49-55. 1985.
79. Lewis, W. J. and Nordlund, D. A. Behavior-modifying chemicals to enhance
natural enemy effectiveness, pp. 89-101. In: M. A. Hoy and G. A. Herzog
(ed.), Biological Control in Agricultural Integrated Pest Management Systems.
Academic Press, Orlando, FL, 589 pp. 1985.
1116 Wolf Prize in Agriculture
90. Hamm, J. J., Styer, E. L., and Lewis, W. J. A baculovirus pathogenic to the
parasitoid Microplitis croceipes (Hymenoptera: Braconidae). J. Invertebr.
Pathol. 52: 189-191. 1988.
91. Herard, F, Keller, M. A., and Lewis, W. J. Rearing Microplitis demolitor
Wilkinson (Hymenoptera: Braconidae) in the laboratory for use in studies of
semiochemical mediated searching behavior. J. Entomol. Sci. 23: 105-111.
1988.
92. Herard, F., Keller, M. A., Lewis, W. J., and Tumlinson, J. H. Beneficial arthropod
behavior mediated by airborne semiochemicals. III. Influence of age and
experience on flight chamber responses of Microplitis demolitor Wilkinson.
J. Chem. Ecol. 14: 1583-1596. 1988.
93. Herard, F., Keller, M. A., Lewis, W. J., and Tumlinson, J. H. Beneficial arthropod
behavior mediated by airborne semiochemicals. IV. Influence of host diet on
host-oriented flight chamber responses of Microplitis demolitor Wilkinson.
J. Chem. Ecol. 14: 1597-1606. 1988.
94. Lewis, W. J., Sonnet, P. E., and Nordlund, D. A. Responses of braconid
parasitoids Microplitis croceipes (Cresson) and M. demolitor Wilkinson
to stereoisomers of kairomone l3-methylhentriacontane. J. Chem. Ecol. l4:
883-888. 1988.
95. Lewis, W. J. and Tumlinson, J. H. Host detection by chemically mediated
associative learning in a parasitic wasp. Nature 331: 257-259. 1988.
96. Noldus, L. P., Lewis, W. J., Tumlinson, J. H., and van Lenteren, J. C. Olfactometer
and wind tunnel experiments on the role of sex pheromones of noctuid
moths in the foraging behavior of Trichogramma spp., pp. 223-228. Illd
International Symposium Guangzhou (China), Nov. 10-15, 1986. INRA Paris,
1988.
97. Nordlund, D. A., Lewis, W. J., and Altieri, M. A. Influence of plant produced
allelochemicals on the host/prey selection behavior of entomophagous insects.,
pp. 65-90. In: P. Barbosa and D. Letourneau (ed.), Novel Aspects of Insect-
Plant Interactions. J. Wiley and Sons, 1988.
98. Keller, M. A. and Lewis, W. J. Behavior modifying chemicals to increase the
efficacy of predators and parasitoids of Heliothis spp, pp. 449-481. In: E. G.
King and R. D. Jackson (ed.), Proceedings, Biological Control of Heliothis:
Increasing the Effectiveness of Natural Enemies, Rekha Printers Pvt. Ltd.,
1989.
99. Lewis, W. J. and Gross, H. R., Jr. Comparative studies on field performance of
Heliothis larval parasitoids, Microplitis croceipes and Cardiochiles nigriceps,
at varying densities and under selected host plant conditions. Fla. Entomol.
72: 6-14. 1989.
100. Nordlund, D. A., Lewis, W. J., and Tumlinson, J. H. Habitat and host location
behavior of Microplitis croceipes. Southwest. Entomol. 12: 39-48. 1989.
1118 Wolf Prize in Agriculture
101. Turlings, T. C. J., Tumlinson, J. H., Lewis, W. J., and Vet, L. E. M. Beneficial
arthropod behavior mediated by airborne semiochemicals. VII. Learning of
host-related odors induced by a brief contact experience with host by-products
in Cotesia marginiventris (Cresson). J. Insect Behav. 2: 217-225. 1989.
102. Zanen, P. O., Lewis, W. J., Carde, R. T., and Mullinix, B. G. Beneficial arthropod
behavior mediated by airborne semiochemicals. VI. Flight responses of female
Microplitis croceipes (Cresson), a braconid endoparasitoid of Heliothis spp.,
to varying olfactory stimulus conditions created with a turbulant jet. J. Chem.
Ecol. 15: 141-168. 1989.
103. Eller, F. J., Tumlinson, J. H., and Lewis, W. J. Intraspecific competition in
Microplitis croceipes (Hymenoptera: Braconidae), a parasitoid of Heliothis
species (Lepidoptera: Noctuidae). Ann. Entomol. Soc. Am. 83: 504-508.
1990.
104. Hamm, J. J., Styer, E. L., and Lewis, W. J. Comparative virogenesis of
filamentous virus and polydnavirus in the female reproductive tract of Cotesia
marginiventris (Hymenoptera: Braconidae). J. Invertebr. Pathol. 55: 357-374.
1990.
105. Lewis, W. J. and Martin, W. R., Jr. Semiochemicals for use with parasitoids:
Status and future. J. Chem. Ecol. 16: 3067-3089. 1990.
106. Lewis, W. J. and Takasu, K. Use of learned odours by a parasitic wasp in
accordance with host and food needs. Nature 348: 635-636. 1990.
107. Lewis, W. J., Vet, L. E. M., Tumlinson, J. H., van Lenteren, J. C., and Papaj,
D. R. Variations in parasitoid foraging behavior: essential element of a sound
biological control theory. Environ. Entomol. 19: 1183-1193. 1990.
108. Noldus, L. P., Lewis, W. J., and Tumlinson, J. H. Beneficial arthropod
behavior mediated by airborne semiochemicals. IX. Differential response of
Trichogramma pretiosum, an egg parasitoid of Heliothis zea, to various
olfactory cues. J. Chem. Ecol. 16: 3531-3544. 1990.
109. Prevost, G. and Lewis, W. J. Heritable differences in the response of the
braconid wasp Microplitis croceipes to volatile allelochemicals. J. Insect Behav.
3: 277-287. 1990.
110. Turlings, T. C. J., Scheepmaker, J. W. A., Vet, L. E. M., Tumlinson, J. H., and
Lewis, W. J. How contact foraging experiences affect preferences for host-
related odors in the larval parasitoid Cotesia marginiventris (Cresson)
(Hymenoptera: Braconidae). J. Chem. Ecol. 16: 1577-1589. 1990.
111. Turlings, T. C. J., Tumlinson, J. H., and Lewis, W. J. Exploitation of herbivore-
induced plant odors by host-seeking parasitic wasps. Science 250:
1251-1253. 1990.
112. Vet, L. E. M., Lewis, W. J., Papaj, D. R., and van Lenteren, J. C. A variable-
response model for parasitoid foraging behavior. J. Insect Behav. 3: 471-490.
1990.
W. Joe Lewis 1119
113. Lewis, W. J., Tumlinson, J. H., and Krasnoff, S. Chemically mediated associative
learning: An important function in the foraging behavior of Microplitis
croceipes (Cresson). J. Chem. Ecol. 17: 1309-1325. 1991.
114. Noldus, L. P., van Lenteren, J. C., and Lewis, W. J. How Trichogramma
parasitoids use moth sex pheromones as kairomones: Orientation behavior
in a wind tunnel. Physiol. Entomol. 16: 313-327. 1991.
115. Tumlinson, J. H. and Lewis, W. J. Chemically mediated foraging behavior of
parasitoids: New implications for biocontrol, pp. 161-178. In: Entomology
Serving Society: Emerging technologies and challenges, Entomological Society
of America. Lanham, MD, 1991.
116. Turlings, T. C. J., Tumlinson, J. H., Eller, F. J., and Lewis, W. J. Larval-damaged
plants: source of volatile synomones that guide the parasitoid Cotesia
marginiventris to the micro-habitat of its host. Entomol. Exp. Appl. 58:
75-82. 1991.
117. Eller, F. J., Tumlinson, J. H., and Lewis, W. J. Effect of host diet and preflight
experience on the flight response of Microplitis croceipes (Cresson). Physiol.
Entomol. 17: 235-240. 1992.
118. Hamm, J. J., Styer, E. L., and Lewis, W. J. Three viruses found in the braconid
parasitoid Microplitis croceipes and their implications in Biological Control
Programs. Biol. Control 2: 329-336. 1992.
119. Sheehan, W., Wackers, F. L., and Lewis, W. J. Discrimination of previously
searched, host-free sites by Microplitis croceipes (Hymenoptera: Braconidae).
J. Insect Behav. 6: 323-331. 1992.
120. Tumlinson, J. H., Turlings, T. C. J., and Lewis, W. J. The semiochemical
complexes that mediate insect parasitoid foraging. Agric. Zool. Reviews
5: 221-252. 1992.
121. McCall, P. J., Turlings, T. C. J., Lewis, W. J., and Tumlinson, J. H. Role of plant
volatiles in host location by the special parasitoid Microplitis croceipes
(Cresson) (Braconidae: Hymenoptera). J. Insect Behav. 6: 625-639. 1993.
122. Prokopy, R. J. and Lewis, W. J. Application of learning to pest management,
pp. 308-342. In: D. R. Papaj and A. Lewis (ed.), Insect Learning: Ecological
and Evolutionary Perspectives. Chapman and Hall, New York, 398 pp. 1993.
123. Ruberson, J. R., Herzog, G. A., and Lewis, W. J. Parasitism of the beet
armyworm, Spodoptera exigua, in south Georgia cotton, 2: 993-997.
Proceedings Beltwide Cotton Conference, 1993.
124. Sheehan, W., Wackers, F. L., and Lewis, W. J. Discrimination of previously
searched, host-free sites by Microplitis croceipes (Hymenoptera: Braconidae).
J. Insect Behav. 6: 323-331. 1993.
125. Takasu, K. and Lewis, W. J. Host-and-food-foraging of the parasitoid
Microplitis croceipes: Learning and physiological state effects. Biol. Control
3: 70-74. 1993.
1120 Wolf Prize in Agriculture
126. Tumlinson, J. H., Lewis, W. J., and Vet, L. E. M. How beneficial parasitic
wasps find plant feeding caterpillars. Sci. Amer. 268: 100-106. 1993.
127. Tumlinson, J. H., Turlings, T. C. J., and Lewis, W. J. Semiochemically mediated
foraging behavior in beneficial parasitic insects. Arch. Insect Biochem. and
Physiol. 22: 385-391. 1993.
128. Turlings, T. C. J., Wackers, F. L., Vet, L. E. M., Lewis, W. J., and Tumlinson,
J. H. Learning of host location cues by insect parasitoids, pp. 51-78. In:
D. R. Papaj and A. Lewis (ed.), Insect Learning: Ecological and Evolutionary
Perspectives. Chapman and Hall, New York, 398 pp. 1993.
129. van Giessen, W. A., Lewis, W. J., Vet, L. E. M., and Wackers, F. L. The influence
of host site experience on subsequent flight behavior in Microplitis croceipes
(Cresson) (Hymenoptera: Braconidae). Biol. Control 3: 75-79. 1993.
130. Lambert, W. R., Herzog, G. A., Ruberson, J. R., and Lewis, W. J. Tobacco
budworm and bollworm managment strategies in the absence of the boll
weevil. Proc. Beltwide Cotton Conf. 2: 794-795. 1994.
131. Lewis, W. J., Sheehan, W., and Tumlinson, J. H. Unraveling the story of how
parasitoids find their hosts, pp. 671-680. In: D. Rosen, F. D. Bennett, and
F. L. Capinera (ed.), Pest Management in the Subtropic: Biological Control -
A Florida Perspective, Intercept Ltd., Andover, Hamshire, UK, 1994.
132. Ruberson, J. R., Herzog, G. A., Lambert, W. R., and Lewis, W. J. Management
of the beet armyworm in cotton: Role of natural enemies. Fla. Entomol. 77:
440-453. 1994.
133. Ruberson, J. R., Herzog, G. A., Lambert, W. R., and Lewis, W. J. Management
of the beet armyworm: Integration of control approaches. Proc. 1994 Beltwide
Cotton Prod. Conf. 2: 857-859. 1994.
134. Soares, G. G., Jr., Lewis, W. J., Strong-Gunderson, J. M., Waters, D. J., and
Hamm, J. J. Integrating the use of MVP Bioinsecticide, a unique BT-based
product, with natural enemies of noctuid pests: A novel concept in cotton
IPM, Proceedings, Second Canberra Bacillus thuringiensis Meeting, 1994.
135. Wackers, F. L. and Lewis, W. J. Olfactory and visual learning and their
interactive influence on host site location by the parasitoid, Microplitis
croceipes. Biol. Control 4: 105-112. 1994.
136. Alborn, H. T., Lewis, W. J., and Tumlinson, J. H. Host-specific recognition
kairmone for the parasitoid Microplitis croceipes (Cresson). J. Chem. Ecol.
21: 1697-1708. 1995.
137. Haney, P. B., Stapel, J. O., Waters, D. J., Lewis, W. J., Diffie, S. K., and Ruberson,
J. R. Dynamics of insect populations in a reducedtillage, crimson clover/
cotton system Part II: Pitfall survey. Proc. 1995 Beltwide Cotton Conference
2: 817-821. 1995.
138. Ruberson, J. R., Lewis, W. J., Waters, D. J., Stapel, J. O., and Haney, P. B.
Dynamics of insect populations in a reduced-tillage, crimson clover/cotton
W. Joe Lewis 1121
system Part 1: Pests and beneficials on plants. Proc. 1995 Beltwide Cotton
Conference 2: 814-817. 1995.
139. Stowe, M. K., Turlings, T. C. J., Loughrin, J. H., Lewis, W. J., and Tumlinson,
J. H. The chemistry of eavesdropping, alarm, and deceit, pp. 23-28.
Proceedings National Acaddemy Science Colloquium, Chemical Ecology: The
Chemistry of Biotic Interaction, 1995.
140. Takasu, K. and Lewis, W. J. Importance of adult food sources to host
searching of the larval parasitoid, Microplitis croceipes. Biol. Control 5:
25-30. 1995.
141. Turlings, T. C. J., Loughrin, J. H., McCall, P. J., Rose, U. S. R., Lewis, W. J., and
Tumlinson, J. H. How caterpillar-damaged plants protect themselves by
attracting parasitic wasps, Proceedings National Academy Science Colloquium,
Self-defense by Plants: Induction and signaling pathways, 1995.
142. Vet, L. E. M., Lewis, W. J., and Carde, R. T. Parasitoid foraging and learning,
pp. 65-101. In: R. T. Carde and W. Bell (ed.), Chemical Ecology of Insects, II.
Chapman & Hall, New York, 1995.
143. Gazit, Y., Lewis, W. J., and Tumlinson, J. H. Arrestment of Telenomus remus
(Hymenoptera: Scelionidae) by a kairomone associated with eggs of its host,
Spodoptera frugiperda (Lepidoptera: Noctuidae). Biol. Control 6: 283-290.
1996.
144. Haney, P. B., Lewis, W. J., and Lambert, W. R. Cotton production and the boll
weevil in Georgia: History, cost of control, and benefits of eradication.
Research Bulletin Number 428. 48 pp. 1996.
145. Haney, P. B., Lewis, W. J., Ruberson, J. R., and Phatak, S. C. Continued studies
of insect population dynamics in crimson clover and refugia/cotton systems.
Part II: Pitfall trap sampling. Proc. Beltwide Cotton Conf. 2: 1115-1119.
1996.
146. Lewis, W. J., Haney, P. B., Ruberson, J. R., and Phatak, S. C. Continued studies
of insect population dynamics in crimson clover and refugia/cotton systems.
Part I: Sweep and whole plant sampling. Proc. Beltwide Cotton Conf. 2:
1108-1114. 1996.
147. Takasu, K. and Lewis, W. J. The role of learning in adult food location by the
larval parasitoid, Microplitis croceipes (Hymenoptera: Braconidae). J. Insect
Behavior 9: 1996.
148. Cortesero, A. M., De Moraes, C. M., Stapel, J. O., Tumlinson, J. H., and Lewis,
W. J. Comparisons and contrasts in host-foraging strategies of two larval
parasitoids with different degrees of host specificity. J. Chem. Ecol. 23(6):
1589-1606. 1997.
149. Haney, P. B. and Lewis, W. J. Year three of insect population dynamics in a
cotton refugia system: Pitfall trap sampling. Proc. Beltwide Cotton Conf. 2:
1127-1129. 1997.
1122 Wolf Prize in Agriculture
150. Lewis, W. J., Haney, P. B., Reed, R., and Walker, A. A total systems approach
for sustainable cotton production in Georgia and southeast: first year results.
Proc. Beltwide Cotton Conf. 2: 1129-1134. 1997.
151. Lewis, W. J., van Lenteren, J. C., Phatak, S. C., and Tumlinson, J. H. A total
system approach to sustainable pest management. Proc. Natl. Acad. Sci. USA
94: 12243-12248. 1997.
152. Reed, R., Phatak, S. C., Page, A., Haney, P. B., and Lewis, W. J. Conservation
tillage in Coffee County cotton. Proc. Beltwide Cotton Conf. 1: 621-622. 1997.
153. Rose, U. S. R., Alborn, H. T., Makranczy, G., Lewis, W. J., and Tumlinson,
J. H. Host recognition by the specialist endoparasitoid Microplitis croceipes.
J. Insect Behav. 10(3): 313-330. 1997.
154. Ruberson, J. R., Phatak, S. C., and Lewis, W. J. Insect population in a cover
crop/strip tillage system. Proc. Beltwide Cotton Conf. 2: 1121-1123. 1997.
155. Stapel, J. O., Cortesero, A. M., De Moraes, C. M., Tumlinson, J. H., and Lewis,
W. J. Extrafloral nectar, honeydew, and sucrose effects on searching behavior
and efficiency of Microplitis croceipes (Hymenoptera: Braconidae) in cotton.
Environ. Entomol. 26(3): 617-623. 1997.
156. Stapel, J. O., Ruberson, J. R., Gross, H. R., Jr., and Lewis, W. J. Progeny
allocation by the parasitoid Lespesia archippivora (Diptera: Tachinidae) in
larvae of Spodoptera exigua (Lepidoptera: Noctuidae). Environ. Entomol.
26(2): 265-271. 1997.
157. De Moraes, C. M., Lewis, W. J., Pare, P. W., Alborn, H. T., and Tumlinson,
J. H. Herbivore-infested plants selectively attract parasitoids. Nature 393:
570-573. 1998.
158. Lewis, W. J., Stapel, J. O., Cortesero, A. M., and Takasu, K. Understanding
how parasitoids balance food and host needs: Importance to biological control.
Biol. Control 11: 175-183. 1998.
159. Stapel, J. O., Waters, D. J., Ruberson, J. R., and Lewis, W. J. Development and
behavior of Spodoptera exigua (Lepidoptera: Noctuidae) larvae in choice
tests with food substrates containing toxins of Bacillus thuringiensis. Biol.
Control 11: 29-37. 1998.
160. De Moraes, C. M. and Lewis, W. J. Analyses of two parasitoids with convergent
foraging strategies. J. Insect Behav. 12: 571-583. 1999.
161. Olson, D. M., , Phatak S. C., and Lewis, W. J. Influence of Nitrogen Levels on
Cotton Plant/Insect Interactions in a Conservation Tillage System, In:
J. E. Hook (ed.), Proceedings of the 22nd Annual Southern Conservation
Tillage Conference for Sustainable Agriculture, July 6-8, 1999. Tifton, GA,
GA Agr. Exp. Stn. Special Pub. 95. 1999.
162. Pare’, P. W., Lewis, W. J., and Tumlinson, J. H. Induced plant volatiles:
Biochemistry and effects on parasitoids, 167-180. In: A. A. Agrawal, S. Tuzun,
and E. Bent (ed.), Inducible plant defenses against pathogens and herbivores:
W. Joe Lewis 1123
174. Cortesero, A.M., J.O. Stapel and W. .J. Lewis. Understanding and manipulating
plant attributes to enhance biological control. Bio.Control 17: 35-49. 2000.
175. Rains, G. C., D. M. Olson, W. J. Lewis and J. H. Tumlinson. Systems
Management In pp-826-828, Encyclopedia of Pest Management, Pimental,
D., ed. Marcel Dekker, Inc. New York, NY, 2002.
176. Meiners, Torsten, Felix Wackers, and W. J. Lewis. The Effect of Molecular
Structure on Olfactory Discrimination by the Parasitoid Microplitis croceipes.
Chem. Senses 27: 811-916. 2002.
177. Wackers, Felix, Bonifay, Claire, Lewis, W. J. Conditioning of Appetitive Behavior
In The Hymenopteran Parasitoid Microplitis croceipes. Entomologia
Experimentalis Et Applicata. 103: 135-238, 2002.
178. Schomberg, H., W. J. Lewis, D. Olson, G. Tillman, P. Timper, S. Phatak, and
D. Wauchope. Conceptual Model for Sustainable Cropping Systems in the
Southeast: Cotton System. In Cropping Systems: Trends and Advances, Special
Edition, J. of Crop Production 8: 307-327, 2003.
179. Meiners, T., Wackers, F., and Lewis, W. J. Associative learning of complex
odours in parasitoid hosts location. Chemical Senses 28: 231-236. 2003.
180. Olson, D. M., Hodges, T. A., Lewis, W. J. Foraging efficacy of a larval parasitoid
in a cotton patch: influence of chemical cues and learning. J. of Insect
Behavior. 16 (5): 613-624, 2003.
181. Olson, D. M. Rains, G. C., Meiners, T., Takasu, K. Tertuliano, M., Tumlinson,
J. H., Wackers, F. L., and Lewis, W. J. Parasitic wasps learn and report
diverse chemicals with unique conditionable behaviors. Chemical Senses 28:
545-549, 2003.
182. Takasu, K., and Lewis, W. J. Learning of host searching cues by the larval
parasitoid, Microplitis croceipes. Entomologia Experimentalis Et. Applicata
108: 77-86, 2003.
183. Rains, G. C. Olson, D. M., Lewis, W. J. Sustainable Agriculture: Definition
and Goals. In pp. 1187-1190, Encyclopedia of Crop and Soil Science,
Agropedia, Marcel Dekker, New York, NY. 2004.
184. Tertuliano, M. Olson, D. M., Rains, G. C., and Lewis, W. J. Influence of
handling and conditioning protocol on learning and memory of Microplitis
croceipes. Entomologia experimentalis Et. Applicata 110: 165-175, 2004.
185. Rains, B. C. J. K. Tomberlin, J. K., D’Alessandro, M. D., Lewis, W. J. Limits of
Volatile Chemical Detection of a Parasitoid Wasp, Microplitis croceipes, and
an Electronic Nose: A Comparative Study. Transactions of the ASAE, 47:
2145-2152. 2004.
186. Olson, Dawn M., Takasu, Keiji, and Lewis, W. J. Interaction surrounding
adult parasitoid food needs in a multitrophic system. In: Plant-provided Food
Supplements in Plant-Carnivore Mutualism (F. Wackers, van Rijin, P. C. J.,
and Bruin, J. Eds.), Cambridge University Press. 2005.
W. Joe Lewis 1125
187. Tomberlin, J.K., Tertuliano, M., Rains, G.C., Lewis, W. J. Conditioned Microplitis
croceipes Cresson (Hymenoptera: Braconidae) detect and respond to 2,
4-DNT: Development of a biological sensor. J. Forensic Sci. 0: 1187-1190,
2005.
188. Rose, U. S. R., Lewis, J., Tumlinson, J. J. Extrafloral nectar from cotton
(Gossypium hirsutum) as a food source for parasitic wasps. Functional Ecology.
20: 67-74. 2006.
189. Olson, D. M., Takasu, K., Lewis, W. J. Interactive-Web of factors of governing
effective natural enemy foraging behavior: Overview of food resources as a
critical component. In: Proceedings of the Second International Symposium
on Biological Control of Arthropods, Davos, Switzerland. P. 389-397,
September, 2005.
190. Rains, G. C., Utley, S. L., Lewis, W. J. Behavioral monitoring of trained insects
for chemical detection. Biotechnology Progress, 22: 2-8, 2006.
191. Wackers, Felix, Bonifay, Claire, Vet, Louise, Lewis, W. J. Gustatory response
and appetitive learning in microplitis croceipes in relation to sugar type and
concentration. Animal Biology, 56: 193-203, 2006.
1126 Wolf Prize in Agriculture
W. Joe Lewis 1127
1128 Wolf Prize in Agriculture
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1130 Wolf Prize in Agriculture
Perspective
ABSTRACT A fundamental shift to a total system ap- In this report, we argue that the central weakness in how we
proach for crop protection is urgently needed to resolve think about pest management as a component of agricultural
escalating economic and environmental consequences of com- systems has not been addressed. We must go beyond replacing
bating agricultural pests. Pest management strategies have toxic chemicals with more sophisticated, biologically based
long been dominated by quests for ‘‘silver bullet’’ products to agents and re-examine the entire paradigm around the ther-
control pest outbreaks. However, managing undesired vari- apeutic approach, including how and why those therapeutics
ables in ecosystems is similar to that for other systems, are used. Truly satisfactory solutions to pest problems will
including the human body and social orders. Experience in require a shift to understanding and promoting naturally
these fields substantiates the fact that therapeutic interven- occurring biological agents and other inherent strengths as
tions into any system are effective only for short term relief components of total agricultural ecosystems and designing our
because these externalities are soon ‘‘neutralized’’ by coun- cropping systems so that these natural forces keep the pests
termoves within the system. Long term resolutions can be within acceptable bounds. Recent discoveries in multitrophic
achieved only by restructuring and managing these systems in interactions (5) together with renewed emphasis on broader
ways that maximize the array of ‘‘built-in’’ preventive based ecosystem management (6) indicate powerful prospects
strengths, with therapeutic tactics serving strictly as backups for this direction. Although we address the subject primarily
to these natural regulators. To date, we have failed to incor- from a perspective of arthropod pests, similar cases can
porate this basic principle into the mainstream of pest man- generally be made for other pests [see Cook et al. (7) for
agement science and continue to regress into a foot race with background information important to related views for other
nature. In this report, we establish why a total system ap- pests].
proach is essential as the guiding premise of pest management
and provide arguments as to how earlier attempts for change Premise of a Revised Approach
and current mainstream initiatives generally fail to follow this
principle. We then draw on emerging knowledge about mul- The underlying principle of our position is that components of
titrophic level interactions and other specific findings about agricultural ecosystems interact, and, through a set of feedback
management of ecosystems to propose a pivotal redirection of loops, maintain ‘‘balance’’ within functional f luctuating
pest management strategies that would honor this principle bounds. Furthermore, therapeutic interventions into these
and, thus, be sustainable. Finally, we discuss the potential systems are met by countermoves that ‘‘neutralize’’ their
immense benefits of such a central shift in pest management effectiveness [see Flint and van den Bosch (8) and Cook and
philosophy. Baker (9) for an elegant discussion of this point]. We are taught
this basic principle from our earliest training in ecology but
often overlook it in practice for various reasons, including our
The therapeutic approach of killing pest organisms with toxic
tendency in science to divide things into specialized parts, i.e.,
chemicals has been the prevailing pest control strategy for over
to apply a reductionist approach. The basic principle for
50 years. Safety problems and ecological disruptions continue
managing undesired variables in agricultural systems is similar
to ensue (1), and there are renewed appeals for effective, safe,
to that for other systems, including the human body and social
and economically acceptable alternatives (2). Considerable
systems. On the surface, it would seem that an optimal
effort has been directed toward such alternatives, and new
corrective action for an undesired entity is to apply a direct
technology has been implemented and is still emerging. How-
external counter force against it. However, there is a long
ever, the major trend has been toward the use of modern history of experiences in medicine and social science where
chemistry and molecular biology to replace traditional pesti- such interventionist actions never produce sustainable desired
cides with less hazardous chemicals or nontoxic biologically effects. Rather, the attempted solution becomes the problem
based products; but these means are still therapeutics. Thus, [See Waltzlawick et al. (10) for a discussion of this subject with
the classic treadmill effect in pursuit of remediation of the coverage of underlying mathematical principles.] We find vivid
symptoms persists (2) while tolls due to pests grow higher by examples to this end in the problems of addiction as a
some estimates. Crop losses due to arthropods, diseases, and consequence of the use of drugs for treatment of pain or
weeds, though disputed by some as a valid measure, have mental distress and black market crime as a repercussion to the
increased on a world basis from 34.9% in 1965 (3) to 42.1% in use of prohibition as an intended solution for alcoholism. Thus,
1988–1990 (4) despite the intensification of pest control. as a matter of fundamental principle, application of external
corrective actions into a system can be effective only for short
The publication costs of this article were defrayed in part by page charge
payment. This article must therefore be hereby marked ‘‘advertisement’’ in
Abbreviations: IPM, Integrated Pest Management; Bt, Bacillus thu-
accordance with 18 U.S.C. §1734 solely to indicate this fact.
ringiensis.
© 1997 by The National Academy of Sciences 0027-8424兾97兾9412243-6$2.00兾0 †To whom reprint requests should be addressed. e-mail: wjl@tifton.
PNAS is available online at http:兾兾www.pnas.org. cpes.peachnet.edu.
12243
12244 Perspective: Lewis et al. Proc. Natl. Acad. Sci. USA 94 (1997)
term relief. Long term, sustainable solutions must be achieved of products used. From this ‘‘product formulation’’ perspective
through restructuring the system so that inherent forces that and from our existing infrastructure, the major emphasis in
function via feedback mechanisms such as density dependence augmentation schemes becomes focused on how to produce and
are added and兾or function more effectively. transport a large number of natural enemies at a low cost. Less
The foundation for pest management in agricultural systems emphasis is placed on how natural enemies function and how we
should be an understanding and shoring up of the full com- can promote their natural effectiveness.
posite of inherent plant defenses, plant mixtures, soil, natural In keeping with the historical therapeutic-based attitude and
enemies, and other components of the system. These natural existing infrastructure, most concentrated efforts for biological
‘‘built in’’ regulators are linked in a web of feedback loops and control appear to be directed toward the ‘‘rear and release’’
are renewable and sustainable. The use of pesticides and other augmentation, followed by importation and thirdly by conserva-
‘‘treat-the-symptoms’’ approaches are unsustainable and tion. This order of priorities should be reversed. First, we need to
should be the last rather than the first line of defense. A pest understand, promote, and maximize the effectiveness of indige-
management strategy should always start with the question nous populations of natural enemies. Then, based on the knowl-
‘‘Why is the pest a pest?’’ and should seek to address underlying edge and results of these actions, we should fill any key gaps by
weaknesses in ecosystems and兾or agronomic practice(s) that importation. Finally, therapeutic propagation and releases should
have allowed organisms to reach pest status. be used as a backup to these programs when necessary.
IPM. Throughout our quest for alternative pest control
Attempts for Change: No Real Change measures, the IPM concept has by far received the most
attention as a comprehensive pest management approach. IPM
Throughout the debate on alternative methods for controlling has had a varied history, has been defined in many ways, and
pests, various ideas have been expressed and new approaches has been implemented under an array of different connota-
have emerged. Three subject areas, Biological Control, Inte- tions. The term was first used as ‘‘integrated control’’ by
grated Pest Management (IPM), and Biotechnology, have Bartlett (11) and was further elaborated on by Stern et al. (12)
achieved particular importance in our quest for better pest in reference to the concept of integrating the use of biological
control strategies. and other controls in complementary ways. The term was later
Biological Control. Biological control has a long history of broadened to embrace coordinated use of all biological, cul-
use in pest management and has gained renewed interest tural, and artificial practices (13). Subsequently, under the
because of problems encountered with the use of pesticides. term ‘‘IPM,’’ various authors have advocated the principle of
The term ‘‘biological control’’ has been used, at times, in a incorporating the full array of pest management practices
broad context to encompass a full spectrum of biological together with production objectives into a total systems ap-
organisms and biologically based products including phero- proach. See Flint and van den Bosch (8) for a comprehensive
mones, resistant plant varieties, and autocidal techniques such and ecologically based discussion of this concept and the
as sterile insects. The historical and more prevalent use of this potential benefits of its implementation.
term is restricted to use of natural enemies to manage popu- The principles discussed by Flint and van den Bosch (8) are,
lations of pest organisms. in our opinion, solid and on target. They make a thorough case
Biological control has been spectacularly successful in many for a comprehensive long term pest management program
instances, with a number of pest problems permanently resolved based on knowledge of an ecosystem that weighs economic,
by importation and successful establishment of natural enemies. environmental, and social consequences of interventions.
These importation successes have been limited largely to certain However, as translated into practice, IPM has been primarily
types of ecosystems and兾or pest situations such as introduced a monitoring program in which thresholds are established and
pests in perennial ecosystems. On the other hand, this approach chemicals are used only on an as-needed basis. Much less
has met with limited success for major pests of row crops or other emphasis has been placed on understanding and promoting
ephemeral systems. In these situations, the problem is often not inherent strengths within systems to limit pest populations
the lack of effective natural enemies but management practices through use of approaches such as landscape ecology. In other
and a lack of concerted research on factors that determine the words, IPM programs have been operated with pesticide
success or failure of importation attempts in the specific agro- management objectives rather than pest management objec-
ecosystem setting. Thus, importation programs, to date, are tives. We hasten to add that their use has been of major benefit
largely a matter of trial and error based on experience of the and has greatly reduced the quantity of pesticides used.
individual specialists involved. Furthermore, activities remain underway to refocus IPM
Conservation of natural enemies received more attention as toward the achievement of its full objectives (14, 15). However,
part of a cultural management approach before the advent of our point is that, again, the tendency has been to remain
synthetic pesticides. Since that time, this realm of biological centered on a monitor and treat-the-symptoms approach vs.
control has been neglected. The term ‘‘conservation’’ tends to the more fundamental question of ‘‘Why is the pest a pest?’’
limit one’s vision to a passive approach of acknowledging that Biotechnology. Although biotechnology is not a pest man-
natural enemies are valuable and should be harmed no more agement approach as such, we include it because it is receiving
than necessary. It is important that we develop a more active major emphasis and is being geared to provide a wave of new
approach that seeks to understand natural enemies and how products for pest management. In fact, many seem to view
they function as a part of the ecosystem and to promote their biotechnology as an innovative means for providing safe and
effectiveness by use of habitat management (landscape ecol- effective tools that will essentially resolve pest management
ogy) and other cultural management approaches. problems. Major technological advances in chemistry, bio-
Augmentation through propagation and release of natural chemistry, behavior, neurophysiology, molecular genetics, and
enemies is an area of biological control that has received much genetic engineering have resulted in an array of biorational
attention in recent years. These efforts include research on in vitro products and materials that are less toxic and hazardous to
and in vivo mass rearing technology and on transport and release humans and the environment than conventional pesticides.
methodology for area-wide population suppression and for field- These products include genetically engineered plants for stron-
to-field therapeutic treatments. Although the development of this ger resistance to pests, plants, and natural enemies with high
technology is valuable, it is an extension of the treat-the- tolerance to pesticides and sophisticated formulations and
symptoms paradigm. In principle, natural enemies used in these delivery methods for biopesticides, semiochemicals, and other
methodologies are biopesticides, and the general approaches new tools. The biorational兾biologically based materials pro-
differ from conventional pesticidal applications only in the kind vided are potentially valuable advancements that have an
1132 Wolf Prize in Agriculture
Perspective: Lewis et al. Proc. Natl. Acad. Sci. USA 94 (1997) 12245
appropriate place in modern pest management. However, the a total ecosystem level. Therefore, the approaches should
strategy for development and use of these ‘‘high tech’’ tools has focus on harnessing inherent strengths within ecosystems and
been dominated by a continued search for ‘‘silver bullet’’ be directed more toward bringing pest populations into ac-
solutions that can be easily deployed in a prescription-like ceptable bounds rather than toward eliminating them (Fig. 1).
manner to remediate pest outbreaks or to exclude the pest’s These solutions would avoid undesirable short term and long
presence. As spectacular and exciting as biotechnology is, its term ripple effects and would be sustainable. Moreover, for
breakthroughs have tended to delay our shift to long term, adoption of such approaches, they must reasonably meet
ecologically based pest management because the rapid array of production demands and be cost-competitive on the short
new products provide a sense of security just as did synthetic term. We suggest three lines along which approaches can be
pesticides at the time of their discovery in the 1940s. Also, developed: (i) ecosystems management; (ii) crop attributes and
industry focuses on using genetic manipulation and other tech- multitrophic level interactions; and (iii) therapeutics with
niques to increase the virulence and host range of biopesticides minimal disruptions. However, with all of these approaches, it
instead of designing them as complements to natural strengths. is important to keep in mind the objective of balance vs. undue
Thereby, the manipulated pathogens and the crops engineered to selective pressure by any single tactic. Recent experiences with
express toxins of pathogens are simply targeted as replacements insect pest management for cotton in the southeastern United
for synthetic pesticides and will become ineffective in the same States will be used for key examples in the discussion.
way that pesticides have. It will be unfortunate if these powerful Ecosystem Management. Understanding and managing an
agents are wasted rather than integrated as key parts of sustain- ecosystem within which we farm is the foundation upon which
able pest management systems. all the farming strategies, including pest management, should
be designed. This foundation has become the victim of reduc-
New Direction tionist approaches. Because of political and funding channels,
scientific teams typically are assembled around commodities
The four major problems encountered with conventional across geographical areas. Therefore, the informational base
pesticides are toxic residues, pest resistance, secondary pests, relative to a particular crop as an interactive component of a
and pest resurgence. The latter three of these are fundamental farming ecosystem is very limited. For example, cotton spe-
consequences of reliance on interventions that are both dis- cialists focus their interactions toward other cotton specialists,
ruptive and of diminishing value because of countermoves of often within their own discipline, across the cotton belt.
the ecological system. Therefore, a mere switch to nontoxic However, both vegetable and cotton production are increasing
pesticides, such as microbials or inundative releases of natural in the same area and sometimes on the same farms in the
enemies, although helpful in reducing environmental contam- southeastern United States. These crops share many of the
ination and safety problems, still does not truly address the same pests and natural enemy fauna. Therefore, pest manage-
ecologically based weakness of the conventional pest control ment practices on one crop can directly or indirectly affect the
approach. Such tools used in this manner, whether chemical, other. A redirection of pest management is needed to incor-
biological or physical, are extensions of the conventional porate year-round soil, weed, cropping, water, and associated
approach that leaves us in a confrontation with nature. Also, practices at farm and community levels and to consider the
this operational philosophy tends to promote the development effects of these practices on the overall fauna, nutritional state,
and adoption of the more disruptive products because, within and balance of local ecosystems (16).
this paradigm, they work better than softer, less obtrusive Recent studies demonstrate that such a redirection would be
materials. highly fruitful. For example, problems with soil erosion have
What, then, would represent a meaningful fundamental shift resulted in major thrusts in use of winter cover crops and
in our pest management strategy? Furthermore, what should conservation tillage. Preliminary studies indicate that cover
be the components of such a strategy, and how can we crops also serve as a bridge兾refugia to stabilize natural enemy兾
crystallize this strategy into programs that result in effective pest balances and relay these balances into the crop season (17,
and lasting pest management systems? Clearly, the central 18). Crimson clover and other legumes, into which cotton can
foundation should be approaches that appreciate the interac- be strip tilled in the Southeast, appear to be good winter and
tive webs in ecosystems and seek solutions with net benefits at spring reservoirs for predators and parasitoids of cotton pests
FIG. 1. Illustration of a shift to a total system approach to pest management through a greater use of inherent strengths based on a good
understanding of interactions within an ecosystem while using therapeutics as backups. The upside-down pyramid to the left reflects the unstable
conditions under heavy reliance on pesticides, and the upright pyramid to the right reflects sustainable qualities of a total system strategy.
W. Joe Lewis 1133
12246 Perspective: Lewis et al. Proc. Natl. Acad. Sci. USA 94 (1997)
(19, 20). The green cloverworm in clover serves as a good cultivars, and their incidental loss, while breeding for other
alternate winter and spring host for the parasitoid Cotesia traits, can be prevented in the future.
marginiventris that limits subsequent outbreaks of armyworms Crop plants also provide vital food resources for certain key
and loopers in the cotton (21). Also, aphid, thrips, and natural enemies. Floral and extra floral nectaries, for example,
budworm兾bollworm populations in clover appear to provide provide necessary food for foraging parasitoids. Extra floral
reservoirs for establishing earlier balances between these pests nectar increases the attraction, efficiency, and retention of the
and their natural enemy guilds. On the other hand, when fields key parasitoids C. marginiventris, Microplitis croceipes, and
are fallow during winter and spring, natural enemy buildups Cardiochiles nigriceps, important to the control of armyworms
cannot begin until a crop is available. Integrating appropriate and bollworms兾budworms in crops such as cotton (34, 35).
cover crops with conservation tillage can have a number of However, extra floral nectar also serves as food for certain
agronomic benefits: reduced soil erosion, enhanced levels of pests such as the adult moths of the caterpillar pests just
organic matter, improved soil drainage and moisture retention, mentioned. Based on information regarding the role of the
restoration of important nutrients, and weed control (20, 22, 23) extra floral nectar as food for moths, nectariless cotton
while restoring and strengthening natural pest control. Other varieties were released a few years ago without regarding their
preventive measures including crop rotations, avoiding large scale importance as a food for natural enemies of cotton pests. These
monocropping, leaving unsprayed strips, and planting field mar- facts emphasize the need to broaden our base of information
gins with appropriate year-round refugia for natural enemies will upon which we design pest management strategies.
contribute to prevention of pest outbreaks (24–26). Also, there is a rapidly expanding body of knowledge about
The growing of clover and兾or encouragement of certain similar signaling that enables injured plants to produce toxins
weeds along field margins and other unplanted areas can also and antifeedants that are directed specifically toward herbi-
provide important refugia for developing natural enemy兾pest vores. For example, feeding activities of certain caterpillars on
balances during a cropping season. For example, two common the leaves of tomatoes and potatoes induce a systemic pro-
weeds in the southeast, fleabane and horsetail, are important duction of protease inhibitors expressed throughout the plant
hosts for plant bugs and their natural enemies. In fact, they are that interfere with the digestion process and feeding behavior
preferred over cotton by the insects, and data indicate that of insects (36).
these plants act as effective decoys to coax plant bugs away Even greater than our limited knowledge of the mechanisms
from cotton (19). Serious infestations of plant bugs occur regulating these important plant attributes is the void in our
primarily where cotton is planted ‘‘ditch bank to ditch bank,’’ knowledge of how factors like soil properties, nutrition, and兾or
along with clean cultivation apparently caused by exclusion of water stress affect their expression. Inadequate availability of
such preferred alternate host plants. a key soil element for example could make a major difference
Obviously landscape ecology practices exert a variety of
in the effectiveness of one or more of a plant’s interactions with
desired or undesired effects on cropping systems (27). Thus, it
herbivores or natural enemies, thereby influencing a plant’s
is vital that we assemble appropriate teams to elucidate
vulnerability to herbivore damage in a major way. Greater
interactions at the ecosystem level to establish the knowledge
understanding of the factors that regulate these interactions in
base for ecologically based pest management systems.
cropping systems can allow us to deal with plant health at an
Crop Attributes and Multitrophic Level Interactions. Con-
entirely different level.
sideration of crop plants as active components of multitrophic
There is a tendency within the traditional paradigm to use
level interactions is crucial to a total systems approach to pest
management. We have known vaguely for a long time that toxins, attractants, or other plant attributes as products and to
plant traits have important impacts on both herbivores and intervene in ways that are out of harmony with natural system
their natural enemies. But, again, the reductionist approach interactions. For example, we identify, synthesize, and formu-
has caused us to manipulate plant traits in ways detrimental to late herbivore toxins and natural enemy attractants as sprays
long term balance in the cropping systems. to kill herbivores and lure the natural enemies, respectively.
Recent discoveries of tritrophic level interactions among Also, we breed and engineer plants for constitutive expression
plants, herbivores, and parasitoids兾predators have demon- of traits in ways that maximize immediate deterrence of pests
strated how tightly interwoven these components are and or attraction of natural enemies without regard to pest density
illustrate the importance of multitrophic perspectives for or plant damage. Natural systems provide evidence that this is
effective and sustainable pest management strategies. Plants not always an appropriate approach for plant defense. In the
have long been known to possess toxins and other chemicals case of the protease inhibitor in tomato and potato cited above,
that serve to discourage herbivore feeding. The discipline of these materials are constitutively expressed in the fruit but only
host–plant resistance directed toward breeding plants resistant induced by damage in leaves (37). We suggest that this system
to pest attack was developed around such knowledge and has been selected in nature because it is the most durable
contributed greatly to pest management. Recent studies also strategy. A system of constitutive expression in fruit but only
show, however, that plants play an active and sophisticated role inducible in leaves experiencing damage by feeding insects
in their defense against insect activities, and their defense provides maximum protection of the fruit. Leaves serve as a
responses often are customized for certain, interactive, mul- decoy alternative for feeding by caterpillars but possess a
titrophic situations (5). For example, some plants respond to mechanism that limits feeding damage. This strategy also
insect herbivory by releasing volatile chemical cues that attract provides host兾prey resources that allow participation by a
predators and parasitoids that, in turn, attack the herbivores plant’s parasitoid兾predator allies. We must observe and con-
(28–30). These volatiles are released only in response to sider natural systems when developing strategies for novel
herbivore damage, not by mechanical damage similar to her- traits such as a gene for producing Bacillus thuringiensis (Bt)
bivory, and are released from the entire plant (31, 32). This toxin, i.e., plant engineering [see Gould (38) for an excellent
effect enables the natural enemy to distinguish infested plants reference in this regard]. For example, cotton cultivars with a
from uninfested neighbors. For example, cotton fed on by beet full constitutive expression of Bt toxin have been introduced
armyworm larvae releases terpenoids that attract the parasi- commercially. This practice amounts to a continuous spraying
toid C. marginiventris. Furthermore, a certain naturalized of an entire plant with the toxin, except the application is from
variety of cotton releases ⬇10 times more of these chemicals inside out. Various methods for resistance management, in-
in response to the insect herbivore damage than do commercial cluding pest兾natural enemy refugia and limiting acreage
lines (33). By understanding the mechanisms governing such planted with a cultivar, are being used. However, we urge more
important defense attributes, they can be restored to domestic concerted efforts toward breeding and engineering plants with
1134 Wolf Prize in Agriculture
Perspective: Lewis et al. Proc. Natl. Acad. Sci. USA 94 (1997) 12247
traits such as tissue-specific and damage-induced chemical We must remember—our primary objective in pest man-
defenses that work in harmony with natural systems. agement is not to eliminate a pest organism but to bring it into
Genetic engineering and other such technologies are pow- acceptable bounds. The role of therapeutics is not to replace
erful tools of great value in pest management. But, if their natural systems. Rather, their role is to serve as complements
deployment is to be sustainable, they must be used in con- while the system is temporarily out of balance. From that
junction with a solid appreciation of multitrophic interactions perspective, it is clear that interventions that interfere with the
and in ways that anticipate countermoves within the systems. restoration of balance are counterproductive. Waage (39)
Otherwise, their effectiveness is prone to neutralization by suggests that biopesticides could form the ‘‘methadone of
resistance in the same manner as with pesticides. IPM,’’ helping agroecosystems to recover from the habit of
Therapeutics. Therapeutics have a valuable role in ecolog- calendar spraying while we are redesigning and nurturing them
ically based pest management strategies, but they should be to a more self-renewing capacity.
viewed as backups rather than as primary lines of defense.
Also, therapeutics should be recognized as potentially disrup- Potential Benefits
tive and used as unobtrusively as possible. The key principle is
that they should be geared toward bringing a pest organism The benefits of a total system approach would be immense,
into acceptable bounds with as little ecological disruption as directly to farming and indirectly to society. The approach
possible. Synthetic products, natural products, and living or- takes into account impacts on our natural resources such as the
ganisms can be effective as therapeutics, and the fact that a preservation of flora and fauna, quality and diversity of
product is natural and兾or nontoxic does not necessarily mean landscape, and conservation of energy and nonrenewable
it is less disruptive than synthetic products. The important resources. Long term sociological benefits would also emerge
thing is to work as much in harmony as possible with the in areas of employment, public health, and well being of
system’s inherent defenses. persons associated with agriculture (42, 43).
A wide array of therapeutic products are available, and more In The Netherlands, prototypes of various multidisciplinary,
are being developed with modern technology. A vast arsenal arable farming systems have been evaluated on a semi-
of natural products identified from plants, insects, and micro- practical scale (44). In 1979, a national experimental farm for
organisms is being synthesized and formulated for use as the development and comparison of alternative farming sys-
biopesticides. Semiochemicals such as sex pheromones and tems was set up in Nagele (one of the ‘‘polders’’). The size of
natural enemy attractants can be used as baits and lures to the farm was 72 hectares (almost 300 acres). Among other
disrupt pest activity and promote natural enemy presence. studies, integrated and conventional farming practices were
Pathogens, parasitoids, as reared in vivo or in vitro, are compared for seed potatoes, dry peas, carrots, onions, sugar
available and are being touted as therapeutic tools. All of these beets, and winter wheat. Crop protection and other manage-
organisms and兾or their by-products are important biofriendly ment practices with the integrated approach followed the basic
alternatives to toxic, broad spectrum, conventional pesticides. principles discussed herein.
Still, our primary pest management tactic should be maximi- Over a 15-year period, pesticide use on these integrated
zation of built in pest reduction features of an ecosystem. farms was reduced over 90% (Fig. 2). They found that pesti-
Therapeutic tools should be used as secondary backups. cides, and fertilizers, can be decreased through implementa-
Overreliance on them will return pest management strategies tion of alternative practices based on intensified knowledge of
to a treadmill situation (Fig. 1). the ecosystem. Artificial fertilizers are replaced by organic
Another problem is the tendency to seek therapeutics that manure and effective use of crop residues. Insect, weed, and
give us the quickest effect. Sales of biological insecticides disease problems are reduced through natural control by the
amount to about $110 million annually, and Bt is the main enriched natural enemy fauna, the use of weed-competitive or
product ($90 million). Generally, microbial organisms work disease- and pest-resistant varieties (with an emphasis on
slowly relative to synthetic pesticides. Therefore, industry has durable systems for resistance), reduction of nitrogen fertili-
as first priority formulation of microbials to obtain faster kill zation, and judicious use of chemical pest control based on
and is less interested in long term pest reduction effects. Thus, careful population sampling and decision thresholds. Results
the role that microbials could play in orchard and forest pest from these demonstration farms have been so encouraging that
management, as well as in programs like control of grasshop- implementation of integrated farming is being enforced by the
pers in Sahelian, Africa, is neglected (39).
Retarded development of pests may be more desirable than
quick kill in certain situations. For example, Bt products are
considered unacceptable for controlling beet armyworms in
cotton because of their slow killing action. Yet, some studies
indicate that a slow kill may be more preferable when exam-
ined from a larger perspective. As indicated above, C. mar-
giniventris is a key parasitoid for managing the beet armyworm
and interventions should avoid disrupting this natural enemy.
Beet armyworm larvae intoxicated by sublethal dosages of
MVP (Mycogen, San Diego) (a Bt-derived biopesticide) ex-
perience retarded development and feeding and are subject to
higher parasitism than nontreated beet armyworm larvae (40).
In other words, an effective, nondisruptive way to manage a
moderate beet armyworm outbreak may be to retard its
development and damage while giving the parasitoids time to
work, thereby strengthening the parasitoids’ effect during
subsequent generations. A similar effect was reported earlier
for Bt and a parasitoid of gypsy moths (41). A quick kill may FIG. 2. Average use of pesticides (kilogram active ingredient兾
provide more immediate results but destroys a resource for hectare兾year) in conventional and integrated farming demonstrations
parasitoids and limits their presence with subsequent genera- in The Netherlands (1986–1990); after Wijnands and Kroonen–
tions of pests, thus leading to resurgence. Backbier (43). ha, hectare.
W. Joe Lewis 1135
12248 Perspective: Lewis et al. Proc. Natl. Acad. Sci. USA 94 (1997)
Dutch Ministry of Agriculture to reduce environmental pol- 14. Board on Agriculture, National Research Council, Committee on
lution and to create a firmer basis for survival of agriculture in Pest and Pathogen Control Through Management of Biological
the longer term. Control Agents and Enhanced Natural Cycles and Processes
Yields were somewhat lower on the demonstration farms but (1996) Ecologically Based Pest Management: New Solutions for a
New Century (National Academy Press, Washington, DC).
were compensated for by cost reduction through lower pesti- 15. van Lenteren, J. C., Benuzzi, M., Nicoli, G. & Maini, S. (1992)
cide and fertilizer inputs. Thus, the net short term profits of the in Biological Control and Integrated Crop Protection: Toward
demonstration farms were equal to those of the conventional Environmentally Safer Agriculture, eds. van Lenteren, J. C., Minks,
farms. We emphasize the short term economic aspect of A. K. & de Ponti, O. M. B. (Purdoc, Wageningen, The Nether-
sustainable farming because immediate profitability figures, lands), pp. 77–89.
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unsound methods as well as its array of inherent strengths that lett, G. B. (Wiley, New York), pp. 347–387.
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but understand and work more in harmony with nature’s 29. Dicke, M., Sabelis, M. W., Takabayashi, J., Bruin, J. & Posthu-
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We are grateful to Karen Idoine and Drs. Charlie E. Rogers (now 32. Röse, U. S. R., Manukian, A., Heath, R. R. & Tumlinson, J. H.
deceased), G. T. Fincher, John Garcia, Bryan Beirne, Thomas Eisner, (1996) Plant Physiol. 111, 487–495.
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1136 Wolf Prize in Agriculture
STING OPERATION
HOW A BUG EXPERT IN TIFTON TRAINS WASPS TO FIGHT IN
THE WAR ON TERROR
BY LUKE DITTRICH, ILLUSTRATION BY CELIA JOHNSON
One day, when Joe Lewis was 11 or 12 years old, he snatched one of his father’s
roosters and smeared it with smut from a wash pot until its neck feathers were
smudgy and dark. Joe’s grandfather had told him that roosters recognize one
another by the coloration of their neck feathers, so Joe had decided to find out
what would happen if he messed with that coloration. This particular rooster was
the top cock in the local pecking order, the one all the other roosters habitually
deferred to in matters of food and females. When the boy returned the besmirched
bird to its flock, what happened next was so exciting that he repeated the process
many times in the weeks that followed.
“He’d go to take charge and take back his domain,” Joe remembers, “and all
these other roosters would go, ‘Who are you?’ That was one of my earliest scientific
studies. I could spend all day watching rooster fights!” Joe Lewis is now 62 years
old, and this is a story about one of his more recent scientific studies. This is also a
story about screaming plants, chocoholic wasps and why the Department of Defense
thinks educated insects might protect us from terrorists.
Joe grew up on 4.5 acres of cotton in South Mississippi, a place he now calls
“the back end of the world.” His father was a share cropper who did not know
how to read or write. The family lived close to the soil, as farmers say. A mule
pulled the plow. Geese cut the grass. Dogs helped the hunt. Joe plucked the fiber
from the bolls. Life on the cotton patch was a complex balance of chores and
processes partitioned between nature and man.
I. Joe has learned many things by studying bugs. Strangely enough, the bugs
that he studies have also learned many things from Joe.
The cotton had enemies. If you had been there, in those fields, and you had knelt
in the dirt, and you had looked closely at the underbellies of the plants, the
enemies would have revealed themselves. You might, for example, have seen a
small caterpillar the color of faded parchment, cutting at a leaf with serrated jaws,
moving as slowly as a minute hand, ingesting healthy plant matter at one end and
ejecting useless brown waste out the other. If you had watched long enough, the
enemies of the plant’s enemies would have revealed themselves too. A tiny wasp,
even smaller than the caterpillar, might have lit upon the worm’s back and impaled
it with its ovipositor.
W. Joe Lewis 1137
The two kinds of bugs, worms and wasps, had a common enemy as well. Every
afternoon, Joe’s father would tie a red handkerchief around his face, haul a sack
of dust out into the field and coat his crops with DDT and other poisons, tainting
wasp, worm and human alike. When Joe left the cotton patch, he went to Mississippi
State University. One of the first books he read there was Rachel Carson’s Silent
Spring, a popular environmentalist work that excoriated chemical pesticides like
those Joe’s father used. Carson recommended an alternative, something called
biological control, which, rather than killing your crop’s enemies with artificial
dust, would kill them with their own natural enemies. If Joe’s father had employed
such methods, he might have, for example, dealt with his bollworm problem by
sprinkling his cotton plants with wasps, rather than DDT. The wasps would then
have done what they do naturally: lay voracious offspring in the bollworms’ bellies,
stunting their growth and shortening their lives. The concept was not new. Chinese
farmers purchased nests of mercenary ants to kill pests in their citrus groves as far
back as the third century, and such methods eventually spread around the world,
becoming ubiquitous by the 1800s. By the 20th century, however, the cheapness
and relative effectiveness of chemical pesticides had almost made biological control
obsolete. Joe found Carson’s ideas hugely appealing. He particularly liked the idea
that they might offer men like his father an alternative to the killing dust. As Joe
soon learned, however, biological control was a process as primitive and unrefined
as it was ancient. Humans, for all their complexity and smarts, knew very little
about effectively harnessing the swarms of insects that surrounded, outnumbered,
helped and hindered them. For example, everyone knew wasps killed bollworms,
but nobody knew how a tiny wasp could find a tiny bollworm in the middle of a
4.5-acre cotton patch. If we could figure out what was guiding the wasps, maybe
we could exploit that knowledge to help the wasps hunt, making them stronger
allies. Until we learned such things, though, biological control methods were unlikely
to ever replace cheap and dirty chemicals.
So Joe decided to study bugs.
II. When he was a boy, back in the back end of the world, he knew only a
little about the caterpillars he called bollworms and the tiny black wasps
that preyed on them. Now he knows a lot more.
Dr. Joe Lewis is a big man with a broad, handsome face, floppy grey hair and a sort
of exuberant politeness that probably serves him well in his role as vice mayor of
Tifton, a small city in South Georgia surrounded by thousands of acres of cotton
and peanuts. Politics is a part-time thing for Joe. His main job is with the United
States Department of Agriculture’s Agriculture Research Service, where he works
as a behavioral entomologist at the USDA’s Crop Protection and Management
Research Laboratory in Tifton. When he was a boy, back in the back end of the
world, he knew only a little about the caterpillars he called bollworms and the tiny
black wasps that preyed on them. Now he knows a lot more. These days, he calls
the bollworms Helicoverpa zea and the wasps Microplitis croceipes, and he writes
papers about them with lofty titles like “Herbivore-Infested Plants Selectively Attract
Parasitoids.”
Today, when he describes a field of cotton, or a field of any other crop,
he sounds like he’s describing a single living organism. His descriptions are
anthropomorphic. The cotton plants are the body, the bollworms are a virus, the
wasps are white blood cells, and every part of the field is reliant upon every other
part in an intricate and delicate balance. Joe has learned many things by studying
bugs. Strangely enough, the bugs that he studies have also learned many things
from Joe. Here is one thing Joe has learned by studying bugs: Plants scream.
When a bollworm slashes through the chlorophyllic flesh of a cotton plant, for
example, the plant pumps a combination of 12 chemical compounds out through
its pores.
Those chemicals are its scream, and only bugs hear it. If a wasp who is hungry
for bollworms senses such a distress signal, it reacts, following the signal to its
source, attacking the worm, defending the plant. The screams are very specific.
The one just described might be translated into English as: “I am a cotton plant,
and a bollworm is eating me!” Had the cotton plant been attacked by an aphid, the
signal would have been different, and the wasp, who has no use for aphids, would
have ignored it. In that case, a passing aphid-starved ladybug might instead respond
to the plant’s scream and feast.
The most important thing Joe has learned from bugs is that bugs can learn
from Joe.
This was a surprise — not just to Joe but to the entire scientific community,
which had long thought bugs to be hard-wired creatures that basically did
everything by instinct. In the late 1980s, however, Joe and his former Mississippi
State University classmate and long-time USDA colleague, Dr. J.H. Tumlinson,
discovered that bugs, in fact, have quite malleable minds and are constantly learning
and adapting to their changing circumstances. For example, if a female parasitic
W. Joe Lewis 1139
wasp encounters a bollworm on a stalk of corn, she will lay her egg in the worm
and, while doing so, her antennae will energetically sample the cornstalk. When
she flies off, the odors of the cornstalk and the satisfaction of egg-laying will be
newly linked in her mind. The next time she detects cornstalk odors, she will track
them to their source to see if she can find another host.
This is called “associative learning.” It is exactly the same process that caused
dogs to salivate when Pavlov rang his tuning fork, and it is a process that, before
Joe came along, everyone thought was exclusive to mammals and other relatively
large-brained creatures.
When Joe learned that bugs learn, he published a paper in the scientific journal
Nature detailing his findings. The paper described a simple experiment in which
Joe demonstrated he could teach a wasp to love vanilla. He did this by exposing a
wasp to the smell of vanilla while it examined its naturally beloved bollworm
feces. After this exposure, the wasp would become a vanilla-hunting fiend, flying
straight toward the scent in a wind tunnel. To make sure this particular species of
wasp didn’t have some strange innate predisposition to vanilla, Joe repeated the
experiment using a different scent. Soon, his trained wasps were flying just as
fixedly toward the scent of chocolate.
Within the Department of Defense, there is something called the Defense
Advanced Research Projects Agency, or DARPA. It used to be known as ARPA,
which is why the Internet, which DARPA invented, used to be known as the ARPANET.
DARPA’s mission, in its own words, is to “prevent technological surprise from
harming our national security by sponsoring revolutionary, high-payoff research
that bridges the gap between fundamental discoveries and their military use.”
Many DARPA-developed technologies — stealth fighters and global positioning
satellites, for instance — have already changed the world. Many other technologies
still in development — performance-enhancing exoskeletons, cognition-enhancing
neural implants — will probably change the future. An important part of what
DARPA does consists of keeping tabs on the latest breakthroughs in every scientific
field. If DARPA hears of a new idea or theory that might have some sort of military
application, it approaches the scientists behind the idea.
That is why, in the summer of 1999, DARPA approached Joe Lewis and asked
whether he and his chocolate and vanilla-loving wasps would like to become
defense contractors.
Humans are very bright, but our senses are very dull. Thousands of species of
nonhuman creatures see better than us, hear better than us and detect smells
better than us. We have always recognized the superior sensitivity of certain animals,
and exploited it, utilizing these animals as what organizations like DARPA would
call “biological sensors.” Dogs are the most familiar example. The first domesticated
dogs were basically organic burglar alarms. They could be counted on to bark at
an intruder long before we could sense the stranger’s approach. Eventually, we
1140 Wolf Prize in Agriculture
employed dogs for more sophisticated detection work. Starting in the seventh century
A.D., we began training them to actively track other animals and humans. Starting
in the last century, we also trained them to sniff out bombs, bongs and bodies.
Even with the recent invention of computerized “electronic noses” — devices
that cost about $10,000 and attempt to replicate biological olfactory systems in
silicon-dogs still possess the best readily available chemical-detecting technology
on the planet. Our most sophisticated electronic noses are only about a tenth as
sensitive as the nose of a trained canine. But although detection dogs are useful
and effective, they are also big and expensive. They require years of training,
experienced handlers and are impossible to use surreptitiously.
DARPA wanted Joe to investigate whether his trained wasps might, in essence,
be smaller, faster, cheaper bloodhounds.
When DARPA first approached him, Joe was a little skeptical. “We knew we
could train [wasps] to natural odors like vanilla and chocolate, but we didn’t know
whether we could train them to these totally unnatural things that had no part in
their natural history, like explosives and nerve gases.”
“But anyway, we said, ‘We’ll look into it.’”
“Do you want to smell it?”
Dr. Glen Rains, an agricultural engineer at the University of Georgia who has
worked closely with Dr. Lewis in his wasp-training work, holds up a little brown
bottle. The label indicates it comes from a chemical manufacturing company called
Aldrich and contains something called cadaverine. Cadaverine is just what you
might guess it is: a chemical compound released by human bodies when they
decay. Believe it or not, it hardly smells at all.
Cadaverine is one of the dozens of compounds that Joe has trained his wasps
to seek out. In fact, once he started on his DARPA work, Joe quickly discovered that
there doesn’t appear to be a single chemical compound that his wasps can’t learn
to love. Before long, he was churning out TNT-sniffing wasps, corpse-sniffing
wasps and, yes, nerve gas–sniffing wasps. Best of all, his wasps appeared to have
olfactory systems just as acutely sensitive as dogs and were able to detect quantities
of any given chemical compound in ratios as low as four parts to a billion.
Once he’d demonstrated that his wasps could, in fact, become bloodhounds,
Joe’s next problem was figuring out how to harness them.
The way DARPA saw things was a little simplistic, Joe says. He recalls that his
DARPA liaison wondered why they couldn’t just release a few thousand bomb-
sniffing wasps and follow them until they clouded around the head of, for example,
someone with explosives hidden under his clothes. “We said, ‘Well, wasps don’t do
that.’ The thing about parasitic insects is, if you put them in a room, or any
unnatural environment, they’re distracted, and they’ll just fly to the light.”
What Joe needed was the wasp equivalent of a bloodhound’s leash —something
that would keep the insects under control and undistracted as they sniffed out
whatever it was they were trained to sniff out.
W. Joe Lewis 1141
That’s it. If the wasps detect the chemical they have been trained to detect, they
will cease their aimless milling and cluster around a hole in the bottom of the
Wasp Hound. The hole is right above the fan, which sucks air inside. The computer
program analyzes the Web cam’s overhead view of the clustering wasps. When
they cluster thickly enough, it sets off an alarm, indicating the presence of the
chemical in question.
With properly trained wasps, the Wasp Hound can detect the merest whiff of
TNT in a pile of suitcases. A different batch of wasps might pick up a smidgen of
cadaverine emanating from beneath a field of wildflowers. Other wasps might
notice a few molecules of aflatoxin, a dangerous toxin, in an otherwise robust crop
of peanuts.
The Wasp Hound is about as compact as the best currently existing electronic
nose, more effective and only about 2 percent as expensive. Dr. Rains sees a sweet
sort of back-tracking in the superiority of wasp-driven chemical sensors over
silicon-driven ones.
“With technology, you’re always trying to get away from the way things used to
be, but as we go further and further, we kind of start going back. Look at the
medical field: Lots of doctors are going back to using maggots to help clean wounds!
So you know, things kind of come back around. Nature’s had millions of years to
develop systems that are as efficient as possible.”
Will bugs soon be battling bad guys?
Maybe.
DARPA might choose to enlist Joe’s trained wasps in the war on terror. In the
field, Wasp Hound–like devices could feasibly sniff out caches of explosives or
send out warnings at the first hint of nerve gas. Now that DARPA has jump-started
1142 Wolf Prize in Agriculture
his research and gleaned what’s possible, Joe’s not even sure he’d know if the
government decided to pursue future military applications. “They might step in
again. We may or may not know. If they want to do something more, they’d
probably want to classify it and do all of it in-house.”
In the meantime, Joe and Dr. Rains and their respective institutions, the USDA
and the University of Georgia, have applied for a patent on the Wasp Hound. They
don’t plan to produce and market a commercial version of the device themselves
but, rather, would like an outside company to lease the patent and take over from
there. There are still problems to work out to make the technology practical. The
most pressing is how to go about mass-producing trained wasps. The current
training process is done one wasp at a time and takes about 20 minutes to get a
cartridge-filling quantity of five wasps ready to go. They also have to be retrained
every 48 hours or so until the end of their lives, which typically comes about 21
days after they’re born. Apart from the problem of mass producing these short-
lived worker wasps, there’s also the inevitable inertia that always attends the birth
of a completely new and unknown sort of product.
“We’re doing stuff in an area where there’s no industry that currently exists,”
Joe says. “It’s not like developing a new pesticide. There’s no infrastructure in
place that takes invertebrate organisms and uses them for devices for chemical
detection. There’s an incredible need in the whole area. But there’s a lack of
infrastructure.”
Joe would just as soon leave the birthing of a new industry and all the associated
headaches to someone else. He’d rather get back to doing what he does best:
learning about bugs. And now that he’s learned just about everything he can about
wasps and proven that they can, in theory at least, double for dogs, he’s decided to
try teaching a new bug some new tricks.
Usually when Joe’s father finished dusting the cotton plants with DDT, he’d go
back to the house and sprinkle a little pile of the poison on the kitchen windowsill
to keep mosquitoes at bay. DDT has since been banned, but as anyone who’s ever
seen a digital watch’s plastic casing melt after an application of a DEET-based
repellent knows, we still engage in potentially hazardous chemical warfare against
mosquitoes all the time. A lot of people wish there were safer ways to fight them.
That’s why, today, if you visit Joe’s laboratory, you’re more likely to find
mosquitoes flying around in his wind tunnel than wasps. Mosquitoes, Joe discovered
just last year, have malleable, trainable minds too, though you have to substitute
blood for bollworm feces when you’re teaching them. Joe envisions several mosquito-
devastating scenarios growing out of his work. For example, you could use bait
stations to train a small number of mosquitoes to be drawn to a certain scent, then
count on the insects’ natural “aggression” behavior —their tendency to follow one
another when in pursuit of food — to give other nearby mosquitoes a nose for the
same scent. By lacing these bait stations with chemicals that sterilize mosquitoes,
you could nip an entire community in the bud.
W. Joe Lewis 1143
Joe has just begun his work with mosquitoes, and he’s hesitant to divulge much
more about it, but the basic principles are the same as his work with wasps. “Once
you can find out what they’re doing and play games with them,” he explains,
“there are all sorts of ways you can intervene and alter them.”
A half-century ago, a boy from the back end of the world grabbed a rooster
and smeared its neck with smut. He had begun playing games with the creatures
he shared his cotton patch with. He hasn’t stopped yet.