Atlas of Plant Viruses

Atlas
of
Plant inViruses
Volume I
Authors
R. I. B. Francki, Ph.D. Robert G. Milne, Ph.D.

Department of Plant Pathology Institute of Applied Plant Virology
Waite Agricultural Research Institute
Editor National Research Council
University of Adelaide Turin
Glen Osmond Italy
South Australia
T. Hatta, Ph.D.
Aburahi Laboratories
Shionogi & Co., Ltd.
Shiga
Japan
(gp)
CRC Press
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Library of Congress Cataloging in Publication Data
Francki, R. I. B., 1930-

Atlas of plant viruses.
Includes bibliographies and indexes.

1 . Plant viruses--Atlases. I. Milne, Robert G.
II. Hatta, T. III. Title.
QR351.F73 1985 632’.8 84-6999
ISBN 0-8493-6501-5 (v. 1)
ISBN 0-8493-6502-3 (v. 2)
A Library of Congress record exists under LC control number: 84006999
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PREFACE
Electron microscopy has played an important role in the development of virology, con-
tributing significantly to an understanding of the molecular biology and pathology of viruses
and to the development of virus taxonomy and diagnostics. Nevertheless, our impression is
that, while electron microscopes and their attendant techniques are generally capable of
producing micrographs of high quality, the micrographs presented to journals (and accepted
by them) or published in books, vary in quality from excellent to poor. No publication exists
that assembles a comprehensive collection of plant virus electron micrographs of good
quality, offers a consistent treatment, and backs the visual data with a consistent and com-
prehensive text.
It was with these considerations in mind that we decided to embark on preparing an atlas
of plant viruses. Our aim has been to bring together, in a systematic way, the available
information about the morphology and cytopathology of plant viruses in all the taxonomic
groups recognized by the International Committee on Taxonomy of Viruses (ICTV). We
have also tried to consider some viruses that are quite well known but still unclassified.
The project might arguably have been better tackled by an editor marshalling a group of
authors to write chapters on viruses with which they were especially familiar. By this
procedure each group of viruses would have been treated in the most authoritative way. In
practice, however, such an approach has its pitfalls. Uniformity and coherence are difficult
to attain in such a book, and the near-impossibility of extracting manuscripts from all the
authors on time can mean that certain chapters (written by the promptest authors) are painfully
out of date at the time of publication. Believing that uniformity and coherence are important
in an atlas, we chose the rather amibitious task of preparing the book ourselves. In striving
to be authoritative we have relied heavily on many colleagues for information and material.
Although this book is primarily about the structure of virus particles and infected cells,
we see little virtue in studying structure without function. Hence we have referred to the
results of biochemical experiments where relevant, so that the virus particles described appear
as part of a replicating complex. Similarly, we have tried to portray infected cells as active
rather than static structures.
In Chapter 1 a brief account is given of the development of studies on virus structure,
nucleic acids, cytopathology and taxonomy from their beginnings. We hope that this w i l l
provide a useful background to the chapters that follow. The two authoritative guides to the
taxonomy of plant viruses used are the Reports of the ICTV and the Commonwealth My-
cological Institute and Association of Applied Biologists' CMI/AAB Descriptions of Plant
Viruses, edited by B. D. Harrison and A. F. Murant. It seems important to standardize on
a list of virus abbreviations or sigla that will come to be used unambiguously by all plant
virologists. We have used the list of van Regenmortel published in his book Serology and
Immunochemistry of Plant Viruses (Academic Press, 1982) where possible, and have ex-
tended it.
We have tried to present not only the current state of knowledge about the viruses discussed,
but also the current state of uncertainty (or sheer ignorance) in particular areas. In several
of the chapters, data are included on unclassified viruses with possible affinities to the group
under discussion; in doing this, we hope to stimulate investigations that will clarify the
positions of such viruses.
The bibliographies at the end of each chapter are not exhaustive, but they include references
that we consider the most relevant, and sufficient for the reader who wishes to explore
further.
Many of the electron micrographs in this book concern viruses investigated as a pan of
a research project supported by a Commonwealth Special Research Grant from the Australian
Department of Primary Industry on the "Characterization of Plant Viruses in Australia".
Many others were prepared in the Italian National Research Council Plant Virus Institute in
Turin, Italy (the Istituto di Fitovirologia Applicata del CNR). Other photographs, or virus
preparations from which we prepared photographs, were kindly supplied by colleagues. For
easier comparison, we have presented the particles of at least one example of each virus
group negatively stained in the same material (uranyl acetate) and magnified a nominal (i.e.,
not always accurately calibrated) 300,000 times. This is not to deny the usefulness of other
negative stains or preparative techniques. In selecting the micrographs of virus-infected cells,
we have included large areas of the cells where possible, so as to give a better idea of the
ultrastructural context.
The treatment of some virus groups is more comprehensive than that of others. This is,
we hope, mostly a reflection of the current unevenness of the available data. Perhaps the
chiaroscuro may encourage some readers to do experiments that would help a future atlas
to be more comprehensive.
R. I. B. Francki
R. G. Milne
T. Hatta
AUTHORS
R. I. B. Francki is a Reader in Plant Pathology at the Waite Agricultural Research

Institute of The University of Adelaide, South Australia.
Dr. Francki received his B.Sc., M.Sc., and Ph.D. degrees in Botany from the University
of Auckland in 1955, 1958, and 1961, respectively.
He has published over 100 research papers and a number of reviews in the field of plant
virology. He was Chairman of the Plant Virus Subcommittee of the International Committee
on Taxonomy of Viruses from 1975 to 1981. He is also presently Secretary of the Virology
Division of the International Union of Microbiological Societies.
Dr. Francki is an Associate Editor of the journal, Virology, and of the Journal of General
Virology and is on the Advisory Boards of Current Topics in Vector Research and Micro-
biological Sciences. He has also served as an Associate Editor of Intervirology (from 1976
to 1981).
Dr. Francki is a member of a number of national and international scientific societies,
including the Society for General Microbiology, the Association of Applied Biologists, the
New York Academy of Science, the International Society for Plant Molecular Biology,
American Society for Virology, and the American Phytopathological Society. In 1983 he
was elected a Fellow of the American Phytopathological Society.
Dr. Francki has worked as a visiting researcher in a number of laboratories including the
Department of Plant Biochemistry, University of California at Los Angeles in 1964 to 1965.
He was a Fulbright Scholar at the Department of Agricultural Biochemistry, University of
Arizona, in 1970 and a State Agricultural University Senior Research Fellow in the De-
partment of Virology, Agricultural University, Wageningen, The Netherlands.
R. G. Milne, Ph.D., is a senior researcher with the Italian National Research Council,
Institute of Applied Plant Virology, Turin.
He received his B.A. from Trinity College, Cambridge, in 1956, and his Ph.D. from
Wye College, University of London, in 1960. From 1960 to 1964 he taught virology at the
Botany School, Oxford University, and from 1964 to 1966 did postdoctoral research in
electron microscopy at the Molecular Biology and Virus Laboratory, University of California,
Berkeley. Here he was lucky to have contact with Wendell Stanley, Robley Williams, Heinz
Fraenkel-Conrat, Arthur Knight, and "Geheimrat" Kleinschmidt. Returning to England, he
joined the Plant Pathology Department, Rothamsted Experimental Station, Harpenden, from
1966 to 1970, and then did a year of medical electron microscopy at the Clinical Research
Centre, Northwick Park, Harrow. In 1972 and again from 1974 to 1977 he was Visiting
Fellow of the Italian National Research Council in the Institute of Applied Plant Virology
in Turin. From 1972 to 1974 he was Associate Professor at the Institute of Molecular and
Cell Biology of the University of Strasbourg. He assumed his present position in 1977.
Dr. Milne was born in Tanganyika, and when very small acquired the nickname Bwana
Madudu ("He of the Creepy-Crawlies") due to his interest in large scorpions. As he grew
bigger, the bugs that claimed his attention grew smaller. He is now author or co-author of
about 80 research papers and reviews dealing mainly with thin sectioning, negative staining,
and immunoelectron microscopy of plant viruses. The plant reo-like viruses have been a
major interest, and lately a side-interest in virus taxonomy and nomenclature has grown
uncomfortably large. He has served on the editorial board of the Journal of General Virology
and is currently a member of the Plant Virus Subcommittee of the International Committee
on Taxonomy of Viruses.
T. Hatta, Ph.D., is a head of the Plant Pathology Division, Aburahi Laboratories, of

Shionogi & Co., Ltd., Shiga, Japan.
Dr. Hatta received his B.Ag.Sc. (1968) and M.Ag.Sc. (1970) from the University of
Osaka Prefecture, Japan, and the Ph.D. (1975) from the University of Auckland, Auckland,
New Zealand, where he studied with S. Bullivant and R. E. F. Matthews.
He was Postdoctoral Fellow in the Department of Plant Pathology at the University of
Adelaide from 1975 until he assumed his present position.
He is the author of a number of research and review papers, particularly in the field of
electron microscopy of plant viruses.
ACKNOWLEDGMENTS
We are much indebted to many colleagues who provided illustrations used in this book;
they are acknowledged individually in the text. We particularly thank Drs. G. P. Martelli,
M. Russo, and V. Masenga for preparing a number of electron micrographs especially for
this book. We also thank many colleagues who provided us with information by corre-
spondence and who sent manuscripts prior to publication. We are especially indebted to the
following colleagues who critically read and commented upon sections of the manuscript:
Drs. A. Appiano, D. H. L. Bishop, G. Boccardo, M. Conti, C. P. de Jager, D. C. Gaidusek,
A.-L. Haenni, B. D. Harrison, I. H. Holmes, R. Hull, T. Inouye, A. T. Jones, R. Koenig,
D.-E. Lesemann, V. Lisa, R. Lister, O. Lovisolo, E. Luisoni, G. P. Martelli, R. E. F.
Matthews, M. A. Mayo, G. I. Mink, D. Mossop, S. Pennazio, D. Peters, R. Redolfi, R.
J. Shepherd, and T. Yamamoto.
We thank Mrs. L. Wichman for much of the art work and Messrs. C. J. Grivell and B.
A. Palk for photographic reproductions. The help of Mr. C. J. Grivell was also invaluable
in putting the manuscript together, as was that of Mrs. S. Suter and Mrs. B. Goldsmith in
cheerfully typing and re-typing the manuscript.
Much of the work for this book was supported by a Commonwealth Special Grant from
the Australian Department of Primary Industry. Research at the Waite Agricultural Research
Institute was also supported by grants form the Australian Research Grants Scheme and the
University of Adelaide. Work on the book done in Turin was supported by the Italian
Consiglio Nazionale delle Richerche.
TABLE OF CONTENTS
Volume I
Chapter 1
Introduction 1
I. Virus Particle Structure 1
II. The Viral Genome 4
III. Virus Cytopathology 5
IV. Virus Taxonomy 7
V. Particle Morphology, Cytopathology, and Virus Identification 8
References 12
Chapter 2
Caulimovirus Group 17
I. Members of the Group and Their Relationships 17
II. Virus Structure and Composition 17

A. Particle Structure 17
B. The Viral Genome 21
III. Cytopathology 23
References 29
Chapter 3
Geminivirus Group 33

References 44
Chapter 4
Plant Reoviridae 47
I. Members of the Family and Their Relationships 47
A. Phytoreovirus 47
B. Fijivirus 47

B. Viral Genomes 57
A. Phytoreovirus 57
B. Fijivirus 60
C. Rice Ragged Stunt Virus 63
References 69
Chapter 5
Plant Rhabdoviridae 73

References 97
Chapter 6
Tomato Spotted Wilt Virus Group 101
I. Virus Structure and Composition 101
II. Cytopathology 102
References 110
Chapter 7
Maize Chlorotic Dwarf Virus Group 111
I. Properties of the Particles 111
III. Rice Tungro Virus — a Possible Member of the Group 111
References 116
Chapter 8
Tymovirus Group 117

References 134
Chapter 9
Luteovirus Group 137

References 151
Chapter 10
Sobemovirus Group 153

References 167
Chapter 11
Tobacco Necrosis Virus Group 171
I. Tobacco Necrosis and Other Possibly Related Viruses 171

1. Tobacco Necrosis Virus 171
2. Satellite Virus 172
1. Tobacco Necrosis Virus 172
2. Satellite Virus 172
References 179
Chapter 12
Tombusvirus Group 181

References 196
List of Virus Abbreviations and Taxonomic Groups 199
Index 209
Volume II
Chapter 1
Comovirus Group 1

References 18
Chapter 2
Nepovirus Group 23

C. Satellite RNAs 30
References 35
Chapter 3
Pea Enation Mosaic Virus Group 39
References 44
Chapter 4
Dianthovirus Group 47
I. Members of the Group 47

References 52
Chapter 5
Cucumovirus Group 53

C. Satellite RNA 56
References 65
Chapter 6
Bromovirus Group 69

References 78
Chapter 7
Ilarvirus Group 81

References 90
Chapter 8
Alfalfa Mosaic Virus Group 93
References 100
Chapter 9
Tobamovirus Group 103
A. True Tobamoviruses 103
B. Possible Tobamoviruses 104

1. True Tobamoviruses 110
2. Possible Tobamoviruses 112

References 127
Chapter 10
Hordeivirus Group 133

References 144
Chapter 11
Tobravirus Group 147

References 156
Chapter 12
Potexvirus Group 159

References 169
Chapter 13
Carlavirus Group 173

References 180
Chapter 14
Potyvirus Group 183


A. Cylindrical Inclusions in the Cytoplasm 191
B. Other Cytoplasmic Effects 201
C. Nuclear Inclusions 204
References 212
Chapter 15
Closterovirus Group 219
II. Particle Structure and Composition 222

References 232
Chapter 16
Some Unclassified Viruses 235
I. Oat Blue Dwarf Virus and Possible Relatives 235
II. Phleum Mottle and Similar Viruses 235
III. Carnation Mottle and Some Similar Viruses 236

IV. Viruses with Small Polyhedral Particles and Single-Stranded RNA Genomes,
but No Clear Taxonomic Affinities 237
V. Cacao Swollen Shoot and Structurally Similar Viruses 237
VI. Rice Stripe and Similar Viruses 237
VII. Carrot Mottle and Similar Viruses 243
VIII. Cryptic Viruses 244
IX. Concluding Remarks 248
References 252
List of Virus Abbreviations and Taxonomic Groups 257
Index 267
Volume I 1
Chapter 1
INTRODUCTION
Who first "discovered" viruses or developed the concept "virus" as we understand it

today is perhaps an idle question, as the idea took form over a number of decades. 12 Flower-
breaking in tulips was familiar in the middle of the 16th century, and by 1719 it was well
established, at least in some circles, that flower-breaking in jasmine could be graft-trans-
mitted. 3 In 1883, Mayer1 made a significant contribution when he found that tobacco mosaic
virus (TMV) was sap-transmissible, and Ivanovski 1 in 1892 made another with his discovery
that TMV was small enough to pass a Chamberland filter candle that prevented passage of
ordinary bacteria. Although this discovery was important, Ivanovski did not appear to realize
it, for he at first favored a bacterial toxin as the disease agent, and later suggested that it
was a bacterium.2 It was left to Beijerinck4 at the turn of the century to develop the concept
that viruses differ basically from bacteria. Nevertheless, his choice of the term contagium
vivumfluidum, or living infectious liquid, was unfortunate and probably hindered his ideas
being taken seriously for some time.
In the mid 1930s research activity on viruses gained impetus when TMV was purified
more or less independently in the U.S., Britain, and Australia5"9 and was found to be a
nucleoprotein.6 Various physical and chemical methods were used to investigate the size
and shape of TMV particles. (Particles by now had superseded fluid, but for a long time
arguments, that we now see as empty, continued about whether or not viruses were "living.")
Even before TMV was purified, it was concluded from rather ingenious stream double
refraction experiments that the particles were rod-shaped.10 Subsequently, using crystalline
purified virus preparations, X-ray diffraction studies confirmed that TMV particles were
rod-shaped and that they were made up of regularly arranged, uniform subunits. 6 Further-
more, it was concluded that the particles were about 18 nm in diameter and at least ten
times as long.6 However, it was not until 1939 that the blindfold was finally removed from
the eyes of researchers when a virus was examined in the electron microscope. TMV again
had the distinction of being the first virus to be studied by this technique."
Although of obvious potential, electron microscopy was initially slow to contribute to
virology. In the early days, the cost of the tricky and capricious instrument represented a
disproportionate amount of the modest budgets of virus laboratories. Suitable specimen
preparation methods had also to be developed before full advantage of the electron microscope
could be taken. Today, most virologists, whether researchers or diagnosticians, would con-
sider an electron microscope as one of the most basic instruments in their laboratories.
Without it, they would feel inadequate to detect, identify, or characterize the viruses they
work with as well as the impurities and contaminants they hope to avoid.
I. VIRUS PARTICLE STRUCTURE
Early electron micrographs of isolated virus particles were not very informative because
of the low contrast between particle and background. This is because neither proteins nor
nucleic acids are opaque to the electron beam. However, even without any enhancement of
image contrast, some valuable work was done. Kausche and his colleagues" confirmed the
shape of the TMV particle and later Stanley and Anderson12 obtained accurate size distri-
butions of rods in purified virus preparations. Viruses with isometric particles were also
examined, but the resolution was rather poor and little was achieved other than dispelling
the then-prevailing view based on hydrodynamic data that particles of all viruses were
anisometric.13
2 Atlas of Plant Viruses
Electron microscopy of viruses was revolutionized when Williams and Wyckoff 14 " 17 in-
troduced metal shadowing as a technique for increasing contrast. The method proved excellent
for revealing the shapes and sizes of virus particles, and where the particle surface needs
to be observed, metal shadowing yields unambiguous images uncomplicated by information
coming from deeper layers of the particle. However, much of the fine detail is hidden due
to the layer of metal deposited and the granularity of the resulting surface.
The introduction of negative staining in the 1950s18 20 was an advance in technique as
significant as metal shadowing. The images of virus particles could suddenly begin to be
interpreted in molecular terms.21 The method largely replaced the more laborious and gen-
erally less informative shadow casting, but there are instances where shadowed preparations
can be at least as informative as those negatively stained. Hatta and Francki22 used both
methods to study Fiji disease virus particles, and found that some aspects of these complex
structures were easier to understand from shadowed preparations. The presence of impurities
is also best detected by shadowing.16
Negative staining has proved so useful because it was found to outline virus particles with
nearly structureless electron-dense material, often also penetrating the surface to give contrast
to the morphological subunits. Perhaps nearly as important, it was found to support the
particle, to a greater or lesser extent, against the flattening and distortion experienced during
drying.
Negative staining was introduced at a time when much interest in virus particle structure
had been generated following the purely theoretical considerations on the subject by Crick
and Watson. 23 These workers suggested that since viruses with small particles had limited
amounts of nucleic acid, their protein shells must be built of numerous identical subunits.
There was already evidence that this was so with some viruses from X-ray crystallographic
and chemical data.24 In a paper accompanying that by Crick and Watson,23 it was shown
by X-ray crystallography that the protein shell of the tomato bushy stunt virus particle
consists of subunits arranged with 5:3:2 symmetry.25
Examination of negatively stained particles of many plant, animal, and bacterial viruses
by Home and Wildy 26 caused great excitement. This was especially so for viruses with
isometric particles because what had previously been seen as roughly spherical blobs became
intricate structures belonging to a number of geometrical families. (Though a large particle,
that of tipula iridescent virus, had already been elegantly shown by shadowing to have
icosahedral form.27) With hindsight, it is ironic that the first manuscript fully describing the
negative staining method was rejected by a leading virological journal. The circumstances
of the rejection have been described in a review by Home and Wildy.28 Fortunately, the
editors soon realized the error of their ways and in 1961 the same journal, in expiation,
published an unusually long and interesting paper on virus architecture based on the results
of negative staining.26 The extent to which the negative stain technique contributed to the
understanding of virus structure in a very short period of time can be gauged from a review
written less than 4 years after introduction of the technique.29
The method of Brenner and Home,20 using neutralized solutions of phosphotungstic acid
(PTA) was in fact so successful that it acquired elements of dogma. Many workers, partic-
ularly those in the medical field, used and still use neutral PTA exclusively and somewhat
indiscriminately: this, despite the fact that Hall 18 had originally used PTA at pH 4.6, Huxley
and Zubay30 had introduced uranyl acetate, and Home at various times has emphasized that
other negative stains have merit. As early as 1963 it was reported by Markham in a personal
communication to Home and Wildy29 that some plant viruses disrupt in neutral PTA. It
appears that one type of virus unstable in the stain is that whose stability depends on
electrovalent bonding between protein and RNA. The problem can usually be overcome by
fixing the virus, lowering the pH of the PTA, or using a stain such as unbuffered uranyl
acetate.31-32
Volume I 3
With enveloped viruses, PTA can produce artifacts of another kind in that the particles
become distorted. The cause of this is unknown but it has been suggested that at least with
the Rhabdovirus, lettuce necrotic yellows virus, osmotic and imbibition effects are respon-
sible. 33 The exact shapes assumed by the stained panicles depend largely on the pH of the
PTA. At neutrality the bacilliform particles become bullet-shaped due to the invagination
of the envelope at one end. The effect of the pH of PTA on the particle morphology of
lettuce necrotic yellows virus has been discussed in detail.34-35
It has been our experience that unbuffered uranyl acetate (pH —4.2) is a better general
negative stain for plant viruses and has been used for the majority of the micrographs in
this book. Uranyl formate,36 though otherwise behaving like uranyl acetate, offers a definite
advantage in revealing the fine structure of rod-shaped viruses. However, staining artifacts
have been encountered when uranyl acetate was used to stain Rhabdoviruses. 17 - 38 It should
also be mentioned that there is a potential danger of obtaining positively stained particles
because of the affinity of uranyl ions for nucleic acids. This is not a problem unless staining
times are inordinately long or the support film is very hydrophobic.
In addition to PTA and uranyl acetate, solutions of sodium tungstate and tungstoborate,
lithium tungstate, ammonium molybdate, lanthanum acetate, and uranyl formate have been
used39 as well as methylamine tungstate, sodium silicotungstate, uranyl oxalate, and sodium
zirconium glycollate. The subject of negative staining often provokes fierce brand loyalties
among microscopists, sometimes obscuring the fact that an ideal negative stain has not yet
been discovered. There is good reason to try out new heavy metal salt formulations, but
these should give exceptional results, repeatable in other laboratories, and should not just
be novelties, in order to replace the generally satisfactory portfolio of PTA at neutral and
low pH, uranyl acetate, uranyl formate, and ammonium molybdate.
One of the most serious problems encountered when examining virus particles in the
electron microscope is that biological objects (and their negative-stain images) exposed to
the electron beam are very readily burnt up, with a consequent drastic loss of structural
detail. This was recognized by Williams and Fisher,40 who devised the minimal exposure
technique and showed that using it can lead to significantly improved capture of fine detail.
The procedure is very simple in that part of the specimen near the region of interest, rather
than the actual field to be photographed, is used for focusing and other adjustments. This
leaves the important region unirradiated until everything is ready for the photograph to be
taken. The method presents problems where the subject is unique or occurs rarely on the
support film. In this case the necessary preliminary search can be made at very low illu-
mination. Wrigley and his colleagues41 have recently suggested refinements to the minimal
exposure technique.
Although electron images of negatively stained particles possess high contrast, their res-
olution is not very good when compared to that attainable by modern electron microscopes.
To extract more information, a number of techniques have been developed.
Markham and his colleagues42 showed that it was possible to enhance detail in micrographs
of radially symmetrical objects by a photographic rotation technique. This method is useful
but has sometimes been employed in a rather uncritical way to analyze images of virus
particles, and great care is required to avoid generating artifacts.43 Markham's group44 also
devised an apparatus for analyzing linear repeating features. Both these methods used pho-
tographic superimposition to reinforce periodic information recorded in the micrograph but
not distinguished from the background "noise" by the naked eye. The success of photo-
graphic image enhancement stimulated Klug and Berger45 to apply optical diffraction tech-
niques to electron micrographs. These methods, used in conjunction with computer analysis,
have reached a high degree of sophistication, and it is now possible to construct three-
dimensional images from electron micrographs of particles with radial or longitudinal
symmetry.46-47
One of the requirements of electron micrographs that are to be processed in this way is
the presence of images of orderly arrays of particles or subunits. This severely limits the
extra information that can be gained from, for example, isometric virus particles randomly
distributed on the grid. To surmount this problem, Home and his colleagues48-49 developed
methods for the preparation of crystalline or paracrystalline monolayers of virus particles
on electron microscope grids. Such arrays are ideal for diffraction and computer analysis,
and several viruses have been successfully manipulated to produce excellent reconstructed
images.50 With current developments in computer technology, it is not difficult to imagine
the possibility of direct display of reconstructed images on the computer screen, derived in
real time from arrays imaged in the electron microscope. One small reservation to bear in
mind is that the particle structure will have been distorted during the formation of the close-
packed array. It is clear, for example, that hexagonally packed "isometric" virus particles
have hexagonal contours whereas the same particles, cubically packed, have square contours.
As we have seen, virus particle structure has also been studied by X-ray diffraction.
Unfortunately, X-ray analysis has been restricted to viruses that can be crystallized and are
relatively simple in structure. However, this method has now reached a high degree of
sophistication.51 Recently the technique of small-angle neutron scattering52"54 has permitted
analysis of the structures of virus particles in solution, thus nicely complementing the results
of both X-ray analysis and electron microscopy.
As electron microscopy has extended its capacity to detect the fine structure of virus
particles, so it has made itself indispensable in diagnosis. It is now possible to identify the
particles of many viruses in juice simply and rapidly extracted from infected plants55 or in
purified and concentrated preparations. Use of virus-specific antisera to clump, decorate, or
trap particles on electron microscope grids56 can enormously amplify this already formidable
diagnostic potential. It must be added, however, that electron microscopists will not thank
you for presenting them with hundreds of routine samples to check. In these cases other
methods of virus detection should be used, such as enzyme-linked immunosorbent assay
(ELISA) which is equally sensitive but more suitable for mass screening programs.56
II. THE VIRAL GENOME
When TMV was purified for the first time by Stanley in 1935,5 he concluded that "tobacco
mosaic virus is regarded as an autocatalytic protein." In spite of the fact that RNA was
identified in purified TMV preparations soon afterwards,6-8 prime attention was paid to viral
proteins in virus research for the next 20 years. It was not until the classic experiments of
Fraenkel-Conrat and his colleagues57-58 on the reconstitution of TMV and those of Gierer
and Schramm59 demonstrating that deproteinized TMV RNA was infectious, that more
attention was given to research on viral nucleic acids. This led to our current concept that
virus particles consist of a nucleic acid genome surrounded by protein; the function of the
protein being, at least in the simplest cases, to protect the nucleic acid from the hazards of
nucleolytic enzymes when the virus is outside the host cells.
All plant viruses whose nucleic acids were analyzed before the late 1960s were shown to
contain single-stranded RNA and it was thought that all viruses infecting plants had single-
stranded RNA as opposed to many viruses infecting bacteria or vertebrates, which have
DNA. This "paradigm" was so strong that at least two plant viruses now known to have
DNA genomes were originally reported to contain RNA. In 1963, wound tumor virus was
the first virus shown to contain double-stranded RNA60-61 and in 1968 it was established
that cauliflower mosaic virus contained double-stranded DNA.62 Now, it is also known that
bean golden mosaic virus and a number of related viruses contain single-stranded DNA.63-64
Initially, research progress on viral nucleic acids was rather slow because of the lack of
appropriate techniques, and it is only recently that the pace has quickened. The first significant
Volume I 5
advance was the development of deproteinization techniques yielding undegraded nucleic

acids many of which retained infectivity. 3 ' 59 - 65 This together with advances in methods for
purifying viruses and separating them by density-gradient centrifugation66'67 led to the dis-
covery that in some plant viruses the genetic information was divided between two or more
nucleic acid species.68"72 The introduction of gel electrophoresis for the separation of nucleic
acids73 significantly improved the preparation of highly purified nucleic acids. This in turn
led to the demonstration that a number of plant viruses whose nucleoprotein particles sed-
imented as a single component had, in fact, tripartite RNA genomes.74-75
The use of density-gradient centrifugation and gel electrophoresis for separating virus
particles and their nucleic acids also led to the discovery of satellite viruses76 and satellite
RNAs.77 The RNA molecules of satellites are small and contain insufficient genetic infor-
mation for autonomous replication; they require normal viruses as helpers.78 Satellites exhibit
considerable specificity as to the viruses which will help them.78
Following the development of cell-free protein synthesis systems for the translation of
mRNAs, a number of plant viral RNAs have been translated in vitro.79-80 Initial attempts to
translate TMV RNA in a bacterial cell-free system were unsuccessful,81 but later work with
extracts from wheat-germ and rabbit reticulocytes has led to some understanding of the
function of single-stranded plant viral RNAs that act directly as messengers, i.e., have
positive polarity.80 Although most single-stranded RNA viruses have positive sense, some
are negatively stranded. This is indicated by the lack of infectivity of the RNA and the
presence of transcriptase in the virus particles.82-83
The recent spectacular advances in nucleic acid cloning and nucleotide sequencing meth-
ods84"87 are contributing greatly to our understanding of viral nucleic acid structures and
functions. Already the entire sequences of a number of viral nucleic acids have been de-
termined including that of cauliflower mosaic virus DNA8S and tobacco mosaic virus RNA. 89
III. VIRUS CYTOPATHOLOGY
Investigations on virus-infected plant cells began very soon after the discovery of viruses.
The first record in the literature of cellular pathology due to virus infection goes back to
the turn of the century when Ivanovski90 described abnormalities in TMV-infected tobacco
cells. Although his observations were very accurate, he still concluded that the causal agent
was a bacterium. Ivanovski90 observed both crystals and rather mysterious amorphous bodies
in the cells. It was shown conclusively by electron microscopy, 50 years later, that the
crystals consisted of virus particles.91 However, in spite of extensive studies, we still do not
understand the significance of the amorphous structures which have become known as X-
bodies.
Studies of virus-infected plant cells with the light microscope have continued to this day,
and it has been shown that many viruses induce characteristic inclusion bodies in the cells
they infect. The inclusions have been classified into various types by their morphology and
cellular location. The presence and properties of these inclusion bodies have been used in
diagnostic work and a vast literature has accumulated about them which has been reviewed
from time to time.92"96
For the examination of virus-infected cells in the electron microscope, a whole new
technology of specimen preparation was required because the fixation, embedding, and
section cutting methods used for light microscopy were not applicable. With the very pri-
mitive techniques available in 1950, Black and his colleagues97 were the first to identify
TMV particles in tobacco leaf cells, and a few years later Smith98 observed what appeared
to be crystalline arrays of tomato bushy stunt virus particles in the cells of infected Datura
stramonium.
With the attention of so many workers from various fields directed towards obtaining
sections suitable for electron microscopic examination, progress came relatively quickly.
Development of suitable methods for plant tissues were slower due to the presence of cell
walls and vacuoles in this type of tissue. The cell walls were a problem because of their
hardness and impermeability to fixatives and embedding resins; the vacuoles, due to their
acidic contents. However, by the early 1960s several developments had set the stage for
real progress. Good microtomes became available and the diamond knife was introduced."
Glutaraldehyde 100 had come to the aid of osmium tetroxide' 01 - 102 as a fixative, and the rather
treacherous methacrylates had been replaced by epoxy embedding resins or other suitable
polymers. 103 - 104 At about this time permanganate fixation was more or less discontinued
following the clear demonstration by Shalla 105 that it degraded TMV particles. The main
virtue of this fixative was its excellent preservation of cell membranes; this was achieved
at the expense of destroying virtually everything else, and resulted in a disarmingly simple
picture of cell structure. With care, however, dilute permanganate can be used without
degrading viruses such as TMV and without disrupting the crystalline arrays that may be
disturbed by glutaraldehyde. 106
By the mid 1960s processing of plant tissues for electron microscopy was well advanced,
and investigation of the effects of viruses in plants progressed rapidly. Comparison of the
review in 1966 by Matsui and Yamaguchi 107 with the book by Esau10" published only 2
years later, shows convincingly that the technology had been revolutionized. Over the next
decade plant cells infected by nearly all the known groups of viruses were examined and a
rich harvest of data accumulated. 95 ' 109
At present, our knowledge of the intracellular location of viruses with large particles
(whether enveloped or isometric) and those that are rod-shaped is better than that of viruses
with small polyhedral particles. This is largely due to the difficulty, in thin sections, of
distinguishing small virus particles from cytoplasmic ribosomes unless the virus particles
assume locations or arrays unlike those of the ribosomes.110""4 Even then it can be difficult
to decide if the unusual arrays consist of virus particles or of ribosomes that have taken up
such configurations due to infection.
The problem of distinguishing small polyhedral virus particles from ribosomes in sections
has been approached in several ways. Milne"" induced virus particles to form crystalline
aggregates by water-stressing leaf tissue prior to fixation. Honda and Matsui" 5 devised a
method of detecting particles of cucumber mosaic virus by floating infected leaf-discs on
phosphate buffer, after which cytoplasmic ribosomes were no longer visible. These methods
have the disadvantage that virus particles are identified in cells subjected to abnormal
physiological conditions before fixation.
Hatta and Francki"6 " 7 showed that the particles of a number of viruses could be distin-
guished from ribosomes after digestion of the ribosomal RNA with pancreatic ribonuclease.
Digestion was accomplished after aldehyde fixation, so that virus particles could be observed
in tissues which had been fixed while under normal physiological conditions. Many of the
micrographs included in this atlas are from tissues prepared in this way. Although this
technique is applicable to many viruses with small polyhedral particles containing single-
stranded RNA, the particles of some other such viruses are partially digested, and with these
the technique is less successful." 7
Cytochemical, immunochemical, and autoradiographic studies can be of considerable help
in understanding what normal metabolic processes are affected by virus infection. There is
not much information from these sources about virus-infected plant cells, at least partly
because of the presence of their impermeable walls. Protoplasts have not been altogether
satisfactory because events may differ from those occurring in cells of intact tissues." 812 °
However, after fixation of tissue pieces in glutaraldehyde, the cells can in fact be cut open
without undue penalty. 121 Fixed and sectioned tissue pieces were shown to contain cells
through which the cutting edge had passed but whose content was well preserved. They
Volume I 1
were used in enzyme cytochemical tests for double- and single-stranded RNAs. A similar
approach has been used for immunological studies of virus-infected plant cells.'- 2 1 2 4
IV. VIRUS TAXONOMY
Taxonomy has two aims: to break down a mass of units into named groups of like
individuals and to relate these groups in some kind of hierarchy reflecting their evolution.
Unfortunately for virus taxonomy, there are no fossils, so that evolutionary relationships
are very speculative. For plant viruses, studies on coat protein amino-acid replacements 125
are beginning to fill this void, and comparison of nucleotide sequences may eventually do
better. However, at present evolutionary clues are few and plant virus taxonomy largely
remains a question of collecting individuals into groups whose phylogenetic interrelationships
are obscure.
As early as 1927, Johnson 126 drew attention to the need for a system of classification and
nomenclature of plant viruses. In the four decades that followed, many schemes were
introduced but none proved acceptable. It is now quite obvious that too much was attempted
too soon. Since little was known about the intrinsic properties of the viruses, great weight
was placed on characters such as symptoms and host range, which we now know often
correlate poorly with physical and chemical parameters. A number of historical accounts of
the early attempts to classify viruses have been published. l27 - 129
The health of virus taxonomy took a turn for the better with the establishment of the
International Committee on Nomenclature of Viruses (ICNV) in 1966.13° The name of the
ICNV was changed in 1974 to a more appropriate one, the International Committee on
Taxonomy of Viruses (ICTV), which is active to this day. It is concerned with the taxonomy
of all viruses and has generally enjoyed wide support among virologists.
Within the structure of the ICTV, plant virus taxonomic matters are first handled by the
Plant Virus Subcommittee (PVS) and there are similar subcommittees concerned with viruses
of vertebrates, bacteria, invertebrates, and fungi. The subcommittees have a membership of
experts who draft the taxonomic proposals, which are then placed before the ICTV for
approval. This procedure is designed to ensure that all taxonomic questions are thoroughly
examined before gaining final ratification. The President of the ICTV publishes a report
every three years that has become the standard handbook on virus taxonomy. The fourth
and most recent report was published in 1982.m
Since the inception of the ICNV the taxonomy of plant, vertebrate, bacterial, and inver-
tebrate viruses has progressed more or less in parallel. However, the approach by the PVS
has been a little different from that of the other subcommittees. The PVS has been reluctant
to classify viruses into families, genera, and species, a practice supported by all the other
subcommittees. It has preferred to classify plant viruses into clusters loosely defined as
"groups", and this choice is supported by the majority of plant virologists. It has been
argued that the concept of the family, genus, and species is inappropriate for entities that
do not reproduce sexually and that it is too rigid to be useful in our present state of uncertainty
about plant virus relationships. Introduction of a binomial nomenclature to replace the current
vernacular system is also not looked upon with favor by many plant virologists.112 By using
the more flexible term "group' and vernacular virus names, it is felt that a more practical
system can develop, capable of accommodating small or large changes in the future, without
the embarrassment that has overtaken previous attempts at classification and nomenclature.
However, the rather loose vernacular system of nomenclature and "group" classification
also presents numerous problems.129
The first proposals from the PVS were presented at the meeting of the ICNV in Mexico
City in 1970, and resulted in the approval of 16 groups of plant viruses.'" •' 34 It was recognized
that certain plant viruses also replicate in insects and are very similar to some viruses of
insects and vertebrates. They were assigned as possible members of the then currently
recognized genera Reovirus andRhabdovirus. Subsequently, a further eight plant virus groups
were approved, four in 1975,115J1<1 three in 1978,'" and one in 1981. l 3 1 Also in 1978, the
plant viruses previously considered to be possible members of the genera Reovirus and
Rhabdovirus were included as regular members of the newly established families Reoviridae
and Rhabdoviridae, respectively. 1 " Furthermore, the plant viruses in the Reoviridae were
divided into two new genera, Phytoreovims and Fijivirus.^1 Thus at present, plant viruses
are divided among 24 groups and 2 families (Figure 1), and we shall use this system despite
its inconsistent and hybrid nature. We should also remember that new viruses are continually
being discovered, and that many already known do not seem to fit into any presently
established group or family.
In choosing characters on which to base a classification, the two extreme approaches are
to try to consider every possible character with equal weight (the Adansonian approach) or
to select, deliberately, those characters felt to be the most fundamental, Harrison and his
colleagues,113 when delimiting the first 16 groups of plant viruses, chose essentially an
Adansonian approach; but subsequently there has been a shift towards using more "fun-
damental" characters to delineate groups, reserving others such as host range or serology
of coat proteins to distinguish individual viruses and virus strains within groups. l19 Inspection
of all the characteristics of all the taxons that include plant viruses (Table 1) in fact shows
that only relatively few properties will consistently separate the groups in the same way.
V. PARTICLE MORPHOLOGY, CYTOPATHOLOGY, AND VIRUS

IDENTIFICATION
Particle morphology is one of the more important characters in assigning a virus to a

taxonomic group. Particles of viruses in the Caulimovirus, Geminivirus, Reoviridae, Rhab-
doviridae, tomato spotted wilt virus, and alfalfa mosaic virus groups are so typical that their
members need little further characterization before they can, at least provisionally, be as-
signed to their correct group or family. Most viruses with elongated particles can also be
assigned relatively easily to a recognized group according to their particle morphology and
modal lengths (Table 1).
Viruses with small icosahedral particles are the most difficult to assign to their groups on
the basis of particle morphology, though viruses such as the Tymoviruses do have particles
that are relatively easily recognized. The particles of some other groups can also be distin-
guished from each other. However, for the differences to be considered significant, the
particles should be compared as mixtures on the same specimen grid. There remain some
groups whose particles seem to be indistinguishable. For example, Hatta and Francki 140
failed to differentiate between particles of viruses belonging to the Luteovirus, Sobemovirus,
and tobacco necrosis virus groups; similarly, the particles of Tombusviruses and Diantho-
viruses appeared indistinguishable, as were those of Comoviruses and Nepoviruses.
The possibility of grouping certain viruses by their particle morphology is of enormous
value in diagnostic work. The morphology can often be determined very rapidly by examining
negatively stained preparations that need not be highly purified. Other tests (such as ser-
ological) can then be applied with a much greater economy of effort.
The cytopathic effects induced by some viruses are sufficiently characteristic to be used
for diagnostic purposes at group level.96 For example, the viroplasms induced by viruses
belonging to the Caulimoviruses or the Reoviridae are group specific. Similarly, the cylin-
drical (pinwheel) inclusions induced by Potyviruses are characteristic of that group, and
minor differences in pinwheel structure have also been used in attempts to subdivide the
viruses of the group. l41 - 142 On the other hand, there are numerous cases where the cytopathic
effects induced by viruses within a given group differ significantly. The Tobamoviruses are
Volume I 9
FIGURE 1. Diagram illustrating the current taxonomy of plant viruses approved by the ICTV.131 Names in large
print are of those families or groups approved by the ICTV. Names in small print are of groups which have been
approved by the ICTV but for which no names have been approved; they are all monotypic groups, ds, double-
stranded; ss, single-stranded; ( + ), genomic RNA positive-strand (i.e., containing coding triplets which can be
translated on ribosomes); ( - ) , genomic RNA negative-sense strand (i.e., strands with sequences complementary
to positive-strands which must be transcribed before being translated). (After Francki. 128 - 138 )
an excellent example (see Table 3 in Chapter 9, Volume 2). There are also instances of
very similar cytopathic structures being induced by viruses belonging to very different groups.
Table 1
THE TAXONOMIC GROUPS OF PLANT VIRUSES AND THEIR MAIN CHARACTERISTICS"
Coat protein
Genome segments polypeptides
Particle morphology and sizes and size
Virus group Type member and size (nm) 1 ' (Mr x 10") (Mt x 10')
Caulimovirus Cauliflower mosaic virus Isometric ~50 1 circular, ds-DNA 1, ~42

(CaMV) -4.8
Geminivirus Maize streak virus (MSV) Geminate ~18 x 30 2 circular, ss-DNA 1 , 28—34
0.7—0.8
Reoviridae Wound tumor virus (WTV), C Isometric ~70 10—12, linear. 6—7, 35—160
Fiji disease virus (FDV) ds-RNA 0.3—3.0
Rhabdoviridae Lettuce necrotic yellows vi- Bacilliform (enveloped) 1, linear, ss-RNA ~4 5, 21—170
rus (LNYV) d 135—380 x 45 x 95
Potato yellow dwarf virus
(PYDV)
Tomato spotted wilt virus gp.° Tomato spotted wilt virus Spherical (enveloped) 4, linear, ss-RNA 4, 27—78
(TSWV) Isometric ~80 1.3—2.6
Maize chlorotic dwarf virus gp.c Maize chlorotic dwarf virus Isometric ~30 1 , linear ss-RNA •7
(MCDV) ~3.2
Tymovirus Turnip yellow mosaic virus Isometric ~30 1, linear ss-RNA 1, ~20
(TYMV) ~2.0
Luteovirus Barley yellow dwarf virus Isometric ~30 1 , linear ss-RNA 1, ~24
(BYDV) ~2.0
Sobemovirus Southern bean mosaic virus Isometric ~30 1 , linear ss-RNA 1, ~30
(SBMV) ~1.4
Tobacco necrosis virus gp.1 Tobacco necrosis virus Isometric ~30 1 , linear ss-RNA 1, ~30
(TNV) ~1.4
Tombusvirus Tomato bushy stunt virus Isometric ^34 1 , linear ss-RNA 1, ^40
(TBSV) ~1.5
Comovirus Cowpea mosaic virus Isometric ^30 2, linear ss-RNA 2, ~22 + 42
(CPMV) ~2.4 + 1.4
Nepovirus Tobacco ringspot virus Isometric ~30 2, linear ss-RNA 1 , 55—60'
(TRSV) ~2.8 + 1.3—2.4
Pea enation mosaic virus gp.c Pea enation mosaic virus Isometric ~30 2, linear ss-RNA 1, ~22
(PEMV) A.I + 1.3
i-x.
Dianthovirus Carnation ringspot virus Isometric ~34 2, linear ss-RNA 1 , ~40

(CRSV) -1.5 + 0.5
Cucumovirus Cucumber mosaic virus Isometric ~29 3, linear ss-RNA 1, ~24
(CMV) i-x-1.3 + 1.1 + 0.8
Bromovirus Brome mosaic virus (BMV) Isometric ~29 3, linear ss-RNA 1, ~20
- 1 . 1 + 1.0 + 0.7
Ilarvirus Tobacco streak virus (TSV) Isometric 26— 35 3, linear ss-RNA 1. ~25
/-s- 1 . 1 + 0.9 + 0.7
Alfalfa mosaic virus gp. c Alfalfa mosaic virus (AMV) Bacilliform 28—58 3, linear ss-RNA 1, -24
x 18 - i . l + 0.8 + 0.7
Tobamovirus Tobacco mosaic virus Rigid rod ~300 x 18 1, linear ss-RNA 1. 17—18
(TMV) ~-2.0
Potexvirus Potato virus X virus (PVX) Flexuous rod 480—580 1, linear ss-RNA 1, 18—23
x 13 -2.1
Carlavirus Carnation latent virus (CLV) Flexuous rod 600—700 1, linear ss-RNA 1. ~32
x 13 -2.7
Potyvirus Potato virus Y virus (PVY) Flexuous rod 680—900 1, linear ss-RNA 1 , 32—36
x 11 3 .0—3.5
Closterovirus Beet yellows virus (BYV) Flexuous rod 600—2,000 1, linear ss-RNA 1 , 23—27
x 12 2 .2—4.7
Tobravirus Tobacco rattle virus (TRY) Rigid rod 180—215 x 2, linear ss-RNA 1. ~22
22 and 46—1 14 x 22 -2.4 + 0.6—1.4
Hordeivirus Barley stripe mosaic virus Rigid rod 100—150 3, linear ss-RNA 1, ~21
(BSMV) x 20 -1.0—1.5
" Taxonomic arrangement according to Matthews." 1

b
Measurements are those reported by Matthews111 or determined by the authors from virus preparations negatively stained with uranyl acetate.
0
The plant-infecting members of the family Reoviridae have been classified into two genera, Phytoreovirus and Fijivirus, with wound tumor
virus and Fiji disease virus as respective type members. 111
d
The plant-infecting members of the family Rhabdoviridae have been tentatively subdivided into two subgroups typified by lettuce necrotic
yellows virus and potato yellow dwarf virus. 111
c
Monotypic groups with no approved group names.
' There is evidence that this may be a stable tetramer.'-"
For example, a Tobamovirus such as cucumber green mottle mosaic virus induces vesicu-
lation of mitochondria similar to that induced by galinsoga mosaic virus, an unclassified
virus with small isometric particles.14-1 Furthermore, galinsoga mosaic virus also induces
vesicles on the chloroplast membranes similar to those of all Tymoviruses, an effect con-
sidered diagnostic for that group. 143
At present, cytopathic effects induced only by some viruses provide a satisfactory indi-
cation of the group to which the virus belongs. In particular cases, viruses within a group
can also be distinguished. It must be admitted, though, that many virus groups, and especially
viruses within groups, are not easy to differentiate in this way, even though more comparative
data and new techniques will certainly help in the future.
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64. Goodman, R. M., Geminiviruses, J. Gen. Virol., 54, 9, 1981.
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68. Lister, R. M., Possible relationships of virus specific products of tobacco rattle virus infections, Virology,
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69. Lister, R. M., Functional relationships between virus-specific products of infection by viruses of the
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70. Van Vloten-Doting, L., Kruseman, J., and Jaspars, E. M. J., The biological function and mutual
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75. Van Vloten-Doting, L. and Jaspars, E. M. J., Plant covirus systems: three-component systems, Compr.
Virol., I I , 1, 1977.
76. Kassanis, B., Properties and behaviour of a virus depending for its multiplication on another, J. Gen.
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77. Schneider, I. R., Characteristics of a satellite-like virus of tobacco ringspot virus, Virology, 45, 108, 1971.
78. Murant, A. F. and Mayo, M. A., Satellites of plant viruses, Annu. Rev. Phytopathol., 20, 49, 1982.
79. Davies, J. W., Translation of plant virus ribonucleic acids in extracts from eukaryotic cells, in Nucleic
Acids in Plants, Vol. 2, Hall, T. C. and Davies, J. W., Eds., CRC Press, Boca Raton, Fla., 1979, 1 1 1 .
80. Davies, J. W. and Hull, R., Genome expression of plant positive-strand RNA viruses, J. Gen. Virol.,
61, 1, 1982.
8 1 . Aach, H. G., Funatsu, G., Nirenberg, M. W., and Fraenkel-Conrat, H., Further attempts to characterize
products of TMV-RNA-directed protein synthesis. Biochemistry, 3, 1362. 1964.
82. Francki, R. I. B. and Randies, J. W., RNA-dependem RNA polymerase associated with particles of
lettuce necrotic yellows virus. Virology, 47, 270, 1972.
83. Toriyama, S. and Peters, D., Differentiation between broccoli necrotic yellows virus and lettuce necrotic
yellows virus by their transcriptase activities, J. Gen. Virol., 56, 59, 1981.
84. Sanger, F., Nicklen, S., and Coulson, A. R., DNA sequencing with chain-terminating inhibitors, Proc.
Natl. Acad. Sci. U.S.A., 74. 5463, 1977.
85. Brown, N. L., Nucleic acid sequencing, in Companion to Biochemistry, Vol. 2, Bull, A. T., Lagnado, J.
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GCC), Nucleic Acids Res., 1. 959. 1979.
87. Maxam, A. M. and Gilbert, W., Sequencing end-labeled DNA with base-specific chemical cleavages,
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Volume I 15
88. Frank, A., Guilley, H., Jonard, G., Richards, K,, and Hirth, L., Nucleotide sequence of cauliflower
mosaic virus DNA, Cell, 21, 285. 1980.
89 Goelet, P., Lomonosoff, G. P., Butler, P. J. G., Akam, M. E., Gait, M. J., and Karn, J., Nucleotide
sequence of tobacco mosaic virus RNA, Proc. Nail. Acad. Sci. U.S.A., 79, 5818, 1982.
90. Ivanovski, D., Uber die Mosaikkrankheit der Tabakspflanze, Z. Pflanzenkrankheit, 13, 1, 1903.
91. Steere, R. L. and Williams, R. C., Identification of crystalline inclusion bodies extracted intact from
plant cells infected with tobacco mosaic virus, Am. J. Hot., 40, 81, 1953.
92. Bald, J. G., Cytology of plant virus infections, Adv. Virus Res., 12, 103, 1966.
93. Rubio-Huertos, M., Inclusion bodies, in Principles and Techniques in Plant Virology, Kado, C. I. and
Agrawal, H. O., Eds., Van Nostrand Reinhold, New York, 1972, 62.
94. Christie, R. G. and Edwardson, J. R., Light and electron microscopy of plant virus inclusions, Fla.
Agric. Exp. Stn. Monogr., No. 9, 1977, 150 pp.
95. Martelli, G. P. and Russo, M., Plant virus inclusion bodies, Adv. Virus Res., 21, 175, 1977.
96. Edwardson, J. R. and Christie, R. G., Use of virus-induced inclusions in classification and diagnosis,
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97. Black, L. M., Morgan, C., and Wyckoff, R. W. G., Visualization of tobacco mosaic virus within infected
cells, Proc. Sot: E.-cp. Biol. Med., 73, 119, 1950.
98. Smith, K. M., Apparent crystallization of tomato bushy stunt virus in the cells of infected plants, Virology:
2, 706, 1956.
99. Fernandez-Moran, H., A diamond knife for ultrathin sectioning, Exp. Cell Res., 5, 255, 1953.
100. Sabatini, D. D., Bensch, K., and Barrnett, R. J., Cytochemistry and electron microscopy. The pres-
ervation of cellular ultrastructures and enzymatic activity by aldehyde fixation, J. CellBiol., 17, 19, 1963.
101. Pease, D. C., Histological Techniques for Electron Microscopy, Academic Press, New York, I960, 274
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103. Maalue. O. and Birch-Andersen, A., On the organization of the "nuclear material" in Salmonella
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Nature (London), 178, 803, 1956.
105. Shalla, T. A., Degradation of tobacco mosaic virus by potassium permanganate, Virology: 13, 383, 1961.
106. Warmke, H. E. and Edwardson, J. R., Use of potassium permanganate as a fixative for virus particles
in plant tissues, Virology: 28, 693, 1966.
107. Matsui, C. and Yamaguchi, A., Some aspects of plant viruses in situ. Adv. Virus Res., 12, 127, 1966.
108. Esau, K., Viruses in Plant Hosts, University of Wisconsin Press, Madison, 1968, 225 pp.
109. Maramorosch, K., Ed., The Atlas of Insect and Plant Viruses, Academic Press, New York, 1977, 478
pp.
110. Milne, R. G., Electron microscopy of leaves infected with sowbane mosaic virus and other small polyhedral
viruses, Virology, 32, 589, 1967.
1 1 1 . Milne, R. G., Plant viruses inside cells, Sri. Prog. (Oxford), 55, 203, 1967.
1 1 2 . Crowley, N. C., Davison, E. M., Francki, R. I. B., and Owusu, G. K., Infection of bean root-meristems
by tobacco ringspot virus, Virology: 39, 322, 1969.
1 1 3 . Lawson, R. H. and Hearon, S., Subcellular localization of chrysanthemum aspermy virus in tobacco and
chrysanthemum leaf tissue, Virology: 41, 30, 1970.
1 1 4 . Hatta, T., Recognition and measurement of small isometric virus particles in thin sections, Virology: 69,
237, 1976.
1 1 5 . Honda, Y. and Matsui, C., Electron microscopy of cucumber mosaic virus-infected tobacco leaves showing
mosaic symptoms, Phytopathology, 64, 534, 1974.
116. Hatta, T. and Francki, R. I. B., Enzyme cytochemical method for identification of cucumber mosaic
virus particles in infected cells, Virology: 93, 265, 1979.
117. Hatta, T. and Francki, R. I. B., Identification of small polyhedral virus particles in thin sections of plant
cells by an enzyme cytochemical technique, J. Ultrastruct. Res., 74, 116, 1981.
118. Otsuki, Y., Takbe, I., Honda, Y., and Matsui, C., Ultrastructure of infection of tobacco mesophyll
protoplasts by tobacco mosaic virus, Virology: 49, 188, 1972.
1 1 9 . Gigot, G., Kopp, M., Schmitt, C., and Milne, R. G., Subcellular changes during isolation and culture
of tobacco mesophyll protoplasts, Protoplasma, 84, 31, 1975.
120. Foglein, F. J., Nyitrai, A., Gulyas, A., Premecz, G., Olah, T., and Farkas, G. L., Crystalline inclusion
bodies are degraded in protoplasts isolated from TMV-infected tobacco leaves, Phytopathoi. Z., 86, 266,
1976.
121. Hatta, T. and Francki, R. I. B., Enzyme cytochemical identification of single-stranded and double-
stranded RNAs in virus-infected plant and insect cells. Virology, 88. 105. 1978.
122. Shalla, T. A. and Amici, A., The distribution of viral antigen in cells infected with tobacco mosaic virus
as revealed by electron microscopy, Virology, 31, 78, 1964.
123. Langenberg, W. G. and Schlegel, D. E., Autoradiography with 125I-labeled antibodies as a means of
localizing TMV antigen in plant cells, Virology, 32, 167, 1967.
124. De Zoeten, G. A., Cytology of virus infection and virus transport, in Encyclopedia of Plant Physiology,
Vol. 4, Heitefuss, R. and Williams, P. H., Eds., Springer-Verlag, Berlin, 1976, 129.
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International Committee on Nomenclature of Viruses, Prog. Med. Virol., 9, 476, 1967.
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Nomenclature of Viruses, Monogr. Virol., 5, 1, 1971.
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on Taxonomy of Viruses, Intervirology, 12, 129, 1979.
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Diseases, Kurstak, E. and Marusyk, R., Eds., Marcel Dekker, New York, 1982, 239.
139 Hamilton, R. I., Edwardson, J. R., Francki, R. I. B., Hsu, H. T., Hull, R., Koenig, R., and Milne,
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mosaic virus, J. Gen. Virol., 64, 687, 1983.
Volume 1 17
Chapter 2
CAULIMOVIRUS GROUP
The group is unique in that its members are the only known plant viruses with double-
stranded DNA genomes.' They have polyhedral particles, about 50 nm in diameter containing
about 17% DNA. The DNA consists of a circular molecule of about 8000 base pairs (bp).
The capsids are constructed from a single species of polypeptide of MT about 42 x 10\
Caulimoviruses are transmitted by aphids in a semipersistent manner and some can also
be transmitted experimentally by mechanical inoculation. Seed transmission has not been
reported. Host ranges of the individual viruses of the group are restricted to only a few plant
species.2
I. MEMBERS OF THE GROUP AND THEIR RELATIONSHIPS
Cauliflower mosaic virus (CaMV), the type member, has been studied more extensively
than any of the other members (Table 1) and hence most of the discussion here will be about
this virus. However, it seems that all the viruses listed in Table 1 are bona fide members
of the group. With the Caulimoviruses, particle morphology and the cell inclusions induced
are so characteristic that they are sufficient criteria for identification of members of the
group. 25
Some members have been shown to be antigenically related to each other. There is good
evidence that CaMV, carnation etched ring virus (CERV), dahlia mosaic virus (DMV), and
strawberry vein banding virus (SVBV) are all antigenically interrelated.6~9 It is interesting
to note that in spite of the very close serological relationship between CaMV and DMV,
the two viruses do not appear to have any common plant hosts.6-7 Many of the viruses listed
in Table 1 have not been studied serologically and for those that have, a variety of methods
have been used. An interesting picture of antigenic relationships among the Caulimoviruses
may well emerge when they are all compared using similar techniques. However, serological
data on the Caulimoviruses, especially those indicating close relationships, should be in-
terpreted with caution because their coat proteins can undergo proteolysis during extraction
and purification of the virus 10 " and it has been shown that changes in the antigenic properties
due to proteolysis of one CaMV isolate were of similar magnitude to those between different
undegraded CaMV isolates.12
Another approach to the comparative study of Caulimoviruses is to examine their DNAs.
Some significant differences in the size, conformation, and other properties of CaMV, CERV,
figwort mosaic virus (FiMV), and mirabilis mosaic virus (MiMV) DNAs have already been
established. 16J9 - 20 Comparisons of the DNAs of CaMV isolates by restriction endonuclease
mapping have also revealed a considerable amount of variation among the 33 isolates
studied.25 It is interesting that the differences in the restriction endonuclease maps of CaMV
isolates could not be correlated with differences in the biological properties of the isolates.
II. VIRUS STRUCTURE AND COMPOSITION
A. Particle Structure
Caulimovirus particles appear roughly spherical in negatively stained preparations, with
little visible structural detail (Figure 1). The reported diameter of the particles ranges from
about 42 to 50 nm depending on the staining method used.13 However, particles in some
preparations become flattened and appear much larger (Figure 2). Sedimentation studies
indicate that the hydrated diameter of CaMV particles is 57 nm with a sedimentation coef-
Table 1
SOME PROPERTIES OF MEMBERS AND PROBABLE MEMBERS OF THE
CAULIMOVIRUS GROUP
Semipersistent Presence of Presence of Presence of

aphid characteristic characteristic double-stranded
Virus transmission cell inclusions particles DNA genome Ref.
Cauliflower Yes Yes Yes Yes 13

mosaic virus (CaMV)
Carnation Yes Yes Yes Yes 14
etched ring virus
(CERV)
Dahlia mosaic Yes Yes Yes Yes 15, 16
virus (DMV)
Strawberry Yes Yes Yes Yes 17
vein banding virus
(SVBV)
Mirabilis Yes Yes Yes Yes 18, 19
mosaic virus (MiMV)
Figwort Unknown Yes Yes Yes 19, 20
mosaic virus (FiMV)
Petunia vein Unknown Yes Yes Unknown 21
clearing virus
(PeVCV)
Blueberry red Unknown Yes Yes Unknown 2">
ringspot virus
(BRRV)
Cassava vein Unknown Yes Yes Unknown 23
mosaic virus
(CaVMV)
Plantago virus 4 Unknown Yes Yes Unknown 24
(P1V4)
Thistle mottle Unknown Yes Yes Yes 20
virus (ThMoV) 20
Horseradish Unknown Yes Yes Yes 24
latent virus (HLaV)
ficient of 208S. From these values an Mr of about 22,8 x 106 for the CaMV particle was
calculated by using Svedberg's equation. 26
In some electron micrographs, Caulimovirus particles appear to have a core about 20 nm
in diameter (Figures 1A and 2). This together with early analyses of CaMV protein by
polyacrylamide gel electrophoresis suggested the presence of several proteins associated with
CaMV particles.27 30 However, it now appears that CaMV particles are much simpler in
structure, having a single shell and only one major protein. 10 - 31 The "core" is probably a
central hole in the particle. Neutron diffraction studies indicate that the CaMV particle
consists of a protein shell lined with DNA with the center remaining empty. 32
Analyses of CaMV coat protein by polyacrylamide gel electrophoresis (PAGE) reported
by five different groups of workers have varied in the number and molecular weight of
polypeptides detected.10-27"30 Tezuka and Taniguchi 27 detected only two polypeptides of MT
33 and 68 x 103, whereas Brunt and his colleagues29 observed in their gels as many as ten
bands, some of which they considered to be degradation products or aggregates, ranging in
size from Mr of 27 to 100 x 103. Variation in the reported molecular weights was probably
due to differences in techniques and marker proteins used. However, the differences in
numbers and relative amounts of the polypeptides detected were more difficult to explain.
Al Ani and colleagues'" concluded from their work that the major structural protein of CaMV
is of M, about 42 x 103 (42K protein). By peptide mapping, they demonstrated homologies
Volume I 19
FIGURE 1. A, Purified cauliflower mosaic virus (CaMV); B, carnation etched ring virus (CERV). Uranyl acetate
staining. Bars = 100 nm.
FIGURE 2. A, Purified CaMV, the particles of which have flattened during preparation for electron microscopy;
B and C illustrate the flattening of the particles when viewed on an intact specimen grid membrane (B) and one
that has broken and curled (C). Uranyl acetate staining. Bars = 100 nm. (Courtesy of Mr. C. M. Clay, National
Vegetable Research Station. Wellesbourne, England.)
between 42K. and some of the smaller components which they concluded to be degradation
products of the 42K protein. Similarly they detected homologies between components of
the two larger proteins of Mr 49 and 55 x 10' (49K and 55K, respectively) which they
considered to be aggregation products of 42K and the smaller protein components. Hahn
Volume I 21
and Shepherd11 reported that the two larger protein components of coat protein preparations
of Mr about 44 and 58 x 103 (probably corresponding to the 42 and 55K protein components
detected by Al Ani and colleagues 10 ) were phosphorylated. They suggested that the larger
is probably the primary coat protein gene product and the smaller a degradation product of
it.This suggestion is supported by subsequent experiments in which virus was pulse labeled
in vivo. 33
Du Plessis and Smith34 observed that concanavalin A attached to CaMV particles and
interfered with antibody binding indicating that sugar residues were located at the surface
of the particles. Thus it appears that CaMV coat protein is both glycosylated and phospho-
rylated." 34 The significance of this is not clear, but it is known that phosphorylated proteins
are common in animal DNA viruses and may play some regulatory role in capsid formation.''
Highly basic polypeptides are also characteristic of some DNA viruses, and in the case of
CaMV, the region of the coat polypeptide with a high lysine content29-35 may interact with
the DNA in the virus particle.
B. The Viral Genome

There seems little doubt that all the viruses listed in Table 1 have double-stranded DNA
genomes although most of the available detailed information is confined to the DNA of
CaMV. The possibility of using Caulimoviruses as vehicles for genetic engineering of plants
has generated considerable interest in their genomes, and CaMV has been used as a model
system.36"39
CaMV DNA can occur in various forms including linear, circular, and supercoiled mol-
ecules.40-41 The circular forms are not covalently closed due to the presence of single-strand
breaks in both strands at specific locations.19-20-42-43 The biological significance of these
various DNA forms is not clear. However, it has been shown that the unusual secondary
structure of CaMV is not essential for infectivity. The linear form of the DNA from CaMV
strain CM4-184 cut with the endonuclease Sal 1 has been shown to be infectious. This
molecule was successfully cloned in the bacterial plasmid pBR322. The cloned DNA had
to be excised from the plasmid to be infectious, but did not need to be ligated into the
circular form.38
CaMV DNA has short discontinuities or gaps at well-defined sites on both strands of the
circular double-stranded molecules42-43 (Figure 3). At each gap there is a triple-stranded
DNA structure with the 5' terminus at a fixed position.44 The 3' end is not fixed and produces
short overlaps of 8 to 43 nucleotides.45 This appears to be characteristic for all strains of
CaMV and other Caulimoviruses.19'20-25 DNA from most strains of CaMV has a single gap
in the transcribed (a) and two in the nontranscribed (3) strand (Figure 3). However, at least
one strain of the virus (CM4-184) has a small deletion in the region of one of the gaps so
that each DNA strand has only one gap.46 FiMV DNA was shown to have four gaps, three
in one strand and one in the other. 19
It has recently been observed that on denaturing CaMV double-stranded DNA (with three
gaps), in addition to the three single-stranded DNA molecules expected, a fourth minor
component 587 nucleotides long (the a, strand) was detected.47 This small DNA has the
same 5' terminus as the a strand and is of the same polarity.47
An unusual feature of CaMV DNA is the presence of several short ribonucleotide sequences
in some DNA molecules.48 These ribonucleotide stretches, which account for less than 1%
of the total nucleic acid of CaMV, may be primers of DNA synthesis which have not been
removed during replication or they could play some role in the generation of the gaps in
the DNA strands.
The entire genomes of three strains of CaMV have now been sequenced; the Cabb-B-S,35
CMI 1841,49 and the D/H50 strains with 8024, 8031, and 8016 bp, respectively. The genetic
organization of all three CaMV strains appears to be the same with six open reading frames
and two intergenic regions (Figure 3).
FIGURE 3. Physical map of the Caulimovirus genome showing the a and p strands of the
double-stranded DNA with its three discontinuities or gaps. The inner segments of the diagram
show the location of the six putative codons (open reading frames); the direction of reading
in the three reading frames is indicated by arrows.
One of the reading frames on CaMV DNA appears to be the coat protein gene, Codon
IV (Figure 3). This frame should code for a protein of M, about 57 X 10' (489 amino acids)
in the case of the Cabb-B-S virus strain.35 This is supported by experiments involving the
in vivo expression of the CaMV genome on plasmid vectors in E. co//. 51 It was demonstrated
that CaMV coat protein antigen was produced by bacteria bearing a plasmid which contained
only a short viral DNA segment derived from the internal position of Codon IV. 5 '
One other CaMV gene product has been identified as a protein of Mr between 62 and 66
x I0 3 . It has been detected in extracts from infected plants and appears to be associated
with the viroplasms. 31 - 52 " 55 The 19S mRNA which appears to be translated into this protein
has been isolated by Odell and Howell52 and the synthesis of a similar RNA has been detected
in isolated nuclei of CaMV-infected leaves.56 Furthermore, the translation product of the
RNA has been shown to react with antibodies specific to the viroplasm protein. Data on the
hybridization of the 19S RNA to restriction endonuclease fragments of CaMV DNA 52 ' 54
indicate that the location of the putative viroplasm protein gene corresponds to Codon VI
indicated in Figure 3. Codon VI has coding capacity for a protein of M, about 61 x 103,35
which is not far short of that estimated for the putative viroplasm protein (M, between 62
and 66 x 10 3 ). 31 - 5254
The genetic information in putative Codon II is not essential for plant infection. It has
been demonstrated that the CM4-184 strain of CaMV has a deletion of 421 bp from the
open region of Codon II, and although the virus replicates normally in plants, it is not
transmissible by aphids.45 Experiments with in vitro constructed genomes from CaMV DNAs
of aphid transmissible and nontransmissible virus strains have shown that Codon II is the
gene for a protein with Mr about 18 x 103 which determines aphid transmissibility. 5s At
Volume I 23
present there are no data to suggest the identity of any virus-specific gene products of the
putative codons I to III, and V (Figure 3).
The primary DNA structures of the three CaMV strains which have been sequenced differ
from one another by only about 5%.49-50 The differences are mainly due to base substitutions
but there are also several deletions and insertions. They are spread uniformly through the
genome except for the two intergenic regions which are more highly conserved. The stability
of these regions is probably due to their probable function in initiation and termination of
transcription. 50 The sequence variation in the coding regions is probably of little importance
as the changes occur predominantly in the third portion of reading frame triplets. 50
CaMV DNA appears to be transcribed in the nucleus56-59-60 and the mechanism by which
this takes place is starting to emerge. 6061 Olszewski and his colleagues60 have isolated
covalently closed circular CaMV DNA; they suggest that transcription takes place on these
minichromosomes which lack the site-specific discontinuities characteristic of the encapsi-
dated DNA. 35 - 49 - 50 Transcription appears to be asymmetric in that only the a-strand is
transcribed.44'64 Four CaMV-specific RNAs have been detected in the polyadenylated RNA
fraction from infected leaves and their locations on the viral genome determined.61-6-1 One
of these RNAs is the 19S mRNA for the virus-induced inclusion body protein already
mentioned. 52 54 The functions of the remaining three RNA transcripts, a major 35S, minor
35S, and 8S RNAs, remain obscure, and the mRNAs specifying polypeptides corresponding
to Codons I, III, and V of CaMV DNA (Figure 3) have not been identified.
From the data available at present, it seems that some features of CaMV replication are
similar to those of retroviruses and hepatitis virus B of vertebrates which involve reverse
transcription steps. The possible involvement of reverse transcription in the replication of
CaMV has recently been discussed elsewhere. 47 - 62
III. CYTOPATHOLOGY
In thin sections, Caulimovirus particles appear spherical, densely stained, and about 40
to 45 nm in diameter. Some are uniformly stained, but most have electron-lucent centers
about 20 nm in diameter (Figure 4). The particles are usually associated with inclusions
typical of Caulimovirus infection, often referred to as viroplasms (Figures 4 and 5A), but
sometimes the particles are scattered in the cytoplasm (arrows in Figure 5A). 3 In a number
of cases, particles have also been seen inside plasmodesmata. 18 - 65 ' 66 Plasmodesmata con-
taining particles (Figure 5, B and C) have been found to be about twice the diameter of
similar structures in healthy plant cells.65 It seems that the particles inside the plasmodesmata
may be in the process of passage from one cell to another (Figure 5, B and C).67-68
Caulimovirus viroplasms vary considerably in size. At early stages of infection they are
usually small granular patches of densely staining material. As infection proceeds the vi-
roplasms grow and develop lacunae (Figure 5A). Virus particles also appear and are either
embedded in the matrix of the viroplasms or aggregate in the lacunae (Figure 6). Viroplasms
are never surrounded by membranes, and ribosomes usually congregate around their pe-
riphery reaching especially high concentrations around those that are smaller and granular
(Figure 6). It has also been observed that numerous Golgi bodies develop near the viroplasms,
often in association with characteristic microvesicles.18-69 In cells infected by some strains
of CaMV, fully developed viroplasms may be as large as 4.5 |xm in diameter.69 Cytochemical
and enzyme-digestion tests indicate that they consist mainly of protein, but also contain
some RNA and DNA.68-70 It is likely that the RNA detected may have been that of the
ribosomes surrounding the viroplasms and the DNA, that of virus particles.
Isolated viroplasms consist almost entirely of protein antigenically unrelated to the viral
coat protein.69-71 Viroplasms from CaMV-infected plants have been found to consist essen-
tially of a single polypeptide species of MT about 55 x 10 3 . 31 -52-55-69 This polypeptide is
FIGURE 4. A, Thin section of CaMV-infected Brassica oleracea (cabbage) leaf cell showing a characteristic
viroplasm containing virus particles; bar = 500 nm. B, CaMV particles embedded in a viroplasm, some showing
characteristic electron-lucent centers: bar = 100 nm.
Volume I 25
FIGURE 5. A, Thin section of CaMV-infected Brassica oleracea (cabbage) leaf cell showing some virus particles
in a small viroplasm and others scattered in the cytoplasm (arrows); bar = 500 nm. B and C, Dahlia mosaic virus
(DMV) particles in the plasmodesmata of infected Dahlia variahilis leaf cells (courtesy of Dr. E. W. Kitajima:
Department of Biology, Fundacao Universidade de Brasilia, Brasilia); arrows in B point to virus particles and that
in C to the plasmalemma lining the plasmadesma. Bar = 100 nm.
FIGURE 6. Thin section of mirabilis mosaic virus (MiMV)-infected Mirabilis jalapa leaf cell showing large and
small viroplasms, some devoid of virus particles. The viroplasms are surrounded by numerous ribosomes. Bar =
500 nm. (Courtesy of Dr. R. Hull, John Innes Institute, Norwich, U . K . )
Volume I 27
significantly smaller than that estimated for the putative viroplasm protein predicted from
nucleotide sequence studies of Codon VI of the CaMV genome as already mentioned. 52 55
Although the viroplasms of all Caulimoviruses are essentially similar, some minor dif-
ferences have been noted among distinct strains of CaMV. 72 - 71 The cytopathological effects
of a number of CaMV strains infecting turnip plants have been compared directly. Differences
were observed in the size of the viroplasms, the frequency of virus particles occurring free
in the cytoplasm as opposed to those in the viroplasms, and the ratio of virus particles to
the amount of viroplasm matrix protein. 72 It was also observed that one of the strains (CM4-
184) which has been shown to have a deletion in its DNA45 induced inclusions in the
chloroplasts. These observations indicate that cytopathological features induced by CaMV
are controlled by its genome. However, one of the CaMV strains induces significantly
different sized viroplasms in two different plant hosts, indicating that the host also has some
control over cytopathological events. 72
Selective incorporation of 3 H-thymidine into viroplasms of CaMV-infected cells has been
demonstrated by electron microscopic autoradiography, suggesting that these inclusions are
the sites of virus DNA replication. 74 - 75 Favali and her colleagues75 also demonstrated that,
initially, significant radioactivity was incorporated into the nuclei indicating that they are
also sites of viral DNA synthesis. Biochemical studies have also indicated that the nucleus
is the site of viral DNA replication.56-59 Furthermore, it was observed that whereas nuclei
from CaMV-infected leaves synthesized viral DNA in vitro, isolated viroplasms did not.59
These data suggest that perhaps viral DNA is synthesized only in the nuclei but that it is
assembled into virus particles in the viroplasms.
Caulitnovirus particles and viroplasms have also been observed occasionally in the nuclei
of infected cells (Figure 7). For example, CERV particles and viroplasms have been detected
in the nuclei of Saponaria vaccaria leaf cells, but not those of Dianthus caryophyllus.1'
Similarly, CaMV particles and viroplasms have been observed in the nuclei of infected
cabbage leaf cells (Figure 7),76 but apparently not in the nuclei of other plant species infected
with the virus. 70 - 72 Hence the ability of viroplasms and virus particles to invade nuclei of
infected cells may be controlled by the host plant species.
In addition to the typical viroplasms described above and illustrated in Figures 4 through
7, another type of inclusion has been observed in cells infected for relatively long periods73-76
(Figure 8). These inclusions are electron-lucent and contain occasional virus particles scat-
tered in the matrix (Figure 8). Cytochemical tests indicate that they contain some RNA. 73
Although their function and significance are obscure at present, it has been suggested that
they may be storage sites of degraded material rather than sites of metabolic activity. 73
In addition to the appearance of virus particles and virus-specific inclusions in Cauli-
movirus-infected cells, some other cytopathological effects have also been observed. For
example, Conti and his colleagues77 noted the formation of fingerlike protrusions of the cell
walls in CaMV-infected cells. These were later shown to be plasmodesmata-related struc-
tures.78 Autoradiographic studies using 3H-glucose and 3 H-phenylalanine demonstrated that
only the 3H-glucose was incorporated into the protrusions, indicating that the structures were
not lignified. 78
Of the structural changes observed in plant cells on infection with Caulimoviruses, only
the appearance of the characteristic virus particles and viroplasms seem to be universal
features observed by all workers with all known Caulimoviruses. These features are good
diagnostic markers for this group of viruses.
FIGURE 7. Thin section of CaMV-infected Brassica oleracea leaf cells, showing in A a viroplasm (P) in the
nucleus and virus particles (arrows) in the cytoplasm; and in B some virus particles in the nucleus (arrows) and a
viroplasm (P) in the cytoplasm. Bars = 500 nm.
Volume I 29
FIGURE 8. Thin section of CaMV-infected Brassica oleracea leaf cell showing a typical viroplasm (P) containing
virus particles and an electron-lucent inclusion (L) characteristic of late stage of infection; a few virus particles
can be seen scattered in the electron-lucent inclusion. Bar = 500 nm.
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of Plant Viruses, No. 182, 1977.
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16. Richins, R. D., and Shepherd, R. J., Physical maps of the genomes of dahlia mosaic virus and mirabilis
mosaic virus — two members of the caulimovirus group, Virology, 124, 208, 1983.
17. Frazier, N. W. and Converse, R. H., Strawberry vein banding virus, CMI/AAB Descriptions of Plant
Viruses, No. 219, 1980.
18. Brunt, A. A. and Kitajima, E. W., Intracellular location and some properties of Mirabilis mosaic virus,
a new member of the cauliflower mosaic group of viruses, Phytopath. Z., 76, 265, 1973.
19. Hull, R. and Donson, J., Physical mapping of the DNAs of carnation etched ring and figwart mosaic
viruses,/. Gen. Virol., 60, 125, 1982.
20. Donson, J. and Hull, R., Physical mapping and molecular cloning of caulimovirus DNA, J. Gen. Virol.,
64, 2281, 1983.
21. Lesemann, D. and Casper, R., Electron microscopy of Petunia vein-clearing virus, an isometric plant
virus associated with specific inclusions in Petunia cells, Phytopathology, 63, 1118, 1973.
22. Kim, K. S., Ramsdell, D. C., Gillett, J. M., and Fulton, J. P., Virions and ultrastructural changes
associated with blueberry red ringspot disease, Phytopathology, 71, 673, 1981.
23. Kitajima, E. W. and Costa, A. S., Particular esferoidais associadas ao virus do mosaico das nerumas da
mandioca, Bragantia, 25, 211, 1966.
24. Shepherd, R. J., personal communication, 1983.
25. Hull, R., Structure of the cauliflower mosaic virus genome. III. Restriction endonuclease mapping of thirty-
three isolates, Virology, 100, 76, 1980.
26. Hull, R., Shepherd, R. J., and Harvey, J. D., Cauliflower mosaic virus: an improved purification
procedure and some properties of the virus particles, J. Gen. Virol., 31, 93, 1976.
27. Tezuka, N. and Taniguchi, T., Structural protein of cauliflower mosaic virus, Virology, 48, 297, 1972.
28. Kelly, D. C., Cooper, V., and Walkey, D. G. A., Cauliflower mosaic virus structural proteins, Microbios,
10, 239. 1974.
29. Brunt, A. A., Barton, R. J., Tremaine, J. H., and Stace-Smith, R., The composition of cauliflower
mosaic virus protein, J. Gen. Virol., 27, 101, 1975.
30. Hull, R. and Shepherd, R. J., The coat proteins of cauliflower mosaic virus, Virology, 70, 217, 1976.
3 1 . Al Ani, R., Pfeiffer, P., Whitechurch, O., Lesot, A., Lebeurier, G., and Hirth, L., A virus specified
protein produced upon infection by cauliflower mosaic virus, Ann. Virol., 131E, 33, 1980.
32. Chauvin, C., Jacrot, B., Lebeurier, G., and Hirth, L., The structure of cauliflower mosaic virus: a
neutron diffraction study, Virology, 96, 640, 1979.
33. Hahn, P. and Shepherd, R. J., Evidence for a 58-kilodalton polypeptide as precursor of the coat protein
of cauliflower mosaic virus. Virology, 116, 480, 1982.
34. Du Plessis, D. H. and Smith, P., Glycosylation of the cauliflower mosaic virus capsid polypeptide,
Virology, 109, 403, 1981.
35. Frank, A., Guilley, H., Jonard, G., Richards, K., and Hirth, L., Nucleotide sequence of cauliflower
mosaic virus DNA, Cell, 2 1 , 285, 1980.
36. Hohn, T., Richards, K., and Lebeurier, G., Cauliflower mosaic virus on its way to becoming a useful
plant vector. Curr. Top. Microbiol. Immunol., 96, 193, 1982.
37. Szeto, W. W., Hamer, D. H., Carlson, P. S., and Thomas, C. A., Jr., Cloning of cauliflower mosaic
virus (CLMV) DNA in Escherichia coli, Science, 196, 210, 1977.
Volume 1 31
38. Howell, S. H., Walker, L. L., and Dudley, R. K., Cloned cauliflower mosaic virus DNA infects turnips
(Brassica rapa), Science, 208, 1265. 1980.
39. Howell, S. H., Plant molecular vehicles: potential vectors for introducing foreign DNA into plants, Annu.
Rev. Plant PhysioL, 33, 609, 1982.
40. Hull, R., The DNA of plant DNA viruses, in Nucleic Acids in Plants, Vol. 2, Hall, T. C. and Davies, J.
W., Eds., CRC Press, Boca Raton, Fla., 1979, 3.
41. Menissier, J., de Murcia, G., Lebeurier, G., and Hirth, L., Electron microscopic studies of the different
topological forms of the cauliflower mosaic virus DNA: knotted encapsidated DNA and nuclear minichro-
mosome, EMBO J., 2, 1067, 1983.
42. Volovitch, M., Drugeon, G., and Yot, P., Studies on the single-stranded discontinuities of the cauliflower
mosaic virus genome, Nucleic Acids Res., 5, 2913, 1978.
43. Hull, R. and Howell, S. H., Structure of the cauliflower mosaic virus genome. II. Variation in DNA
structure and the sequence between isolates, Virology, 86, 482, 1978.
44. Hull, R., Covey, S. N., Stanley, J., and Davies, J. W., The polarity of the cauliflower mosaic virus
genome, Nucleic Acids Res., 1, 669, 1979.
45. Richards, K. E., Guilley, H., and Jonard, G., Further characterization of the discontinuities in cauliflower
mosaic virus DNA, FEBS Lett., 134, 67, 1981.
46. Howarth, A. J., Gardner, R. C., Messing, J., and Shepherd, R. J., Nucleotide sequence of naturally
occurring deletion mutants of cauliflower mosaic virus, Virology, 112, 678, 1981.
47. Guilley, H., Richards, K. E., and Jonard, G., Observations concerning the discontinuous DNAs of
cauliflower mosaic virus, EMBO J., 2, 277, 1983.
48. Hull, R. and Shepherd, R. J., The structure of cauliflower mosaic virus genome, Virology, 79, 216,
1977.
49. Gardner, R. C., Howarth, A. J., Hahn, P., Brown-Leudi, M., Shepherd, R. J., and Messing, J.,
The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by Ml3 mp7 shotgun
sequencing, Nucleic Acids Res., 9, 2871, 1981.
50. Balazs, E., Guilley, H., Jonard, G., and Richards, K., Nucleotide sequence of DNA from an altered-
virulence isolate D/H of the cauliflower mosaic virus, Gene, 19, 239. 1982.
5 1 . Daubert, S., Richins, R., Shepherd, R. J., and Gardner, R. C., Mapping of the coat protein gene of
cauliflower mosaic virus by its expression in a prokaryotic system, Virology, 122, 444, 1982.
52. Odell, J. T. and Howell, S. H., The identification, mapping and characterization of mRNA for P66, a
cauliflower mosaic virus-coded protein, Virology, 102, 349, 1980.
53. Odell, J. T., Dudley, R. K., and Howell, S. H., Structure of the 19S RNA transcript encoded by the
cauliflower mosaic virus genome, Virology, 1 1 1 , 377, 1981.
54. Covey, S. N. and Hull, R., Transcription of cauliflower mosaic virus DNA. Detection of transcripts,
properties and location of the gene encoding the virus inclusion body protein, Virology, 1 1 1 , 463, 1981.
55. Shockey, M. W., Gardner, C. O., Melcher, U., and Essenberg, R. C., Polypeptides associated with
inclusion bodies from leaves of turnip infected with cauliflower mosaic virus, Virology, 105, 575, 1980.
56. Guilfoyle, T. J., Transcription of the cauliflower mosaic virus genome in isolated nuclei from turnip leaves,
Virology, 107, 71, 1980.
57. Xiong, C., Muller, S., Lebeurier, G., and Hirth, L., Identification by immunoprecipitation of cauliflower
mosaic virus in vitro major translation product with a specific serum against viroplasm protein, EMBO J.,
1, 971, 1982.
58. Woolston, C. J., Covey, S. N.,Penswick, J. R., and Davies, J. W., Aphid transmission and a polypeptide
are specified by a defined region of the cauliflower mosaic virus genome, Gene, 23, 15, 1983.
59. Ansa, O. A., Bowyer, J. W., and Shepherd, R. J., Evidence for replication of cauliflower mosaic virus
DNA in plant nuclei, Virology, 121, 147, 1982.
60. Olszewski, N., Hagen, G., and Guilfoyle, T. J., A transcriptionally active, covalently closed minichro-
mosome of cauliflower mosaic virus DNA isolated from infected turnip leaves, Cell, 29, 395, 1982.
61. Guilley, H., Dudley, R. K., Jonard, G., Balazs, E., and Richards, K. E., Transcription of cauliflower
mosaic virus DNA: detection of promoter sequences, and characterization of transcripts, Cell. 30, 763,
1982.
62. Hull, R. and Covey, S. N., Does cauliflower mosaic virus replicate by reverse transcription?, Trends
Biochem. Sci., 8, 119, 1983.
63. Dudley, R. K., Odell, J. T., and Howell, S. H., Structure and 5'-termini of the large and 19s RNA
transcripts encoded by the cauliflower mosaic virus genome, Virology, 117, 19, 1982.
64. Howell, S. H. and Hull, R., Replication of cauliflower mosaic virus and transcription of its genome in
turnip leaf protoplasts, Virology, 86, 468, 1978.
65. Kitajima, E. W. and Lauritis, J. A., Plant virions in plasmodesmata, Virology, 37, 681, 1969.
66. Lawson, R. H. and Hearon, S. S., Ultrastructure of carnation etched ring virus-infected Saponaria vaccaria
and Dianthus caryophyllus, J. U/trastruct. Res., 48, 201, 1973.
67. Petzold, H., Elektronemikroskopische Untersuchungen an von Dahlienmosaikvirus infi/.ierten Pflan/en,

Phytopathol. Z., 63, 201, 1968.
68. Kitajima, E. W., Lauritis, J. A., and Swift, H., Fine structure of zinnia leal tissues infected with dahlia
mosaic virus. Virology, 39, 240, 1969.
69. Shepherd, R. J., Richins, R., and Shalla, T. A., Isolation and properties of the inclusion bodies of
cauliflower mosaic virus, Virology, 102, 389, 1980.
70. Martelli, G. P. and Castellano, M. A., Light and electron microscopy of the intracellular inclusions of
cauliflower mosaic virus, J. Gen. Virol., 13, 133, 1971.
71. Lawson, R. H. and Hearon, S. S., Carnation etched ring virus inclusions: serology and ultrastructure of
alkaline-treated inclusions, Phytopathology, 70, 327, 1980.
72. Shalla, T. A., Shepherd, R. J., and Petersen, L. J., Comparative cytology of nine isolates of cauliflower
mosaic virus, Virology, 102, 381, 1980.
73. Xiong, C., Balazs, E., Lebeurier, G., Hindelang, C., Stoeckel, M. E., and Porte, A., Comparative
cytology of two isolates of cauliflower mosaic virus, J. Gen. Virol., 61, 75, 1982.
74. Kamei, T., Rubio-Huertos, M., and Matsui, C., Thymidine-'H uptake by x-bodies associated with
cauliflower mosaic virus infection, Virology, 37, 506, 1969.
75. Favali, M. A., Bassi, M., and Conti, G. G., A quantitative autoradiographic study of intracellular sites
for replication of cauliflower mosaic virus. Virology, 53, 115, 1973.
76. Hatta, T., unpublished results, 1981.
77. Conti, G. G., Vegetti, G., Bassi, M., and Favali, M. A., Some ultrastructural and cytochemical obser-
vations on Chinese cabbage leaves infected with cauliflower mosaic virus, Virology, 47, 694, 1972.
78. Bassi, M., Favali, M. A., and Conti, G. G., Cell wall protrusions induced by cauliflower mosaic virus
in Chinese cabbage leaves: a cytochemical and autoradiographic study, Virology, 60, 353, 1974.
Volume I 33
Chapter 3
GEMINIVIRUS GROUP
Viruses belonging to this group are called Geminiviruses because they have unique, paired
or twinned particles about 18 x 30 nm. 1 ' 3 They are also unique in being the only group of
plant viruses known to possess circular, single-stranded DNA genomes which in at least
some cases are bipartite. Each virus particle appears to consist of a single molecule of DNA
of Mr between 0.7 and 0.8 x 106 encapsidated by a shell consisting of two incomplete
icosahedra built from a single species of polypeptide of MT between 27 and 34 x 10\
Some Geminiviruses are transmitted by whiteflies and others by leafhoppers in a persistent
manner, but the viruses do not appear to replicate in their vectors and are not seed transmitted.
A few of the viruses can be transmitted experimentally by mechanical inoculation. Each of
the Geminiviruses appears to have a relatively narrow host range.' 3
A number of plant diseases suspected of being caused by viruses and transmitted by

whiteflies4 and leafhoppers5 were studied for many years without the causal agents being
identified. Eventually, isometric particles 18 to 20 nm in diameter and usually occurring in
pairs were isolated from maize streak-diseased plants. 6 Subsequently, similar particles have
been isolated from plants affected by a number of other diseases. 1 - 2 At present, there are
five apparently distinct Geminiviruses, whose nucleic acids and coat proteins have been
characterized (Table 1). The particles of another eleven viruses have been identified as
geminate (Table 2). These viruses have not yet been studied in detail and some may be
sufficiently closely related to be considered strains of the same virus. Already there is
evidence that tobacco yellow dwarf virus (TYDV) is antigenically closely related to bean
summer death virus (BSDV). 7 Also, it has been suggested that tobacco leaf curl virus (TLCV)
and tomato yellow dwarf virus (ToYDV) are similar and may be identical.8 It has been
reported that beet curly top virus (BCTV) is serologically related to TYDV and BSDV,
although the degree of relationship is not clear.7 Similarly, there appear to be distant ser-
ological relationships between cassava latent virus (CaLV) and BGMV and between CaLV
and BCTV.9
It has been suggested that abutilon mosaic, which is transmitted by the whitefly Bemisia
(abaci, is caused by a Gemini virus, 30 - 31 but the evidence does not seem strong enough at
present to warrant inclusion of the disease agent in the Geminivirus group. The precise cause
of the African cassava mosaic disease is not clear, but CaLV may well be implicated. 1 ' 32 - 33
Most purified preparations of Geminiviruses contain predominantly geminate particles
about 18 x 30 nm with varying numbers of polyhedral particles about 18 nm in diameter
(Figure l). 7 - 17 - 34 - 35 In some Geminivirus preparations, occasional triplet particles have also
been seen.36 Initially it was thought that perhaps the single polyhedral particles were the
complete infectious entities and that the geminate particles were a preparative artifact. 10 - 34 37
However, later work with chloris striate mosaic virus (CSMV) supported the view that the
geminate particles are the complete units and that polyhedral particles are degradation prod-
ucts.38 This now appears to be generally accepted.1-2
Hatta and Francki38 observed numerous geminate particles both in crude leaf extracts and
in purified preparations of CSMV stained with uranyl acetate or prepared by freeze-drying
Table 1
SOME PROPERTIES OF GEMINIVIRUSES
Mrof Mrof
ssDNA coat protein
Virus Vector species (x 10") ( x 103) Ref.
Bean golden mosaic vi- Bemisia tabaci 0.80 27.4 10—12

rus (BGMV) (whitefly)
Cassava latent virus Unknown 0.80 34.0 13—15
(CaLV)
Chloris striate mosaic vi- Nesoclutha pa/lida 0.71 28.0 16
rus (CSMV) (leafhopper)
Maize streak virus (MV) Cicadulina mbila 0.71 28.0 13, 17, 18
(leafhopper)
Tomato golden mosaic Bemisia tabaci 0.79 27.9 19, 20
virus (ToGMV) (whitefly)
Table 2
VIRUSES WITH GEMINIVIRUS-LIKE PARTICLES
Virus Vector Ref.
Bean summer death virus (BSDV) Orosius argentatus 7

(leafhopper)
Beet curly top virus (BCTV) Circulifer tenellus 21, 22
(leafhopper)
Euphorbia mosaic virus (EuMV) Bemisia tabaci 23
(whitefly)
Mungbean yellow mosaic virus (MYMV)" B. tabaci 24
(whitefly)
Paspalum striate mosaic virus (PSMV) Nesoclutha pallida 25, 26
(leafhopper)
Tobacco leaf curl virus (TLCV) B. tabaci 8
(whitefly)
Tobacco yellow dwarf virus (TYDV) O. argentatus 7
(leafhopper)
Tomato yellow dwarf virus (ToYDV) B. tabaci 8
(whitefly)
Tomato yellow leaf curl virus (ToYLV)" B. tabaci 27
(whitefly)
Tomato yellow mosaic virus (ToYMV) a B. tabaci 28
(whitefly)
Wheat dwarf virus (WDV) Psammotettix 29
alienus
(leafhopper)
" Identification of virus as a Geminivirus only by cytopathological exami-

nation of infected plants by electron microscopy.
and shadow casting. They also showed, by tilting thin sections of virus-infected cells in the
electron microscope, that many of the virus-like particles in the nuclei were geminate, and
not simply superimposed isometric particles. Thin sections of pelleted purified virus gave
similar results.
Proteins from preparations of the five viruses listed in Table 1 and those of TYDV and
BSDV7 have yielded single major polypeptides of Mr ranging from 27 to 34 x 103 when
estimated by polyacrylamide gel electrophoresis under denaturing conditions. However, some
investigators also detected small amounts of additional protein components. The minor
component detected in preparations of CSMV was thought to be a dimer of the major
Volume I 35
FIGURE I. A, Purified chloris striate mosaic virus (CSMV); B, maize streak virus (MSV). Uranyl acetate
staining. Bar = 100 nm.
polypeptide, 39 but the origin and significance of the minor proteins detected in preparations
of BGMV, TYDV, and BSDV are obscure. 7 1 2
Studies on the fine structure of CSMV indicate that each geminate particle is constructed
FIGURE 2. Two selected particles of CSMV (A and B, left) on which recognizable capsomeres have been
identified (center), fitted to a model of the Geminivinis particle (right). The model consists of two incomplete T
= 1 icosahedra, each with I I capsomeres. as proposed by Hatta and Hrancki." 1
from two incomplete icosahedra with T = 1 surface lattice symmetry and has a total of 22
capsomeres (Figure 2). 3 X Data indicating that each geminate particle of CSMV and BGMV
contains a single molecule of DNA 12 - 1 '' confirm the conclusion that the native CSMV and
BGMV particles are geminate.
B. The Viral Genome

Geminivirus particles contain single-stranded circular DNAs of M, between 0.7 and 0.8
x 106 (Table 1). In most DNA preparations from Geminiviruses, there are also some linear
molecules."" 13 - 35 " 39 However, only the circular molecules appear to be infectious. No evi-
dence has been found for the covalent bonding of protein to the DNA and it seems that all
the nucleotides of the circular DNA molecules are linked by covalent bonds. 12 Restriction
endonuclease maps of circular and linear molecules are indistinguishable indicating that the
linear molecules are nicked circular ones.40
Initially MSV was thought to have a bipartite single-stranded RNA genome."* This led
to the suggestion that the geminate particle morphology was adapted for packaging and
delivery of both nucleic acid molecules at the same infective site. 18 The idea that the
Geminivirus particle is an adaptation for delivery of the nucleic acid molecules is now
untenable if, as discussed above, each geminate particle accommodates only a single molecule
of DNA. 12 39 Recent studies on restriction mapping of BGMV DNA suggest the existence
of two types of DNA molecules with similar molecular weights but distinct nucleotide
Volume I 37
Table 3
ELECTRON MICROSCOPIC DETECTION OF GEMINIVIRUS PARTICLES IN
INFECTED PLANTS AND THEIR CYTOPATHOLOGICAL EFFECTS
Virus particles
Virus particles
Random Crystalline observed in
Virus aggregates aggregates Fibrillar rings cytoplasm Ref.
Bean golden mosaic virus Yes Yes Yes None detected 53

(BGMV)
Beet curly top virus (BCTV) Yes Yes Yes None detected 45^7
Chloris striate mosaic virus Yes None detected None detected Occasionally 35, 38
(CSMV) detected
Euphorbia mosaic virus Yes None detected Yes None detected 48
(EuMV)
Maize streak virus (MSV) None reported Yes None reported None reported 17
Mungbean yellow mosaic vi- Yes Yes Yes Yes (also in 24
rus (MYMV) vacuole)
Paspalum striate mosaic virus None detected Yes None detected None detected 25. 26
(PSMV)
Tobacco leaf curl virus Yes Yes None reported None reported 8
(TLCV)
Tomato yellow dwarf virus Yes Yes None reported None reported 8
(ToYDV)
Tomato yellow leaf curl virus Yes None detected Yes None reported 27
(ToYLV)
Tomato yellow mosaic virus Yes None detected Yes Yes 28
(ToYMV)
Wheat dwarf virus (WDV) Yes Yes None reported None reported 44
sequences. Infectivity-dilution curves of DNA preparations on bean protoplasts also support

the view that two DNA molecules are required for infectivity. 40
Two DNA molecules similar in size but differing in base sequences have also been detected
in preparations of CaLV15 and tomato golden mosaic virus (ToGMV), 41 - 42 suggesting that
their genomes may also be bipartite. Compelling evidence that the genome of a Geminivirus
is bipartite comes from recent work by Stanley43 who cloned both molecules of CaLV15 and
showed that both were required for infectivity.
The complete nucleotide sequences of the two DNA molecules from CaLV with 2779
and 2724 nucleotides, respectively, 15 have been compared and shown to have a stretch of
about 200 nucleotides in common. 15 It seems that this region may be involved with the
origin of DNA replication. The potential coding capacity has been investigated and a number
of putative codons located on each of the DNA molecules.15 One of the codons appears to
have the information for a protein with similar amino composition to that of the coat
protein. 15 - 43
III. CYTOPATHOLOGY
In all the cases studied, Geminivirus particles accumulate in the nuclei of infected cells
(Table 3). However, there are some instances where particles have also been observed in
the cytoplasm (Table 3). It has been suggested that these particles were extruded from the
nuclei through breaks in the nuclear envelope,28 but whether such breaks are direct results
of virus infection is unknown.
The particles of most Gemini viruses are confined to phloem cells, but some also appear
to invade other tissues. For example, extensive electron microscopic and immunochemical
studies on several species of BCTV-infected plants indicate that the virus is confined to the
phloem and the virus-induced hyperplastic phloem cells. 45 " 4749 - 5 " Immunochemical studies
on CaLV-infected plants detected numerous nuclei containing the virus, but some cells of
other tissues were also shown to be infected. 9 In contrast, CSMV particles were detected
by electron microscopy in all cell types of infected Chloris gayana leaves except the epidermal
and possibly the xylem tracheid cells. 35 MSV probably also invades other tissues besides
the phloem, as it is acquired by the vector, like CSMV, after brief probes inthemesophyll. 5 1 - 5 2
Particles of some Geminiviruses accumulate in the nucleus as randomly arranged aggre-
gates such as those in Chloris gayana leaf cells infected with CSMV (Figure 3).* In other
instances, the nuclear aggregates consist of crystalline arrays of particles such as those in
Paspalum dilatatum infected with paspalum striate mosaic virus (PSMV) (Figure 4). In many
cases particles accumulate in such large numbers as to occupy much of the nuclear volume,
displacing the chromatin to the periphery. 35
Kim and his colleagues53 studied the sequence of structural changes in the nuclei of French
bean leaves inoculated with BGMV. The nucleoli increased in size and then segregated into
granular and fibrillar regions (Figure 5). The fibrillar material then condensed into rings of
various sizes, sometimes several to a nucleus (Figures 5 through 7), and finally, virus particles
appeared in the nucleoplasm (Figure 8). Cytochemical investigations indicated that, in seg-
regated nucleoli, the granular material was composed mainly of ribonucleoprotein, whereas
the fibrillar material was mainly deoxyribonucleoprotein.53 Similar structural changes have
been observed in plants infected by a number of other Geminiviruses (Table 3), but their
significance in the process of virus multiplication remains to be elucidated.
At present there is little known about the mechanism of Geminivirus replication. 41 - 54 55
However, virus-specific double-stranded DNA assumed to be the replicative form has been
isolated from plants infected with BGMV, 54 and several virus-specific single- and double-
stranded DNAs have been detected in ToGMV-infected plants. 41
* Figures 3 through 8 follow text, beginning on page 39.

Volume I 39
FIGURE 3. A, Cell of Chloris gayuna leaf infected with CSMV showing virus particles (V) in the nucleus; bar
= 500 nm. B, Part of the CSMV-infected nucleus showing difference between appearance of virus particles (V)
and chromatin (Ch.). Bar = 100 nm.
FIGURE 4. A. Part of a cell of Paspalum dilatation infected with paspalum striate mosaic virus (PSMV) showing
crystalline arrays of virus particles in the nucleus: bar = 500 nm. B, Part of the PSMV-infected nucleus showing
details of crystalline arrays of particles; bar = 500 nm. C, Details of PSMV particle crystals. Bar = 100 nm.
Volume I 41
FIGURE 5. Granular (G) and fibrillar (F) regions of the nucleolar region in leaf cells of Phaseolus vulgaris
infected with bean golden mosaic virus (BGMV). Bar = 500 nm. (Courtesy of Dr. K. S. Kim, Department of
Plant Pathology, University of Arkansas, Fayetteville.)
FIGURE 6. Small fibrillar rings (presumably sectioned hollow spheres) scattered in the nucleoplasm of a Phaseolus
vulgaris leaf cell at an early stage of infection with BGMV. Bar = 500 nm. (Courtesy of Dr. K. S. Kim. Department
of Plant Pathology, University of Arkansas, Fayetteville.)
Volume I 43
FIGURE 7. Large fibrillar ring (F) in the nucleoplasm of a leaf cell of Phaseolus vulgaris infected with BGMV:
the ring is as large as the nucleolus ( N l ) . Bar = 500 nm. (Courtesy of Dr. K. S. Kim, Department of Plant
Pathology, University of Arkansas, Fayetteville.)
FIGURE 8. Granular (G) and fibrillar (F) regions in the nucleoplasm of a BGMV-infected Phaseolus vulgaris
leaf cell also containing virus particles (V). Bar = 500 nm. (Courtesy of Dr. K. S. Kim, Department of Plant
Pathology, University of Arkansas, Fayetteville.)
REFERENCES
1 . Goodman, R. M., Geminiviruses: the single-stranded DNA viruses of plants, in Handbook of Plant Virus
Infections and Comparative Diagnosis, Kurstak, E., Ed., Elsevier, Amsterdam, 1981, 879.
2. Goodman, R. M., Geminiviruses, J. Gen. Virol, 54, 9, 1981.
3. Bock, K. R., Geminivirus diseases in tropical crops, Plant Dis., 66, 266, 1982.
4. Bird, J. and Maramorosch, K., Viruses and virus diseases associated with whiteflies, Adv. Virus Res.,
22, 55, 1978.
Volume I 45
5. Maramorosch, K. and Harris, K. F., Eds., Lea/hopper Vectors and Plant Disease Agents, Academic
Press, New York, 1979, 654 pp.
6. Bock, K. R., Guthrie, E. J., and Woods, R. D., Purification of maize streak virus and its relationship
to viruses associated with streak diseases of sugarcane and Panicum maximum, Ann. Appl. Biol., 77, 289,
1974.
7. Thomas, J. E. and Bowyer, J. W., Properties of tobacco yellow dwarf and bean summer death viruses,
Phytopathology, 70, 214, 1980.
8. Osaki, T. and Inouye, T., Resemblance in morphology and intranuclear appearance of viruses isolated
from yellow dwarf diseased tomato and leaf curl diseased tobacco, Ann. Phytopathol. Soc. Jpn., 44, 167,
1978.
9. Sequeira, J. C. and Harrison, B. D., Serological studies on cassava latent virus, Ann. Appl. Biol., 101,
33, 1982.
10. Goodman, R. M. and Bird, J., Bean golden mosaic virus, CMI/AAB Descriptions of Plant Viruses, No.
192, 1978.
11. Reisman, D., Ricciardi, R. P., and Goodman, R. M., The size and topology of single-stranded DNA
from bean golden mosaic virus, Virology, 97, 388, 1979.
12. Goodman, R. M., Shock, T. L., Haber, S., Browning, K. S., and Bowers, G. R., The composition
of bean golden mosaic virus and its single-stranded DNA genome, Virology, 106, 168, 1980.
13. Harrison, B. D., Barker, H., Bock, K. R., Guthrie, E. J., Meredith, G., and Atkinson, M., Plant
viruses with circular single-stranded DNA, Nature {London), 270, 760, 1977.
14. Bock, K. R., Guthrie, E. J., and Meredith, G., Distribution, host range, properties and purification of
cassava latent virus, a geminivirus, Ann. Appl. Biol., 90, 361, 1978.
15. Stanley, J. and Gay, M. R., Nucleotide sequence of cassava latent virus DNA, Nature (London), 301,
260, 1983.
16. Francki, R. I. B. and Hatta, T., Chloris striate mosaic virus, CMI/AAB Descriptions of Plant Viruses,
No. 221, 1980.
17. Bock, K. R., Maize streak virus, CMIIAAB Descriptions of Plant Viruses, No. 133, 1974.
18. Bock, K. R., Guthrie, E. J., and Meredith, G., RNA and protein components of maize streak and
cassava latent viruses, Ann. Appl. Biol., 85, 305, 1977.
19. Matyis, J. C., Silva, D. M., Oliveira, A. R., and Costa, A. S., Purifac.ao e morfologia do virus do
mosaico dourado do tomateiro, Summa Phyopathol., 1, 267, 1975.
20. Hamilton, W. D. O., Sanders, R. C., Coutts, R. H. A., and Buck, K. W., Characterization of tomato
golden mosaic virus as a geminivirus, FEMS Microbiol, Lett., 11, 263, 1981.
21. Thomas, P. E. and Mink, G. I., Beet curly top virus, CMI/AAB Descriptions of Plant Viruses, No. 210,
1979.
22. Bennett, C. W., The curly top disease of sugar beet and other plants, Am. Phytopathol. Soc. Monogr.,
No. 7, American Phytopathology Society, St. Paul, Minnesota, 1979.
23. Matyis, J. C., Silva, D. M., Oliviera, A. R., and Costa, A. S., Aspectos morfoligicos do virus do
mosaico da Euphorbia. V. Coloquio Brasiliero de Microscopia Electronica, 1976, 56 pp.
24. Thongmeearkom, P., Honda, Y., Saito, Y., and Syamananda, R., Nuclear ultrastructural changes and
aggregates of virus-like particles in mungbean cells affected by mungbean yellow mosaic disease, Phyto-
pathology, 71, 41, 1981.
25. Grylls, N. E., Leafhopper vectors and the plant disease agents they transmit in Australia, in Lea/hopper
Vectors and Plant Disease Agents, Maramorosch, K. and Harris, K. F., Eds., Academic Press, New York,
1979, 179.
26. Francki, R. I. B. and Hatta, T., unpublished results, 1981.
27. Rosso, M., Cohen, S., and Martelli, G. P., Virus-like particles in tomato plants affected by the yellow
leaf curl disease, /. Gen. Virol, 49, 209, 1980.
28. Lastra, R. and Gil, F,, Ultrastructural host cell changes associated with tomato yellow mosaic, Phyto-
pathology, 71, 524, 1981.
29. Lindsten, K., Wheat dwarf — an old disease caused by a unique and earlier unknown virus, Vdxtskydds-
notiser, 44, 54, 1980.
30. Jeske, H. and Werz, G., Ultrastructural and biochemical investigations on the whitefly transmitted Abutilon
mosaic virus (AbMV), Phytopathol. Z., 97, 43, 1980.
31. Jeske, H. and Werz, G., Cytochemical characterization of plastidial inclusions in Abutilon mosaic-infected
Malva parriflora mesophyll cells, Virology, 106, 155, 1980.
32. Horvat, F. and Verhogen, M., Cytologica) modifications and presence of virus-like particles in cells of
Nicotiana benthamiana (Domin) and Manihot utilissima (Pohl) infected with the geminivirus isolated from
cassava infected with the cassava African mosaic disease, Parasitica, 37, 119, 1981.
33. Walter, B., Isolation and purification of a virus transmitted from mosaic-diseased cassava in the Ivory
Coast, Plant Dis., 64, 1040, 1980.
34. Goodman, R. M., Bird, J., and Thongmeearkom, P., An unusual virus-like particle associated with
golden yellow mosaic of beans, Phytopathology, 67, 37, 1977.
35. Francki, R. I. B., Hatta, T., Grylls, N. E., and Grivell, C. J., The particle morphology and some other
properties of chloris striate mosaic virus, Ann. Appl. Rial., 91, 51, 1979.
36. Magyarosy, A. C., Beet curly top virus from phloem exudate of Amsinckia doug/asiana, Ann. Appl. Biol.,
96, 295, 1980.
37. Goodman, R. M., Infectious DNA from a whitefly-transmitted virus of Phaseolus vulgaris, Nature (London),
266, 54, 1977.
38. Hatta, T. and Francki, R.I. B., The fine structure of chloris striate mosaic virus,'Virology, 92, 428,
1979.
39. Francki, R. I. B., Hatta, T., Boccardo, G., and Randies, J. W., The composition of chloris striate
mosaic virus, a Geminivirus, Virology, 101, 233, 1980.
40. Haber, S., Ikegami, M., Bajet, N. B., and Goodman, R. M., Evidence for a divided genome in bean
golden mosaic virus, a Geminivirus, Nature (London), 289, 324, 1981.
41. Hamilton, W. D. O., Bisaro, D. M., and Buck, K. W., Identification of novel DNA forms in tomato
golden mosaic virus infected tissue. Evidence for a two component viral genome, Nucleic Acids Res., 10,
4901, 1982.
42 Bisaro, D. M., Hamilton, W. D. O., Coutts, R. H. A., and Buck, K. W., Molecular cloning and
characterisation of the two DNA components of tomato golden mosaic virus, Nucleic Acids Res., 10, 4913,
1982.
43. Stanley, J., Infectivity of the cloned geminivirus genome requires sequences from both DNAs, Nature
(London), 305, 643, 1983.
44. Tomenius, K. and Oxelfelt, P., Preliminary observations of virus like particles in nuclei of cells of wheat
infected with the wheat dwarf disease, Phytopathol. Z., 101, 163, 1981.
45. Esau. K. and Hoefert, L. L., Particles and associated inclusions in sugarbeet infected with the curly top
virus, Virology, 56, 454, 1973.
46. Esau, K., Virus-like particles in nuclei of phloem cells in spinach leaves infected with the curly top virus,
J. U/trastruct. Res., 61. 78, 1977.
47. Esau, K. and Magyarosy, A. C., Nuclear abnormalities and cytoplasmic inclusion in Amsinckia infected
with the curly top virus, J. Ultrastruct. Res., 66, 1 1 , 1979.
48. Kim, K. S. and Flores, E. M., Nuclear changes associated with euphorbia mosaic virus transmitted by
the whitefly, Phytopathology, 69, 980, 1979.
49. Mumford, D. L. and Thornley, W. R., Location of curly top virus antigen in bean, sugarbeet, tobacco
and tomato by fluorescent antibody staining, Phytopathology, 67, 1313, 1977.
50. Esau, K. and Magyarosy, A. C., A study of the source of virus in the phloem exudate appearing in leaves
of Amsinckia infected with the beet curly top virus, Plant, Cell Environ., 3, 425, 1980.
51. Storey, H. H., Investigations of the mechanism of the transmission of plant viruses by insect vectors. 11.
The part played by puncture in transmission, Proc. R. Soc. London Ser. B, 125, 445, 1938.
52. Grylls, N. E., A striate mosaic virus disease of grasses and cereals in Australia, transmitted by the Cicadellid
Nesoclutha obscura, Aust. J. Agric. Res., 14, 143, 1963.
53. Kim, K. S., Shock, T. L., and Goodman, R. M., Infection of Phaseolus vulgaris by bean golden mosaic
virus: ultrastructural aspects. Virology, 89, 22, 1978.
54. Ikegami, M., Haber, S., and Goodman, R. M., Isolation and characterization of virus-specific double-
stranded DNA from tissues infected by bean golden mosaic virus, Proc. Natl. Acad. Sci. U.S.A., 78, 4102,
1981.
55. Howarth, A. J. and Goodman, R. M., Plant viruses with genomes of single-stranded DNA, Trends
Biochem. Sci., 7, 180, 1982.
Volume I 47
Chapter 4
PLANT REOVIRIDAE
The family was originally erected around reovirus, which infects man and other mammals
and whose name was derived from the description "respiratory enteric orphan virus";
"orphan" because at the time no disease had been found associated with the particles. Later
it became clear that Reoviruses had obvious affinities with some viruses infecting vertebrates,
insects, and plants. Thus the ICTV came to approve the inclusion of plant viruses with
Reovirus-like particles in the family Reoviridae, 1 in spite of the objections of many plant
virologists to the classification of plant viruses into families and genera (see Chapter 1).
Bearing in mind this problem, we will follow here the classification approved by the ICTV.'
The plant Reoviridae have double-shelled particles with icosahedral symmetry, 65 to 70
nm in diameter, containing 10 to 12 segments of double-stranded RNA. Their vectors, either
leafhoppers or planthoppers, transmit the viruses propagatively, and hence they can also be
considered as insect viruses. Each of the viruses infects only a limited number of host plant
species. None appear to be transmissible by mechanical inoculation (except rarely by needle
puncture) or through the seed.2 5
I. MEMBERS OF THE FAMILY AND THEIR RELATIONSHIPS
The family is divided into six genera, two of which, Phytoreovirus and Fijivirus, include
viruses which infect both plants and insects (Table 1). In addition, there is one other virus,
rice ragged stunt virus (RRSV), that is not classified in either genus. 4 • 5
A. Phytoreovirus
The three viruses in this genus, wound tumor (WTV), rice dwarf (RDV), and rice gall
dwarf (RGDV) all have a similar capsid structure (Figure 1), 12 double-stranded RNA
genome segments of similar sizes (Figure 2), and are all vectored by leafhoppers, but are
not serologically interrelated. 612
The symptoms and host ranges of these viruses are of interest and worth discussing briefly.
The natural host range of WTV is unknown but, experimentally, it can be made to infect a
wide range of dicotyledons.6 In contrast, RDV is confined to the Gramineae,7 and this may
well also be true of RGDV" though its host range has not been widely tested.
The symptoms produced by the three viruses are quite different. WTV-infected plants
may at first show only vein enlargement, but tumors develop where plants are wounded and
where lateral root initials break through the pericycle. 6 - 13 - 14 RDV is the only virus among
the plant Reoviridae that does not induce neoplasia; instead, plants are stunted and develop
chlorotic flecks.7 RGDV, on the other hand, induces symptoms very similar to those found
in Fijivirus-infected plants, namely stunting, dark green coloring, leaf distortion, enation or
gall development on the veins of leaves and leaf sheaths, and suppression of flowering. 12
B. Fijivirus
The viruses in this genus (Figure 3) possess particles that are morphologically indistin-
guishable and all genomes consist of 10 double-stranded RNA segments of similar sizes
(Figure 2). All the viruses are vectored by planthoppers and induce similar characteristic
symptoms in plants, as described above for RGDV.
The viruses can, at present, be classified according to the serological relationships of their
inner shells (Figure 4) and the relative sizes of the genomic RNAs (Figure 2). 4 - 5 - 1 5 1 7
The serology is complicated by the fact that the double-stranded RNA within the particles
Table 1
SOME PROPERTIES OF CURRENTLY RECOGNIZED GENERA WITHIN THE
REOVIRIDAE"
Particle morphology
No. of
double-stranded Spikes
RNA genome
Genus Hosts segments Shell A B
Reovirus Mammals and birds 10 Double +

Orbivirus Mammals, insects, and other 10 Double -
arthropods
Rotavirus Mammals 10 Double - -
Phytoreovirus Higher plants and insects 12 Double -
Fijivirus Higher plants and insects 10 Double + +
Cytoplasmic Insects 10 Single plus matrix + +
polyhedrosis protein
Data compiled from Matthews. 1
is itself immunogenic and elicits antisera of fairly low liter that react with double-stranded
polyribonucleotides of differing origin and base sequence.18 21 A second complication is that
the capsids contain at least six polypeptides22 so that the antigenic possibilities are numerous.
The antisera generally available are those against the stable B-spiked subviral particles
(SVPs), though with Fiji disease virus (FDV) and maize rough dwarf virus (MRDV) antisera
reacting with the outer shells have also been prepared.23-24 Fortunately, the weak reactions
due to double-stranded RNA can be eliminated by absorbing the sera, or discounted by
directly observing the reacted particle using immunoelectron microscopy.24"30
The conclusion reached from serological and other studies28 3I is that the viruses can be
divided into three unrelated subgroups (Figure 4). The sole representative of subgroup I is
FDV, which appears to be unrelated to any other Fijivirus in its inner shell and B spikes.
Moreoever, an antiserum reacting with the outer shell of FDV did not react with that of
MRDV, pangola stunt virus (PaSV), or oat sterile dwarf virus (OSDV).24 In subgroup II,
MRDV, rice black-streaked dwarf virus (RBSDV) and PaSV are closely related, but not
identical in their inner shell and B spike antigens; cereal tillering disease virus (CTDV) is
also closely related, but tests for any possible differences have not yet been done (Figure
4). Subgroup III contains OSDV, arrhenatherum blue dwarf virus (ABDV) and lolium
enation virus (LEV), whose inner shells and B spikes appear serologically indistinguishable
(Figure 4), although critical tests for minor differences have not been done. The outer shells
of MRDV and OSDV also appear to be unrelated.
As seen in Figure 2 and discussed in more detail later, the genome patterns of the
Fijiviruses, derived by polyacrylamide gel electrophoresis, also form three clusters that
correspond with those derived from serology. It is clear, therefore, the FDV, MRDV, and
OSDV are distinct viruses. Historically, and because of geographical separation and differ-
ences in the main host plant infected, RBSDV, PaSV, and CTDV have been considered as
viruses distinct from MRDV. 32 35 CTDV probably has the weakest claim to separate status
and should be considered as a strain of MRDV. ABDV and LEV are so similar to OSDV
in all properties studied that they should be considered isolates of that virus.36
The plant Reoviridae so far studied all contain double-stranded RNA inside icosahedrally
Volume I 49
FIGURE I . Purified Phytorecn-inix preparations. A. Wound tumor virus (WTV); B, rice dwarf virus (RDV):
potassium phosphotungstate stain. Bar -- 100 nm. (Courtcs\ ol I. Kiniura. National Institute of Agrobiological
Resources. Tsakuba City, Japan.)
FIGURE 2. Sizes of the individual double-stranded RNA genomes of the members of the genera Phytoreovirus
and Fijivirus. RDV, rice dwarf virus; WTV, wound tumor virus; RGDV, rice gall dwarf virus; FDV, Fiji disease
virus; MRDV, maize rough dwarf virus; RBSDV, rice black-streaked dwarf virus; PaSV, pangola stunt virus;
OSDV, oat sterile dwarf virus; ABDV, arrhenatherum blue dwarf virus; and RRSV, rice ragged stunt virus.
(Modified after Francki and Boccardo.4)
constructed protein shells whose diameter has variously been measured as between 45 and
59 nm. Upon these shells is built a partial or complete second protein layer. Within this
framework, three morphologies are found, that of Phytoreovirus (Figure 1), that of Fijivirus
(Figures 3 and 5A through H and S), and that of RRSV (Figure 5L through R). However,
only with Fijivirus is there a generally agreed interpretation of the structures observed. The
type of surface lattice from which the outer shell is constructed is not known with certainty
for any plant Reo-like virus, and very little is known of the structure of the inner shell.
WTV and RDV particles appear to share a common morphology, and look very similar
in the electron microscope (Figure 1). Nevertheless, measurements of the diameters of the
supposedly intact particles (IPs) and the inner shells or SVPs, and interpretations of the
surface structures visible have varied widely (Table 2). The morphology of RGDV particles
is somewhat similar to that of WTV and RDV, but co-imaging of mixed preparations has
not been done, so the true extent of the resemblance is uncertain.
With WTV, the evidence seems good for the presence of an amorphous protein layer that
is easily lost and that lies external to the outer shell." This layer would be analogous to
that found in particles of the genus Orbivirus.^ Such a layer has not been reported for RDV
Volume I 51
FIGURE 3. Purified maize rough dwarf virus (MRDV). Uranyl acetate stain. Bar = 100 nm.
FIGURE 4. Serological relationships among members of the genus Fijivirus. Solid lines indicate positive ser-
ological reaction; broken lines indicate reported tests where no serological cross-reactions were detected. Arrowheads
at both ends of the lines indicate that antisera to both viruses were used. A single arrow indicates that antiserum
to only one virus was used, the one from which the arrowhead ponts. FDV, Fiji disease virus; MRDV, maize
rough dwarf virus; OSDV, oat sterile dwarf virus; RBSDV, rice black-streaked dwarf virus: PaSV, pangola stunt
virus; CTDV, cereal tillering disease virus; ABDV, arrhenatherum blue dwarf virus; and LEV, lolium enation
virus. (After Francki and Boccardo.4)
or RGDV, but negatively stained preparations of all Phytoreovirus particles do strikingly

resemble those of Orbivirus ,43 Whether the T = 9 or T = 3 surface lattice is correct for
the outer shell requires further study, though the most recent evidence suggests a T = 9
structure of 92 holes surrounded by 180 subunits.42 The outer shell of Phytoreovirus particles,
unlike that of Fijivirus particles, remains intact after mounting in neutral PTA.
Within the genus Fijivirus, only the particles of FDV and MRDV have been studied in
detail.16-27-44-45 There is close agreement both on the results and their interpretation. Work
on other members of the genus suggests that their particles are also very similar32-"-36 and
hence that the Fijiviruses are morphologically homogeneous (Table 3). Nevertheless, the
intact particles differ somewhat in their tendency to break down into SVPs; OSDV particles
are rather resistant, MRDV particles intermediate, and FDV particles, the most fragile.45-46
The intact Fijivirus particle (IP) is a slightly angular icosahedral structure with an A spike
at each of its twelve vertices (Figures 3 and 5A and I). Loss of the A spikes and parts of
the outer shell (Figure 5B and C) reveal an icosahedral inner shell (subviral particle, or
SVP) with B spikes also attached to each vertex (Figure 5D). The B spikes are in effect
Volume 1 53
FIGURE 5. Particle structure of a Fijivirus (maize rough dwarf virus, MRDV) and of rice ragged stunt virus
(RRSV), as seen in negatively stained preparations. A, Intact MRDV particle showing A spikes (arrowheads); B
through D, images of MRDV particles showing successive loss of elements of the outer shell to give a subviral
particle (SVP) with B spikes (arrowheads in D). Uranyl acetate stain. E through H, Models representing particles
in A through D, respectively. 1, Similar particle to that in A but fixed in glutaraldehyde and stained in neutral
potassium phosphotungstate (PTA). Note that apparent diameter of particle in I is greater than in A and the A
spikes (arrowheads) are a different shape. J, A smooth SVP of MRDV (devoid of B spikes shown in D); such
particles are produced when intact or B-spiked SVPs are mounted, unfixed in PTA. K, Individual detached B
spikes in face view (upper row) and profile (lower row). L through P, Particles of RRSV stained in uranyl acetate
showing the typical B spikes which appear broadened at the base as shown in Q and R; they differ from the B
spikes of Fijivirus particles shown in S. It is not clear yet whether particles of RRSV like those shown in L through
P are intact or subviral particles. All bars = 20 nm.
Table 2
MEASUREMENTS AND INTERPRETATIONS OF PHYTOREOVIRUS PARTICLE
STRUCTURE
Diameter (nm) of
Outer
amorphous Outer Inner Interpretation of outer shell
Virus layer shell shell structure Ref.
Wound tumor virus 73 63 59 — 37

(WTV)
WTV — 66 35^0" — 38
WTV — 70 T = 3; 32 morphological 6
subunits
WTV — 72—78.5 — 39
WTV — 70 T = 9; 92 morphological 40
subunits
Rice dwarf virus — 70 T = 3; 32 morphological 2, 7, 41
(RDV) subunits
RDV — 70 T = 9; 92 holes surrounded by 42
180 morphological subunits
Rice gall dwarf virus — 65 45" — 10
(RGDV)
Not clear whether the authors referred to the inner shell or the nucleic acid core.
Table 3
STRUCTURES AND THEIR
DIMENSIONS IN A TYPICAL
FIJIVIRUS PARTICLE"
Structure Measurement Size (nm)
Intact particle (IP) Diameter 65—70

A spikes Width 10.5—11
Height 9—11
B spikes Width 11—13.5
Height 8
Inner shell Diameter 50—57
" Modified from Milne and colleagues44 and Hatta

and Francki.45
resistant units of the outer shell; they are hollow, and each is built of five subunits (Figure
5K). The viral RNA may extend from the SVPs,27 presumably through the hollow B spikes
(Figure 6). Each B spike rests on a base-plate which forms part of the inner shell. The inner
shell is the most strongly built part of the particle. Apart from the base-plate, it shows little
evidence of morphological subunits (Figure 5J) and its structure is not understood. A proposed
model of the Fijivirus particle is presented in Figure 7.
The morphology of the IP of RRSV is not yet clear. A particle resembling an IP of
Fijivirus has not been observed in negatively stained preparations despite investigation in
several laboratories.47'50 Instead, particles have been observed that resemble Fiijivirus SVPs
in form and size except that the RRSV B spikes are broader at the base50 (Figure 5L through
S). It seems possible that the RRSV IP lacks a complete outer shell and so somewhat
resembles particles of the genus Cypovirus but without the A spikes.51-52
Volume I 55
FIGURE 6. Fiji disease virus (FDV) subviral particles (SVPs) prepared by the cytochrome c spreading and rotary-
shadowing technique, showing extruding strands of double-stranded RNA compared to those negatively stained
with uranyl acetate (insets). All micrographs taken at the same magnification. Bar = 100 nm.
FIGURE 7. A scale model of the Fijivirus particle, showing its A and B spikes. A
part of the particle outer shell (O) and one of the A spikes has been removed to expose
the inner particle core (C) and structure of the B spikes. Bar = 20 nm. (From Hatta,
T. and Francki, R. I. B., Virology, 76, 797, 1977. With permission.)
Some data on the polypeptides composing the capsids of plant Reoviridae are shown in
Table 4. Seven polypeptides have been separated from the WTV capsid37 and the presence
of corresponding gene products has been deduced from in vitro and in vivo transcription
and translation experiments53 (Table 4). Two of the capsid proteins (II and IV) could be
removed using trypsin or chymotrypsin. There was then no loss of infectivity, but the particles
appeared in the electron microscope to have lost an amorphous outer layer while retaining
the structural characteristics of the outer shell. Peptides VI and VII were lost if the particles
were sedimented in CsCl gradients, and electron microscopy indicated that the particles were
stripped down to the inner capsids. From these results Reddy and MacLeod" suggested that
peptides II and IV formed the outermost layer of the particle, peptides VI and VII made up
the remainder of the outer capsid, and peptides I, III, and V constituted the inner shell.
Table 4
THE CAPSID POLYPEPTIDES OF
SOME PLANT REOVIRIDAE
Estimated MT x 10-'
Peptides WTV 37 RDV54 MRDV 22
I 160" 155" 193" 139"

II 131 130 152 126
III 118 108 131 123
IV 96 ~78 110 111
V 58 57 62 97
VI 36 42 46 64
VII 35 41.5 45 —
11
Determined by polyacrylamide-gel electrophoresis.22 " 54
b
Determined from in vitro and in vivo transcription and
translation experiments. 53
Earlier, Lewandowski and Traynor38 had detected only four polypeptides with M, of about
156, 122, 63, and 44 x 103 in their WTV preparations, but they had used CsCl density-
gradient centrifugation in their purification procedure. It seems that the first three of their
polypeptides correspond to polypeptides I, III, and V in Table 4. The fourth may correspond
to either VI or VII, or perhaps both as these are of similar size and may not have been
resolved under the conditions used.
Nakata and his colleagues54 detected seven polypeptides in preparations of RDV, with
molecular weights similar to those reported for WTV (Table 4). As with WTV, polypeptides
II and IV could be removed with chymotrypsin. These two polypeptides and polypeptides
VI and VII were also lost from particles sedimenting through CsCl.
Thus, the stable inner shells of Phytoreovirus particles, containing the genomic double-
stranded RNA, appear to consist of polypeptides I, III, and V, while the remaining poly-
peptides are used to build the outer shell. However, it should be remembered that this picture
is based only on PAGE studies. Some of the larger polypeptides detected could be aggregates
of smaller ones, or the smaller ones partial digests of the larger ones, as has been found in
studies of the protein coat of, for example, cauliflower mosaic virus (Chapter 2).
Currently available information on the proteins of Fijivirus particles is more fragmentary
due to difficulties in preparing adequate samples of the particles and their subfractions.
Boccardo and Milne22 separated six'polypeptides by PAGE from MRDV preparations con-
taining high proportions of IPs, three (I, II, and III) from preparations of MRDV SVPs with
B spikes, and only two (I and II) from SVPs devoid of B spikes (Table 4).
Van der Lubbe and his colleagues27 separated three polypeptides from preparations of
FDV SVPs with B spikes, but the protein composition of the IPs could not be investigated
because of their instability.45 The three SVP polypeptides appeared to be of Mr 145 to 150,
125 to 129, and 36 to 39 X 103. The two larger polypeptides could correspond to two of
the polypeptides I to III of MRDV (Table 4), but the third appears to be much smaller than
any of the six polypeptides isolated from MRDV.
With WTV, it has been shown that two enzyme activities are associated with the inner
shell of the particle. An RNA-dependent RNA polymerase (transcriptase) synthesizes a full
length single-stranded RNA copy (presumably the messenger RNA) of each of the 12 double-
stranded RNA genome segments.55'57 The second enzyme methylates the 5' ends of each of
the 12 single-stranded RNA transcripts.58-59 Purified RDV particles also contain a transcrip-
tase, but its location within the virus particles was not determined.60
Transcriptase activity presumably associated with Fijivirus particles was not detected in
Volume I 57
purified preparations of FDV SVPs probably because of damage to the particles during
extraction and purification. 61 The enzyme was, however, detected in freshly prepared extracts
from galls of FDV-infected sugarcane leaves, though not in extracts of healthy plants. 6 ' The
activity sedimented at the same rate as the SVPs, but was detectable only during the first
15 to 20 min of incubation. Although the single-stranded RNA products annealed to FDV
genomic RNA, they were a heterogeneous mixture of low-molecular-weight molecules. This
was not altogether surprising as the reaction took place in relatively crude extracts that must
have contained ribonucleases and other degradative enzymes.
B. Viral Genomes
Figure 2 compares the numbers and estimated molecular weights of the genomes of the
plant Reoviridae that have been studied (see References 8, 10, 15, 17, 29, 30, 33, 36, and
50). It can be seen that the RNAs of the Phytoreoviruses, Fijiviruses, and of RRSV are
quite distinct. The genome of LEV has not been investigated, but is likely to be indistin-
guishable from that of OSDV and ABDV, because LEV resembles these viruses very closely
in all other properties investigated.26'36 Although the fully functional genome of WTV consists
of 12 segments, Reddy and Black62-63 showed that long culture of isolates by vegetative
propagation in crimson clover caused deletions within certain segments, or even loss of
complete segments. Such isolates partially or totally lost their ability to be transmitted by
vectors or to replicate in vector cell monolayers. Nine of 16 such mutants were shown to
have deletions in the largest RNA segment (segment 1) and the others were in segments 4
and 10. Mutants that had completely lost segments 2 or 5 could still replicate in sweet
clover, producing characteristic tumors.
WTV appears to have a genome strategy similar to that of Reovirus. In WTV-infected
vector cell monolayers, 12 polypeptides corresponding to the 12 genome segments have
been identified.53 The twelve mRNAs responsible for producing these polypeptides have also
been detected in vivo. 53 The other plant Reoviridae can be expected to have similar strategies,
though no data on the question are so far available.
III. CYTOPATHOLOGY
The histopathology and cytopathology of the plant Reoviridae have been reviewed peri-
odically. 2 ' 4 ' 16 - 6466 The particles of all member viruses, with the exception of RRSV, 49 have
a similar appearance in thin sections of infected cells (compare Figures 8 and 9). They are
about 70 nm in diameter, with a densely stained core about 45 nm in diameter that represents
the location of the viral RNA. 67 The mature virus particles are always found in the cytoplasm
and may or may not form crystalline arrays. Cells infected with the plant Reoviridae (and
indeed with all members of the Reoviridae)4 contain large cytoplasmic inclusions bodies or
viroplasms. Cells infected with the plant Reoviridae can also contain tubular structures in
the cytoplasm, at least on some occasions and in some hosts.
A. Phytoreovirus
RDV differs from all the other plant Reoviridae in not inducing cell hyperplasia. RDV-
infected plants are stunted and develop chlorotic patches, like Gramineae infected by a wide
variety of viruses. The parenchyma cells as well as the phloem tissues contain virus particles
and viroplasms.68'70 WTV and RGDV appear to be confined to neoplastically developed
phloem-derived tissue.71
Cells infected by each of the three viruses have a similar appearance, and the general
features are also similar whether the cells are those of plant or insect hosts.2-3'"-68-70-71"74
Viroplasms consisting of fibrillar and amorphous material, unbounded by any membrane
and about the size of nuclei, occur in the cytoplasm. Embedded in the viroplasms are dense
FIGURE 8. A, Thin section of a Fiji disease virus (FDV)-infected sugarcane gall cell showing virus particles
scattered in the cytoplasm and the edge of a viroplasm with darker (D) and lighter (L) staining regions. Bar =
500 nm. B, High magnification of thin sectioned FDV particles. Bar = 100 nm.
Volume I 59
FIGURE 9. A, Thin section of a rice ragged stunt virus (RRSV) infected rice gall cell showing virus particles
scattered throughout the viroplasm and a few particles in the cytoplasm (arrows). Bar = 500 nm. B and C show
high magnifications of sectioned particles of RRSV and oat sterile dwarf virus (OSDV), respectively, at the same
magnification. Bar = 50 nm.
particles, about 50 nm in diameter, that are probably immature virus particles. On cytological
grounds, it is generally considered that the viroplasms are the sites of virus synthesis.75
Most of the studies on the structure of Phytoreovirus-mfected cells were done in the 1960s
using techniques less sophisticated than at present available. The published micrographs are
therefore not very informative, and Phytoreovirus cytopathology could well do with
reinvestigation.
B. Fijivirus
The histopathology and cytopathology of cells infected by each of the Fijiviruses are
similar, and have been reviewed regularly.2 ~ 4 - 16 - 76 - 77
Histologically, the neoplastic growths induced by Fijiviruses are derived from the phloem,
though xylem tracheids may also be involved. Only the meristematic tissues respond to
infection by abnormal division and expansion of cells, and after a short while the enations
or galls cease growth. This has been recently documented for FDV78-79 and appears to differ
from the behavior of tumors induced by WTV, which can continue growing for much longer
periods.6-13-14-80 The difference in tumor growth characteristics may well be related to the
physiological differences between dicotyledonous and monocotyledonous plants.
MRDV and OSDV have been observed to induce cell wall proliferations sometimes
enclosing virus particles,81"84 and lysosome-like bodies have also been described in MRDV-
infected plant cells.82
The cytopathology of the viroplasms induced by Fijiviruses has been widely stud-
ie(j 2-4,16.64-66.82-86 when seen in thin sections, the viroplasms are separated into zones con-
taining two different kinds of material (Figure 10). The lighter-staining areas (Figure 11 A)
consist of intertwined strands that fit the description of the kinky filaments found in Reovirus-
infected cells.87 89 The darker areas contain structures that are annular in profile and about
55 nm in diameter and seem to be empty immature inner shells of the virus particles (Figure
11B). Other similar particles with electron-dense cores are likely to be the same shells
charged with nucleic acid (Figures 10 and 12).
Mature particles are seen in the cytoplasm outside the viroplasms (Figures 8 and 10
through 12). With some viruses such as MRDV, the particles can aggregate into crystalline
arrays (Figure 13), which however are not seen in cells infected with viruses such as
pj-)y 78.9o.9i yne consistent presence of viroplasms in infected cells and the fact that they
appear to contain maturation stages of the virus has suggested, as with WTV, that they are
the sites of virus synthesis. This suggestion has been strengthened by autoradiographic studies
showing that 3 H-uridine is first seen to be incorporated into viroplasms and only later into
the virus particles.84-92 Further support has come from a study of the viroplasms in FDV-
infected cells, using light and electron microscopy in conjunction with cytochemical ex-
periments.78 This work confirmed previous observations that the viroplasms consist mainly
of protein and RNA93-94 and indicated that the more densely staining areas of the viroplasm
contained the bulk of the viroplasm RNA, which is double-stranded.
The "kinky" filaments that make up the lighter areas of the viroplasm, and are also found
to some extent in the dense areas together with the inner shells, appear to be about 7 nm
wide in thin sections (Figure 11 and Figure 14). Milne95 suggested that they were the same
as the helical filaments about 13 nm wide found by Hatta and Francki45 in uranyl acetate-
stained FDV preparations (Figure 15) and subsequently also detected in preparations of all
other Fijiviruses examined. 95 This suggestion is supported by the finding that in freeze-
etched preparations of viroplasms, the filaments in situ are about 13 nm in diameter and
resemble those seen in negative stain.78
The discrepancy in the diameter of the filaments when seen in thin sections and when
otherwise observed (Figure 15) is not yet explained, except that fixation may cause shrinkage
or staining may be incomplete in sections. The function of the filaments is not known. Even
Volume I 61
FIGURE 10. A, Thin section of a sugarcane gall cell infected with FDV showing a prominent viroplasm and
numerous virus particles in the cytoplasm. Bar = 500 nm. B, Higher magnification of sectioned FDV particles
in the cytoplasm. Bar = 100 nm.
FIGURE 1 1 . A, Thin section of a sugarcane gall cell infected with FDV showing the lightly staining region of
the viroplasm with "kinky" filaments (L) and a part of the cytoplasm with scattered complete virus particles. B.
Thin section of the same cell as in A, but showing the dense-staining region of the viroplasm (D) with apparently
empty subviral particles and a part of a lightly staining region of the viroplasm (L). Both regions are adjoining
the cytoplasm rich in ribosomes. Bars = 500 nm.
Volume I 63
their composition is uncertain, though they appear to be made of protein. Those found in
MRDV-infected cells do not react with antiserum to the MRDV SVPs.46
The tubules found in most Fijivirus-'mfected cells (Figures 12 and 14*) also appear to be
a common, though not a universal, feature of infection by members of the Reoviridae, as
they or similar structures have been found with all family members except cytoplasmic
polyhedrosis virus. 4 The tubules may be very frequently found, as with MRDV, OSDV,
and some related viruses, x '- 84 - 86 - 9497 but with FDV they are encountered only very rarely in
the infected insect and not at all in infected plant cells.98 It seems likely that the tubules are
a consequence of infection rather than a vital part of its mechanism.
The tubules are found only in the cytoplasm (Figures 12 and 14), may contain rows of
mature virus particles (Figure 16B), and may be incompletely closed, forming scrolls.77 K 2 8 5 S 6 W
They have been shown by cytochemical methods to consist of protein,94 and in transverse
section their walls appear three-layered (Figure 14B).16 The fine structure of the tubule wall
has been resolved both in thin sections and in uranyl acetate stained preparations; in the
latter it appears as a square lattice repeating every 4 nm (Figure 16A) both with MRDV 16
and OSDV.86 The tubules from MRDV- and OSDV-infected cells did not react with their
respective anti-SVP sera,16-36 but preliminary experiments suggest that the MRDV-related
tubules may react with an antiserum containing antibodies to the outer shell of the virus. 100
C. Rice Ragged Stunt Virus

The cytopathology of RRSV has been investigated in three different laboratories,47 4 9 1 0 1 1 < ) 2
but some details are still not agreed upon.50 The virus multiplies in the insect vector 101 - 102
as well as the plant host (Figure 9), inducing typical Fijivirus-like hyperplasia in the latter,
and forming viroplasms in all infected cells. Tubules containing virus particles have been
seen only in the insect host.
Evidence has been presented that the genome of RRSV is smaller than that of other
Reoviridae, containing only 8 (not 10 or 12) double-stranded RNA segments. 103 However,
more recent studies indicate that there are 10 as in the Fijiviruses.IM There is also some
evidence from negative staining of RRSV-infected plant extracts that the RRSV particle
lacks a complete outer shell and resembles the B-spiked SVPs of Fijivirus or the particles
less their A spikes.49-'04 It was found that the virus-like particles had the same appearance
and dimensions whether they were inside the viroplasms or outside them, free in the cy-
toplasm (Figure 9). This would imply the absence of the final maturation stage undergone
by Fijiviruses, in which they acquire the complete outer shell and move out from the
viroplasm to the cytoplasm.49
Whether images representing Fijivirus-like IPs are present or not in the micrographs of
Hibino101 and Hibino and his colleagues102 is difficult to establish, but they may perhaps be
present in their micrographs of RRSV-infected planthopper cells. A very careful high-
resolution study is required to settle this point, which has some importance in deciding the
taxonomic position of the virus.
* Figures 12 through 15 follow text, beginning on page 64.

FIGURE 12. Thin section of a gall cell from an oat plant infected with OSDV showing a large viroplasm consisting
of a light-staining region (L) surrounded by a darker-staining area (D) containing densely staining virus particles.
Arrows indicate longitudinally sectioned tubules. Bar = 500 nm.
Volume I 65
FIOURK 13. A. Thin section of a root cell of maize infected with MRDV showing a typical viroplasm with light
( L ) and dark (D) stained regions and virus particles in the cytoplasm, most of which are forming a crystal (Cr).
Bar = I (J.H1. B. Higher magnification of a cytoplasmic crystal formed from MRDV particles. Bar = 500 nm.
(Courtesy of Dr. A. Appiano. Instituto di Fitovirologia Applicata, Torino. Italy.)
FIGURE 14. A, Thin section of a portion of a gall cell from a maize plant infected with MRDV showing the
edge o f a viroplasm with "kinky" filaments of the light-staining zone (L) and a dark-staining zone (D) with suhviral
particles. In the cytoplasm, complete virus particles can he seen as well as tubules sectioned transversely (t) and
forming open scrolls (o). B, Cytoplasmic tubules in the cytoplasm of an OSDV-infected cell cut in transverse
section and showing their three-layered structure (arrows). Bars = 100 nm.
Volume I 67
FIGURE 15. A and B, Negatively stained and shadowed preparations, respectively, of FDV subviral particles
and filaments thought to he the same as the "kinky" filaments in a thin section of a RRSV-infected cell. Bars =
100 nm.
FIGURE 16. A, Intact and subviral particles of MRDV and a cytoplasmic tubule from an infected cell, stained
with uranyl acetate. B, Thin section of cytoplasmic tubules in a cell infected with MRDV: some tubules contain
intact particles. Bars = 100 nm.
Volume I 69
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In vitro transcription of wound tumour virus RNA by virion-associated RNA transcriptase, Virology, 80,
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Volume I 71
64. Shikata, E., Cytopathological changes in leufhopper vectors of planl viruses, in Leafhopper Vectors and
Plant Disease Agents, Maramorosch. K. and Harris, K. F.. Eds., Academic Press. New York, 1979, 309.
65. Harris, K. F., LealTioppers and aphids as biological vectors: vector-virus relationships, in Leu/hopper
Vectors and Plant Disease Agents, Maramorosch. K. and Harris, K. P.. Eds.. Academic Press. New York,
1979. 217.
66. Shikata, E., Rice viruses and MLOs. and leafhopper vectors, in Leufltop/>er Vectors and Plain Disease
Agents, Maramorosch, K. and Harris. K. F.. Eds.. Academic Press, New York, 1979, 515.
67. Hatta, T. and Francki, R. I. B., En/.yme cytochemical identification of single-stranded and double-
stranded RNAs in virus-infected plant and insect cells. Virology, 88, 105, 1978.
68. Fukushi, T., Shikata, E., and ki in lira. I., Some morphological characters of rice dwarf virus. Virolog\,
18, 192. 1962.
69. Hirai, T., Suzuki, N., Kimura, I., Nakazawa, M., and Kashiwagi, Y., Large inclusion bodies associated
with virus diseases of rice. Phytopathology. 54, 367, 1964.
70. Shikata, E., Electron microscopic studies in plant viruses. ./. Few. Agric. Hokkaido Univ., 55, 1, 1966.
71. Shikata, E. and Maramorosch, K., An electron microscopic study of plant neoplasia induced by wound
tumor virus, J. Natl. Cancer lust., 36, 97. 1965.
72. Shikata, E., Orenski, S. W., Hirumi, H., Mitsuhashi, J., and Maramorosch, K., Electron micrographs
of wound-tumour virus in an animal host and in a plant tumour. Virology, 23, 441, 1964.
73. Shikata, E., Electron microscopic studies of rice viruses, in Proc. Svmp. Virus Diseases of the Rice Plant.
Los Banos, Philippines, Johns Hopkins Press, Baltimore, 1969, 223.
74. Granados, R. R., Ward, L. S., and Maramorosch, K., Insect viremia caused by a plant-pathogenic
virus: electron microscopy of vector hemocytes. Virology, 34, 790. 1968.
75. Shikata, E. and Maramorosch, K., Electron microscopy of wound tumor virus assembly sites in insect
vectors and plants. Virology, 32, 363. 1967.
76. Harpaz, I., Maize rough dwarf. A Plant/topper Virus Disease Affecting Maize, Rice, Small Grain and
Grasses, Israel Universities Press. Jerusalem, 1972, 251 pp.
77. Martelli, G. P. and Russo, M., Plant virus inclusion bodies. Adv. Virus Res., 21, 175, 1977.
78. Hatta, T. and Francki, R. I. B., Development and cytopathology of virus-induced galls on leaves of
sugarcane infected with Fiji disease virus, Physiol. Plant PathoL, 19, 337, 1981.
79. Hatta, T., Boccardo, G., and Francki, R. I. B., Anatomy of leaf galls induced by some Reoviridae and
by wallaby ear disease. Physiol. Plant Pathol.. 20. 43. 1982.
80. Black, L. M., Physiology of virus-induced tumors in plants, mHandbuchder Pjlanzenphysiologie, Ruhland,
W., et al., Eds., Vol. 152, 1965. 236.
81. Bassi, M., Favali, M. A., and Appiano, A., An autoradiographic study of the cell wall modifications
induced by maize rough dwarf virus in the host cells, Riv. Patol. Veg., 10, 19. 1974.
82. Appiano, A. and Lovisolo, O., Ultrastructure of maize roots infected with maize rough dwarf virus and
presence of virus particles in vacuoles with lysosomal activity, Microbiologica, 2, 37, 1979.
83. Break, J. and Kralik, O., Cytopathic effects of oat sterile dwarf virus in enation cells of oat leaves, Ada
Virol.. 24, 346, 1980.
84. Favali, M. A. and Lotti, M., Oat leaves infected with oat sterile dwarf virus (OSDV): an ultrastructural
and autoradiographic study, Microbiologica, 4, 435. 1981.
85. Shikata, E. and Kitagawa, Y., Rice black-streaked dwarf virus: its properties, morphology and intracellular
localization. Virology, 77, 826. 1977.
86. Milne, R. G., Electron microscopy of thin sections of Italian ryegrass infected with both ryegrass cryptic
virus and oat sterile dwarf virus, Microbiologica, 3, 333. 1980.
87. Dales, S., Gomatos, P. J., and Hsu, K. C., The uptake and development of reovirus in strain L cells
followed with labelled viral ribonucleic acid and ferritin-antibody conjugates, Virology, 25, 193, 1965.
88. Murphy, F. A., Coleman, P. G., Harrison, A. K., and Gary, G. W., Colorado tick fever virus: an
electron microscopic study. Virology, 35, 28, 1968.
89. Walker, E. R., Friedman, M. H., and Olson, N. O., Electron microscopic study of an avian reovirus
that causes arthritis, J. Ultrastruct. Res., 41, 67, 1972.
90. Hatta, T. and Francki, R. I. B., Anatomy of virus induced galls on leaves of sugarcane infected with
Fiji disease virus and the cellular distribution of virus particles, Physiol. Plant Pathol., 9, 321, 1976.
91. Hatta, T., Francki, R. I. B., and Home, R. W., Intracellular arrays of Fiji disease virus particles with
five-fold symmetry, Micron, 12, 349, 1981.
92. Favali, M. A., Bassi, M., and Appiano, A., Synthesis and migration of maize rough dwarf virus in the
host cell: an autoradiographic study. J. Gen. Virol., 24, 563, 1974.
93. Giannotti, J. and Monsarrat, P., Etude histologique et histochimique des tumeurs foliaires des Cannes a
sucre atteintes de la maladie de Fidji, An Epiphyt., 19, 707, 1968.
94. Bassi, M. and Favali, M. A., Electron microscopy of maize rough dwarf virus assembly sites in maize.
Cytochemical and autoradiographic observations, J. Gen. Virol., 16, 153, 1972.
95. Milne, R. G., Structure of the viroplasm of the niai/e rough dwarf-like viruses. Ann. Ph\topathoL, 9,
333. 1977.
96. Milne, R. G., Kempiak, G., Lovisolo, O., and Miihle, E., Viral nature of Arrhenatherum blue dwarf
(Blauver/wergung von Arrhenatherum elatius). Phyltiput/wl. Z., 79, 315, 1974.
97. Lesemann, D., and Hulh, W., Nachweis von mai/e rough dwarf virus — ahnlichen Fartikeln in Hnationen
von Lolium — Pflan/en aus Deutschland. Pliytit/xilhol. Z., 82. 246, 1975.
98. Hatta, T., unpublished observations, 1981.
99. Giannotti, J. and Milne, R. G., Pangola stunt virus in thin sections and in negative stain. Virology, 80,
347. 1977.
100. Boccardo, G. and Milne, R. G., unpublished observations, 1982.
101. Hibino, H., Rice ragged stunt, a new virus disease occurring in tropical Asia. Rev. Plain Prou-c. Res.,
12. 98, 1979.
102. Hibino, H., Saleh, N., and Roechan, M., Reovirus-like particles associated with rice ragged stunt diseased
rice and insect vector cells. Ann. Phylopalhol. Soc. Jpn., 45, 228, 1979.
103. Boccardo, G. and Milne, R. G., Electrophoretic fractionation of the double-stranded RNA genome of
rice ragged stunt virus, Intervirology, 14, 57, 1980.
104. Kawano, S., Uyeda, I., and Shikata, E., Particle structure and double stranded RNA of rice ragged stunt
virus. J. Fac. AKric. Hokkaido Univ.. 61, 408, 1984.
Volume I 73
Chapter 5
PLANT RHABDOVIRIDAE
This group of viruses, infecting both plants and insects, has been classified by the ICTV
in the family Rhabdoviridae. The family also includes viruses of vertebrates and other
invertebrates. 1 The Rhabdoviridae were established around vesicular stomatitis virus (VSV)
and its close relatives. The family name is derived from the Greek word rhabdos, meaning
rod, in recognition of the rod-shaped particles of its members. Two genera within the family
Rhabdoviridae have been established, Vesiculovirus and Lyssavirus, with VSV and rabies
as the respective type members, but the family members infecting plants have not been
officially classified into genera.
Plant rhabdoviruses have bacilliform or bullet-shaped particles 50 to 95 nm in diameter
and 200 to 500 nm long, sedimenting at about 1000S. The particles are enveloped and each
consists of at least four structural proteins and a single molecule of negative-sense, single-
stranded RNA of Mr about 4 X 106, which can be transcribed by a particle-associated
trauscriptase. The viruses are transmitted by aphids, leafhoppers, lacebugs, or mites, but
never by more than one of these types of vectors. The viruses are propagative in their vectors
and most are transmitted to a restricted range of plant species. There appear to be no reports
of seed transmission. The plant members of the family have been reviewed periodically,2'7
and all the members of the Rhabdoviridae have been comprehensively reviewed in an
extensive monograph.8
At first sight there appear to be very numerous plant rhabdoviruses and the list increases
continually. 1 " 6 However, the majority of the viruses described have not been investigated in
any detail, and many have been categorized purely on the basis of rhabdovirus-like particles
having been observed in plant cells or in leaf-dip preparations from diseased plants. Recent
lists of plant rhabdoviruses have included nearly 70 names. K5 ' 6 Thirty of the viruses described
have been transmitted experimentally, either by mechanical inoculation or with their insect
vectors; they are listed in Table 1 together with some of their properties. A number of these
viruses may eventually prove to be sufficiently closely related to be treated as strains of the
same virus. For example, barley yellow striate mosaic virus (BYSMV) from Italy9 and
northern cereal mosaic virus (NCMV) from Japan10 have many properties in common and
may well be the same virus. Few successful attempts have been made to compare rhabdo-
viruses serologically.4-5
It has been suggested that the plant Rhabdoviridae can be divided into two subgroups
according to the properties of their proteins, the kinetics of their transcriptase activities, and
the site of maturation of their particles.6-47 The subgroups would correspond with the two
existing genera of the animal Rhabdoviridae. Thus, in a proposed subgroup I, broccoli
necrotic yellows virus (BNYV), lettuce necrotic yellows virus (LNYV), Sonchus virus
(SonV), and perhaps some others show similarities to the genus Vesiculovirus. The particles
of viruses in this subgroup are located in the cytoplasm, contain one envelope M protein,
and possess transcriptase activity readily detectable in vitro. In the proposed subgroup II,
viruses such as eggplant mottled dwarf virus (EMDV), potato yellow dwarf virus (PYDV),
sonchus yellow net virus (SYNV), and sowthistle yellow vein virus (SYVV) appear to have
affinities with the genus Lyssavirus. Particles of the viruses in this subgroup accumulate in
the perinuclear space, possess two M proteins (M, and M2) and have low in vitro transcriptase
activity. This idea will doubtless be the stimulus for further experiments and comparisons,
74 At lets of Plant Viruses
Table 1
MEMBERS OF THE PLANT RHABDOVIRUS GROUP"
In vivo Particle
Mechanical site of dimensions
Virus Vector transmission assembly (nm) Ref.
Barley yellow striate Laodelphax striatellin; — Cyt., vpl. 330 x 45" 9

mosaic virus (planthopper)
(BYSMV)
Beet leaf curl virus Piesma quadratum Nuc. 225—350 x 80" 11
(Beta virus 3) (BLCV) (lace bug)
Broccoli necrotic yel- Bre\'U'or\ne brassiccie + Cyt. (plant), 297 x 65" 12
lows virus (BNYV) (aphid) nuc. (aphid)
Carrot latent virus Semiaphis heraclei Nuc. 220 x 70b 13
(CaLV) (aphid)
Cereal chlorotic mottle Nesotiutha pallida Nuc. 230 x 64" 14
virus (CeCMV) (leafhopper)
Coffee ringspot virus Brevipdlpu.f phocnicis + Nuc. 178—224 x 59—76" 15
(CRV) (mite)
Colocasia bobone dis- Torophagus proser- - Nuc. 300— 335b 16, 17
ease virus (CBDV) pine (leafliopper)
Cow parsnip mosaic vi- — + Nuc. 265 x 90" 18
rus (CoPMV)
Cynara virus (CV) — + Cyt. 260 x 75" 19
Digitaria striate virus Sogatella kolophon Cyt. ca. 45 diam" 20
(DSV) (planthopper) 280 x 55J
Eggplant mottled dwarf — + Nuc. 220 x 66h 21
virus (EMDV)
Finger millet mosaic vi- Sogalella longifurci- Nuc. 285 x 80" 22
rus (FMMV) fera (planthopper)
Gomphrena virus (GV) — + Nuc. 230—250 x 75b 23
Ivy vein clearing virus — + Nuc. 325 x 55" 24
(1VCV)
Lettuce necrotic yellows Hyperom\:us lactucae + Cyt. 360 x 52" 25
virus (LNYV) (aphid)
Lucerne enation virus Aphis craccirora Nuc. 250 x 82—89" 26
(LuEV) (aphid)
Maize mosaic virus Peregrinus maiiJis Nuc. 240 x 48" 27
(MMV) (leafhopper)
Maize sterile stunt virus Peregrinus maidis , Cyt. 255—300 x 45" 28
(MMSV) Sogatella kolophon
(leafhopper)
Melilotus latent virus — + Nuc. 300—350 x 80b 29
(MeLV)
Northern cereal mosaic Laoik'lphax striutellus — Cyt., vpl. 300—350 x 60" 10
virus (NCMV) and others
(planthoppers)
Oat striate mosaic virus Graminella nigrifrons Nuc. 400 x 100" 30
(OSMV) (leafhopper)
Parsley rhabdovirus + Cyt. 214 x 87J 31
(PRV)'
Pelargonium vein clear- — + Nuc. 250 x 70" 32
ing virus (PVCV)
Pisum virus (PV) — + Cyt. 240 x 45" 33
Pittosporum vein yel- — + Nuc. 245 x 80" 34
lowing virus (PVYV)
Potato yellow dwarf vi- Aceratogallia sanguin- + Nuc. 380 x 75" 35
rus (PYDV) olenta, Agallia con-
stricta and others
(leafhoppers)
Raphanus virus (RV) — + Cyt. 250—300 x 50—70" 36
Volume I 75
Table 1 (continued)
MEMBERS OF THE PLANT RHABDOVIRUS GROUP"
In vivo Particle
Mechanical site of dimensions
Virus Vector transmission assembly (nm) Ref.
Raspberry vein chlorosis Aphis idaei (aphid) Cyt. 430 x 65" 37

virus (RVCV)
Rice transitory yellow- Nepholettix apicalis — Nuc. (plant). 180—210" x 94 38
ing virus (RTYV) and others cyt. (hopper)
(leafhoppers)
Sonchus yellow net vi- Aphis coreopsidis + Nuc. 75—80 diam" 39
rus (SYNV) (aphid) 248 x 94"
Sonchus virus (SonV) — + Cyt. 270—350 x 45—57" 40
Sowthistle yellow vein Hyperomyzus lactucae — Nuc. 220 x 80h 41
virus (SYVV) (aphid)
Strawberry crinkle virus Chaetosiphon fragae- Cyt. 190—380 x 69" 42
(StCV) folii and others
(aphids)
Wheat chlorotic streak Laodelphax striatel/us Cyt. 355 x 55" 43
virus (WCSV) (planthopper)
Wheat rosette stunt vi- Laodelphax striatellus Cyt. 320—400 x 40—54" 44
rus (WRSV) (planthopper) 290—370 x 54—64"
American wheat striate Endria inimica Cyt. 250 x 75" 45
mosaic virus (leafhopper)
(AWSMV)
Winter wheat (Russian) Psammotettix striatus — 260 x 60b 46
mosaic virus and others
(WWMV) (leafhoppers)
* List includes all those viruses which have been transmitted experimentally.
b
Measurements were made on particles in thin sections of plant cells.
c
In a previous publication, 5 this virus was referred to as parsley latent virus (ParL V). This was confusing
because a virus with small polyhedral particles and obscure taxonomic affinity had been described previously
by that name (see Volume 2, Chapter 16, Table 3).
cl
Measurements were made on isolated particles after negative staining.
but cannot be properly evaluated at present for lack of sufficient hard data.
Much can also probably be done to clarify some interrelationships among the rhabdoviruses
using serology. This approach has been neglected with this group of viruses, although
preliminary studies indicate that LNYV, SYVV, SYNV, and EMDV are antigenically dis-
tinct. 48 In similar studies serological relationships have been detected between American
wheat striate mosaic virus (AWSMV) and oat striate mosaic virus (OSMV), and between
maize mosaic virus (MMV) and BYSMV. However, the degree of relationship remains
obscure.49
Rhabdovirus-like particles have often been observed by electron microscopy without any
other information being reported (Table 2), and in these cases it is uncertain whether or not
the viruses are new. All in all, considering Tables 1 and 2, it seems very likely that there
are far fewer individual rhabdoviruses than recorded names.
In a number of cases, rhabdovirus-like particles devoid of envelopes have been detected
in diseased plants. One such particle type, that of orchid fleck virus (OFV), has been
transmitted experimentally.50 Similar particles have been observed in diseased plants of
several orchid species51"55 and in citrus.56 At present, their relationship to rhabdoviruses is
obscure. It may be of interest that Ammar and his colleagues57 found rather similar virus-
like particles in the tissues of the planthopper Javesella pellucida, though there was no
evidence that the particles were transmissible to plants.
Table 2
RHABDOVIRUS-LIKE PARTICLES DETECTED IN PLANTS"
Plant Species Disease Symptoms Plant Species Disease Symptoms
Ananas comosus Chlorotic leaf streak Lolium sp. No clear symptoms

Alropa belladonna Mosaic Lupinus polyphyllus Vein chlorosis
Cajanus cujan Tissue proliferation Manihot esculenta Latent
Callislephus chinensis Chlorosis Phalaenopsis sp. Chlorotic spots
Chondrilla juncea Stunting Phmtago lanceolata Mottle
Chrysanthemum sp. Vein chlorosis Pogostemon patchouli Mottle
Cucumis melo Leaf variegation Ranunculus repens —
Dendrobium hybrid Leaf streak Saintpaulia sp. Leaf necrosis
Euonymus japonica Fasciation Sambucus sp. Vein clearing
Festuca gigantea Leaf streak Sarracenia purpurea —
Gerhera sp. Symptomless Th\mus X citriodorus Leaf chlorosis
Holcus lanatus h
Trifo/ium incarnatum Mosaic
Iris germanicci Leaf stripe Trifolium repens Enations
Laburnum anagvroides Vein chlorosis Triticum aestivum Chlorotic spots
Laclia sp. Red spotting Vigna sinensis Mosaic
Zea mays Mosaic
Compiled from data already reviewed. 1 •"
h
Not specified.
Discussions of plant rhabdovirus structure are found in reviews by Peters and Schultz 58
and Hull, 59 as well as in the more general reviews already cited. 2 8
Rhabdoviruses have the largest and most complex particles of all known plant viruses,
containing about 70% proteins, 25% lipids, 4% polysaccharides, and 1% RNA. Although
the particles often appear bullet-shaped in negatively stained preparations (Figure 1), it is
now generally accepted that the particles of nearly all rhabdoviruses are naturally bacilliform
(Figure 2). This conclusion is based on studies of particle morphology in thin sections of
infected cells and in negatively stained specimens prepared by methods designed to preserve
particle integrity. Nonetheless, the problems of preparative artifacts"-60 make it very hard to
determine the true morphology and dimensions of the particles. Generally, those who stain
rhabdoviruses with neutral phosphotungstate see obviously damaged or bullet-shaped par-
ticles, whereas those using unbuffered uranyl acetate (pH 4.2 to 4.5) may see bacilliform
or pleomorphic particles (Figures 3 to 5) or the nucleocapsids stripped of their envelopes. 2
A great variety of other negative stains could be used, but it seems that for rhabdoviruses
there is no single stain that is totally satisfactory. Fixation with osmium vapors"1 or glutaralde-
hyde62'66 has been used, and bifunctional cross-linking agents67 should also be tried. It should
be noted that fixatives do not always "fix," but can introduce their own array of artifacts.
More realistic images and consistent (if only relative) measurements have probably been
obtained from particles in thin sections (Figure 6), but here shrinkage during fixation and
embedding can be a problem. For example, the dimensions of MMV have been reported as
240 x 48 and 300 X 75 nm, from two different laboratories27-68 using different techniques.
Despite these problems, some kind of portrait of the rhabdovirus particle may be drawn.
A typical rhabdovirus particle such as that of LNYV (Figure 7) consists of an outer envelope
enclosing a long strand of nucleoprotein that is helically wound in about 40 turns to form
a parallel-sided tube with a pitch of 4.0 to 4.5 nm. The envelope has projections 6 to 10
nm long and is acquired from modified host cell membranes during particle maturation.
With LNYV and many other rhabdoviruses,61-69 7 I the structure of the envelope appears as
Volume I 77
UGL'RH I. Partially purified preparation ol hnkxoli nccrotic \ o l l i m \ viru> ( U N Y V ) nosialivcly slaincil \\iih
neutral ammonium molyhdalo usinj; 0.05'/; huoilracin as a woitiiii; agent. Bar = KM) nm. (Courtesy ol Mr. C'. M.
Cla\. National Vegetable Research Station. Wolleshourne. England.)
|-'KiL'RK 2. Sowihisllo yellow \oin virus i S V \ ' \ ' ) allot fixation w i t h silularaklohulo and ncsalixc siainint! uilli
uranyl accialo. Some of llic pailiolos lui\o hcon ili-.lorlcil In llaiu-nin;; a-, shown In iho apparcnlly ihick ciuoln|V->.
Bar - 100 nin. (Courtesy of Dr. h'. \V. Kilujinia. IVparlnicnl of liiolo;j\. l-'undacuo l.'nixorsiUaJo do Brasilia.
Brasilia, and Dr. ['). Pclcrs. Dcpurtnioni of Virology. Auriailtural l'ni\crsiiy. Wa^cninycn. 'Iho Nolhorlaikls.i
Volume I 79
FIGURE 3. Selected lettuce necrotic yellows virus (LNYV) particles distorted during negative staining with uranyl
acetate. Bar = 100 nm. (Courtesy of Drs. T. C. Chambers and B. S. Wolanski, School of Botany. University of
Melbourne. Melbourne, Australia.)
FIGURE 4. A. Bi//are structure formed from LNYV particles stained with potassium phosphotungstate, pH 6.5).
B. Apparently undistorted L N Y V particle stained with uranyl acetate. C. LNYV particles stained in neutral potassium
phosphotungstate. showing invaginations at one or both ends of the particles. D, Barley yellow striate mosaic virus
( B Y S M V ) particles fixed with glutaraldehyde and stained with uranyl acetate, showing undistorted particles pen-
etrated ( l e f t ) and unpenetrated ( r i g h t ) by the stain. Bar = 100 nm. (A and C courtesy of Dr. S. B. Wolanski: B
courtesy of Dr. T. C. Chambers. School of Botany, University of Melbourne. Melbourne. Australia.)
Volume I 81
FIGURE 5. Possible origin of aberrant lettuce necrotic yellows virus particles by various methods of negative
staining. (From Francki, R. I. B., Adv. Virus Res., 18, 257, 1973. With permission.)
a hexagonal lattice; though with some viruses such as BYSMV, for example, repeating
structures on the envelope are not always clearly seen. 72
The nucleoprotein helix is responsible for the typical cross-striations seen on rhabdovirus
particles that have been penetrated by negative stain (Figures 1 through 4). The precise
arrangement of the nucleocapsid strand and the structure of the envelope at the ends of the
particles is not clear and a number of possibilities have been suggested. 2 58-59 It seems that
in some cases the intact nucleocapsid is bullet shaped, with one end domed and the other
truncated, whereas the envelope, when undamaged, is bacilliform (i.e., rounded at both
ends). A good example of this type of structure is seen in thin sections of SYVV 71 (Figure
8A). In some other cases it appears that two nucleocapsids are enclosed inside one envelope
so that the two domed ends face outward and the truncated ends abut in the middle of the
particle (Figure 8B). Such an arrangement is also seen in Figure 3 of the paper by Vela and
Rubio-Huertos74 and that by Peters.73
It has been pointed out previously that where data are available, the particles of plant
rhabdoviruses appear to be longer but not thinner than those of vertebrate rhabdoviruses. 2 - 4
Available sedimentation data also support the view that plant rhabdovirus particles are larger.4
However, the reported sizes of the plant and vertebrate viral RNAs are similar, as are the
molecular weights of the structural proteins2-4 (Table 3). It has been suggested that these
apparently conflicting sets of data might be reconciled by supposing that vertebrate rhab-
dovirus particles consist of one bullet-shaped nucleocapsid surrounded by an envelope whereas
most plant rhabdovirus particles consist of two nucleocapsids enveloped together (Figure
9). However, this view only receives partial support from the results of thin sectioning. 3
Rhabdoviruses have high sedimentation coefficients but low buoyant densities due to the
envelopes containing lipid (Table 3). The particles can be dissociated by detergents in two
ways. Nonionic detergents such as Triton X-100® or Nonidet P40® remove the envelope to
expose a more-or-less intact nucleocapsid which is still infectious.60 On the other hand, ionic
detergents such as sodium dodecyl sulfate dissociate both the envelopes and nucleocapsids
FIGURE 6. Thin section of pelargonium vein clearing virus (PVCV) in the perinuclear space of an infected
Nicotiana benthamiana leaf cell. Most of the particles in the longitidunal section appear to be bacilliform: others
in the transverse section show the various particle layers. Bar = 100 nm. (Courtesy of Drs. G. P. Martelli and
M. Russo, Istituto di Patologia Vegetale, Universita degli Studi, Bari, Italy.)
Volume 1 83
FIGURE 7. Model of a rhabdovirus particle cut through

the middle to expose the internal structures. Bar = 10 nm.
(After Francki. R. 1. B. and Randies. J. W., in Rhabdo-
V;>M.«'.V, Vol. 3. Bishop, D. H. L., Ed., CRC Press, Boca
Raton, Fla., 1980, 135. With permission.)
to yield noninfectious RNA and a number of proteins (Figure 10). 77 Dissociation studies
accompanied by protein analyses using PAGE have shown that the rhabdovirus envelope
contains either two or three proteins. All the viruses contain a glycosylated protein (G
protein) which makes up the projections on the surface of the envelope. Some plant rhab-
doviruses contain one and others two envelope matrix proteins (M proteins) (Figure 7 and
Table 4). Although the viral envelope buds from cellular membranes during particle assem-
bly, the M protein(s) appear to be virus specified. The nucleocapsids contain two to three
proteins; one, the N protein, which is associated with the nucleic acid, forms the characteristic
coiled strand. The other two proteins (NS and L) are present in only small amounts and
their exact locations are obscure. There is a fair range in the moicular weights of the proteins
from different viruses (Table 4).
B. The Viral Genome

Rhabdoviruses contain only one species of single-stranded RNA of M, about 4 X 10''
which is not infectious (Table 3). The lack of infectivity indicates that the RNAs have
negative sense — a conclusion supported by the detection of RNA-dependent RNA poly-
merase (transcriptase) activity associated with the virus particles. At present this enzyme
has been detected only in preparations of LNYV1*4"87 and BNYV. S Searches for the enzyme
in preparations of PYDV, SYVV, and SYNV have been unsuccessful. m The reason for this
is not understood, but could be due to a number of factors which have been discussed in
detail elsewhere.4-89 The difficulties of detecting transcriptases associated with the plant
rhabdoviruses may parallel the experiences with some vertebrate rhabdoviruses such as
rabies. Here, efforts to demonstrate enzyme activity were fruitless until low levels were
finally demonstrated.90-"1
The mechanism of cell infection by plant rhabdoviruses is not well understood, but it is
FKiUKI: X. A. Thin section ol' S Y VV particles in the pcrinuclcur space of a Soiitiius olcraceus leaf cell showing
that many of (he particles have bullet-shaped micleocapsids (arrows) (courtesy of Dr. L. L. Hoefen. Agricultural
Research Service. U.S. Department of Agriculture. Salinas. Calif.). B, Rhatxloxirus-like particles in a cell of
diseased Trifuliiun inairmiiiim. showing what appear to he two nucleocapsids within each viral envelope (courtesy
of Dr. M. Kuhio-lluertos. Jaime l-'erran Institute of Microbiology, Madrid. Spain). Bars = 500 nm.
Volume I 85
Table 3
PHYSICAL PROPERTIES OF THE BETTER CHARACTERIZED
PLANT RHABDOVIRUSES
Lettuce Potato Sonchus American wheat Broccoli

necrotic yellow yellow net striate mosaic necrotic
yellows virus dwarf virus virus virus yellows virus
Property (LNYV) 25 - 76 - 77 (PYDV) 3578 (SYNVr-66 (AWSMV)4"'-8" (BYNVp
Dimension of virus particles (nm)

In sections 360 x 52 380 x 75 —" 250 x 75 297 x 64
In negative stain 221 x 66 — 250 x 94 245 x 75 275 x 75
Sedimentation coefficients,, iJ 945 880 1045 875 874
Buoyant density (gm/cm 1 ) 1.20 1.17 1.18 1.22 1.19
Nucleocapsids
Sedimentation 260 250 250 — —
coefficient(S 20 J
Infectivity Yes Yes Yes — —
Ribonucleic acid
Sedimentation 43 45 44 32 —
coefficients^ )
Strandedness Single Single Single Single —
Infectivity No No No No —
M, ( x 10") 4.2 4.6 4.4 2.2 —
" Data not available.
FIGURE 9. Diagrammatic representation of possible structural dif-

ferences in the assembly of animal (e.g., vesicular stomatitis virus,
VSV) and plant rhabdoviruses (e.g., lettuce necrotic yellows virus,
LNYV). (From Francki. R. I. B. and Randies, J. W., in Rhabdo-
viruses, Vol. 3, Bishop, D. H. L., Ed.. CRC Press, Boca Raton. Fla..
1980, 135. With permission.)
interesting to note that the in vitro transcription of a virus such as LNYV appears to be very
similar to that of VSV. 84 - 86 - 89 Thus it seems likely that the genome strategies of the two
viruses are similar; that of VSV is well known. 8 Jackson and his colleagues have demonstrated
that SYNV-infected plants contain RNAs with base sequences complementary to the virus
and of similar sizes to those in VSV infected cells. 9 -- 93
III. CYTOPATHOLOGY
When cells of plants infected with rhabdoviruses are examined in thin sections, numerous
characteristic particles sectioned at various angles are readily detected. Particles of many of
FIGURE 10. Diagrammatic representation of the disassembly of let-

tuce necrotic yellows virus (LNYV) with nonionic and ionic detergents.
(Modified from Francki and colleagues. 5 )
Table 4
THE STRUCTURAL PROTEINS OF SOME PLANT
RHABDOVIRUSES
Virus L G N NS M, M2 Ret.
Potato yellow dwarf +a 78 56 -h 33 22 81

virus (PYDV)
Sowthistle yellow '> 83 60 44 36 82
vein virus (SYVV)
Sonchus yellow net + 77 64 44 39 66
virus (SYNV)
Eggplant mottled •> 83 61 27 21 47
dwarf virus
(EMDV)
Lettuce necrotic 170 71 56 38 19 47
yellows virus
(LNYV)
Sonchus virus (SonV) 170 72 55 38 19 47
American wheat 145 92 59 25 83
striate mosaic virus
(AWSMV)
" Detected but molecular weight not determined.

h
Not detected.
the viruses accumulate in the perinuclear space (Figures 6, 8, and 11), whereas those of
others accumulate in vesicles of the endoplasmic reticulum (Figures 12 and 13). In most
cases examined, as Peters6 has pointed out, the sites of maturation and accumulation of the
particles in vectors appear to be the same as in the plant hosts. However, BNYV seems to
accumulate in the perinuclear space in insects but mainly in the cytoplasm in plants. 94 The
reverse has been reported for rice transitory yellowing virus (RTYV). 95 With a few of the
Volume 1 87
FIGURE 1 1 . A. Thin section of a nucleus of a Nicotiana labticum leaf cell infected with eggplant mottled dwarf
virus (EMDV) showing virus particles accumulating in the perinuclear space. Bar = 500 nm. B, EMDV particles
budding (arrows) from the inner nuclear membrane to be released into the perinuclear space. Bar = 100 nm.
(Courtesy of G. P. Martelli and M. Russo. Istitituto de Patologia Vegetale, Universita degli Studi. Bari. I t a l y . )
FIGURE 12. A, Thin section of a Nicotianu clevelandii leaf cell infected with LNYV showing particles in vesicles
of the endoplasmic reticulum, from which some of them are budding (arrows). Bar = 500 nm. B, B N Y V particles
budding (arrows) from the endoplasmic reticulum. Bar = 100 nm. (Courtesy of R. G. Garrett. Victorian Plant
Research Institute, Melbourne. Australia.)
Volume I 89
FIGURE 13. A, Thin section odiNicotianu clevelundii leaf cell infected with LNYV showing numerous particles
in vesicles of the entioplasmic reticulum and a region with virus-specific thread-like structures (T). Bar = 500
nm. B, Higher magnification of the cytoplasmic thread-like structures (T) adjoining a nucleus. Bar = 100 nm.
"cytoplasmic" rhabdoviruses such as BYSMV, the particles accumulate in plant cells around
the edges of granular inclusions (Figure 14) usually referred to as viroplasms 9 (Table 1).
The viroplasms associated with BYSMV infection are composed largely of protein but are
apparently not the sites of viral RNA synthesis or accumulation. 96
Virus particles which accumulate in the perinuclear spaces are often seen budding from
the inner nuclear membrane (Figure 11), and nucleocapsids are sometimes seen in the
nucleoplasm. It appears that the nucleocapsids acquire their envelopes from the inner nuclear
membrane while passing from the nucleoplasm into the perinuclear space (Figure 1 IB). In
some instances the particles accumulate in such large numbers that the perinuclear inclusions
can be easily seen by light microscopy. 97 Besides the distortions in the perinuclear spaces
caused by the accumulation of virus particles, internal nuclear disorganization may also take
place involving the disappearance of chromatin, swelling of the nucleolus, and appearance
of viroplasm-like structures. 98 SYNV is one of the rhabdoviruses whose particles accumulate
in perinuclear spaces.97 Biochemical studies with this virus have shown that RNA with base
sequences complementary to the viral RNA is present in association with both free and
membrane-bound polyribosomes from cytoplasmic fractions, but no complementary viral
RNA was detected in isolated nuclei. 91 This suggests that virus-specific proteins synthesized
in the cytoplasm are transported into the nucleus for assembly into virus particles.
Particles of those viruses that accumulate in the cytoplasm appear to bud from membranes
of the endoplasmic reticulum (Figures 12 and 13). However, it seems that at early stages
of infection by at least some of these viruses, the nucleus is also involved in events leading
to virus synthesis. For example, by sampling the first systemically infected leaf at various
times after inoculation with LNYV, Wolanski and Chambers99 followed the incorporation
of 'H-uridine into the infected cells by autoradiography at the light microscope level. At
early stages of infection, incorporation occurred mainly in the nucleus, but later most of the
label was detected in the cytoplasm. Electron microscopy showed that nuclei underwent
significant changes during the early stages of infection. The outer nuclear membrane de-
veloped "blisters" containing small vesicles and, later, occasionally virus particles also
appeared there (Figure 15). With BYSMV, 'H-uridine incorporation and electron microscope
autoradiography produced evidence for viral RNA synthesis in the cytoplasm only, even at
early stages, whereas RNA synthesis in the nucleus was depressed.96
It would appear that at early stages of infection, most rhabdoviruses are assembled in the
perinuclear space; those that will later accumulate in the cytoplasm acquire their envelopes
from the outer nuclear membrane which is continuous with the endoplasmic reticulum (Figure
16). On the other hand, those which will accumulate largely in the perinuclear space acquire
their envelopes from the inner membrane of the nuclear envelope. MMV particles budding
from endoplasmic reticulum in direct continuity with the nuclear envelope can be seen in
Figure 16 of the review by Martelli and Russo. 1
In plants infected with several rhabdoviruses, such as LNYV, melon variegation virus
(MVV), and BYSMV, whose particles accumulate in the cytoplasm, long thread-like struc-
tures have been observed in the cytoplasm, sometimes forming extensive masses (Figure
13). The exact structure and function of these is unknown, but it seems that they may be
unenveloped nucleocapsid materials. 100
A number of plant rhabdoviruses have been studied by electron microscopy in their insect
vectors, in which they undoubtedly multiply, and the evidence for this has been discussed
elsewhere. 2 - 4 These viruses are therefore just as much insect as plant viruses, though there
is no evidence that the vector insects suffer any disease.
Several viruses whose particles resemble those of rhabdovirus nucleocapsids, such as those
ol'OFV (Figure 17*). have been detected in infected plant cells. 50 Wl The particles have been
* Figures 17 through 19 1'ollow (e\t, on pages 94 through 96.

Volume 1 91
FIGURE 14. A, Thin section of a barley leaf cell infected with barley yellow striate mosaic virus (BYSMV)
showing a granular inclusion (viroplasm, Vp) and virus particles, some of which appear to be in the process of
budding from the viroplasm (arrows). Bar = 500 nm. B, Higher magnification of BYSMV particles budding
(arrows) from the viroplasm. Bar = 100 nm. (Courtesy of D. A. Appiano, Istituto di Fitovirologia Applicata, C.
N. R., Torino, Italy.)
FIGURE 15. A, Thin section of a Nicotiana clevelandii leaf cell infected with LNYV showing the development
of perinuclear blisters containing small vesicles and sometimes virus particles. The blisters are formed by distension
of the outer nuclear membrane. Bar = 500 nm. B, Higher magnification of a perinuclear blister containing vesicles
induced by LNYV infection. Bar = 100 nm.
Volume 1 93
FIGURE 16. Diagrammatic representation of the synthesis and cellular distribution of a rhabdovirus such as
LNYV or maize mosaic virus (MMV). (Modified after Francki. 2 )
detected both in nuclei and cytoplasm (Figures 18 and 19), and in some cases shown to be
associated with electron-lucent viroplasms. Many particles are usually seen butting up against
the inner nuclear membrane or cytoplasmic membranes, making indentations in the mem-
brane but without becoming enveloped (Figures ISA and 19). Regions of the nuclear mem-
brane where a number of particles converge often protrude outwards to form "blebs". It
seems that some of these blebs may pinch off from the nuclei to form rounded "spokewheel"
structures which are liberated into the cytoplasm (Figure 18B).
FIGURE 17. A anil B. Purified preparations of virus-like particles, negatively stained with ammonium molybdate,
isolated from diseased Dcntlrohiiiin phalacnopsis. The particles resemble nucleocapsids ol rhabdoviruses. I5ars =
100 nm. (Courtesy of D.-li. Lesemann. Institut fiir Viruskrankheiten der Pllanzen. Braunschweig. West Germany.)
Volume 1 95
I-'IUUKE \X. A. Thin section of an orchid leal'cell inl'ecled with orchid fleck virus showing typical virus particles
associated with membranes in the cytoplasm (Cm) and the inner nuclear membrane (Nm). B. Thin section ol" a
cell as in A. but showing "spokewheel" structures formed from membranes and virus panicles in the cytoplasm.
Bars = 500 nm.
FIGURE 19. A, Preparation as in Figure 18, showing details of the interaction of the nucleocapsids and the inner
nuclear membrane. B, A "spokewheel" structure within a sectioned nucleus. Bars = 100 nm.
Volume I 97
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24. Castellano, M. A. and Rana, G. L., Transmission and ultrastructure of ivy vein clearing virus infections,
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la luzerne (LEV). Premiers resultats, Ann. Phytopathol., 5, 441, 1973.
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30. Milbrath, G. M. and Jedlinski, H., Electron microscopy and purification of oat striate mosaic virus, a
new member of the rhabdovirus group, Proc. Am. Phytopathol. Soc., 3, 253, 1976.
31. Tomlinson, J. A. and Webb, M. J. W., Virus diseases of parsley. Null. Vef>. Res. Stti. Ann. Kept, fur
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32. Di Franco, A., Kus.so. M., and Martelli, G. P., Isolation and some properties of pelargonium vein clearing
virus. Phytopathol. Mctliterr.. 18, 41, 1979.
33. Caner, J., July, J. R., and Vincente, M., Caracteristicas de um rhabdovirus isolads de plantas do crvilha.
Summa Phytopathol., 2, 264, 1976.
34. Rana, G. L. and Di Franco, A., Mechanical transmission of Pittosporum vein clearing virus. Phytopathol.
Mediterr., 18, 48, 1979.
35. Black, L. M., Potato yellow dwarf virus, CMI/AAB Descriptions of Plant Viruses, No. 35. 1970.
36. Kitajima, E. W. and Costa, A. S., Rhabdovirus-like particles in tissues of five different plant species.
Fir. Bnixii. 4, 55, 1979.
37. Jones, A. T., Murant, A. F., and Stace-Smith, R., Raspberry vein chlorosis virus. CMI/AAB Descriptions
38. Shikata, E., Rice transitory yellowing virus, CMIIAAB Descriptions of Plant Viruses, No. 100, 1972.
39. Jackson, A. O. and Christie, S. R., Sonchus yellow net virus, CMIIAAB Descriptions of Plant Viruses.
No. 205, 1979.
40. Vega, J., Gracia, O., Rubio-Huertos, M., and Feldman, J. M., Transmission of a bacilliform virus of
sow thistle: mitochondria modifications in the infected cells, Phvtopathol. Z., 85. 7. 1976.
41. Peters, D., Sowthistle yellow vein virus, CMI/AAB Descriptions of Plant Viruses. No. 62. 1971.
42. Sylvester, E. S., Frazier, N. W., and Richardson, J., Strawberry crinkle virus. CMIIAAB Descriptions
43. Signoret, P.-A., Giannotti, J., and Alliot, B., Particules de type viral chez Triticum durum Desf. presentant
des symptomes de stries chlorotiques, Ann. Phytopathol., 4. 45, 1972.
44. Anon., Nature of the pathogen of wheat rosette stunt disase, Scientia Sin., 22, 573, 1979.
45. Sinha, R. C. and Behki, R. M., American wheat striate mosaic virus, CMI/AAB Descriptions of Plant
Viruses, No. 99. 1972.
46. Razvjaskina, G. M. and Poljakova, G. P., Electron microscopic study of the winter wheat mosaic virus.
Plant Virology Proc. 6th Conf. Czech. Plant Virol. 1967, Czechoslovak Academy of Science. Prague.
1969. 129.
47. Dale, J. L. and Peters, D., Protein composition of the virions of the five plant rhabdoviruses, Iniervirology.
16, 86, 1981.
48. Peters, D., personal communication, 1977.
49. Jackson, A. O., personal communication, 1978.
50. Doi, Y., Chang, M. U. and Yora, K., Orchid fleck virus, CMIIAAB Descriptions of Plant Viruses. No.
183, 1977.
51. Duvel, D. and Peters, K. R., Virusahnliche partikeln in Dendrobium antennatum Ldl., Ganenwelt. 7 1 ,
52. 1971.
52. Petzold, H., Der elektronenmikroskopische Nachweis eines bacilliformen Virus an blattfleckenkranken
Dendrobien, Phytopathol. Z., 70, 43, 1971.
53. Lesemann, D. and Begtrup, J., Elektronenmikroskopischer Nachweis eines bazilliformen Virus in Plui-
laenopsis, Phytopathol. Z., 71, 257, 1971.
54. Kitajima, E. W., Blumenschein, A., and Costa, A. S., Rodlike particles associated with ringspot symp-
toms in several orchid species in Brazil, Phytopathol. Z., 81, 280, 1974.
55. Lesemann, D. and Doraiswamy, S., Bullet-shaped virus-like particles in chlorotic and necrotic leaf lesions
of orchids, Phytopathol. Z., 83, 27, 1975.
56. Kitajima, E. W., Miiller, G. W., Costa, A. S., and Yuki, W., Short, rod-like particles associated with
Citrus leprosis, Virology, 50, 254, 1972.
57. Ammar, E.-D., Milne, R. G., and Watson, M. A., Virus-like particles in the plant hopper Javesella
pellucida Fab., J. Gen. Virol., 6, 315, 1970.
58. Peters, D. and Schultz, M. G., A model of rhabdovirus morphogenesis, Proc. K. Ned. Akad. Wet. Ser.
C, 78, 172. 1975.
59. Hull, R., The structure of tubular viruses, Adv. Virus Res., 20, 1, 1976.
60. Francki, R. I. B. and Randies, J. W., Composition of the plant rhabdovirus lettuce necrotic yellows
virus in relation to its biological properties, in Negative Strand Viruses, Vol. 1. Mahy, B. W. J. and Barry,
R. D., Eds., Academic Press, London, 1975, 223.
61. Wolanski, B. S. and Francki, R. I. B., Structure of lettuce necrotic yellows virus. II. Electron microscopic
studies on the effect of pH of phosphotungstic acid stain on the morphology of the virus, Virologv, 37,
437, 1969.
62. MacLeod, R., An interpretation of the observed polymorphism of potato yellow dwarf virus. Virologv,
34, 771. 1968.
63. Peters, D. and Kitajima, E. W., Purification and electron microscopy of sowthistle yellow vein virus,
Virology, 41, 135, 1970.
Volume I 99
64. Ahmed, M. E., Sinha. R. C., and Hochsler, R. M., Purification and some morphological characters of
wheat striate mosaic virus. Virology, 41, 768, 1970.
65. Russo, M. and Martelli, G. P., A study of the structure of eggplant mottled dwarf virus, Virology, 52,
39, 1973.
66. Jackson, A. O. and Christie, S. R., Purification and some physicochemical properties of sonchus yellow
net virus, Virolog\, 77, 344, 1977.
67. Boccardo, G. and Milne, R. G., Enhancement of the immunogenicity of the maize rough dwarf virus
outer shell with the cross-linking reagent dithiobis(succinimidyl)propionate, J. Virol. Meth., 3, 109, 1981.
68. Martelli, G. P., Russo, M., and Malaguti, G., Ultrastructural aspects of maize mosaic virus in the host
cells. Phytopathol. Mediterr., 14, 140, 1975.
69. Hills, G. J. and Campbell, R. N., Morphology of broccoli necrotic yellows virus. J. Ultrastruct. Res.,
24, 134, 1968.
70. YVolanski, B. S. and Chambers, T. C., Structure of lettuce necrotic yellows virus. HI. Electron micro-
scopical studies of the viral envelope, Virology, 47, 656, 1972.
71. Greber, R. S., Cereal chlorotic mottle virus — a rhabdovirus of Gramineae in Australia transmitted by
Nesocluthu pallida (Evans), Aust. J. Agric. Res., 30, 433, 1979.
72. Conti, M., Vector relationships and other characteristics of barley yellow striate mosaic virus (BYSMV),
Ann. Appl. Bio/., 95. 83, 1980.
73. Steinkamp, M. P. and Hoefert, L. L., Annulate lamellae in phloem cells of virus-infected Sonchus plants,
J. Cell Bio/.. 74, 1 1 1 , 1977.
74. Vela, A. and Rubio-Huertos, M., Bacilliform particles within infected cells of Trifolium incarnatum,
Phytopathol. Z., 79, 343, 1974.
75. Peters, K.-R., Orchid viruses: a new rhabdovirus in Lae/ia red leafspots, J. Ultrastruct. Res., 58, 166,
1977.
76. Chambers, T. C., Crowley, N. C., and Francki, R. I. B., Localization of lettuce necrotic yellows virus
in host leaf tissue, Virology, 27, 320, 1965.
77. Francki, R, I. B. and Randies, J. W., Some properties of lettuce necrotic yellows virus RNA and its I'H
vitro transcription by virion-associated transcriptase, Virology, 54, 359, 1973.
78. Reeder, G. S., Knudson, D. L., and MacLeod, R., The ribonucleic acid of potato yellow dwarf virus,
Virology, 50, 301, 1972.
79. Sinha, R. C., Sehgal, O. P., and Thottappilly, G., Effect of temperature on infectivity and some physico-
chemical properties of purified wheat striate mosaic virus, Ph\topathol. Z., 84, 300, 1975.
80. Sinha, R. C., Harwalkar, V. R., and Behki, R. M., Chemical composition and some properties of wheat
striate mosaic virus, Phytopathol. Z.. 87, 314, 1976.
81. Knudson, D. L. and MacLeod, R., The proteins of potato yellow dwarf virus, Virology, 47, 285, 1972.
82. Ziemiecki, A. and Peters, D., The proteins of sowthistle yellow vein virus: characterization and location,
/. Gen. Virol., 32, 369. 1976.
83. Trefzger-Stevens, J. and Lee, P. E., The structural proteins of wheat striate mosaic virus, plant rhabdovirus,
Virology, 78, 144. 1977.
84. Francki, R. I. B. and Randies, J. W., RNA-dependent RNA polymerase associated with particles of
lettuce necrotic yellows virus. Virology, 47, 270, 1972.
85. Randies, J. W. and Francki, R. I. B., Infectious nucleocapsid particles of lettuce necrotic yellows virus
with RNA-dependent RNA polymerase activity, Virology, 50, 297, 1972.
86. Toriyama, S. and Peters, D., In vitro synthesis of RNA by dissociated lettuce necrotic yellows virus
particles,/. Gen. Virol.. 50, 125, 1980.
87. Toriyama, S. and Peters, D., Differentiation between broccoli necrotic yellows virus and lettuce necrotic
yellows virus by their transcriptase activities, J. Gen. Virol., 56, 59, 1981.
88. MacLeod, R., personal communication, 1983.
89. Francki, R. I. B. and Peters, D., The interference of cytoplasmic membrane-bound material from plant
cells with the detection of a plant rhabdovirus transcriptase. J. Gen. Virol., 41, 467, 1978.
90. Kawai, A., Transcriptase activity associated with rabies virion, J . Virol., 24, 826, 1977.
91. Flamand, A., Delagneau, J. F., and Bussereau, F., An RNA polymerase activity in purified rabies
virions.y. Gen. Virol.. 40, 233, 1978.
92. Milner, J. J. and Jackson, A. O., Sequence complementarity of sonchus yellow net virus RNA with
RNA isolated from the polysomes of infected tobacco, Virology. 97. 90, 1979.
93. Milner, J. J., Hakkaart, M. J. J., and Jackson, A. O., Subcellular distribution of RNA sequences
complementary to Sonchus yellow net virus RNA, Virology, 98, 497, 1979.
94. Garrett, R. G. and O'Loughlin, G. T., Broccoli necrotic yellows virus in cauliflower and in the aphid,
Brevicoryne brassicae L., Virology, 76, 653, 1977.
95. Chen, M.-J. and Shikata, E., Electron microscopy and recovery of rice transitory yellowing virus from
its leafhopper vector, Nephotenix cincticeps. Virology, 47, 483, 1972.
96. Bassi, M., Barbieri, N., Appiano, A., Conti, M., D'Agostino, G., and Caciagli, P., Cytochemical and
autoradiographic studies on the genome and site(s) of replication of barley yellow striate mosaic virus in
barley plants, J. Submicros. Cytol., 12, 201, 1980.
97. Christie, S. R., Christie, R. G., and Edwardson, J. R., Transmission of a bacilliform virus of sowthistle
and Bidens pilosa, Ph\topalholog\, 64, 840, 1974.
98. Martelli, G. P. and Russo, M., Plant virus inclusion bodies, Adv. Virus Res., 21, 175, 1977.
99. Wolanski, B. S. and Chambers, T. C., The multiplication of lettuce necrotic yellows virus, Virology,
44, 582, 1971.
100. Conti, M. and Plumb, R. T., Barley yellow striate mosaic virus in the salivary glands of its planthopper
vector Laodelphax striatellus Fallen, J. Gen. Virol., 34, 107, 1977.
Volume I 101
Chapter 6
TOMATO SPOTTED WILT VIRUS GROUP*
Tomato spotted wilt virus (TSWV) is the sole member of the group. 1 The particles of
this virus are roughly spherical, enveloped, and about 85 nm in diameter. They are quite
different from those of any other plant virus, although the structure and cytopathology does
resemble that of some members of the Bunyaviridae, a family of viruses infecting vertebrates
and also multiplying in their arthropod vectors.1 The virus has a single-stranded RNA
genome, probably consisting of three components of uncertain polarity and MT about 2.7,
1.7, and 1.1 x 106. The particles contain at least four distinct proteins. TSWV is vectored
by several species of thrips in a persistent manner although it has not been shown to multiply
in the vector.2 The virus can also be transmitted mechanically to a wide variety of plant
species.3"5 Seed transmission has been reported,4 but requires confirmation.
I. VIRUS STRUCTURE AND COMPOSITION
Several groups of workers have investigated the structure and composition of TSWV, but
there are still some unresolved inconsistencies in the results.6"10 This is undoubtedly due to
the difficulties of working with a virus that is structurally complex and extremely unstable
in vitro. 1
TSWV particles are about 85 nm in diameter (Figures 1 and 2) and sediment at 520 to
530S. 3 -"-' 2 TSWV envelopes can be removed by non-ionic detergents to leave what appear
to be strands of nucleoprotein. This material has very low infectivity when compared to
intact virus and contains only one protein of M, about 27 x 103.6"9
The intact TSWV particles appear to contain at least four major proteins (I to IV), with
approximate Mr of 78, 58, 52, and 27 x 10\ respectively.8'9 Proteins I, II, and III can be
removed by incubation with bromelein. This treatment causes only a slight drop in infectivity
of the virus, but removes the outer layer of the particle leaving a smooth envelope.9 This,
together with the results of the in vitro iodination experiments of Tas and colleagues,8
indicates that protein IV is located inside the envelope and the other three proteins outside
it. There does not appear to be a membrane matrix protein as there is with the majority of
enveloped viruses.13 Mohamed and his colleagues6 reported that all four major proteins of
TSWV are glycosylated, whereas Tas and his colleagues8 detected carbohydrate in proteins
I, II, and III, but not IV. Since proteins I, II, and III seem to be associated with the spikes
on the virus particle envelope, glycosylation might be expected, but if protein IV is complexed
with the RNA, then glycosylation would be surprising.
At present it is not clear if TSWV particles contain three or four distinct RNA species,
and their exact molecular weights are also uncertain. Van den Hurk and colleagues7 detected
four RNA species of Mr about 2.7, 1.9, 1.7, and 1.3 x 106, but later work in the same
laboratory detected only three species of Mr 2.7, 1.7, and 1.1 x 106.10 On the other hand,
Mohamed9 detected four RNAs of MT about 3.6, 2.8, 2.0, and 1.3 x 106.
There is agreement that RNA preparations from TSWV are not infectious.7-9 This suggests
that the virus is negative-stranded, and yet a claim to the contrary has been made by Verkleij
and colleagues.10 In a wheat germ cell-free system and a messenger-dependent reticulocyte
lysate, these workers detected a number of products, some of which were precipitated by
anti-TSWV serum and had the electrophoretic mobility of the TSWV protein IV. More data
are needed to resolve whether TSWV RNA has positive or negative polarity.
* Figures follow text, beginning on page 104.

II. CYTOPATHOLOGY
The very characteristic enveloped particles of TSWV, about 85 nm in diameter, are easily
identified in thin sections of infected cells (Figure 2), and have always been found inside
membranous cisternae in the cytoplasm. 14 " 20 Amorphous or granular densely staining patches
in the cytoplasm (viroplasms) occur regularly (Figure 3). Tubular membranes characteristic
of infection have also been observed in the cytoplasm (Figure 4A). No abnormalities have
been seen in the nuclei of infected cells, but Mohamed 21 found small peripheral vesicles in
the chloroplasts of infected cells. However, his data suggested that they were a secondary
result of infection. Francki and Grivell 18 found bundles of long filaments with densely staining
cores in the cytoplasm of cells from plants inoculated 3 weeks previously (Figure 4B). The
significance of these filaments was not clear, and they have not been reported by other
workers who, however, have generally not studied such late stages of infection. Ie 19 observed
roughly spherical particles 28 to 33 nm in diameter in TSWV-infected cells, but the particles
did not appear to be those of a virus or to be consistently related to TSWV infection.
The TSWV particle is complex and unique among plant viruses (Figures 1 and 5B), and
determining its method of maturation not only has intrinsic interest but would shed light on
the taxonomic position of the virus. Two attempts have been made to study the viral
maturation sequence by thin sectioning and electron microscopy,17 19 and each has led to
different conclusions.
Milne 17 studied TSWV development in both systemic and local lesion hosts. The systemic
host was tomato and the inoculated leaves were sampled every 5 to 8 hr until mature virus
had appeared. The local lesion hosts were Nicotiana clevelandii, N. glutinosa, Vigna sinensis,
and Chenopodium amaranticolor, and in these, expanding lesions were sectioned at various
radii that represented different stages of infection. In both systemic and local lesion hosts
the same maturation stages were found, though most of the time either no particles or
completed particles were seen, suggesting that the intermediate stages were passed through
rapidly.
The main point of interest was that double-enveloped (DE) particles (Figures 3A, 5A,
and 6) were found in the viroplasm areas, though at an early stage of infection only. They
appeared to arise directly from condensing lumps of viroplasm (Figure 5A) or to be budded
from regions of paired parallel membrane (Figure 6A through C). The DE particles were
not enclosed in membranous cisternae but their inner portions closely resembled the mature
(M) particle in appearance and size (Figure 5B). It was suggested that the final maturation
stage occurred when several DE particles coalesced, their outer membranes forming the
cisterna and their inner portions being released inside as M particles. Hatta and Francki22
have also obseved DE particles in infected cells, together with the viroplasm and M particles
such as those in Figure 3A.
Ie19 studied TSWV in systemically infected Tropaeolum majus and Nicotiana tabacum
Samsun NN and in the expanding local lesions formed in the cotyledons of Cucumus sativus.
He agreed that M particles were probably derived from condensing lumps of viroplasm that
in certain cases were seen to have a 5-nm striation (Figure 3B) that may have represented
the nucleocapsid. However, he did not observe DE particles or budding configurations, and
suggested that M particles may have been formed directly, with the enveloping cisterna
being completed later. However, there is little support for this last suggestion in the published
micrographs.
Further very careful work is necessary to resolve the problem of how the TSWV particle
matures. We suggest that the sequence described by Milne 17 may be correct and that other
workers have not described the early rather transient stages found by him. At least the DE
particles require explanation, having been found independently in two different laborato-
ries. 17 - 22 Budding and release of M particles into cytoplasmic cisternae would be normal if
Volume I 103
Table 1
A COMPARISON OF SOME PROPERTIES OF TOMATO SPOTTED WILT
VIRUS AND MEMBERS OF THE BUNYAVIRIDAE
Property TSWV Bunyaviridae
Particle morphology
Shape Quasispherical Quasispherical
Diameter 85 nm 80—110 nm
Helical nucleocapsid, easily No No
observable
Envelope Present Present
External spikes Present Present
Location of mature particles Aggregated within intracellular Aggregated within intracellular
membranous cisternae membranous cisternae
Maturation By budding, within cytoplasmic By budding within cytoplasmic
cisternae cisternae
Nucleic acid
Type Single-stranded RNA Single-stranded RNA
Polarity Uncertain Negative
No. of segments 3 (or possibly 4) 3
Proteins of virus particle
Nucleocapsid protein Present Present
Glycosylated? Uncertain No
Other proteins 3 3
Glycosylated? All All
Vector: Arthropod Arthropod
TSWV were related to the Bunyaviridae. For example, the budding sequence of the bun-
yavirus, sandfly fever virus, 23 appears to resemble closely that of TSWV illustrated here in
Figure 6. TSWV resembles members of this family more closely than any other group of
plant or animal viruses (Table I). 24 28 This has been discussed more fully elsewhere.29
Interesting isolates of TSWV have recently been derived from normal virus cultures after
continuous passage by mechanical inoculation of host plants in the glasshouse. 10 - 11 The
isolates are defective in that infected cells produce the amorphous inclusions characteristic
of infection, but no virus particles.29 The defect seems to be caused by a deletion in the
viral RNA segment of intermediate size, preventing synthesis of at least one of the proteins
which forms the outer layer of the particle.10
FIGURE 1. Partially purified tomato spotted wilt virus (TSWV). Uranyl acetate stain. Bar = 100 nm. (Courtesy
of Dr. T. S. le. Department of Virology. Agricultural University, Wageningen, The Netherlands.)
Volume I 105
FICJUKI: 2. Thin section ol 'ISWY-inlected Dutiini Mniinoiiiuin leal" cell showing the mature virus particles
within eytoplasmic mcinhranous eislernae. Bar = 500 nm.
FIGURE 3. A, Thin section of a TSWV-infected Datum stramonium leaf cell showing a characteristic viroplasm.
Double-enveloped (DE) particles (arrows) are also seen. Bar = 500 nm. B, Dense patches of a viroplasm enlarged
to show fine structure with a 5-nm repeat. Bar = 100 nm.
Volume I 107
FIGURIi 4. Thin sections ol'TSWV-inlecied Datura stramonium leat cells. A. Membranous lubules characteristic
of inlcclion. B. Bundles of densely stained filaments of unknown significance but characteristic of infection by
some TSWV isolates. Bars = 100 mn.
FIGURE 5. Thin sections of TSWV-infected tomato leaves. A, Relatively early stage (60 hr after inoculation)
showing double-enveloped (DE) particles embedded in diffuse viroplasm. No mature particles are seen. B. Late
stage (1 week after inoculation) showing mature particles in cytoplasmic membranous cisternae. Bars = 100 nm.
Volume I 109
FIGURE 6. Thin sections of TSWV-infected tomato leaves at relatively early stages of infection (55 to 80 hr
after inoculation). A. Paired parallel membranes enclosing electron-dense material. One paired membrane (arrow)
has the appearance of budding to form a double-enveloped particle. B, Bud-like structures possibly representing
the formation of double-enveloped particles. C, A double enveloped particle in equatorial section. D, Two double-
enveloped particles; the outer envelope of the lower particle can be interpreted as expanding to release the maturing
particle into the resulting cisterna. The end result of this budding process would be mature particles as seen in
Figure 5B. All micrographs are at the same magnification. Bar = 100 nm.
REFERENCES
2. Paliwal, Y. C., Occurrence and localization of spherical virus-like particles in tissues of apparently healthy
tobacco thrips, Frankliniellafusca, a vector of tomato spotted wilt virus, /. Invert. Pathol., 33, 307, 1979.
3. Best, R. J., Tomato spotted wilt virus, Adv. Virus Res., 13, 65, 1968.
4. le, T. S., Tomato spotted wilt virus, CMl/AAB Descriptions of Plant Viruses, No. 39, 1970.
5. Francki, R. I. B. and Hatta, T., Tomato spotted wilt virus, in Handbook of Plant Virus Infections and
Comparative Diagnosis, Kurstak, E., Ed., Elsevier, Amsterdam, 1981, 491.
6. Mohamed, N. A., Randies, J. W., and Francki, R. I. B., Protein composition of tomato spotted wilt
virus. Virology, 56, 12, 1973.
7. Van den Hurk, J., Tas, P. W. L., and Peters, D., The ribonucleic acid of tomato spotted wilt virus, J.
Gen. Virol., 36, 81, 1977.
8. Tas, P. W. L., Boerjan, M. L., and Peters, D., The structural proteins of tomato spotted wilt virus, J.
Gen. Virol., 36, 267, 1977.
9. Mohamed, N. A., Isolation and characterization of subviral structures from tomato spotted wilt virus, J.
Gen. Virol., 53, 197, 1981.
10. Verkleij, F. N., De Vries, P., and Peters, D., Evidence that tomato spotted wilt virus RNA is a positive
strand, /. Gen. Virol., 58, 329, 1982.
1 1 . Van Kamnien, A., Henstra, S., and le, T. S., Morphology of tomato spotted wilt virus, Virology, 30,
574, 1966.
12. Joubert, J. J., Hahn, J. S., von Wechmar, M. B., and van Regenmortel, M. H. V., Purification and
properties of tomato spotted wilt virus, Virology, 57, 11, 1974.
13. Lenard, J. and Compans, R. W., The membrane structure of Hpid-containing viruses, Biochim. Biophys.
Ada, 344, 51, 1974.
14. le, T. S., An electron microscope study of tomato spotted wilt virus in the plant cell, Neth. J. Plant Pathol.,
70, 114, 1964.
15. Kitajima, E. W., Electron microscopy of Vira-Cabeca virus (Brazilian tomato spotted wilt virus) within
the host cell, Virology, 26, 89, 1965.
16. Milne, R. G. and de Zoeten, G. A., A comparison of some methods of preparation of thin sections of
virus-infected leaves for electron microscopy, J. U/trastruct. Res., 19, 398, 1967.
17. Milne, R. G., An electron microscope study of tomato spotted wilt virus in sections of infected cells and
in negative stain preparations, J. Gen. Virol., 6, 267, 1970.
18. Francki, R. I. B. and Grivell, C. J., An electron microscope study of the distribution of tomato spotted
wilt virus in systemically infected Datura stramonium leaves, Virology, 42, 969, 1970.
19. le, T. S., Electron microscopy of developmental stages of tomato spotted wilt virus in plant cells, Virology,
43, 468, 1971.
20. le, T. S., Tomato spotted wilt virus in the anthers of Tropaeolum majus, Neth. J. Plant Pathol., 79, 249,
1973.
21. Mohamed, N. A., Some effects of systemic infection by tomato spotted wilt virus on chloroplasts of
Nicotiana tabacum leaves. Physiol. Plant Pathol., 3, 509, 1973.
22. Hatta, T. and Francki, R. I. B., unpublished data, 1981.
23. Smith, J. F. and Pifat, D. Y., Morphogenesis of sandfly fever viruses (Bunyaviridae family), Virology,
121. 61, 1982.
24. Murphy, F. A., Harrison, A. K., and Tzianabos, T., Electron microscopic observations of mouse brain
infected with bunyamwera group arboviruses. J. Virol., 2, 1315, 1968.
25. Holmes, I. H., Morphological similarity of bunyamwera supergroup viruses. Virology, 43, 708, 1971.
26. Murphy, F. A., Harrison, A. K., and Whitfield, S. G., Bunyaviridae: morphologic and morphogenetic
similarities of bunyamwera serologic supergroup viruses and several other arthropod-borne viruses, Inter-
virology, 1, 297, 1973.
27. Bishop, D. H. L., Calisher, C. H., Casals, J., Chumakov, M. P., Gaidamovich, S. Ya., Hannoun,
C., Luov, D. K., Marshall, I. D., Oker-Blom, N., Pettersson, R. F., Porterfield, J. S., Russell, P.
K., Shope, R. E., and Westaway, E. G., Bunyaviridae, Intervirology, 14, 125. 1980.
28. Clerx, J. P. M., Casals, J., and Bishop, D. H. L., Structural characteristics of Nairoviruses (Genus
Nairm'irus. Bunyaviridae), /. Gen. Virol., 55. 165. 1981.
29. Milne, R. G. and Francki, R. I. B., Should tomato spotted wilt virus be considered as a possible member
of the family Bunyaviridae?, Intervirology, 22, 72, 1984.
30. le, T. S., A sap transmissible, defective form of tomato spotted wilt virus, J. Gen. Virol., 59, 387. 1982.
31. Verkleij, F. N. and Peters, D., Characterization of a defective form of tomato spotted wilt virus, J. Gen.
Virol., 64, 677, 1983.
Volume! Ill
Chapter 7
MAIZE CHLOROTIC DWARF VIRUS GROUP
Maize chlorotic dwarf virus (MCDV) is at present the sole member of the group 1 and has
not been studied very extensively. It has polyhedral particles about 30 nm in diameter and
the genome consists of one single-stranded RNA molecule of Mt about 3.2 x 106. MCDV
is unusual in that it is transmitted by leafhoppers in a semipersistent manner, but not by
mechanical inoculation. 2 - 3 The virus is confined to some species within the family Gramineae
and there are no reports of seed transmission. 2
I. PROPERTIES OF THE PARTICLES
Little is known about the structure of MCDV particles, other than that they are polyhedral
when negatively stained, and measure about 30 nm in diameter (Figure 1). It has been
suggested that the capsid is made up of two polypeptides of M, about 18 and 30 x 10' and
that a third polypeptide may also be present. 3 However, further work is needed to substantiate
this.
MCDV particles differ from those of other known plant viruses of similar size and gross
morphology in that they sediment faster (180S) and are denser (buoyant density in CsCl is
about 1.51). MCDV particles each contain one molecule of RNA of M, about 3.2 x 106
as estimated by PAGE and sedimentation in sucrose density gradients. 4
II. CYTOPATHOLOGY
Isometric particles, assumed to be those of MCDV, have been observed embedded in

electron-dense granular inclusions in the cytoplasm of phloem tissue cells (Figure 2) and
occasionally also in those of the mesophyll. 5 - 6 Although most of the virus particles appear
to be associated with the inclusions (Figure 2), individual particles were sometimes also
observed in the vacuoles. 6 MCDV-infected cells containing virus particles and granular
inclusions also sometimes contained fibrillar material, accumulations of vesicles and par-
ticulate aggregates, and laminate inclusions of unidentified composition.6
III. RICE TUNGRO VIRUS — A POSSIBLE MEMBER OF THE GROUP
It has been suggested that rice tungro virus (RTV) may be sufficiently similar to MCDV
to be classified in the same group.1 Galvez7 purified a virus with small polyhedral particles,
30 to 33 nm in diameter sedimenting at 175S from rice tungro-diseased plants and found
the preparations infectious when tested by a leafhopper assay. More recently two types of
particles have been found to be associated with rice tungro disease; these are a small
polyhedral particle about 30 nm in diameter and a bacilliform particle about 35 nm wide
and 150 to 350 nm long8'10 (Figure 3). The bacilliform particles resemble those associated
with cacao swollen shoot virus, cacao mottle leaf virus, and colocasia virus 2 which are as
yet unclassified. " J2 Experiments with rice tungro disease indicated that symptoms char-
acteristic of tungro disease in rice were caused by the bacilliform particles and that the
polyhedral particles, when present, intensified the disease.8'9
It is interesting to note that in cells of rice plants infected with tungro disease, particles
28 to 30 nm in diameter have also been observed, embedded like the particles of MCDV,
in electron-dense granular material.13'14 However, Favali and her colleagues14 also observed
bacilliform "tubules" about 30 nm in diameter and 250 nm long (Figure 4C and D). Further
FIGURE 1. Partially purified preparation of mui/c chlorotic dwarf virus ( M C D V ) . L'rany] acetate stain. Bar =
100 nm. (Courtesy of Dr. O. E. Bradlute. Department of Plant Pathology. Ohio State University, Wooster, Ohio.)
work is needed to determine the exact etiology of rice tungro disease and the taxonomic
positions of what appear to be two viruses associated with it.
Volume I 113
FIGURE 2. A thin section ot a cell from the phloem ot a maize plant mtected with MCDV showing an inclusion
containing virus particles. Bar = 500 nin. (Courtesy of Dr. K. F. Harris, Department of Entomology, Texas A
& M University. College Station. Texas.)
FIGURE 3. Purified preparation of virus-like particles isolated from plants with tungro disease. Uranyl acetate
stain. Bar = 100 nm.
Volume I 115
FICiURK 4. A. Thin scclion of a rice leaf cell infected wilh lungro disease, showing a virus-specific inclusion
(I) containing polyhedral virus-like panicles (I'). Bar =• 500 nm. B. Crystalline array of polyhedral virus-like
panicles in a cell vacuole of a uingro-infecled rice leaf. Bar - 100 nm. C. Thin section as in A. showing virus-
specific inclusions ( I ) , polyhedral virus-like particles (Pi. and hacilliform virus-like particles (ti). Bar = 500 nm.
IX The bacilliform particles enlarged. Bar - 100 nm. (Courtesy of Dr. M. A. Favali. Institute of Botanical
Science. Milan, llalv.l
REFERENCES
mittee on Taxonomy of Viruses. Intervirology, 17, I. 1982.
2. Gingery, R. E., Bradfute, O. E., Gordon, D. T., and Nault, L. R., Maize chlorotic dwarf virus. CM//
AAB Descriptions of Plant Viruses. No. 194. 1978.
3. Gingery, R. E., Gordon, D. T., Nault, L. R., and Bradfute, O. E., Mai/e chlorotic dwarf virus, in
Handbook of Plant Virus Infections and Comparative Diagnosis, Kurstak, E. Ed., Elsevier, Amsterdam.
1981, 19.
4. Gingery, R. E., Properties of maize chlorotic dwarf virus and its ribonucleic acid. Virology. 73. 3 1 1 ,
1976.
5. Pirone, T. P., Bradfute, O. E., Freytag, P. H., Lung, M. C. Y., and Poneleit, C. G., Virus like
particles associated with a leafhopper-transmitted disease of corn in Kentucky, Plant Dis. Rep.. 56. 652,
1972.
6. Harris, K. R. and Childress, S. A., Cytology of maize chlorotic dwarf virus infection in corn, J. Trap.
Plant Dis., in press.
7. Galvez, G. E., Purification and characterization of rice tungro virus by analytical density-gradient cen-
trifugation. Virology. 35, 418. 1968.
8. Hibino, H., Roechan, M., and Sudarisman, S., Association of two types of virus particles with penyakit
habang (tungro disease) of rice in Indonesia, Phytopathology, 68, 1412, 1978.
9. Hibino, H., Saleh, N., and Roechan, M., Transmission of two kinds of rice tungro-associated viruses
by insect vectors. Phytopathology. 69, 1266, 1979.
10. Milne, R. G., Boccardo, G., and Ling, K. C., Electron microscopy of particles of rice tungro virus
complex. Int. Rice Res. Newslett., 6:5, 13, 1981.
1 1 . Brunt, A. A., Cacao swollen shoot virus. CMIIAAB Descriptions of Plant Viruses, No. 10, 1970.
12. James, M., Kenten, R. H., and Woods, R. D., Virus-like particles associated with two diseases of
Coloctisia esculenta (L.) Schott. in the Solomon Islands, J. Gen. Viral., 21, 145, 1973.
13. Galvez, G. E., Shikata, E., and Mian, M. S. A., Transmission and electron microscopy of a rice tungro
virus strain. Phytopathol. Z.. 70. 53, 1971.
14. Favali, M. A., Pellegrini, S., and Bassi, M., Ultrastructural alterations induced by rice tungro virus in
rice leaves. Virology, 66, 502, 1975.
Volume 1 117
Chapter 8
TYMOVIRUS GROUP
Tymoviruses1 derive their name from the type member, turnip yellow mosaic virus (TYMV),
one of the most extensively studied of plant viruses. Members of the group have small
isometric particles about 29 nm in diameter. All members have at least two particle classes
constructed from a single species of polypeptide of Mr about 20 x 10'. The surface properties
of protein shells of the two types are indistinguishable, but while one shell contains no
nucleic acid, the other encapsidates about 37% RNA. The genome of each member virus
consists of a single species of positive-sense, single-stranded RNA of Mr about 2 X 106.
The group forms a network of serologically interrelated viruses. Many members are
transmitted by beetles and all are readily transmitted by mechanical inoculation, each to a
relatively narrow range of plants. Seed transmission has only been reported for andean potato
latent virus (APLV). They all induce characteristic cytopathological effects in the chloroplasts
of infected cells. 2 4
The Tymoviruses form a homogeneous group. Their physical properties are similar, as
can be seen from the data summarized in Table 1. Other characteristics common to all the
viruses in the group are the high cytidine contents of their RNAs and some of the cyto-
pathological effects they induce. The homogeneity of the group is also evident from ser-
ological data, which indicate that each member is related to at least one other. This has
been established by very thorough serological studies with 14 of the viruses listed in Table
I. 5 - 6 The results are presented in Figure 1. Three other viruses, physalis mosaic (PhMV),
erysimum latent (ErLV) and peanut yellow mottle (PYMV), have also been shown to be
related to a number of Tymoviruses. 7 9
Figure 1 shows that a number of Tymoviruses are very closely related serologically, some
being separated by SDIs of only one. Differences between the strains of some viruses such
as APLV are in fact as large as those separating other variants described as distinct viruses. 24
The validity of their separate virus status has been discussed elsewhere.4
The relationships among Tymoviruses have also been investigated by using coat protein
amino acid composition data.25 The classification obtained, although showing some simi-
larities to that constructed from serological studies, also showed some significant differences,
making overall correlation rather poor. So far, little has been done to investigate relationships
among Tymoviruses based on nucleotide sequence homologies. However, RNA-RNA hy-
bridization data failed to detect any significant homology between the RNAs of APLV and
eggplant mosaic virus (EMV). 26 This is surprising because the two viruses are closely related
serologically.27-28 All this indicates that in our present state of knowledge a Tymovirus is
relatively easy to identify as such, but it is much more difficult to decide to which of the
established viruses it should be assigned or, indeed, if it can be assigned to any one of them.
Maize rayado fino virus (MRFV) is interesting in that its particles are remarkably similar
to those of the Tymoviruses 29 31 (Table 2). However, the virus does not seem to be sero-
logically related to any of the Tymoviruses and differs biologically in that it is transmitted
persistently by leafhoppers, in which it appears to replicate.32-33 Furthermore, MRFV is not
transmissible by mechanical inoculation.29 It infects only a narrow range of host plants within
the Gramineae.29 These unique properties of MRFV make its taxonomic position obscure
at present.
Table 1
MEMBERS OF THE TYMOVIRUS GROUP AND THEIR
PROPERTIES
^20w M, of
M, of RNA coat protein
Virus T B x 10" x 10' Ref.
Turnip yellow mosaic 53—54 116—117 210 20.1 10. I I

virus (TYMV)
Kennedya yellow 54 110 ~l.9 20.5 12
mosaic virus
(KYMV)
Clitoria yellow vein 50 109 — •' 20.0 13
virus (CYVV)
Cacao yellow mosaic 49 108 — — 14
virus ( C Y M V )
Desmodium yellow 54 114 2.0 — 15
mottle virus
(DYMV)
Okra mosaic virus 42 106 — 20.0 16
(OkMV)
Wild cucumber mos- 53 119 — 20.2 17
aic virus (WCMV)
Scrophularia mottle 54 116 2.1 21.6 18
virus (ScrMV)
Plantago mottle virus — — — — 19
(P1MV)
Ononis yellow mos- 55 115 — — 20
aic virus (OYMV)
Belladonna mottle vi- 53 113 2.0 20.3 21
rus (BeMV)
Dulcamara mottle vi- 55 115 — — 20
rus (DuMV)
Andean potato latent 55 115 — 20.2 20. 22
virus (APLV)
Eggplant mosaic vi- 53 111 2.0—2.5 20.0—20.5 23
rus (EMV)
Physalis mosaic virus 50 112 — — 7
(PhMV)
Erysimum latent vi- 59 113 2.0 21.7 8
rus (ErLV)
Peanut yellow mottle 52 110 — — 9
virus (PYMV)
a
Not determined.
Nearly all our knowledge of Tymovirus structure and composition is based on TYMV,
which has been more thoroughly investigated than any other plant virus except tobacco
mosaic. 10 -" Data on other Tymoviruses is much less detailed, but as the group is so ho-
mogeneous, much of the discussion of TYMV should also be relevant to the other group
members.
Particles of Tymoviruses can be seen in electron micrographs of negatively stained prep-
arations to be about 29 nm in diameter and to consist of 32 morphological subunits arranged
with icosahedral symmetry (Figure 2). The particle structure, easily recognized by routine
Volume I 119
FIGURE I . Antigenic relationships among Tymoviruses based on serological differentiation indices (SD1) in
reciprocal tests after Koenig.'' (The SDI is defined as the number of twofold dilution steps separating homologous
and heterologous tilers.) The numbers indicate SDI values between the viruses whose full names are listed in
Table I .
high-resolution electron microscopy, is an almost foolproof guide to identifying members

of the Tymovirus group. The diameter of the TYMV particle, determined by X-ray dif-
fraction, is about 28 nm14 and the hydrodynamic particle diameter was shown to be 29.5
nm using diffusion measurements from laser-beam spectroscopy.1s The close correspondence
of these values with measurements in the electron microscope indicate that there is little
shrinkage of the particles during drying. A model of TYMV structure has been proposed
(Figure 3B); this model consists of 180 banana-shaped protein subunits whose complete
primary structure of 189 amino acids has been determined. 16 These subunits are clustered
into 20 hexamers and 12 pentamers and arranged with T = 3 lattice symmetry." The model
agrees well with a three-dimensional image reconstruction of a particle from electron
micrographs1" (Figure 3A).
Tymovirus preparations always consist of at least two centrifugal components, an infec-
tious bottom (B) and a noninfectious top (T) component sedimenting at about 115 and 54S,
respectively (Table 1). Although the T particles of TYMV are devoid of RNA, those of
three other Tymoviruses, EMV, wild cucumber mosaic virus (WCMV), and okra mosaic
virus (OkMV), have been shown to contain small amounts of RNA. About 90% of this was
host tRNA, and the rest was subgenomic, coat protein mRNA. 16
The structure of Tymovirus T particles is identical to that of the B particles except for
the absence of the RNA. Artificial TYMV empty shells have been prepared by incubating
virus preparations in 1 M KC1 at pH 11.5 at 30°C.40 This causes the shells to swell by about
Table 2
COMPARISON OF THE PHYSICAL PROPERTIES OF MAIZE RAYADO FINO
AND TURNIP YELLOW MOSAIC VIRUSES
Maize rayado fino virus Turnip yellow mosaic virus

Property (MRFV)29"" (TYMV) 10 -"
Virus Particles
Diameter (nm) 27—33 28—29
Capsid structure T = 3 with 32 clearly visible T = 3 with 32 clearly visible
capsomeres capsomeres
M, of coat polypeptide ( x 101) 21—22 20.1
Sedimentation coefficient (S,,,J
T 47—54 53—54
B 120—124 116—117
Buoyant density in CsCl (g/cm 1 )
T 1.27—1.28 1.28
B 1.43—1.46 1.40
Viral Genome
M, of ss-RNA ( X 10") 2.0—2.4 2.0
Base ratios
C 31.4 38.3
G 23.1 17.2
A 16.8 22.4
U 28.7 22.1
4% and allows the RNA to escape in a degraded form. 41 - 42 Empty protein shells can also
be obtained by freezing B particles, but under these conditions the extruded RNA appears
to remain intact. 43 Release of RNA by both methods involves the loss of a few coat protein
subunits. 42 - 44
The stability of empty protein shells indicates that Tymovirus particles depend largely on
protein-protein interactions for their integrity. The exact distribution of the RNA in B particles
is not known. Finch and Klug" considered that it was folded in such a way as to be intimately
associated with all the morphological subunits of the protein shells. However, neutron small-
angle scattering data indicate that there is relatively little penetration of the protein shell by
RNA, 4 5 although the presence of the RNA does appear to affect the conformation of the
viral protein.4'1'48 The protein-RNA interactions in TYMV appear to involve hydrogen bonds
between the nucleotide amino and protein carboxyl groups. 46
In addition to the B and T components, particles of intermediate densities have been
detected in preparations of some Tymoviruses by isopycnic density-gradient centrifugation
in CsCl. 19 - 49 With TYMV, as many as 12 nucleoprotein particles have been separated ranging
in densities from 1.294 to 1.430 g c m ~ \ 4 1 These particles have been shown to contain
various sized subgenomic RNAs. Most of them appear to be natural products of TYMV
replication, although at least one is an artifact of centrifugation in CsCl.50
In highly purified preparations of TYMV, Johnson and Markham 51 detected polyamines
apparently absent from healthy plants. They showed that most of the polyamines were bound
to the RNA. Beer and Kosuge 52 identified the polyamines as spermidine and a much smaller
proportion of spermine. Their exact amounts varied from preparation to preparation, but
they accounted for only about 1% of the virus by weight. This was calculated to be sufficient
to neutralize about 2(Wc of the phosphate groups of the RNA. Polyamines are known to be
efficient in preventing the unfolding of TYMV RNA 51 and although their function in TYMV
particles has not been established, they may serve to neutralize the charged phosphate groups
of the viral RNA and so maintain a compact structure within the particle. More recently,
Cohen and Greenberg54 demonstrated that spermidine is associated with TYMV RNA before
packaging in particles and is essentially nonexchangeable.
Volume I 121
FIGURE 2. Purified turnip yellow mosaic virus (TYMV) stained in uranyl acetate. Most of the particles are
oriented so that they are viewed down the twofold axes (a). Some particles are sufficiently well resolved so that
their subunits appear to have central holes (arrows). Bar = 100 nm.
FIGURE 3. A, Three-dimensional image of a TYMV particle reconstructed from an electron micrograph; B,

Diagramatic representation of the outer surface of the TYMV particle. (A, from Mellema, J. E. and Amos, L. A.,
J. Mol. Biol., 72. 819, 1972; B, from Finch, J. T. and Klug, A., J. Mol. Biol., 15, 344, 1966. Copyright:
Academic Press, Inc. (London) Ltd. With permission.)
B. The Viral Genome

The largest RNA molecule found inside TYMV B particles is of Mr about 2 x 106 and
is infectious by itself. 55 In addition, TYMV preparations contain substantial amounts of
subgenomic RNA molecules of M, about 0.25 x 106 which can function as coat protein
mRNA in vitro; they can be encapsidated in the minor nucleoprotein components and also
in the B particles. 56 Both RNAs are capped at the 5' ends, the genomic RNA 57 with m7Gppp5G
. . . and the subgenomic coat protein mRNA 58 with m 7 GpppA . . . . The TYMV RNA
3' ends consist of a tRNA-like sequence which accepts valine in vitro59"61 and in vivo. 62 - 63
Similar in vitro valine-accepting activity has been reported for the RNAs of four
other Tymoviruses including cacao yellow mosaic virus (CYMV), EMV, OkMV, and
WCMV. 64 67 The significance of the tRNA-like structure is not understood, but it could play
a regulatory role in viral RNA replication or in protein synthesis. 6 "
The entire base sequence of TYMV coat protein mRNA is known, and consists of 695
nucleotides, excluding the cap.58 From its 5' end, this mRNA contains a stretch of 19
untranslated nucleotides followed by 567 nucleotides of the coat protein cistron. At the 3'
end there are 109 untranslated nucleotides, of which about 86 form the tRNA-like structure. 69
The coat protein mRNA sequences occupy the 3' terminal portion of the genomic RNA. 70
The coat protein cistron of the full-length, infective TYMV RNA is not translated in vitro.
The subgenomic coat protein mRNA isolated from TYMV preparations is, however, a very
efficient messenger.71 73 Since this messenger, as a separate piece of RNA, is not mandatory
for infectivity, it must be generated in vivo by genome cleavage, by particle replication of
the negative genome-length RNA strand, or by some other mechanism.
TYMV genomic RNA has been translated in vitro into two products of Mr about 195
(195K) and 150 x 103 (150K). The longer protein appears to undergo posttranslational
cleavage to yield polypeptides of M, about 120 and 78 x 103 (120K and 78K, respectively),
and it is interesting to note that the 150K product does not seem to be similarly cleaved.74 75
Both the 195K and 150K primary translation products possess common amino acid se-
quences,76 and their synthesis begins at a unique initiation site located in the 5' terminal
region of the genomic RNA. 72 It has also been observed that in the presence of yeast amber
suppressor tRNA, a still larger protein of Mr about 210 x 103 (210K) is synthesized in vitro
to the disadvantage of the 195K protein. It appears to be a "readthrough" product. 77 The
available TYMV RNA sequence data and a comparison of the tryptic peptides of the 195K,
Volume I 123
FIGURE 4. Physical map of the Tymovirus genome based on data from TYMV. Only the genomic RNA and
the coat protein mRNA are shown; the other subgenomic RNAs of uncertain structure and function are not
included.* 73
210K, and coat proteins suggest that the noncoding region between the 195K and coat protein
genes is only 14 nucleotides long. 78
A series of subgenomic TYMV RNAs isolated from virus preparations have been translated
in a rabbit-reticulocyte in vitro system. 73 The products overlap with one another and with
the translation products of the full-length RNA. These RNAs seem to share a common
translation initiation site near their 5' ends.
A model of the TYMV genome based on current knowledge is presented in Figure 4.
The additional subgenomic RNAs detected56-73 have not been included as their detailed
structures and possible functions are unknown. The 195K protein, its cleavage products and
the 150K protein detected in in vitro translation systems have not yet been shown to be
produced in vivo, and their functions are also not known.
III. CYTOPATHOLOGY
In infected cells, Tymoviruses induce a number of very characteristic cytological changes,

the most prominent of which are associated with the chloroplasts2 79 8<) (Figures 5 through
8). The cytopathology of TYMV has been studied much more extensively than that of the
other members of the group and it appears that with minor variations, all Tymovirus-infected
cells undergo essentially similar changes.80
FIGURE 5. A, Thin section of a Brassica pekinensis (Chinese cabbage) leaf mesophyll cell infected with TYMV
showing the swollen and rounded chloroplasts with numerous peripheral vesicles; bar = 500 nm. B, Higher
magnification of chloroplast peripheral vesicles showing their double-membraned structure and fibrillar material
inside the vesicles (arrows); bar = 100 nm.
Volume I 125
FIGURE 6. A. Thin section of a Brassicu pekinensis (Chinese cabbage) leal mesophyll cell infected with TYMV
showing the characteristic chloroplast vesicles and virus particles (V) between the two chloroplasts. B, Selected
vesicles to show their double-membraned structure and their necks opening (arrows) into the cytoplasm. Bars =
100 nm.
FIGURE 7. Thin section of a Pisum sativum (garden pea) leaf mesophyll cell infected with kennedya yellow
mosaic virus (KYMV) at a relatively late stage of infection showing rounded chloroplasts with large vacuoles and
numerous peripheral vesicles in the chloroplasts. Bar = 500 nm.
Volume I 127
FIGURE 8. A, Thin section of Nicotiana gtutinosa leaf mesophyll cell infected with eggplant mosaic virus (EMV)
showing chloroplasts with peripheral vesicles and virus particles (arrow) near some vesicles between two adjoining
chloroplasts; bar = 500 nm. B, Higher magnification of the region between the two adjoining chloroplasts indicated
by an arrow in A, showing the double-membraned chloroplast vesicles and densely stained fibrillar material inside
(arrows); bar = 100 nm.
Early effects of virus infection involve the chloroplasts whose membranes develop small
peripheral vesicles*1"86 (Figures 5 through 8). The vesicles are formed by the invagination
of both chloroplast membranes, and their necks remain open so that their contents are in
contact with the cytoplasm (Figure 6). Fibrillar material reminiscent of nucleic acid has been
seen in the vesicles81 (Figures 5B and 8B), and both biochemical and autoradiographic
evidence indicate that the vesicles are the sites of TYMV RNA replication. 87 " 89 Thus, the
fibrillar material is probably the virus-specific replicative form double-stranded RNA.
In the case of TYMV, the cytological events following infection have been followed in
some detail. 9<) - 91 They have been arbitrarily divided into seven stages as follows:
1. Small peripheral vesicles appear on the chloroplast surface without any significant
changes to their size or shape (compare Figure 9A and B).
2. The chloroplasts swell and assume roughly spherical outlines (Figure 5).
3. More peripheral vesicles develop on the chloroplasts and form clusters (Figures 5, 6
and 9C). Also at about this time, prominent endoplasmic reticulum membranes develop
adjoining the clumps of chloroplast vesicles (Figure 10A). These membranes appear
to be smooth on the side of the chloroplast surface but have ribosomes attached on
the side facing the bulk of the cytoplasm. The membranes probably develop just prior
to the clusters of vesicles and appear to be interconnected with others to form a network
surrounding the surface of the chloroplast. 9 "
4. The endoplasmic reticulum associated with vesicle clusters on chloroplasts disappears
to form electron-lucent regions proximal to the clusters.
5. The chloroplasts clump, usually so that the spaces between them are the electron-
lucent zones (Figures 5 and 9D).
6. Spaces between the clumped chloroplasts become more densely staining and virus
particles start to accumulate there (Figures 8 and 10B).
7. Numerous virus particles accumulate between the chloroplasts (Figure 6A). This is
the mature infection which has been described by some authors. 81 - 8 '
During very late stages of infection when the leaves show signs of senescence, the
cytoplasm and chloroplasts become vacuolated and disorganized, with numerous particles
scattered throughout the cytoplasm and vacuoles (Figure 11). The quantity of virus particles
that are present can be gauged by the large crystalline aggregates that are formed when the
fully infected cells are water stressed (Figure 12).
Freeze-etching experiments have revealed that at about stage 3 of TYMV infection, small
protein particles appear in the outer chloroplast membrane near the vesicles. The size of the
particles corresponds to that expected for coat protein pentamers and hexamers. 79 The earliest
detected virus particles appear in the cytoplasm near the vesicles (Figures 6, 8, and 10B).
On the basis of these observations it has been suggested that viral RNA is synthesized in
the vesicles and is encapsidated as it emerges into the cytoplasm. 79 - 90
Empty protein shells, whose synthesis is so characteristic of Tymovirus infection, have
been detected in the cytoplasm of infected cells, but more usually they accumulate in the
nuclei (Figure 13), sometimes forming large crystalline array s.79-80-85 With some viruses
such as OkMV, the empty shells reach especially high concentrations in the nuclei. 92 The
reason why virus particles preferentially accumulate in the cytoplasm and empty shells in
the nuclei is not understood. However, there is an indication from biochemical studies that
the empty shells in the nucleus are assembled there using protein subunits transported from
the cytoplasm. 92 It seems that viral coat protein can probably move freely through the
endoplasmic reticulum and the nuclear envelope. It appears to be synthesized in excess of
the amount necessary to encapsidate the viral RNA. Thus the viral RNA extruded from the
chloroplast vesicles into the cytoplasm would encounter coat protein subunits and most
Volume I 129
FIGURE 9. A. Thin section ol'a chloroplast from an uninl'cclcd lirassiai pekinfiisis (Chinese cabbage) leaf cell.
B. C. and D, chloroplasls Irom similar plants as in A but at stages I . 4. and 5 of infection with TYMV (see text
for detailed description). Bars = 500 nm.
FIGURE 10. Thin sections of chloroplasts from Braxsica pekincnsis (Chinese cabbage) leaf cells infected with
TYMV. A. Clump of peripheral vesicles of a chloroplast with characteristic endoplasmic reticulum in the adjoining
cytoplasm (arrows); the cell was at stage 3 of infection as described in the text. B, Aggregates of virus particles
(v) and ribosomcs (r) in the cytoplasm adjoining a clump of peripheral vesicles of a chloroplast at stage 4 of
infection as described in the text (tissue was water-stressed before fixation for electron microscopy to aggregate
the virus particles" 11 ). Bars = 100 nm.
would become encapsidated. Excess coat protein migrating into the nucleus would not
encounter any viral RNA and would then polymerize into empty protein shells.
Lesemann*" observed that in cells infected by some Tymoviruses the mitochondria were
sometimes swollen and assumed irregular shapes. Some vacuolation was observed which
probably arose through swelling of single mitochondria! cristae. Some of these mitochondrial
vesicles contained what appeared to be empty protein shells, but nothing resembling virus
particles was seen. With clitoria yellow vein virus (CYVV), small vesicles like those induced
in chloroplasts were also seen in the mitochondria. They were bounded by a double membrane
and contained fibrillar material reminiscent of nucleic acid,*" but their significance remains
to be determined.
Volume I 131
FIGURE 1 1 . Thin section of a Brassica pekinensis (Chinese cabbage) leaf mesophyll cell at a late stage of
infection with TYMV (20 days after inoculation) showing the rounded chloroplasts with peripheral vesicles and
large vacuoles; numerous virus particles are scattered throughout the apparently structureless cytoplasm. Bar =
500 nm.
FIGURE 12. Thin section of a Braxsica pekinensis (Chinese cabbage) leaf mesophyll cell at a late stage of
infection with TYMV (20 days after inoculation). The cell had been plasmolyzed prior to fixation to aggregate the
virus particles. Bar = 500 nm.
Volume I 133
FIGURE 13. Thin section of a part of a Phaseolus vulgaris (French bean) leaf mesophyll cell infected with
desmodium yellow mottle virus (DYMV) showing virus particles in the cytoplasm and predominantly empty protein
shells in the nucleus. Bar = 100 nm.
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region of turnip yellow mosaic virus genomic RNA, Virology, 119, 193, 1982.
79. Hatta, T. and Matthews, R. E. F., Sites of coat protein accumulation in turnip yellow mosaic virus-
infected cells. Virology, 73, I , 1976.
80. Lesemann, D.-E., Virus group-specific and virus-specific cytological alterations induced by members of
the Tymovirus group, Phytopathol. Z., 90, 315, 1977.
81. Chalcroft, J. and Matthews, R. E. F., Cytological changes induced by turnip yellow mosaic virus in
Chinese cabbage leaves. Virology, 28, 555, 1966.
82. Gerola, F. M., Bassi, M., and Guissani, G., Some observations on the shape and localization of different
viruses in experimentally infected plants, and on the fine structure of the host cells. III. Turnip yellow
mosaic virus in Brassica chinensis L., Caryologia, 19, 457. 1966.
83. Ushiyama, R. and Matthews, R. E. F., The significance of chloroplast abnormalities associated with
infection by turnip yellow mosaic virus, Virology, 42, 293, 1970.
84. Allen, T. C., Subcellular responses of mesophyll cells to wild cucumber mosaic virus, Virology, 47, 467,
1972.
85. Moline, H. E., Ultrastructure of Datura stramonium leaves infected with the physalis mottle strain of
belladonna mottle virus. Virology, 56, 123, 1973.
86. Hatta, T., Bullivant, S., and Matthews, R. E. F., Fine structure of vesicles induced in chloroplasts of
Chinese cabbage leaves by infection with turnip yellow mosaic virus, J. Gen. Virol., 20, 37, 1973.
87. Lafleche, D. and Bove, J. M., Sites of incorporation of tritiated uridine into the cells of the foliar
parenchyma of healthy or turnip yellow mosaic virus infected Brassica chinensis, C. R. Acad. Sci. Paris,
266, 1839, 1968.
88. Lafleche, D. and Bove, J. M., Turnip yellow mosaic virus: cellular site of viral RNA replication. Physio/.
Veg.. 9, 487, 1971.
89. Lafleche, D., Bove, C., Dupant, G., Mouches, C., Astier, T., Gamier, M., and Bove, J. M., Site of
viral RNA replication in the cells of higher plants. TYMV (turnip yellow mosaic virus) — RNA synthesis
on the chloroplast outer membrane system, Proc. Fed. Eur. Biochem. Soc., 72, 43, 1972.
90. Hatta, T. and Matthews, R. E. F., The sequence of early cytological changes in Chinese cabbage leaf
cells following systemic infection with turnip yellow mosaic virus, Virology, 59, 383, 1974.
91. Matthews, R. E. F., Plant Virology, 2nd Ed., Academic Press, New York, 1981, 897 pp.
92. Marshall, B. and Matthews, R. E. F., Okra mosaic virus empty protein shells in nuclei. Virology, 110,
253, 1981.
Volume I 137
Chapter 9
LUTEOVIRUS GROUP*
The name of this large group of viruses is derived from the Latin word luteus, meaning
yellow, in recognition of the yellowing symptoms that many members induce in their plant
hosts. The viruses have polyhedral particles with reported diameters between 25 and 30 nm
and sedimentation coefficients of about 115S. The capsids are constructed from a single
species of polypeptide of Mr about 24 X 103 and their genomes appear to consist of single
molecules of positive-sense, single-stranded RNA of M, about 2.4 x 10''.
All the viruses of the group seem to be confined to the phloem tissues of the host plants.
They are transmitted by aphid vectors in a persistent but not propagative manner. None of
the viruses has been transmitted by mechanical inoculation and there are no records of seed
transmission. 1 2
The large number of viruses that have been listed as members, probable members, and
possible members of the group 2 - 3 may not give a true picture of the membership, which will
probably need considerable modification in the future. Luteoviruses have in the past been
named and differentiated purely on biological properties, mainly by the hosts they infect
and their vector specificities. These are poor criteria for deciding if isolates should be
considered as distinct viruses or strains of the same virus. Attempts at sophisticated studies
of their physical, chemical and serological properties have been frustrated by inability to
obtain sufficient purified virus. This is undoubtedly due to the confinement of the viruses
to cells of the phloem which makes what little virus the plants contain difficult to ex-
tract.4"9 It is only now that improved methods for purification are being developed, such as
the use of tissue-macerating enzymes to release cell-bound virus. 10 l 3 In the meantime we
are forced to make lists, such as those in Tables 1 through 3, which in the future will almost
certainly be shown to contain errors.
Some of the viruses listed in Table 1 may be sufficiently closely related to be considered
strains of the same virus. For example, it has been shown that in immunodiffusion tests, an
antiserum to soybean dwarf virus (SDV) with a homologous liter of 1/1024 reacted with
subterranean clover red leaf virus (SCRLV) to a liter of 1/512. This suggests thai SCRLV
and SDV may have to be considered as strains of the same virus, especially now that their
host ranges also appear to be very similar, 31 and that they are transmitted by the same aphid,
Aulacorthum so/an/.26-32 Similarly, it has been shown lhal potato leafroll virus (PLRV) and
tomato yellow top virus (TYTV) (Table 2) reacted with each other's antisera to the ho-
mologous liters.33 This suggests that these Iwo viruses may also have to be considered as
strains of the same virus.
Many other antigenic relalionships recently reviewed by Rochow and Duffus 2 have been
detected among viruses listed in Tables 1 and 2. However, some of the data were based on
serum neutralization of infectivily in aphid vectors34 and more recently by immunoelectron
microscopy (IBM),35 — tests which provided qualitalive ralher lhan quanlilative results.
It is also becoming apparent that certain isolates considered to be variants of the same
virus based on their biological properties, are antigenically unrelated or only very distantly
related and thus should be considered to be distincl viruses. For example, Ihe slrains and
isolates al presenl grouped under the names barley yellow dwarf virus (BYDV) and beet
western yellows virus (BWYV) may eventually need revision. The MAV (Macrosiphum
* Figures follow text, beginning on page 142.

Table 1
DEFINITIVE MEMBERS OF THE LUTEOVIRUS GROUP AND SOME OF
THEIR PROPERTIES"
Virus confined Mr of M, of coat

Virus to phloem RNA protein
Virus SM. tissues ( x 10"| (x 10') Ref.
Barley yellow dwarf 115—120 Yes 2.0 24.5 14— 17

virus (BYDV) h
Beet western yellows 62' and 116 Yes 1.85 24.0 18—20
virus (BWYV)
Potato leafroll virus 112—127 Yes ~2.0 26.3 11. 21—24
(PLRV)
Tobacco necrotic dwarf 115 Yes 2.0 25.7 25
virus (TNDV)
Soybean dwarf virus d
Yes — 22.0 26
(SoyDV)
Subterranean clover red 114 Yes — 22.6 9, 12, 13, 27
leaf virus (SCRLV)
Pea (bean) leafroll vi- — Yes 2.4 30—35 28, 29
rus (PeLRV)
Carrot red leaf virus 104 Yes — — 8, 1 1 , 30
(CaRLV)
" All viruses included in the table have small polyhedral particles 26 to 30 nm in diameter and are transmitted
by aphids in a persistent manner.
b
This includes rice giallume virus (RGV).
c
The 62S component was shown to be devoid of RNA.
d
Not determined.
Table 2
OTHER MEMBERS OF THE LUTEOVIRUS GROUP BASED ON
SEROLOGICAL EVIDENCE
Virus Serologically related to:2'29
Beet mild yellowing virus ( B M Y V ) BWYV, BYDV, SoyDV

Malva yellows virus (MYV) BWYV, BYDV, SoyDV
Turnip yellows virus (TYV) BWYV, BYDV, SoyDV
Legume yellows virus (LYV) BWYV, BYDV, SoyDV, PLRV
Solanum yellows virus (SYV) BWYV
Tomato yellow top virus (TYTV) PLRV.SCRLV
ave«ae-transmitted) and RPV (Rhopalosiphum padi-lransmitted) isolates of BYDV do not

cross-react in immunodiffusion tests with each other's antisera. They have different aphid
specificities"-38 and they induce different cytopathological effects. 19 On these grounds they
should probably be considered as different viruses. On the other hand, the RPV isolate of
BYDV is clearly related serologically to BWYV31' and should therefore perhaps be consi-
dered a strain of that virus. The MAV and PAV (transmitted by M. avenae, R. padi, and
Schizaphis graminum] isolates of BYDV are also serologically related to BWYV, though
more distantly, and the distinction between BYDV and BWYV is further blurred by the
finding that BWYV is not restricted to dicotyledonous plants but can also infect Gramineae.-
It has been suggested that pending the development of a better classification, the terms
"Group I" and "Group II" might be used for BYDV isolates. 19 - 40 Group I would contain
isolates MAV, PAV, and SGV (S. graw/mwj-transmitted), while "Group II" would contain
isolates RPV, RMV (R. ma/'dw-transmitted), and rice giallume virus. These groups would
follow established serological differences, together with the cytopathological differences
Volume I 139
Table 3
VIRUSES THAT MAY BELONG TO THE LUTEOVIRUS GROUP"
Virus Aphid vector11
Banana bunchy top Pentalonia nigronervosa

Beet yellow net Myzus persicae
Celery yellow spot Hyadaphis foeniculi
Cotton anthocyanosis Aphis gossypii
Filaree red leaf Acyrthosiphon pelargonii
Ground nut rosette assistor Aphis craccivora
Milk-vetch dwarf Aphis craccivora
Physalis mild chlorosis Myzus persicae
Physalis vein blotch Myzus persicae
Raspberry leaf curl Aphis rubicola
Strawberry mild yellow edge Chaetosiphon fragaefolii
Subterranean clover stunt Aphis craccivora
Tobacco vein distorting Myzus persicae
Tobacco yellow net Myzus persicae
Tobacco yellow vein assistor Myzus persicae
Tomato yellow net Myzus persicae
* Data modified from Rochow and Duffus. 2 The viruses have been characterized only with
respect to their biological properties.
b
Transmitted in a persistent manner.
observed between the isolates6-19 4I discussed further in Section III. The proposed Group II
isolates share some serological and cytopathological characters with BWYV. Such grouping
would recognize that clear differences exist within isolates which are currently referred to
as "barley yellow dwarf virus" without making definitive taxonomic commitments that
might prove premature.
The present confusion concerning the taxonomy of Luteoviruses can raise problems in
their field identification, and hence in control of the diseases they cause. For example, in
the past, viruses transmitted in a circulative manner and causing leaf-rolling symptoms in
potatoes have been considered as isolates of PLRV. However, recently it has been shown
that BWYV can also infect potato plants, causing symptoms like those of PLRV. 42 Since
the two viruses appear to be serologically only remotely related, 2 - 42 using an antiserum to
one of the viruses in potato screening programs might well be misleading if infection by
the other were also present.
The reported diameter of negatively stained Luteovirus particles varies considerably. For
example, PLRV particles stained in phosphotungstate were reported by Peters21 to have a
diameter of about 24 nm, while Jayasena and his colleagues9 estimated them to be about
30 nm when stained in uranyl acetate. The lowest estimate of a Luteovirus particle diameter
is about 20 nm from phosphotungstate-stained preparations of BYDV. 14 This is almost
certainly an underestimate because the same report quotes a diameter of 30 nm from prep-
arations of shadowed particles. It seems that the spread of values reported reflects the
preparative methods used and errors in calibration of magnification rather than real differences
in size.
When stained in uranyl acetate, particles of SCRLV have hexagonal outlines and diameters
of about 30 nm (Figure 1). In such preparations SCRLV particles were indistinguishable
from those of PLRV 9 and southern bean mosaic virus which belongs to the Sobemovirus
Group (see Chapter 10).
There also appears to be considerable variation in the reported sedimentation rates of the
viruses (Table 1). Although some of this may be significant, 20 some is certainly due to
experimental errors. For example, values of between 112 and 127S have been reported for
PLRV (Table 1). It seems that with the exception of BWYV, 20 the particles of all other
Luteoviruses sediment as a single nucleoprotein component. In addition to a nucleoprotein
component sedimenting at 116S, preparations of BWYV have been shown to contain empty
protein shells sedimenting at 62S.20
Luteoviruses whose coat proteins have been examined appear to contain only a single
polypeptide of Mr between 22 and 30 x 103 (Table 1). However, these rather widely differing
estimates are from determinations by PAGE, and should be reexamined when and where
technical problems permit.
B. The Viral Genome

Those Luteoviruses whose nucleic acids have been examined contain a single species of
RNA of M, about 2 x 10" (Table 1). With tobacco necrotic dwarf virus (TNDV), two minor
components of M, about 0.1 x 106 have also been detected,23 but their significance is at
present obscure.
Until recently, the nature of PLRV nucleic acid was in some doubt. Sarkar43 had claimed
that the virus contained double-stranded DNA but this was difficult to reconcile with the
other properties of PLRV which are typically Luteovirus-like. 21 - 44 Subsequent work in a
number of laboratories leaves little doubt that the PLRV nucleic acid is a single-stranded
RNA of M, about 2 x 106.22 24 PLRV RNA appears to have positive sense as it has been
used successfully to infect plant protoplasts.45 However, in vitro translation of the RNA
yielded a single major polypeptide product of M, about 71 x 103, but nothing of the size
of coat protein. 45
The RNA of PLRV has a genome-linked protein (VPg) of M, about 7 x 103, but contains
no polyadenylate sequence. 45 The VPg does not appear to be essential for infectivity because
treatment of the PLRV RNA with proteinase K did not abolish its ability to infect proto-
plasts.45 It will be interesting to see if these properties of PLRV RNA turn out to be
characteristic of other Luteoviruses.
III. CYTOPATHOLOGY
In plant cells, detection of Luteovirus particles presents a problem because they resemble
cytoplasmic ribosomes when viewed in thin sections. It has been difficult to identify the
virus particles with confidence unless they aggregate into crystalline arrays or assume lo-
cations in the cell uncharacteristic of ribosomes. The problem of Luteovirus particle iden-
tification in thin sections has now been overcome by digesting the RNA of ribosomes in
situ with pancreatic ribonuclease after fixing the tissue to be examined with aldehyde before
post-fixation with osmium. 46 Using this technique it has been possible to detect and identify
Luteovirus particles even if only a few are scattered in the cytoplasm among ribosomes9-46-47
(Figure 2). They are also very readily detected in plasmodesmata (Figures 3A and 4A) and
vacuoles (Figure 5B) of infected cells.
All Luteoviruses that have been studied in thin sections appear to be confined to the
phloem tissues (Table 1). The most extensive cytopathological studies have been those on
BWYV, 5 4 * BYDV. 6 - 1 "- 41 - 4 "- 51 and PLRV. 4 5 2 The other viruses listed in Table 1 apparently
behave similarly, but the reports are less complete. 7 " 9 - 2 * Determining the sequence of cy-
tological events following Luteovirus infection presents considerable problems because of
the restriction in the type of cell that supports virus replication and because the viruses are
Volume I 141
not mechanically transmissible. All plant inoculations have to be done with the aphid vectors.2
Esau and Hoefert 5 - 48 examined thin sections of successively older leaves of sugar beet
systemically infected with BWYV; they assumed these to be representative of tissues infected
at various stages and interpreted, perhaps rather freely, the sequence of events during infection
as follows. They observed that virus-associated changes first occurred in cells next to infected
sieve elements and later in cells further from them. They suggested that virus particles pass
from the sieve elements, into which they are presumably introduced by the aphid vector by
way of plasmodesmata, to phloem parenchyma cells including companion cells. Particles
in plasmodesmata have also been observed in thin sections of cells infected with PLRV 47 - 52
(Figure 3A) and carrot red leaf virus (CaRLV)47 (Figure 4A).
Later on in infection, double-membraned vesicles were observed scattered in the cytoplasm
of BWYV-infected cells 5 48 (Figure 6A). Similar structures have also been observed in cells
infected with BYDV, CaRLV, PLRV, and SCRLV 947 52 (Figures 3B and 5A). In the case
of BWYV, similar vesicles have been observed in the perinuclear spaces of infected cells
(Figures 6B and 7). Esau'and Hoefert 548 suggested that these vesicles first develop in the
endoplasmic reticulum and then migrate towards the nucleus to form perinuclear vesicles
(Figure 6B). Soon after the appearance of vesicles in the perinuclear space, virus particles
appeared in the nucleus. 5 48 Initially the particles were seen close to the nucleolus, but later
became scattered throughout the nucleoplasm and sometimes formed crystalline aggregates
(Figure 8). Particles also appeared scattered in the cytoplasm and these were assumed to
have been released from the nucleus. 5 The cytoplasm of BWYV infected cells also contains
areas of densely convoluted membranes similar to those observed in plant cells infected with
PLRV (Figure 3B). The function of these membranes is obscure. They degenerate at late
stages of BWYV infection as do the cytoplasm and nuclei. 5 A very similar sequence of
events following infection with PLRV has also been postulated.52
It has been observed that, in PLRV-infected cells, the cristae of the mitochondria become
dilated and vesicles containing either fibrillar or opaque material appear in the cytoplasm.52
In cells infected with CaRLV, mitochondria also undergo morphological changes;47 they
become distended, misshapen and sometimes develop large vacuoles (Figures 4B and 9).
Vesicles containing fibrillar material also are formed at the mitochondrial periphery (Figure
9). The significance of these changes is obscure but it seems likely that the fibrillar material
within the vesicles is double-stranded RNA, possibly replicative form RNA.
Gill and Chong 6 - 19 - 41 examined oat plants infected with a number of isolates of BYDV,
which they divided on the basis of cytopathological differences into two categories. Isolates
of one, including the RPV isolate,36 produced cytopathological effects similar to those
induced by BWYV. The other isolates, including PAV and MAV, 36 behaved differently in
two respects. Firstly, the nuclei became distorted and accumulated large amounts of densely
staining material; secondly, virus particles accumulated in the cytoplasm early in infection,
rather than in nuclei and nucleoli. These differences in cytopathology correlate with the
serological differences between the isolates.36 The correlation has led to the suggestion
already discussed, that BYDV isolates should be divided into two groups.
It is not known why Luteoviruses do not invade cells outside the phloem. However, it
has been shown that protoplasts from tobacco mesophyll cells can be infected by and can
support the replication of TNDV.53
FIGURE 1. Purified preparations of A, barley yellow dwarf virus (BYDV) and B subterranean clover red leaf
virus (SCRLV), both stained in uranyl acetate and at the same magnification. Bar = 100 nm.
Volume I 143
FIGURE 2. Thin section of phloem tissue of Trijiiliiiin .\iibierriineiini (subterranean clover), showin;; a transfer
cell (above) whose cytoplasm contains numerous scattered virus particles and a phloem parenchyma cell (below)
apparently devoid of virus panicles. (Tissue cleared of ribosomes by RNase treatment.) Bar = 500 nin.
FIGURE 3. A, Thin section of two adjoining phloem tissue cells of a potato leal'roll virus (PLRV)-infected
Ph\salis floridaiia plant showing virus particles (arrows) near and in a plasmodesma. (Tissue cleared of ribosomes
by RNase treatment.) B. Thin section of a phloem parenchyma cell of PLRV-infected P. floridana showing
cytoplasmic virus-induced vesicles containing t'ibrillar material (arrows) reminiscent of nucleic acid and patches
of convoluted membranes (in). (Tissues cleared of ribosomes bv RNase treatment.) Bars = 500 nm.
Volume I 145
FIGURE 4. A. Thin scclion of two adjoining phloem tissue cells of Daiicus ctirotti (carrot) infected with carrot
red leaf virus (CaRLV). showing virus particles in the cytoplasm and plasmodesmata (arrows). (Tissue cleared of
ribosomes by RNase treatment.) B. Thin section of mitochondria in the cytoplasm of a D. caroia (carrot) phloem
leaf cell infected with CaRLV. showing vesicles with fibrillar material (arrows). (Tissue cleared of ribosomes by
RNase treatment.) liars = 100 nm.
FIGURE 5. A, Thin section of a leaf phloem parenchyma cell of P. floridana infected with PLRV, showing
groups of virus particles and virus-induced cytoplasmic vesicles containing fibrillar material. (Tissue cleared of
ribosomes by RNase treatment.) B, Thin section of a leaf phloem cell of P. floridana infected with PLRV, showing
crystalline arrays of virus particles in the vacuole. (Tissue cleared of ribosomes by RNase treatment.) Bars = 500
nm.
Volume I 147
FIGURE 6. Thin sections of /J<?/o vulgaris (sugar beet) leaf phloem cells infected with beet western yellows virus
(BWYV), showing virus-induced vesicles containing fibrils. A, Vesicles located in the cytoplasm; B, one vesicle
in the cytoplasm and one in the perinuclear space (arrow). Bars = 500 nm. (Courtesy of Dr. K. Esau, Department
of Biological Science, University of California, Santa Barbara.)
FIGURE 7. Thin section of a Beta vulgaris (sugarheet) leaf phloem cell infected with BWYV. showing char-
acteristic virus-induced vesicles in the perinuclear space (arrows). Bar = 500 nm. (Courtesy of Dr. K. Esau.
Department of Biological Science, University of California. Santa Barbara.)
Volume I 149
FIGURB 8. A, Thin section of Beta vulgaris (sugarhect) leaf phloem eell infected with BWYV, showing aggregates
of virus particles (arrows) in the nucleus. B, Higher magnification of section in A, showing details of virus particles
in the nucleus. Bars = 500 nm. (Courtesy of Dr. K. Esau, Department of Biological Science, University of
California. Santa Barbara.)
FIGURE 9. Thin section of a leaf phloem parenchyma cell of D. carota (carrot) infected with CaRLV, showing
an abnormal mitochondrion with a central vacuole and peripheral vesicles containing fibrillar material (arrows).
(Tissue cleared of ribosomes by RNase treatment.) Bar = 500 nm.
Volume I 151
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48. Esau, K. and Hoefert, L. L., Ultrastructure of sugarbeet leaves infected with beet western yellows virus.
J. Ultrastruct. Res., 40, 556, 1972.
49. Gill, C. C. and Chong, J., Vascular cell alterations and predisposed xylem infection in oats by inoculation
with paired barley yellow dwarf viruses. Virology, 114, 405, 1981.
50. Jensen, S. G., Occurrence of virus particles in the phloem tissue of BYDV-infected barley. Virology, 38,
83, 1969.
5 1 . Aurici, A., Faoro, F., and Tornaghi, R., Intracellular crystallisation in roots of Leersiu ory:oides of a
strain of barley yellow dwarf virus. Micron, 10, 5, 1979.
52. Shepardson, S., Esau, K., and McCrum, R., Ultrastructure of potato leaf phloem infected with potato
leafroll virus, Virology, 105, 379, 1980.
53. Kubo, S. and Takanami, Y., Infection of tobacco mesophyll protoplasts with tobacco necrotic dwarf
virus, a phloem-limited virus, J. Gen. Virol., 42, 387, 1979.
Volume I 153
Chapter 10
SOBEMOVIRUS GROUP*
The Sobemovirus group is named after southern bean mosaic virus (SBMV), the type
member. 1 The viruses have stable polyhedral particles about 30 nm in diameter with T =
3 symmetry built from identical protein subunits of Mr about 30 x 10' which sediment at
about 115S. Their genomes consist of a single molecule of linear positive-sense, single-
stranded RNA of Mr about 1.4 x 106.
Sobemoviruses are transmitted by beetles, through seeds, and mechanically, each to a
narrow range of plant species.2
At present, the ICTV recognizes only two definite members in the group, 1 SBMV 1 and
turnip rosette virus (TRoV). 4 Four other viruses have been listed by Matthews' as possible
members: blueberry shoestring virus (BSSV), S cocksfoot mottle virus (CoMV), 6 rice yellow
mottle virus (RYMV), 7 and sowbane mosaic virus (SoMV). B The last three of these viruses
together with SBMV and TRoV have been considered definite members of the group by
Sehgal, 2 and Forster and Jones9 have included lucerne transient streak virus (LTSV) as a
member of the group. No serological relationships have been reported between any of these
viruses and further data are needed to establish beyond doubt that they are indeed
Sobemoviruses.
A number of other viruses have some properties sufficiently similar to SBMV to be
considered possible members of the Sobemovirus group (Table 1). Cynosurus mottle virus
(CyMoV) has been shown to be serologically distantly related to CoMV; 10 this observation
was confirmed by Catherall and reported as a private communication to Sehgal. 2 However,
this relationship was not easily detected by Paul and his colleagues." These workers studied
the serological relationships between a number of viruses isolated from Gramineae including
CoMV and CyMoV which Hull 1 2 suggested should be included in a group with phleuin
mottle virus as type member. It would appear that viruses such as CoMV on the one hand
and phleum mottle virus on the other, may not be as different as envisaged by Hull, 1 2 and
the possible affinities of both viruses should be considered with members of the Sobemovirus
group. However, more data are needed before firm taxonomic proposals are made.
Recently, three new viruses, those of velvet tobacco mottle (VTMoV), 13 solanum nodi-
florum mottle (SNMV), 14 - 15 and subterranean clover mottle (SCMoV), have been shown to
share a number of properties characteristic of Sobemoviruses. However, the RNA comple-
ments of these viruses are unlike those of SBMV or TRoV since in addition to single-
stranded RNAs of Mr about 1.5 X 106, they also encapsidate small circular viroid-like
RNAs of M, about 1.2 X 10V- 15 - 16 A close serological relationship has been demonstrated
between VTMoV and SNMV. 1 - 1 - 17 A viroid-like RNA has also been detected in the particles
of LTSV, 18 and this was shown to be serologically very distantly related to SCMoV. 1 "
Except for the encapsidation of viroid-like RNAs by the particles of VTMoV, SNMV,
LTSV, and SCMoV, these viruses seem remarkably like Sobemoviruses. 22 The significance
of the encapsidated viroid-like RNAs is not altogether clear at present. With VTMoV and
SNMV it has been reported that both the large virus-like RNA and the small circular viroid-
like RNA are required for infectivity. 21 However, with LTSV it has been shown that the
* Figures I through 8 follow text, beginning on page 159.

Table 1
SOME BIOLOGICAL AND PHYSICAL PROPERTIES OF SOBEMOVIRUSES
AND
OTHER POSSIBLY RELATED VIRUSES
Physical properties
Virus Viral R N A M r
Transmission by particles ( X 10")
Viral oat
p in Virus- Viroid- protein
Virus Vector Seed S*. CsCl like like M, ( x 10') Ref.
Southern bean mosaic Beetles Yes 115 1.36 1.4 Absent 29 3

virus (SBMV)
T u r n i p rosette virus Beetles — •' 112 1.34 1.4 Absent 30 4
(TRoV)
Blueberry shoestring Beetles No 120 1.39 1.4 — 30 5
virus (BSSV)
Cocksfoot mottle Beetles No 118 1.37 1.4 — 32 6,19
virus (CoMV)
Rice yellow mottle Beetles No 109 1.37 1.4 — 31 7,20
virus (RYMV)
Sowbane mosaic virus Leaf'miner, Yes 113 1.35 1.3 Absent 31 8
(SoMV) leaf-
hopper
Cynosurus mottle Aphid — 113 1.40 1.5 — 31 10,19,21
virus (CyMoV)
Velvet tobacco mottle Mirid No 115 1.37 1.5 0.12 37 13,17
virus (VTMoV) (33, 31.5)"
Solanum nodiflorum Beetles No 115 1.37 1.5 0.12 31 14,15,17
mottle virus (SNMV)
Lucerne transient streak — No 113 1.37 1.4 0.10 34(30) 9,18
virus (LTSV)
Subterranean clover — — 115 — 1.5 0.14 or 29 (26) 16
mottle virus (SCMoV) 0.10
" Not determined.

b
Values in brackets indicate molecular weights of minor components detected in preparations of coat proteins
by PAGE.
viroid-like RNA is not essential for infectivity and that it appears to be a satellite RNA. 24 25
Until more is known about the exact relationship of the viroid-like RNAs to these viruses,
they should best be considered as possible members of the Sobemovirus group.
It is interesting to note that while the majority of viruses listed in Table 1 are transmitted
by insects with biting mouth parts (e.g., beetles), BSSV has been transmitted in one ex-
periment by the blueberry aphid Masonaphis pepperi.^ CyMoV has also been reported to
be transmitted by the aphid Rhopalosiphum padi.2' Liriomyza langei (a leafminer fly) and
Circulifer tenellus (the beet leafhopper) have also been reported as vectors of SoMV;8-26
however, these reports need confirmation.
SBMV, on which more vector work has been done, can be transmitted without an in-
cubation period by its chrysomelid beetle vector. The virus appears to enter the haemolymph
and can continue to be transmitted by the insect for some days after feeding on infected
plants. Insects injected with virus into the haemocoel can also become infective. 27
At present we consider it likely that all the viruses listed in Table 1 belong to the
Sobemovirus group. However, further discussion here will mainly concern SBMV because
so much more is known about it than about any of the other viruses.
Volume I 155
Particles of SBMV stained in uranyl acetate appear hexagonal with no obvious structural
detail (Figure 1). They measure 30 nm in diameter, which agrees well with determination
from X-ray diffraction data. 28 X-ray diffraction studies have also revealed that each SBMV
capsid consists of 180 polypeptides arranged with T = 3 quasi-symmetry surrounding the
viral RNA. 28 " 31 Neutron small-angle scattering data indicate that the SBMV particle has a
tightly packed outer protein shell, with the RNA and about 15% of the protein in the interior
of the particle. 32 - 31 The protein penetrates as deeply as the RNA into the particle center.33
SBMV particles are constructed from a single protein species of M, about 29 x 103. The
sequence of the 260 amino acids of the cowpea strain coat protein has been determined. 34
The 87 carboxy-terminal arnino acids of the bean strain of SBMV have also been deduced
from RNA sequencing data. 35 In the capsid there appear to be three different types of quasi-
equivalent subunit with different conformations but the same covalent structure, designated
as the A, B, and C subunits. 31 The A subunits cluster around the fivefold axis and sets of
B and C subunits form the quasi-sixfold vertices.
Data on the size of the coat protein subunit, determined from its amino acid sequence
(A/ r ~28.2 x 10 3 ), 34 and the size of the RNA (Mr ~1.4 x 106),35 have led to the deduction
that the SBMV particle must be of Mr about 6.5 x 10". This is very close to the values of
between 6.5 and 6.6 x 106 derived from physical data.36-37
The integrity of the SBMV particle is dependent on divalent metal ions and the function
of both calcium and magnesium has been studied extensively. 30 - 38 41 On removal of these
ions under mildly alkaline conditions, the particles swell, 40 - 42 increasing in diameter by about
7%. 30 In this state they are susceptible to dissociation 43 or enzymatic degradation. 44 Atomic
absorption spectroscopic studies indicated that 120 Mg 2 + and 80 Ca 2+ are firmly bound to
each native SBMV particle, 40 whereas Hull 45 detected 280 Ca2 + and 380 Mg2 + per particle.
From studies involving the dissociation of SBMV and TRoV particles with EDTA, it was
concluded that the particles are stabilized by three types of bonds, one involving the divalent
cations, one being pH dependent, and one due to a salt link between protein and RNA. 45
The work of Abdel-Meguid and colleagues46 indicates that the Ca 2+ are located on the quasi-
threefold axes between subunits A, B, and C.
When SBMV particles swell in response to high pH and chelation of divalent cations,
the highly basic amino-terminal coat protein arm is vulnerable to trypsin. 44 - 47 On the loss
of this fragment of 61 residues whose function it is to penetrate deep into the virus particle
and stabilize its quasi-sixfold arrangement, 33 T = 3 particles can no longer be reassembled.
However, T = 1 particles can be formed from the residual polypeptides of A/r about 22 x
10' in the absence of RNA. 4 *- 49 It has been shown that if the termini can be protected from
excision, by proteolytic enzyme inhibitors. T = 3 particles can be reassembled in vitro from
the cowpea strain of SBMV coat protein and RNA under carefully controlled ionic
environments. 4 ''
Relatively little is known about the particle structure of the other viruses listed in Table
1. but it has been shown that the particles of SoMV (Figure IB), VTMoV (Figure 1C),
SNMV, LTSV (Figure ID), and SCMoV appear indistinguishable from those of SBMV
when preparations are mixed together, stained in uranyl acetate, and viewed in the electron
microscope. 22 It is difficult to conclude whether particles of the other viruses listed in Table
1 are distinguishable from those of SBMV because of the variable quality of micrographs
published by different authors. Also, the stabilities of the other viruses have not been studied
in detail although a number of them have been shown to dissociate in EDTA. 1 - 4 2 5 0 This
indicates that their particle integrity may also be dependent on factors similar to those
stabilizing SBMV.
B. The Viral Genome

Infectious RNA of SBMV consists of a single component of M r about 1.4 x 10V5-51 A
protein (VPg) of A/r about 12 x 103 is covalently bound to the 5' end of this RNA, but
there is no poly A tract at the 3' end. 52 The VPg is essential for infectivity. 53 SBMV particles
also encapsidate a subgenomic RNA of Mr about 0.38 x 106 which has been shown to have
coat protein mRNA activity in vitro. 5I - 54 - 55 The 3' terminal 90 residues and its 5' terminal
VPg are indistinguishable from those of the genomic RNA. 56
The genomic RNA can be translated in vitro to yield three proteins of Mr 105, 75, and
14 x 103, but no coat protein. 51 - 54 The base sequences of the coat protein mRNA are present
near its 3' end; hence it exists as a silent cistron,56 at least as regards in vitro translation.
The genome of TRoV, like that of SBMV, consists of a single RNA molecule of A/r about
1.5 x 106 with a VPg at its 5' end, but no poly A tail at the 3' end. 57 The virus also
encapsidates a subgenomic mRNA. The genomic RNA when translated in vitro produced
polypeptides of Mr about 105, 57, 35, and 30 x 103, the smallest one being the coat
protein. 57
Whereas no low-molecular-weight RNAs have been detected in particles of SBMV, TRoV,
and SoMV, 13 - 58 circular single-stranded RNAs from 324 to 388 bases long occur in those
of VTMoV, SNMV, LTSV, and SCMoV. '3.15.16.18.22 of these four viruses with low-molec-
ular-weight components, VTMoV is the most extensively studied. 22
RNA isolated from VTMoV particles can be separated by polyacrylamide-gel electro-
phoresis into five fractions with apparent Mrs of about 1.5 x 106 (RNA 1), 0.63 x 106
(RNA la), 0.25 x 10" (RNA Ib), 0.16 x 106 (RNA 2), and 0.12 x 10" (RNA 3). RNAs
la and Ib were shown to be fragments of RNA 1, la probably being the coat protein
mRNA. 22 RNAs 2 and 3 have base sequences indistinguishable from each other, but with
no detectable homology to RNA I. 5 9 RNA 2 appears to be a circular form of RNA 3. 13 - 59
The native conformation of both RNAs 2 and 3 is partially double stranded and highly
reminiscent of viroid RNAs. 59
It has been reported that VTMoV and SNMV require both RNA 1 and RNA 2 for
infectivity. 23 Furthermore, the interaction of the two essential RNAs appears to be highly
specific because heterologous mixtures of RNA 1 and RNA 2 failed to infect plants. 23 On
the other hand, LTSV has been shown to require only RNA 1 for biological activity and
RNA 2 appears to behave as a satellite RNA. 25 Moreover, RNA 2 of LTSV appears fairly
unspecific, having been shown to be capable of replication in the presence of RNA 1 from
either SNMV 24 of SoMV60 although field isolates or SoMV have never been found to contain
viroid-like RNAs. The apparently different nature of the viroid-like RNAs of VTMoV and
SNMV from that of LTSV is puzzling at present and more information is needed for an
understanding of their functions.
Attempts to translate the viroid-like RNAs of VTMoV and LTSV in vitro have been
unsuccessful, 22 - 61 and hence in this respect they behave like the viroids.62 However, it has
been demonstrated that LTSV RNA 1 is translated in vitro to yield polypeptides of Mr about
105, 78, and 33 x 103, the smallest of which was identified as the coat protein.61 Fur-
thermore, it was shown that a subgenomic RNA of Mr about 0.35 x 106 was a more efficient
messenger for coat protein.61 Thus, the translational strategy of LTSV seems to be similar
to those of SBMV and TRoV. 51 - 54 - 57
The RNAs of BSSV, CoMV, RYMV, and CyMoV (Table 1) have not been studied in
any depth. Although no viroid-like RNAs have been reported, a thorough search does not
appear to have been made.
III. CYTOPATHOLOGY
The cytopathology of all the viruses listed in Table 1 except apparently that of TRoV,
Volume I 157
Table 2
CYTOPATHOLOGICAL EFFECTS OF SOBEMOVIRUSES AND OTHER
APPARENTLY RELATED VIRUSES
Presence in
Presence of virus particles in cytoplasm of
Virus Nucleus Cytoplasm Vacuole Fibrils Tubules Ref.
Southern bean mosaic Yes" Yes" Yes" Yes No 22, 65—67, 69

virus (SBMV)
Blueberry shoestring No Yes Yes No Yes" 63
virus (BSSV)
Cocksfoot mottle Yes Yes Yes No No 70
virus (CoMV)
Rice yellow mottle No Yes" Yes" Yes Yes 20
virus (RYMV)
Sowbane mosaic virus Yes Yes Yes Yes No 22, 68
(SoMV)
Cynosurus mottle No Yes Yes Yes No 19, 64
virus (CyMoV)
Velvet tobacco mottle Yes Yes Yes Yes No 13, 22
virus (VTMoV)
Solanum nodiflorum Yes Yes Yes Yes Yes" 14, 22
mottle virus (SNMV)
Lucerne transient Yes Yes Yes Yes No 22
streak virus (LTSV)
Subterranean clover Yes Yes Yes Yes No 16, 22
mottle virus
(SCMoV)
" Virus particles sometimes crystallize into inclusions. In the case of SBMV, the cowpea strain induced
crystals whereas the type strain did not.
b
Tubules also present in the nuclei.
has been investigated to varying degrees. Particles of all the viruses listed in Table 2 have
been detected in the cytoplasm (Figures 2 through 8) and vacuoles (Figure 5) of infected
cells. With the exception of BSSV, RYMV, and CyMoV, particles have also been observed
in the nuclei (Figures 2 and 5 through 8). In the case of BSSV and RYMV, it is not clear
if a careful search was made for particles in the nuclei,20-63 but in spite of some effort to
find them, none were detected in nuclei of CyMoV-infected cells.64 In a few instances,
particles have been seen in crystalline arrays in the cytoplasm and vacuoles.20-65'67 Milne68
observed crystals of SoMV particles in cells of leaves, but only when the leaves had been
wilted (Figure 4C). Particles of most of the viruses accumulate in large numbers without
crystallizing (Figures 2 through 5). With some viruses such as SCMoV, the particles ag-
gregate even in cells containing relatively few particles (Figure 8). In sections of SoMV-
infected cells, dense circular areas in the cytoplasm composed of virus particles in a dense
matrix have been observed68 (Figure 4B). No particles have been detected in either chlo-
roplasts or mitochondria of cells infected with any of the viruses listed in Table 2.
Cells infected with a number of the viruses listed in Table 2 contain fibrils dispersed in
the cytoplasm, mainly as patches in the ground cytoplasm (Table 2 and Figures 3 through
5), but some are also enclosed in vesicles of the endoplasmic reticulum (Figures 2B, 3, and
6B). The nature of this material is uncertain but it resembles nucleic acid68 and some evidence
has been reported for it being double-stranded replicative form RNA. 22 At least three of the
viruses, BSSV, RYMV, and SNMV, also induce the development of characteristic tubules
which usually aggregate into bundles. l4 - 20 - 22 In the case of SNMV-infected cells, these
bundles accumulate in both cytoplasm and nuclei 14 - 22 (Figure 6); their origin and function
remain unknown. It was suspected in the case of SNMV that they could have resulted from
infection by a second virus, but this seems unlikely because cells of plants infected with
preparations of virus purified by isopycnic banding in CsCl and shown to contain only
SNMV particles still contained the tubules. Similarly, cells of plants infected with mixtures
of RNA 1 and RNA 2 purified by two cycles of PAGE again contained the tubules. 71
Volume I 159
FICiURE I . Purified virus particles. A, Southern hean mosaic virus (SBMV); B. sowhane mosaic virus (SoMV);
C, velvet tobacco mottle virus (VTMoV): D. lucerne transient streak virus (LTSV). All preparations stained in
uranyl acetate and photographed at the same magnification. Bar = 100 nm.
FIGURE 2. Thin sections of Phaseohis vulgurix (French bean) leal'cells infected with SBMV (tissue cleared of
ribosomes by RNase treatment). A. Virus particles scattered throughout the cytoplasm and nucleus: B. heavier
concentration of virus particles in the nucleus and a virus-specific vesicle with fibrils (arrow) in the cytoplasm.
Bars = 500 nm.
Volume I 161
FIGURE 3. Material as in Figure 2 showing virus particles scattered in the cytoplasm and virus-specific fibrils
in the ground cytoplasm (arrowheads) and in cytoplasmic vesicles (arrows). Bar = 500 nm.
FIGURE 4. Thin sections of Chenopodium qiiinoa leaf cells infected with sovvbanc mosaic virus (SoMV). A,
Virus-specific fibrils in the ground cytoplasm; B. dense cytoplasmic matrix containing virus particles (tissues
cleared of ribosomes by RNase treatment in both A and B). Bars = 500 nm. C, SoMV particles crystallized in
the cytoplasm of a C. amtiranticolor leaf cell after wilting. Bar = 100 nm.
Volume I 163
FIGURE 5. Thin section of a Nicoliana clevclandii leaf cell infected with velvet tobacco mottle virus (VTMoV)
showing virus particles in the cytoplasm, nucleus and vacuole, and fibrils in the ground cytoplasm (arrows). (Tissue
cleared of ribosomes by RNase treatment.) Bar = 500 nm.
FIGURE 6. Thin sections of Nirotiana clerrlandii leaf cells infected with solanuni nodiflorum mottle virus
(SNMV) showing virus panicles anil virus-specific tubules in the nucleus (A) and cytoplasm (B). Virus specific
fibrils in cytoplasmic vesicles (arrows) can also he seen in (B). (Tissue cleared of ribosomes by RNase treatment.)
Bars = 500 nm.
Volume I 165
FIGURE 7. Thin section of a Chenopodium quinoa leaf cell infected with lucerne transient streak virus (LTSV)
showing virus particles scattered in the nucleus and cytoplasm. (Tissue cleared of ribosomes by RNase treatment.)
Bar = 500 nm.
FIGURE 8. Thin section of a leaf of Trifolium subterraneum (subterranean clover) infected with subterranean
clover mottle virus (SCMoV) showing aggregates of virus particles in the nucleus and cytoplasm. (Tissue cleared
of ribosomes by RNase treatment.) Bar = 500 nm.
Volume I 167
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46. Abdel-Meguid, S. S., Yamane, T., Fukuyama, K., and Rossmann, M. G., The location of calcium
ions in southern bean mosaic virus, Virology, 114, 81, 1981.
47. Tremaine, J. H. and Ronald, W. P, Limited proteolysis of southern bean mosaic virus by trypsin, Virology,
91, 164, 1978.
48. Erickson, J. W. and Rossmann, M. G., Assembly and crystallization of a T = 1 icosahedral particle
from trypsinized southern bean mosaic virus coat protein, Virology, 116, 128, 1982.
49. Savithri, H. S. and Erickson, J. W., The self-assembly of the cowpea strain of southern bean mosaic
virus: formation of T = 1 and T = 3 nucleoprotein particles, Virology, 126, 328, 1983.
50. Forster, R. L. S. and Jones, A. T., Properties of lucerne transient streak virus, and evidence of its affinity
to southern bean mosaic virus, Ann. Appl. Biol., 93, 181, 1979.
51. Rutgers, T., Salerno-Rife, T., and Kaesberg, P., Messenger RNA for the coat protein of southern bean
mosaic virus, Virology, 104, 506, 1980.
52. Ghosh, A., Dasgupta, R., Salerno-Rife, T., Rutgers, T., and Kaesberg, P., Southern bean mosaic virus
has a 5'-linked protein but lacks a 3' terminal poly (A), Nucleic Acids Res., 7, 2137, 1979.
53. Veerisetty, V. and Sehgal, O. P., Proteinase K-sensitive factor essential for the infectivity of southern
bean mosaic virus nucleic acid. Phytopathology, 70, 282, 1980.
54. Salerno-Rife, T., Rutgers, T., and Kaesberg, P., Translation of southern bean mosaic virus RNA in
wheat embryo and rabbit reticulocyte extracts, J. Virol., 34, 51, 1980.
55. Weber, K. A. and Sehgal, O. P., Subgenomic RNAs in virions of southern bean mosaic virus, Phyto-
pathology, 72, 909, 1982.
56. Ghosh, A., Rutgers, T., Ke-Qiang, M., and Kaesberg, P., Characterization of the coat protein mRNA
of southern bean mosaic virus and its relationship to the genomic RNA, J. Virol., 39, 87, 1981.
57. Morris-Krsinich, B. A. M. and Hull, R., Translation of turnip rosette virus RNA in rabbit reticulocyte
lysates, Virology, 114, 98, 1981.
58. Hull, R., personal communication, 1983.
59. Gould, A. R., Studies on encapsidated viroid-like RNA. II. Purification and characterization of a viroid-
like RNA associated with velvet tobacco mottle virus (VTMoV), Virology, 108, 123, 1981.
60. Francki, R. I. B., Chu, P. W. G., and Keese, P. K., The satellite nature of a viroid-like RNA from
lucerne transient streak virus, in Current Communications in Molecular Biology, Cold Spring Harbor
Laboratory, Cold Spring Harbor, N.Y., 1983, 175.
Volume I 169
6 1 . Morris-Krsinich, B. A. M. and Forster, R. L. S., Lucerne transient streak virus RNA and its translation
in rabbit reticulocyte lysate and wheat germ extract. Virology, 128, 176, 1983.
62. Diener, T. O., Viroids: abnormal products of plant metabolism. Ann. Rev. Plant Physiol.. 32, 313, 1981.
63. Hartmann, J. X., Bath, J. E., and Hooper, G. R., Electron microscopy of viruslike particles from
shoestring-diseased highbush blueberry. Vaccinium corymbosum L., Phytopathology, 63, 432, 1973.
64. Mohamed, N. A., An ultrastructural study of three grasses naturally infected with cynosurus mottle virus,
Phytopathol. Z., 100. 121, 1981.
65. Edwardson, J. R., Purcifull, D. E., and Christie, R. G., Electron microscopy of two small spherical
plant viruses in thin sections, Can. J. Bin., 44, 821, 1966.
66. Weintraub, M. and Ragetli, H. W. J., Electron microscopy of the bean and cowpea strains of southern
bean mosaic virus within leaf cells. J. Ultrastruct. Res., 32, 167, 1970.
67. Weintraub, M. and Ragetli, H. W. J., Identification of the constituents of southern bean mosaic virus
in crystals of infected cells, and their distribution within the virion, Virology, 41, 729, 1970.
69. De Zoeten, G. A. and Gaard, G., Possibilities for inter- and intracellular translocation of some icosahedral
plant viruses, J. Cell. Bio!., 40, 814,1969.
70. Chamberlain, J. A. and Catherall, P. L., Electron microscopy of some grasses and cereals infected with
cocksfoot mottle, phleum mottle and cocksfoot mild mosaic viruses, J. Gen. Virol., 30, 41, 1976.
71. Hatta, T., Gould, A. R., and Francki, R. I. B., unpublished results, 1980.
Volume I 171
Chapter 11
TOBACCO NECROSIS VIRUS GROUP*
Tobacco necrosis (TNV) is the only approved member of the group. 1 The virus has
polyhedral particles about 30 nm in diameter sedimenting as a single component of about
118S and containing positive-sense RNA of Mt about 1.4 x 106.2 The viral coat protein
consists of one type of polypeptide of M, about 30 x 10V
TNV is not of great economic importance as it seldom causes diseases other than in tulips,
French beans, and cucumbers. 2 However, it commonly occurs in the roots of many plant
species and it can be transmitted mechanically to leaves, usually resulting in the development
of necrotic local lesions from which the virus does not spread.2 The virus has been studied
extensively as it is one of the few transmitted by a pythiaceous fungus 4 " 6 and because it can
act as a helper for the otherwise noninfectious satellite virus of TNV (SV). 7 - 8
I. TOBACCO NECROSIS AND OTHER POSSIBLY RELATED VIRUSES
A number of TNV strains have been characterized and found to vary considerably in their
biological properties, stability, and serological specificity. 9 - 10 Some strains differ serologi-
cally by SDIs as large as 3 to 5, and eight strains of TNV have been divided into two
serotypes. 10 " This classification has been questioned by Uyemoto and colleagues, 12 who
were unable to detect clear serological boundaries between strains. There is specificity as
to which strains of TNV can support the multiplication of different strains of SV."- 1 2 This
marked diversity among TNV strains suggests that some could have been considered as
distinct viruses.
Cucumber necrosis virus (CuNV) 11 has been considered as a possible member of the
tobacco necrosis virus group, 1 presumably because it produces local lesions on many host
plants and is transmitted by a species of Olpidium.14'15 However, CuNV particles are ser-
ologically unrelated to those of TNV and differ in a number of other properties, 10 - 16 - 17
indicating that the two viruses should not be included in the same group. Some properties
of CuNV are discussed in Chapter 12.
It should also be noted that a different virus of cucumbers reported from Holland was
named cucumber necrosis virus 18 and later shown to be a serotype of TNV. 10
Another virus that may be considered as a possible member of the group is carnation
yellow stripe virus (CYSV), 19 though it is not yet clear whether it might not equally well
be considered a strain of TNV. The virus has reported physical and chemical properties
similar to those of TNV, except that the sedimentation coefficient is apparently only 99 to
105S. There is a single RNA species of Mr about 1.5 x 106, a single coat protein of Mr
about 27 x 10', and the buoyant density of the particle in CsCl is 1.369 g/rn£. The virus
is serologically related to both a South African grapevine strain and a German strain of
TNV. The narrow host range, low sedimentation coefficient, and induction of multivesicular
bodies in infected cells argues for CYSV being regarded as a distinct virus within the TNV
group. 19
/. Tobacco Necrosis Virus
The reported diameter of TNV varies from 26 to 30 nm. 2 When stained in uranyl acetate,
* Figures follow text beginning on page 175.

the particles have hexagonal outlines and diameters of about 30 nm (Figure 1A). Lesnaw
and Reichmann 20 provided evidence that TNV particles consist of icosahedra with T = 3
symmetry. There is no evidence for this from electron microscopy because most published
micrographs, like that in Figure 1A, fail to reveal any surface structure. 21 Reports of other
properties of TNV vary considerably. The particles sediment at 112 to 133S, contain between
18 to 21% RNA, and are of A/,, between 6.3 and 7.6 x 106.2 It is not clear if this variation
is due to experimental conditions, error, or intrinsic differences between virus strains.
The virus particle is built from a single species of polypeptide. however, there was
confusion for some time as to the size of this molecule, with reported Mr values ranging
from 23 to 34 x 103.2 The correct value is probably close to 30 x 103,3 which would give
a particle Mr of about 6.8 x 106.
2. Satellite Virus
The particle structure and composition of SV is better understood than that of its helper,
TNV. When stained with uranyl acetate, SV particles appear as polyhedra about 17 nm in
diameter (Figure IB). This agrees well with the value of 16.8 nm obtained for the particle
diameter in solution when measured by small-angle X-ray methods. 22 The particles contain
about 20% RNA 23 and sediment at between 49 and 58S.2
SV is readily crystallized, and X-ray data on the crystals indicate that the particle has
icosahedral, T = 1 symmetry. 24 The detailed structure of the SV particle has been investigated
by X-ray crystallography at a resolution of 3.0 A. 25 The particle is built from a single
polypeptide species of about Mr 21.6 x 10\ whose entire sequence of 195 amino acids has
been determined both directly2'1 and by deduction from the viral RNA base sequence. 27 From
these data and the M,. of the RNA, which is 0.34 X 106,27 the molecular weight of the SV
particle must be 1.64 x 106; this is close to the value reported by other authors. 2228
B. The Viral Genome

/. Tobacco Necrosis Virus
The TNV genome consists of a single molecule of single-stranded, positive-sense RNA
which sediments at between 24 and 27S.20-29 From these data M, values of 1.32 and 1.57
x 106 were calculated. The lower value is very close to that of 1.3 x 106 obtained by
polyacrylamide-gel electrophoresis.30
TNV RNA has been translated in vitro into the coat protein of Mr about 30 x 103 and
only small amounts of three polypeptides of M, about 63, 43, and 26 x 103.3 Condit and
Fraenkel-Conrat31 have isolated three classes of double-stranded RNA from TNV-infected
plants, one corresponding to a duplex of TNV RNA of M, about 2.6 x 10" and two smaller
molecules of Mr about 1.05 and 0.94 x 106. Oligonucleotide maps of T, RNase digests of
these RNAs indicate that the two smaller species are derived from the 3' end of the genomic
RNA. 3 ' It has also been shown that the smallest RNA can code in vitro for the coat protein.
Thus it appears that TNV RNA contains at least three cistrons, only one of which is translated
efficiently in vitro.
2. Satellite Virus
The complete nucleotide sequence of SV RNA (1239 nucleotides) has been determined
and a secondary structure model of the RNA proposed.27 It appears that SV RNA can be
folded into a transfer-like RNA cloverleaf structure with an anticodon for AUG at residues
30—32 from the 5' terminus. This is followed by an open reading frame for the coat protein,
up to a UAA termination site at residues 618—620, accounting for half the coding capacity
of the RNA. The rest of the molecule does not appear to code for any protein.27
The postulated highly base-paired structure of SV RNA may explain its stability in vivo
when inoculated to plants in the absence of the helper virus. SV RNA free of TNV RNA,
when inoculated to plants, does not replicate; yet if the plants are subsequently reinoculated
Volume I 173
with TNV RNA, both satellite and helper viruses will multiply, even if the time between
inoculation with the two viruses is as long as 10 days. 32
Unlike most eukaryotic mRNAs, including most plant viral RNAs, the 5' ends of both
TNV RNA and SV RNA terminate in ppAGU and lack a cap structure. There is also no 3'
terminal poly-A tail. 31 - 14 Nevertheless, SV RNA can function in vitro as a monocistronic
mRNA coding for SV coat protein both in eukaryotic 3 - 35 - 36 and prokaryotic systems. 16 - 37 - 3x
Although the 5' terminal cap on eukaryotic mRNAs is thought to be involved in the formation
of initiation complexes for translation, 19 SV RNA is not capped before or during translation
in the wheat-germ system.16 Moreover, the addition of a cap to the 5' terminus of SV RNA
does not enhance its translation efficiency. 40
Lesnaw and Richmann 41 consider that the occurrence of AGU at the 5' termini of both
SV and TNV RNAs is not fortuitous. They suggest that this sequence may have a role in
the recognition of the RNA replicase.
III. CYTOPATHOLOGY
Edwardson and his colleagues42 were the first to detect TNV particles in infected tobacco
leaves. It would appear that their isolate was free of SV. They observed TNV particles,
many forming crystalline aggregates, throughout the cytoplasm of necrotic cells. Such cells
had contorted walls and their contents consisted largely of structureless, densely staining
material. No virus was seen in non-necrotic cells.
Sections of non-necrotic cells from the edges of local lesions on Nicotiana X edwardsonii
leaves infected with a TNV isolate devoid of SV were examined by Hatta and Francki. 43
The cells were first cleared of ribosomes by treating fixed tissue with RNase. 43 Numerous
virus particles were found scattered throughout the cytoplasm often in groups (Figures 2
and 3) but never forming crystalline aggregates. The endoplasmic reticulum was prominent,
usually occurring as pairs of membranes stretching over large regions of the cells (Figures
2C and 3). Sometimes patches of electron-dense amorphous material (Figures 2A and 3)
were also seen. The amorphous material appears to be similar to that observed in necrotic
cells examined by Edwardson and colleagues42 and its presence may be an indication that
the cells are on the point of turning necrotic.
Many tonoplasts of TNV-infected cells bore numerous membrane-bound vesicles (Figures
2C and 3) similar to those observed in cells infected by Cucumoviruses 44 (see Chapter 2,
Volume 2), some Tobamoviruses (see Chapter 9, Volume 2), and Potexviruses (see Chapter
12, Volume 2). With Cucumoviruses, it has been suggested that the tonoplast vesicles may
be the sites of viral RNA replication.44
A possible reason why Edwardson and his colleagues42 failed to detect virus particles in
non-necrotic cells surrounding local lesions is that they were unable to distinguish them
from ribosomes, a common problem when dealing with small polyhedral virus particles.43-45
Edwardson and his colleagues42 realized the problem and concluded that they were detecting
TNV particles in necrotic cells because necrosis induced their crystallization, probably due
to cell desiccation. It is well established that many viruses with small polyhedral particles
tend to crystallize in cells due to water-stress.46-47
Kassanis and his colleagues48 examined the cells of French bean leaves infected with the
bean stipple streak strain of TNV and its SV. Non-necrotic tissue was selected for the study.
Cells infected with TNV alone contained defined regions in which virus particles accumulated
though they did not crystallize. The cells also contained prominent cytoplasmic membranes
and structureless, electron-dense material like that shown in Figures 2 and 3.
In cells infected by both TNV and SV (Figure 4), the smaller SV particles usually formed
crystalline arrays, whereas the numerous TNV particles were scattered throughout the cy-
toplasm. Whenever SV was detected in cells, TNV particles were also present. However,
tissues infected by both viruses sometimes had cells that apparently contained only TNV.
This is understandable because TNV can replicate on its own whereas SV needs its TNV
helper. 7
All the studies of TNV and SV in cells have so far been rather superficial and more work
is needed. Of special interest will be the examination of cells infected by the various strains
of TNV to see if there are any significant differences in their cytopathology.
Volume I 175
FIGURE 1. A. Purified tobacco necrosis virus (TNV) and B. satellite virus (SV). stained in uranyl acetate and
photographed at the same magnification. Bar = 100 nm.
FIGURE 2. Thin sections of Nicoiiana x edwunlsonii leaf cells infected with TNV hut not SV. (Tissue cleared
of rihosomes by RNase treatment.) A. Virus particles in clusters and virus-specific amorphous electron-dense
material in the cytoplasm: B. a cluster of virus particles and part of the prominent endoplasmic reticulum: C,
groups of virus particles, and endoplasmic reticulum formed into paired membranes. Bars = 500 nm.
Volume I 177
FIGURE 3. Part of Figure 2C enlarged. Note the small vesicles associated with the tonoplast. Bar = 100 nm.
FIGURE 4. Thin sections of Phaseolus vu/garis (French bean) leaf cells infected with both TNV and SV. A,
Both TNV (arrows) and SV particles (crystallized, indicated by arrowheads) are clearly seen despite the presence
of ribosomes. Bar = 500 nm. B, Crystallized SV particles are seen surrounded by a few scattered TNV particles.
Bar = 100 nm. (Courtesy of Drs. D. A. Vince and R. D. Woods, Rothamsted Experiment Station, Harpenden,
U.K.)
Volume I 179
REFERENCES
2. Uyemoto, J. K., Tobacco necrosis and satellite viruses, in Handbook of Plant Virus Infections and Com-
parative Diagnosis, Kurstak, E.. Ed.. Elsevier, Amsterdam, 1981. 123.
3. Salvato, M. S. and Fraenkel-Conrat, H., Translation of tobacco necrosis virus and its satellite in a cell-
free wheat germ system, Proc. Null. Acad. Sci. U.S.A., 74. 2288, 1977.
4. Teakle, D. S., Transmission of tobacco necrosis virus by a fungus, Olpidium brassicae. Viroloi>\, 18, 224.
1962.
5. Kassanis, B. and MacFarlane, I., Interaction of virus strain, fungus isolate, and host species in the
transmission of tobacco necrosis virus, Virology, 26, 603. 1965.
6. Temmink, J. H. M., Campbell, R. N., and Smith, P. R., Specificity and site of in vitro acquisition of
tobacco necrosis virus by zoospoores of Olpidium brassicae, J. Gen. Virol., 9, 201, 1970.
7. Kassanis, B. and Nixon, H. L., Activation of one tobacco necrosis virus by another. J. Gen. Microbiol.,
25, 459, 1961.
8. Kassanis, B., Portraits of viruses: tobacco necrosis virus and its satellite virus, Intervirology, 15, 57, 1981.
9. Babos, P. and Kassanis, B., The behaviour of some tobacco necrosis virus strains in plants. Virolog\,
20, 498, 1963.
10. Babos, P. and Kassanis, B., Serological relationships and some properties of tobacco necrosis virus strains,
J. Gen. Microbiol., 32, 135, 1963.
1 1 . Kassanis, B. and Phillips, M. P., Serological relationships of strains of tobacco necrosis virus and their
ability to activate strains of satellite virus, J. Gen. Virol., 9, 119, 1970.
12. Uyemoto, J. K., Grogan, R. G., and Wakeman, J. R., Selective activation of satellite virus strains by
strains of tobacco necrosis virus, Virology, 34, 410, 1968.
13. McKeen, C. D., Cucumber necrosis virus. Can. J. Bot., 37, 913, 1959.
14. Dias, H. F., The relationship between cucumber necrosis virus and its vector, Olpidium cucurbitacearum.
Virology, 42, 204, 1970.
15. Dias, H. F. and McKeen, C. D., Cucumber necrosis virus, CMI/AAB Descriptions of Plant Viruses, No.
82, 1972.
16. Dias, H. F. and Doane, F. W., Evidence for lack of relationship between Canadian cucumber necrosis
and tobacco necrosis viruses, Can. J. Bot., 46, 47, 1968.
17. Tremaine, J. H., Purification and properties of cucumber necrosis virus and a smaller top component,
Virology, 48, 582, 1972.
18. Van Koot, Y. and Van Dorst, H. J. M., A new virus disease of cucumbers, Tijdschr. Plantenziekten,
61, 163, 1955.
19. Gallitelli, D., Castellano, M. A., Di Franco, A., and Rana, G. L., Properties of carnation yellow stripe
virus, a member of the tobacco necrosis virus group, Phytopathol. Medit., 18, 31, 1979.
20. Lesnaw, J. A. and Reichmann, M. E., The structure of tobacco necrosis virus. I. The protein subunit
and the nature of the nucleic acid, Virology, 39, 729, 1969.
2 1 . Kassanis, B., Tobacco necrosis virus group, in The Atlas of Insect and Plant Viruses, Maramorosch, K.,
Ed., Academic Press, New York, 1977, 281.
22. Sjoberg, B., A small-angle X-ray investigation of the satellite tobacco necrosis virus, Eur. J. Biochem.,
81, 277, 1977.
23. Reichmann, M. E., The satellite tobacco necrosis virus: a single protein and its genetic code, Proc. Null.
Acad. Sci. U.S.A., 52, 1009, 1964.
24. Strandberg, B., Lentz, P., Kannan, K., Vaara, I., Unge, T., Fridborg, K., Borell, A., Petej, G.,
and Nordman, C., The structure of satellite tobacco necrosis virus: three-dimensional X-ray diffraction
analysis at 10 A resolution, Acta Crystallogr., A31, S46, 1975.
25. Liljas, L., Unge, T., Jones, T. A., Fridborg, K., Lovgren, S., Skoglund, U., and Strandberg, B.,
Structure of satellite tobacco necrosis virus at 3.0 A resolution, J. Mol. Hio/.. 159. 93. 1982.
26. Hendricksson, D., Tanis, R. J., Tashian, R. E., and Nyman, P. O., Amino acid sequence of the coat
protein subunit in satellite tobacco necrosis virus, J. Mol. Biol., 152, 171, 1981.
27. Ysebaert, M., van Emmelo, J., and Fiers, W., Total nucleotide sequence of a nearly full-size DNA copy
of satellite tobacco necrosis virus RNA, J. Mol. Biol., 143, 273, 1980.
28. Reichmann, M. E., Chang, A. Y., Faiman, L., and Clark, J. M., The satellite tobacco necrosis virus
in studies of genetic coding, Cold Spring Harbor Symp. Quant. Biol., 31, 139, 1966.
29. Uyemoto, J. K. and Grogan, R. G., Chemical characterization of tobacco necrosis and satellite viruses,
Virology, 39, 79, 1969.
30. Bishop, D. H. L., Claybrook, J. R., and Spiegelman, S., Electrophoretic separation of viral nucleic
acids on polyacrylamide gels, J. Mol. Biol., 26, 373, 1967.
31. Condit, C. and Fraenkel-Conrat, H., Isolation of replicative forms of 3' terminal subgenomic RNAs of
tobacco necrosis virus, Virology, 97, 122, 1979.
32. Mossop, D. W. and Francki, R. I. B., The stability of satellite viral RNAs in vivo and in vitro, Virology,
94, 243, 1979.
33. Wimmer, E. and Reichmann, M. E., Two 3'-terminal sequences in satellite tobacco necrosis virus RNA,
Nature (London), 221, 1122, 1969.
34. Horst, J., Keith, J., and Fraenkel-Conrat, H., Characteristic two-dimensional patterns of enzymatic
digests of oncorna and other viral RNAs, Nature (London) New Biol., 240, 105, 1972.
35. Klein, W. H., Nolan, C., Lazar, J. M., and Clark, J. M., Translation of satellite tobacco necrosis virus
ribonucleic acid. I. Characterization of in vitro procaryotic and eucaryotic translation products, Biochemistry,
11, 2009, 1972.
36. Leung, D. W., Gilbert, C. W., Smith, R. E., Sasavage, N. L., and Clark, J. M., Translation of satellite
tobacco necrosis virus ribonucleic acid by an in vitro system from wheat germ, Biochemistry, 15, 4943,
1976.
37. Clark, J. M., Chang, A. Y., Spiegelman, S., and Reichmann, M. E., The in vitro translation of a
monocistronic message, Proc. Nad. Acad. Sci. U.S.A., 54, 1193, 1965.
38. Rice, R. and Fraenkel-Conrat, H., Fidelity of translation of satellite tobacco necrosis virus ribonucleic
acid in a cell-free Escherichia coli system, Biochemistry, 12, 181, 1973.
39. Kozak, M., How do eucaryotic ribosomes select initiation regions in messenger RNA?, Cell, 15, 1109,
1978.
40. Smith, R. E. and Clark, J. M., Effect of capping upon the mRNA properties of satellite tobacco necrosis
virus ribonucleic acid, Biochemistry, 18, 1366, 1979.
41. Lesnaw, J. A. and Reichmann, M. E., Identity of the 5'-terminal RNA nucleotide sequence of the satellite
tobacco necrosis virus and its helper virus: possible role of the 5'-terminus in the recognition by virus-
specific RNA replicase, Proc. Nad. Acad. Sci. U.S.A., 66, 140, 1970.
42. Edwardson, J. R., Purcifull, D. E., and Christie, R. G., Electron microscopy of two small spherical
plant viruses in thin sections, Can. J. Bot., 44, 821, 1966.
cells by an enzyme cytochemical technique, J. Ultrastruct. Res., 74, 116, 1981.
44. Hatta, T. and Francki, R. I. B., Cytopathic structures associated with tonoplasts of plant cells infected
with cucumber mosaic and tomato aspermy viruses, J. Gen. Virol, 53, 343, 1981.
45. Hatta, T. and Francki, R. I. B., Enzyme cytochemical method for identification of cucumber mosaic
virus particles in infected cells, Virology, 93, 265, 1979.
47. Ushiyama, R. and Matthews, R. E. F., The significance of chloroplast abnormalities associated with
infection by turnip yellow mosaic virus, Virology, 42, 293, 1970.
48. Kassanis, B., Vince, D. A., and Woods, R. D., Light and electron microscopy of cells infected with
tobacco necrosis and satellite viruses, J. Gen. Virol., 7, 143, 1970.
Volume I 181
Chapter 12
TOMBUSVIRUS GROUP
The Tombusvirus group consists of tomato bushy stunt virus (TBSV), the type member,
and several related viruses. 1 All members have isometric particles about 34 nm in diameter
with rounded outlines, sedimenting at about 135S. Each particle consists of 180 protein
subunits of Mr about 40 X 103 arranged with T = 3 icosahedral lattice symmetry and
containing a single molecule of positive-sense, single-stranded RNA of Mr about 1.5 x
10".
Tombusviruses are stable and readily transmissible by mechanical inoculation and may
be acquired through the soil and water, apparently without the intervention of vectors. There
are no reports of seed transmission. Each can infect a wide range of plants and induce a
variety of symptoms. Characteristic multivesicular bodies are induced in infected cells.2"4
At present, the ICTV recognizes seven viruses as definite members of the group (Table
1). All of these, except for cymbidium ringspot virus (CyRSV)5 and glycine mottle virus
(GMoV), 6 are serologically closely interrelated.7-8 From the serological studies by Rollings
and Stone,7 it seems that differences between some isolates of the viruses are as great as
differences between some of the viruses. This raises the question often posed regarding
viruses in other groups; that is, how to determine which isolates should be considered as
strains of the same virus and which should be regarded as distinct viruses. At present there
does not seem to be a satisfactory solution to the problem.
Although CyRSV appears to be antigenically unrelated to the other six Tombusviruses
listed in Table 1, the physical properties of its particles are like those of the other member
viruses. 5 Furthermore, CyRSV has been shown to induce multivesicular bodies in the cy-
toplasm of infected cells characteristic of Tombusvirus infection. 9 GMoV was recently
described as a possible member of the group. 6 However, its reported properties are so like
those of other Tombusviruses (Table 1) that we have included it as a member of the group.
Several viruses with small isometric particles are in some respects similar to Tombus-
viruses, but are at present not included in this or any other group. Some of these are listed
in Table 2 together with a number of their properties.
Turnip crinkle virus (TCV) is at present considered by the ICTV as a possible member
of the Tombusvirus group.' The particle structure of TCV closely resembles that of TBSV. l 2 2 7
At present the best argument for the exclusion of TCV from the Tombusvirus group is that
the multivesicular inclusions induced by infection are not derived from peroxisomes as with
TBSV, but from mitochondria. 21 It is not at present clear what weighting should be given
to this cytopathological characteristic in determining the taxonomic position of a virus.
Probably it is not of paramount importance, as some viruses, which are known to be related
by most other criteria including their antigenic properties, can have significantly different
cytopathic effects. The Tobamoviruses (see Chapter 9, Volume 2) are a good example.
Particles of the remaining three viruses listed in Table 2 all have single-stranded RNA
genomes of Mr between 1.4 and 1.6 x 106 and coat protein subunits between 36 and 40
x 10\ Considering that most of these values have been obtained from polyacrylamide-gel
electrophoretic data gathered under various conditions and with all the inherent errors of the
method, they are remarkably similar to the values reported for the approved members of
the Tombusvirus group (Table 1). However, none of these viruses has been shown to be
antigenically related to the Tombusviruses.
Table 1
SOME PROPERTIES OF TOMBUSVIRUSES
Presence of
Buoyant multivesicular
density MT of coat bodies
in CsCl M, of RNA protein in peroxisomes
Virus S21). (g/cnr1) (x 10') (x 10') of infected cells Ref.
Tomato bushy stunt 131 — 140 1.39 1.4 (soy Yes 10—13
virus (TBSV) 38.0
(28)
Artichoke mottled 132 1.35 1.5 h
Yes 2, 14
crinkle virus
(AMCV)
Carnation Italian 135 — — Yes 15, 16
ringspot virus
(C1RSV)
Pelargonium leaf curl 136 — — Yes 17, 18
virus (PLCV)
Petunia asteroid mos- 134 1.35 — — 19
aic virus (PAMV)
Cymbidium ringspot 137 1.36 1.7 43.6 Yes 5, 9
virus (CyRSV)
Eggplant mottled 132 — — 41.0 Yes 8
crinkle virus
(EMCV)
Glycine mottle virus 138 — 1.48 39.0 Yes 6
(GMoV) (37.5)
(31.0)
Values in parenthesis are minor components of unknown significance.

h
Not determined.
Saguaro cactus virus (SCV) has also been included as a possible Tombusvirus in the most
recent ICTV report.' However, the lower sedimentation rate28 of the particles of this virus
and failure to induce multivesicular bodies of any kind in infected cells21 indicate that it
may be more like carnation mottle virus (CarMV) which is at present unclassified. Both
these viruses are discussed in Chapter 16, Volume 2 (see Table 2 in that Chapter).
Although particles of all the viruses listed in Table 2 appear similar in some respects,
there are also indications of some significant differences. For example, galinsoga mosaic
virus (GaMV) and TBSV have recently been directly compared with respect to the physical
properties of their particles and cytopathology, and differences were established. 24 The
particles of TBSV and GaMV are indistinguishable in appearance when stained in uranyl
acetate (compare Figures 1A and IB), but their sedimentation coefficients differ significantly.
Their cytopathological effects also differ in that GaMV induces vesiculated inclusions which
develop from mitochondria, unlike those of TBSV but like those of TCV21 (see Section III
below for details). It is interesting that whereas TCV and GaMV induce very similar cy-
topathic effects in plant cells, their particles appear to sediment at different rates (Table 2).
Observations such as these raise difficult taxonomic problems which are unlikely to be
resolved without much more comparative data on all the viruses.
Hibiscus chlorotic ringspot virus (HCRSV) has been shown to have particles indistin-
guishable from those of TBSV and GaMV when uranyl-acetate-stained virus preparations
were viewed in the electron microscope (compare Figures 1A and 1C). However, the virus
has not been extensively studied and cannot be assigned to any group without more data.
Cucumber necrosis virus (CuNV) has been considered as a possible member of the mon-
Volume 1 183
Table 2
SOME PROPERTIES OF VIRUSES WITH SOME SIMILARITIES TO
TOMBUSVIRUSES
Buoyant
density M, of coat
in CsCl M, of RNA protein Cytopathological
Virus S20w (g/cm3) (x 106) (x 10') effects Ref.
Turnip crinkle virus 129 —•' 1.4 (80)" Vesiculation of 12,

(TCV) 38^1 mitochondria 20—2;
(28)
Galinsoga mosaic 118 — 1.55 36.4 Vesiculation of 23. 24
virus (GaMV) mitochondria
Hibiscus chlorotic 118 — 1.55 — — 25
ringspot virus
(HCRSV)
Cucumber necrosis 133 — 1.4 40.3 — 26
virus (CuNV)
" Not determined.

h
Values in parenthesis are minor components of unknown significance.
otypic tobacco necrosis virus group by the ICTV. 1 (See also Chapter 11.) This suggestion
was probably based on its being transmitted by the same vector as tobacco necrosis virus
(TNV), the soil phycomycete fungus Olpidium brassicae ,26 However, the physical properties
of CuNV particles are unlike those of TNV (see Table 2 and Chapter 11), and hence it
seems difficult to imagine that the two viruses are closely related. Unfortunately CuNV has
not been studied extensively, and so at present it is difficult to establish its proper taxonomic
position, but inclusion of CuNV in the Tombusvirus group should be considered.
The structure of the TBS V particle has been extensively studied by both electron microscopy27
and X-ray diffraction. 29 In virus preparations stained with uranyl acetate (Figure 1A), the
particles are about 34 nm in diameter, 30 whereas X-ray studies indicate a maximum diameter
of 33 nm. 3 1 Electron microscopic studies led to the construction of a model consisting of
180 protein subunits with a T = 3 surface lattice. 12 The protein subunits were shown to be
clustered with 90 dimers arranged in rings of five and six. A three-dimensional model of
the particle has been reconstructed from electron micrographs as shown in Figure 2. 27
Protein analyses of TBSV preparations have established that each particle consists of 180
polypeptides of Mr about 41 X 103 (380 amino acid residues) and possibly a small amount
of a polypeptide of Mr about 87 x 10 3 . 12 - 33 It was calculated that there was enough of the
larger polypeptide for each virus particle to contain only one molecule. 33 Its location and
function remain unknown, but Butler 12 suggested that it may be a polymerase, possibly
attached to the viral RNA. This is a suggestion that needs to be substantiated. At present it
cannot be completely ruled out that the protein is not a contaminant in virus preparations.
X-ray crystallographic analysis of TBSV at 2.9 A resolution has yielded interesting results
on the basis of which a model has been constructed (Figure 3).29 34 It was shown that the
particles of TBSV and southern bean mosaic virus (SBMV) of the Sobemovirus group have
many structural features in common. 35 36 However, they appear different in the electron
microscope when stained in uranyl acetate.24
Each TBSV coat protein subunit is folded into two globular parts (the P and S domains)
which are connected by a flexible hinge (h in Figure 3A). In addition, each subunit also
l-'IGURfc I . I'urified preparations of A. tomato bushy .stunt virus (TBSV): B. <;alinsogu mosaic virus (CiaMV):
and C. hibiscus chlorotic ringspot virus (HCRSV). All preparations were stained with uranyl acetate and all
photographs are at the same magnification. Bar = 100 nm.
Volume 1 185
FIGURE 2. Three-dimensional reconstruction of a TBSV particle showing A. a density map

and B. a contour map viewed in approximately the same orientation. In each case only one halt
of the particle is shown. (From Crowther. R. A. and Amos, L. A., Cold Spring Harbor Symp.
Quant. Hiol., 36, 4X9. 1971. With permission.)
has a flexibly linked N terminal of about 80 to 90 amino acids (a in Figure 3A). The subunits
appear to exist in three states, A, B, and C (Figure 3B and 3D). All 60 C-type subunits,
including their N-terminal "tails", are interconnected to form an ordered, cage-like do-
decahedral shell in which the subunits are exactly equivalent and the hinges between the P
and S domains are "up", with the domains at right angles to each other. The 120 A and
B subunits fill in the gaps in this fenestrated structure, taking up quasi-equivalent positions,
with the hinges "down" and the P and S domains facing each other at an obtuse angle.
The N-terminal "tails" of the A and B subunits lie toward the interior of the particle; they
appear less ordered than in the C configuration, and possibly interact with the RNA. That
the N termini are associated with the RNA is supported by data from small-angle neutron-
scattering studies which indicate that the protein is distributed as two concentric shells about
3 nm apart. 37 Most of the RNA appears to be located betwen these shells (Figure 3E).
Evidence from "P nuclear magnetic resonance studies indicates that the RNA is highly
immobile in the particle.' 8
Although TCV is not at present a definite member of the Tombusvirus group (Table 2),
the structure of its particles has been studied extensively and shown to be very similar to
that of TBSV. 12 - 22 - 11 " 33 This seems a persuasive argument favouring its inclusion in the group.
Although the TBSV particle is a very stable structure it can be made to swell by chelation
of cations under mild alkaline conditions; but the transition is reversible.39-4" On raising the
pH above 7 in the presence of EDTA, the particles swell by about 12% in radius although
the radial distribution of RNA and protein remain similar to that of the native particle (Figure
5D). The swelling can be reversed by addition of calcium or by reducing the pH. It appears
that during swelling the particle protein subunit domains undergo no significant internal
configurational changes and that the swelling involves only the flexible joints of the subunits.40
B. The Viral Genome

Infectious RNA isolated from purified TBSV was shown to have a Mr of about 1.55 x
106 estimated from its sedimentation properties and electrophoretic mobility in polyacrylam-
ide gels." Later work by Henriques and Morris 41 showed that the A/r of TBSV RNA varies
with the strain of the virus from 1.55 to 1.65 x 10''. More recently, subgenomic RNA of
Mr about 1 x 10'' was also detected in preparations of TBSV together with a small RNA
(MT M).15 x 10'') which was suspected of being a satellite RNA. 42 RNAs of Mr between
1.3 and 1.7 x 10'' have been isolated from some other Tombusviruses (Table 1) and possible
FIGURE 3. Models of the TBSV particle structure. A, A chemical subunit with the P and S domains, the
interdomain hinge (h). and the N-terminal arm (a). B. Arrangement of the subunits (A, B, and C) in the virus
particle with their three packing environments (center surfaces of the subunit S domains are shaded). The S domains
of the A subunits pack around fivefold axes and those of B and C alternate around the threefold axes (see text for
further details). C, Two states of the TBSV subunit viewed as dimers about strict (s2) and local (q2) twofold axes.
D. Schematic diagram showing a more detailed view of the subunits seen nearly face-view in B with the barrel-
shaped P domains protruding outwards while the more extensive S domains (A, B, and C) make contacts across
all adjacent symmetry axes. E, Model of a transverse section of the internal structure of TBSV derived from neutron
small-angle scattering data: the dark area indicates that occupied by the RNA in which the protein is embedded:
numbers indicate distances from the center of the particle in A. (Figures 3A through D from Harrison. S. C..
Olson, A. J., Schutt. C. E.. Winkler, F. K., and Bricogne. G., Nature (London), 276. 368, 1978. Copyright
Macmillan Journals Ltd. (London). With permission. Figure 3E from Chauvin. C., Witz, J . , and Jacrot, B., J.
Mol. Bio/., 124, 641. 1978. Copyright: Academic Press, Inc. (London) Ltd. With permission.)
members of the group (Table 2). However, there does not appear to be any more detailed
information about their structure.
Henriques and Morris41 isolated two species of virus-specific double-stranded RNA from
TBSV-infected plants of M, about 3.2 x 106 and 1.5 x 10". Both RNAs were shown to
share nucleotide sequences with the viral RNA, and the larger of the two has the expected
A/r of TBSV replicative form. Henriques and Morris41 suggested that the smaller double-
stranded RNA was the replicative form of a subgenomic mRNA of M, about 0.75 X 106
synthesized in infected cells. However, this putative mRNA has not yet been detected in
infected tissues.
Although TCV is similar in many respects to TBSV, it appears to induce the synthesis
of only one detectable double-stranded RNA in infected plants. 41 The Mr of this RNA was
estimated to be about 3.0 x 106 which corresponds to that expected for the viral RNA
replicative form.
Dougherty and Kaesberg 43 examined the RNA composition of TCV preparations and
detected a genomic RNA of M, about 1.4 X 106, a small RNA of Mr about 0.15 x 106,
Volume I 187
FIGURE 3 (continued)
and a heterogeneous collection of RNAs intermediate in size. In vitro translation studies

revealed that the genomic RNA was an efficient messenger for a protein of Mr about 25 x
I0 3 (25K protein), whereas subgenomic RNA fractions translated mainly into proteins of
Mr about 36 x I0 3 (36K protein) and 38 X I0 3 (viral coat protein). In addition a number
of minor products were also detected. Tryptic analyses established that the three major
protein products were distinct and that the minor ones were related to them. These trans-
lational and tryptic analysis data suggest that the gene for the 25K protein is near the 5'
terminus of TCV RNA which can, however, be read through into a protein of MT about 80
X I0 3 . It appears that the 36K protein is translated from a subgenomic RNA lacking the
initiation site of the 25K protein gene. A read-through of this cistron can result in synthesis
of a protein of Mr about 56 x I0 3 whose termination coincides with that of the Mr ^80 x
I0 3 product. It also appears that the coat protein was translated from a subgenomic RNA
lacking both the initiation sites for the 25K and 36K protein genes. The sequence for the
coat protein mRNA is probably located near the 3' terminus of the genomic RNA. 43 Es-
sentially similar results of studies on the translation of TCV RNA were subsequently reported
by Altenbach and Howell. 44
The small RNA of M, about 0.15 x 10" detected in capsids of TCV by Dougherty and
Kaesberg43 is probably the same as that reported by Altenbach and Howell45 who estimated
the M, to be about 0.17 X 10" and showed the RNA to be a satellite. 45 Both groups found
that this small RNA had negligible in vitro messenger activity. 43 - 44
III. CYTOPATHOLOGY
Cells infected by Tombusviruses contain densely staining particles about 26 nm in di-

ameter. 2 46 The particles accumulate in large numbers in the cytoplasm and vacuoles where
they sometimes form crystalline arrays 18 - 47 - 48 (Figure 4). They have also sometimes been
detected in nuclei (Figure 5), chloroplasts, and mitochondria. 2946
Virus particles in the cytoplasm often accumulate within extensions of the tonoplast,
forming bubble-like structures protruding into the vacuole (Figure 4). It has been suggested
FIGURE 4. A, Thin section of a Nicotians Cleveland!/ leaf cell infected with TBSV showing virus particles
scattered in the cytoplasm, some of which protrude into the vacuole. Bar = 500 nm. B, Section from a similar
cell to that in A, but showing particle aggregation near the tonoplast which appears to be ruptured, releasing
particles into the vacuole. Bar = 100 nm. (Courtesy of Drs. M. Russo and G. P. Martelli, Istituto di Patologia
Vegetale, Universita degli Studi, Bari, Italy.)
Volume 1 189
FIGURE 5. Thin section of a Nicoliana clevelandii leaf cell infected with "I'BSV showing virus particles (arrows)
in both nucleus and cytoplasm. Bar = 500 nm. (Courtesy of Drs. M. Russo and G. P. Martelli. Istituto di Patalogia
Vegetale, Universita degli Studi, Bari. Italy.)
that these bubbles become detached from the tonoplast and become scattered within the
vacuole.'*- 47 - 48
Tombusvirus infections may alter the ultrastructure of mitochondria, chloroplasts, and
nuclei to varying extents, and virus particles may or may not be found in these organelles,
depending on the virus, the host, and probably the environmental conditions. 2 For example,
the major organelles in artichoke cells infected with artichoke mottled crinkle virus (AMCV)
are relatively unaffected although the same virus in Chenopodium quinoa produces extensive
damage, including the formation of large membranous inclusions in the nuclei. 14 - 47 Chlo-
roplasts in pelargonium leaves infected with pelargonium leaf curl virus (PLCV) undergo a
form of budding, and also contain free virus-like particles. 18 These features have not been
reported for other Tombusviruses. 3
The most characteristic feature of cells infected with Tombuviruses is the development
of multivesicular bodies in the cytoplasm (Figures 6 through 8). At first it was suggested
that these structures are formed from the membranes of the endoplasmic reticulum and
dictyosomes.4x Later observations led to the conclusion that they were derived from or
associated with the chloroplasts." However, other evidence indicates that the multivesicular
bodies develop from microbodies and retain microbody enzymatic activities. This has been
clearly demonstrated in cells infected by TBSV, eggplant mottled crinkle virus (EMCV),
and CyRSV. 8 - 24 - 4 "- 49
Early in their development, the multivesicular bodies in TBSV infected cells are mem-
brane-bounded structures with peripheral vesicles (Figure 8). The vesicles, often contain
fibrils reminiscent of nucleic acid, resembling the chloroplast vesicles induced by Tymovirus
infection (see Chapter 8) or those associated with tonoplasts in cells infected with Cucu-
moviruses (see Chapter 5, Volume 2), tobacco necrosis virus (see Chapter 11), and some
Tobamoviruses (see Chapter 9, Volume 2). In the case of CyRSV, they have been shown
to contain RNA, some of which is double-stranded. 49 Later, the multivesicular bodies become
vacuolated and the proportion of membranes and vesicles, compared to stroma, increases
(Figures 6 and 7).
The significance of the multivesicular bodies is not clear, though it has been suggested
that they may be involved in virus replication. 1 -4C> They could be the sites of viral RNA
synthesis, and the vesicles could contain the virus-specific double-stranded RNA detected
in infected tissues by Henriques and Morris. 41 A study of the kinetics of local lesion formation
by TBSV in Gomphrena globasa50 indicates that the multivesicular bodies appear some
hours in advance of the first detectable virus particles as would be expected if they play a
precursor role.
Local lesion formation by Tombusviruses in hypersensitive hosts has been discussed by
Martelli and colleagues 2 - 51 and the ultrastructural aspects of local lesion formation by TBSV
in Gomphrena globosa have been studied by Appiano and colleagues.50-52'53 Apart from data
on the multivesicular bodies discussed above, the most interesting finding was that the
lesions did not seem to be limited by the accumulation of barrier substances or by the
interruption of plasmodesmata. Callose deposits around the plasmodesmata occurred after
the appearance of virus particles in the cells; suberin and lignin deposits were detected at
an even later stage, and then only in the intercellular spaces. Limitation of the infection was
therefore ascribed to physiological rather than structural mechanisms.
The cytopathic effects of only a few of the viruses listed in Table 2 have been investigated.
Multivesicular bodies have been observed in plant cells infected with TCV and GaMV.
However, these multivesicular bodies, although in some respects very similar to those of
TBSV-infected cells, have been shown to develop from mitochondria 2 ' 24 (Figures 9 and
10).
Volume 1 191
FRjLIRE 6. Thin section of a Dutura stramonium lea! cell infected with TBSV showing the presence of virus
particles scattered in the cytoplasm and vacuoles and of virus-induced multivesicular bodies in the cytoplasm.
Some of the multivesicular bodies appear to be at early stages of development still retaining the appearance of
modified peroxisomcs (arrows) whereas others show extensive vesiculation and severe disruption. (Tissue cleared
of ribosomes bv RNase treatment.) Bar = 500 nm.
FIGURE 7. Thin section of TBSV-infected Datura stramonium leaf tissue showing a cell with numerous virus
particles and virus-induced multivesicular bodies at various stages of development. (Tissue cleared of ribosomes
by RNase treament.) Bar = 500 nm.
Volume 1 193
FIGURE 8. Thin section of a Nicotiana clevelandii leaf cell infected with TBSV showing a virus-induced
multivesicular body and numerous scattered virus particles in the cytoplasm. The multivesieular body contains a
crystal and numerous vesicles around its periphery with densely staining fibrils (arrows) reminiscent of nucleic
acid. Bar = 100 nm. (Courtesy of Drs. M. Russo and G. P. Martelli, Istituto di Patologia Vegetale. Universita
degli Studi, Bari, Italy.)
FIGURE 9. A. Thin section of a Phaseohix vulgaris (French bean) leaf cell infected with GaMV showing large
virus-induced multivesicular bodies in the cytoplasm. Bar = 500 nm. B. Peripheral vesicles of a multivesicular
body similar to those in A, containing fibrillar material (arrows) reminiscent of nucleic acid. Bar = 100 nm.
Volume I 195
FIGURE 10. A. Thin section of a Phaxcolu.i vulgaris (French bean) leaf cell infected with GaMV showing
numerous virus particles and virus-induced multivesicular bodies scattered in the cytoplasm; a few vesicles can
also be seen on the periphery of the chloroplasts (arrows). (Tissue cleared of ribosomes by RNase treatment.) B.
Thin section of a mitochondrion apparently developing into a multivesicular body in a cell similar to that in A.
Bars = 500 nm.
REFERENCES
2. Martelli, G. P., Russo, M., and Quacquarelli, A., Tombusvirus (Tomato bushy stunt virus) Group, in
The Atlas of Insect and Plant Viruses, Maramorosch, K. Ed., Academic Press, New York, 1977, 257.
3. Martelli, G. P., Tombusviruses, in Handbook of Plant Virus Infections and Comparative Diagnosis,
Kurstak, E., Ed., Elsevier, Amsterdam, 1981, 61.
4. Tomlinson, J. A., Faithful!, E., Flewett, T. H., and Beards, G., Isolation of infective tomato bushy
stunt virus after passage through the human alimentary tract, Nature (London), 300, 637, 1982.
5. Hollings, M., Stone, O. M., and Barton, R. J., Pathology, soil transmission and characterization of
cymbidium ringspot, a virus from cymbidium orchids and white clover (Trifo/ium repens), Ann. Appl. Biol.,
85, 233, 1977.
6. Behncken, G. M. and Dale, J. L., Glycine mottle virus: a possible member of the tombusvirus group,
Intervirology, 21, 159, 1984.
7. Hollings, M. and Stone, O. M., Serological and immunoelectrophoretic relationships among viruses in
the tombusvirus group, Ann. Appl. Biol., 80, 37, 1975.
8. Makkouk, K. M., Koenig, R., and Lesemann, D.-E., Characterization of a tombusvirus isolated from
eggplant, Phytopathology, 71, 572, 1981.
9. Martelii, G. P. and Russo, M., The fine structure of Cymbidium ringspot virus in host tissue. I. Electron
microscopy of systemic infection, J. Ultrastruct. Res., 77, 93, 1981.
10. Martelli, G. P., Quacquarelli, A., and Russo, M., Tomato bushy stunt virus, CMIIAAB Descriptions of
Plant Viruses, No. 69, 1971.
11. Dome, B. and Pinck, L., Molecular weight of tomato bushy stunt virus-RNA, FEBSLett., 12,241, 1971.
12. Butler, P. J. G., Structures of turnip crinkle and tomato bushy stunt viruses. III. The chemical subunits:
molecular weights and number of molecules per particle, J. Mol. Biol., 52, 589, 1970.
13. Appiano, A., Pennazio, S., and Redolfi, P., Cytological alterations in tissues of Gomphrena globosa
plants systemically infected with tomato bushy stunt virus, J. Gen. Virol., 40, 277, 1978.
14. Martelli, G. P. and Russo, M., Electron microscopy of artichoke mottled crinkle virus in leaves of
Chenopodium quinoa Willd., J. Ultrastruct. Res., 42, 93, 1973.
15. Hollings, M., Stone, O. M., and Bouttell, G. C., Carnation Italian ringspot virus, Ann. Appl. Biol., 65,
299, 1970.
16. Martelli, G. P. and Russo, M., Ultrastructure of tomato bushy stunt virus strains in plant tissues, Mik-
robiologija, 9, 177, 1972.
17. Hollings, M. and Stone, O. M., Studies on pelargonium leaf curl virus. II. Relationships to tomato bushy
stunt and other viruses, Ann. Appl. Biol., 56, 87, 1965.
18. Martelli, G. P. and Russo, M., Pelargonium leaf curl virus in host leaf tissues, J. Gen. Virol., 15, 193,
1972.
19. Lovisolo, O., Ambrosino, C., Liberator!, J., and Papa, G., Ricerche sul virus del rachitismo cespuglioso
del pomodoro (tomato bushy stunt virus). I. Differenziazione per via elettrocinetica ed immunochimica del
ceppo "Petunia" del ceppo "BS-3", Atti Accad. Sci. Torino Cl. Sci. Fis. Mat. Nat., 98, 391, 1964.
20. Hollings, M. and Stone, O. M., Turnip crinkle virus, CMIIAAB Descriptions of Plant Viruses, No. 109,
1972.
21. Russo, M. and Martelli, G. P., Ultrastructure of turnip crinkle- and saguaro cactus virus-infected tissues,
Virology, 118, 109, 1982.
22. Golden, J. S. and Harrison, S. C., Proteolytic dissection of turnip crinkle virus subunit in solution,
Biochemistry, 21, 3862, 1982.
23. Behncken, G. M., Francki, R. I. B., and Gibbs, A. J., Galinsoga mosaic virus, CMIIAAB Descriptions
24. Hatta, T., Francki, R, I. B., and Grivell, C. J., Particle morphology and cytopathology of galinsoga
mosaic virus, /. Gen. Virol., 64, 687, 1983.
25. Waterworth, H., Hibiscus chlorotic ringspot virus, CMIIAAB Descriptions of Plant Viruses, No. 227,
1980.
26. Dias, H. F. and McKeen, C. D., Cucumber necrosis virus, CMIIAAB Descriptions of Plant Viruses, No.
82, 1972.
27. Crowther, R. A. and Amos, L. A., Three-dimensional image reconstruction of some small spherical
viruses. Cold Spring Harbor S\mp. Quant. Biol,, 36, 489, 1971.
28. Nelson, M. R., Yoshimura, M. A., and Tremaine, J. H., Saguaro cactus virus, CMIIAAB Descriptions
29. Harrison, S. C., Olson, A. J., Schutt, C. E., Winkler, F. K., and Bricogne, G., Tomato bushy stunt
virus at 2.9A resolution. Nature (London), 276. 368. 1978.
Volume I 197
30. Milne, R. G. and Hatta, T., unpublished results, 1981.

31. Leberman, R. and Finch, J. T., The structures of turnip crinkle and tomato bushy stunt viruses. I. A
small protein particle derived from turnip crinkle virus, /. Mol. Biol., 50, 209, 1970.
32. Finch, J. T., Klug, A., and Leberman, R., The structures of turnip crinkle and tomato bushy stunt
viruses. II. The surface structure: dimer clustering patterns, J. Mol. Biol., 50, 215, 1970.
33. Ziegler, A., Harrison, S. C., and Leberman, R., The minor proteins in tomato bushy stunt and turnip
crinkle viruses, Virology, 59, 509, 1974.
34. Harrison, S. C., Protein interfaces and intersubunit bonding. The case of tomato bushy stunt virus, Biophys.
J., 32, 139, 1980.
35. Abad-Zapatero, C., Abdel-Meguid, S. S., Johnson, J. E., Leslie, A. G. W., Rayment, I., Rossmann,
M. G., Suck, D., and Tsukihara, T., Structure of southern bean mosaic virus at 2.SA resolution, Nature
(London), 286, 33, 1980.
36. Harrison, S. C., Virus crystallography comes of age, Nature (London), 286, 558, 1980.
37. Chauvin, C., Witz, J., and Jacrot, B., Structure of the tomato bushy stunt virus: a model for protein-
RNA interaction, J. Mol. Biol., 124, 641, 1978.
38. Munowitz, M. G., Dobson, C. M., Griffin, R. G., and Harrison, S. C., On the rigidity of RNA in
tomato bushy stunt virus, J. Mol. Biol., 141, 327, 1980.
39. Robinson, I. K. and Harrison, S. C., Structure of the expanded state of tomato bushy stunt virus, Nature
(London), 297, 563, 1982.
40. Kriise, J., Kriise, K. M., Witz, J., Chauvin, C., Jacrot, B., and Tardieu, A., Divalent ion-dependent
reversible swelling of tomato bushy stunt virus and organization of the expanded virion, J. Mol. Biol.,
162, 393, 1982.
41. Henriques, M.-I. C. and Morris, T. J., Evidence for different replicative strategies in the plant tom-
busviruses, Virology, 99, 66, 1979.
42. Hillman, B. and Morris, T. J., Nucleic acids of tomato bushy stunt virus, Phytopathology, 72, 952,
1982.
43. Dougherty, W. G. and Kaesberg, P., Turnip crinkle virus RNA and its translation in rabbit reticulocyte
and wheat embryo extracts. Virology, \ 15, 45, 1981.
44. Altenbach, S. B. and Howell, S. H., In vitro translation products of turnip crinkle virus RNA, Virology,
118, 128, 1982.
45. Altenbach, S. B. and Howell, S. H., Identification of a satellite RNA associated with turnip crinkle virus,
Virology, 112, 25, 1981.
cells by an enzyme cytochemical technique, J. U/trastruct. Res., 74, 116, 1981.
47. Russo, M., Martelli, G. P., and Quacquarelli, A., Studies on the agent of artichoke mottle crinkle. IV.
Intracellular localization of the virus, Virology, 34, 679, 1968.
48. Russo, M. and Martelli, G. P., Ultrastructural observations on tomato bushy stunt virus in plant cells,
Virology, 49, 122, 1972.
49. Russo, M., Di Franco, A., and Martelli, G. P., The fine structure oiCymbidium ringspot virus infections
in host tissues. III. Role of peroxisomes in the genesis of multivesicular bodies, J. Ultrastruct. Res., 82,
52, 1983.
50. Appiano, A., D'Agostino, G., Redolfi, P., and Pennazio, S., Sequence of cytological events during the
process of local lesion formation in the tomato bushy stunt virus- Gomphrena globosa hypersensitive system,
J. Ultrastruct. Res., 76, 173, 1981.
51. Martelli, G. P., Ultrastructural aspects of possible defence reactions in virus-infected plant cells, Micro-
biologica, 3, 369, 1980.
52. Appiano, A., Pennazio, S., D'Agostino, G., and Redolfi, P., Fine structure of necrotic local lesions
induced by tomato bushy stunt virus in Gomphrena globosa leaves, Physiol. Plant Pathol., 11, 327, 1977.
53. Pennazio, S., D'Agostino, G., Appiano, A., and Redolfi, P., infrastructure and histochemistry of the
resistant tissue surrounding lesions of tomato bushy stunt virus in Gomphrena globosa leaves, Physiol.
Plant Pathol., 13, 165, 1978.
Volume I 199
VIRUS ABBREVIATIONS AND TAXONOMIC GROUPS
Virus Abbreviation Group*
Agropyron mosaic virus AgMV Potyvirus

Alfalfa cryptic virus ACV Unclassified
Alfalfa mosaic virus AMV Alfalfa mosaic virus group
Alfalfa temperate virus ATeV Unclassified
Alligatorweed stunting virus AWSV Closterovirus I?
American hop latent virus AHLV Carlavirus
American plum line pattern virus APLPV Ilarvirus
American wheat striate mosiac virus AWSMV Rhabdoviridae
Andean potato latent virus APLV Tymovirus
Andean potato mottle virus APMV Comovirus
Anthoxanthum latent blanching virus ALBV Hordeivirus?
Apple chlorotic leafspot virus ACLV Closterovirus II
Apple mosaic virus ApMV Ilarvirus
Apple stem grooving virus ASGV Closterovirus III
Arabis mosaic virus ArMV Nepovirus
Arracacha virus A AVA Nepovirus
Arracacha virus B AVB Unclassified
Arrhenatherum blue dwarf virus ABDV Reoviridae (Fijivirus)
Artichoke Italian latent virus AILV Nepovirus
Artichoke mottled crinkle virus AMCV Tombusvirus
Artichoke vein banding virus AVBV Nepovirus
Artichoke yellow ringspot virus AYRV Nepovirus
Asparagus stunt virus ASV Ilarvirus
Asparagus virus II AVII Ilarvirus
Bamboo mosaic virus BaMV Potexvirus
Barley stripe mosaic virus BSMV Hordeivirus
Barley yellow mosaic virus BaYMV Potyvirus?
Barley yellow dwarf virus BYDV Luteovirus
Barley yellow striate mosaic virus BYSMV Rhabdoviridae
Barrel cactus virus BaCV Potexvirus
Bean common mosaic virus BCMV Potyvirus
Bean curly dwarf mosaic virus BCDMV Comovirus
Bean golden mosaic virus BGMV Geminivirus
Bean mild mosaic virus BMMV Unclassified
Bean pod mottle virus BPMV Comovirus
Bean rugose mosaic virus BRMV Comovirus
Bean summer death virus BSDV Geminivirus
Bean yellow mosaic virus BYMV Potyvirus
Bean yellow vein banding virus BYVBV Unclassified
Bearded iris mosaic virus BIMV Potyvirus
Beet cryptic virus BCV Unclassified
Beet curly top virus BCTV Geminivirus
Beet leaf curl virus BLCV Rhabdoviridae
* Group or family names are those approved by the ICTV. In the family Reoviridae, the genus into which a
virus is classified is also indicated in parentheses. Question marks indicate uncertainty of the taxonomic status
of the viruses. In the case of the Closterovirus group, we have divided the members into subgroups I to III
(see Chapter 15, Volume II) which are also indicated here.
Beet mild yellowing virus BMW Luteovirus

Beet mosaic virus BtMV Potyvirus
Beet necrotic yellow vein virus BNYVV Tobamovirus?
Beet temperate virus BTeV Unclassified
Beet western yellows virus BWW Luteovirus
Beet yellow stunt virus BYSV Closterovirus I
Beet yellows virus BW Closterovirus I
Belladonna mottle virus BeMV Tymovirus
Bidens mottle virus BiMV Potyvirus
Black raspberry latent virus BRLV Ilarvirus
Blackeye cowpea mosaic virus B1CMV Potyvirus
Blackgram mottle virus BMoV Unclassified
Blueberry red ringspot virus BRRV Caulimovirus
Blueberry shoestring virus BSSV Sobemovirus?
Boletus virus BoV Potexvirus?
Broad bean mottle virus BBMV Bromovirus
Broad bean necrosis virus BBNV Tobamovirus?
Broad bean stain virus BBSV Comovirus
Broad bean true mosaic virus BBTMV Comovirus
Broad bean wilt virus BBWV Comovirus?
Broccoli necrotic yellows virus BNW Rhabdoviridae
Brome mosaic virus BMV Bromovirus
Burdock yellows virus BuYV Closterovirus I
Cacao necrosis virus CNV Nepovirus
Cacao swollen shoot virus CSSV Unclassified
Cacao yellow mosaic virus CYMV Tymovirus
Cactus virus 2 CV2 Carlavirus
Cactus virus X cvx Potexvirus
Canna yellow mottle virus CaYMV Unclassified
Carnation cryptic virus CarCV Unclassified
Carnation etched ring virus CERV Caulimovirus
Carnation Italian ringspot virus CIRSV Tombusvirus
Carnation latent virus CLV Carlavirus
Carnation mottle virus CarMV Unclassified
Carnation necrotic fleck virus CNFV Closterovirus I
Carnation ringspot virus CRSV Dianthovirus
Carnation vein mottle virus CVMV Potyvirus
Carnation yellow stripe virus CYSV Tobacco necrosis virus
group?
Carrot latent virus CaLV Rhabdoviridae
Carrot mottle virus CMotV Unclassified
Carrot red leaf virus CaRLV Luteovirus
Carrot thin leaf virus CTLV Potyvirus
Carrot yellow leaf virus CYLV Closterovirus I
Cassava common mosaic virus CsCMV Potexvirus
Cassava latent virus CaLV Geminivirus
Cassava vein mosaic virus CaVMV Caulimovirus
Cauliflower mosaic virus CaMV Caulimovirus
Celery mosaic virus CeMV Potyvirus
Cereal chlorotic mottle virus CeCMV Rhabdoviridae
Cereal tillering disease virus CTDV Reoviridae (Fijivirus)
Chara corallina virus CCV Tobamovirus?
Volume I 201
Cherry leafroll virus CLRV Nepovirus

Cherry rasp leaf virus CRLV Nepovirus?
Chicory yellow mottle virus ChYMV Nepovirus
Chloris striate mosaic virus CSMV Geminivirus
Chrysanthemum virus B CVB Carlavirus
Citrus crinkly leaf virus CiCLV Ilarvirus
Citrus leaf rugose virus CiLRV Ilarvirus
Citrus tatter leaf virus CiTLV Closterovirus II?
Citrus tristeza virus CTV Closterovirus I
Citrus variegation virus CVV Ilarvirus
Clitoria yellow vein virus CYVV Tymovirus
Clover blotch virus CBV Cucumovirus
Clover primary leaf necrosis virus CPLNV Dianthovirus
Clover yellow mosaic virus C1YMV Potexvirus
Clover yellows virus CYV Closterovirus I
Clover yellow vein virus C1YVV Poty virus
Cocksfoot mottle virus CoMV Sobemovirus?
Cocksfoot streak virus CSV Potyvirus
Coffee ringspot virus CRV Rhabdoviridae
Colocasia bacilliform virus CoBV Unclassified
Colocasia bobone disease virus CBDV Rhabdoviridae
Commelina virus X ComVX Potexvirus
Cow parsnip mosaic virus CoPMV Rhabdoviridae
Cowpea aphid-borne mosaic virus CABMV Potyvirus
Cowpea chlorotic mottle virus CCMV Bromovirus
Cowpea mild mottle virus CPMMV Carlavirus?
Cowpea mosaic virus CPMV Comovirus
Cowpea mottle virus CpMoV Unclassified
Cowpea ringspot virus CPRSV Cucumovirus?
Cowpea severe mosaic virus CPSMV Comovirus
Crimson clover latent virus CCLV Nepovirus
Cucumber green mottle mosaic virus CGMMV Tobamovirus
Cucumber mosaic virus CMV Cucumovirus
Cucumber necrosis virus CuMV Tobacco necrosis virus
group?
Cucumber soilborne virus CSBV Unclassified
Cucumber virus 4 CV4 Tobamovirus
Cucumber yellows virus CuYV Closterovirus II?
Cymbidium mosaic virus CybMV Potexvirus
Cymbidium ringspot virus CyRSV Tombusvirus
Cynara virus CV Rhabdoviridae
Cynosurus mottle virus CyMoV Sobemovirus?
Dahlia mosaic virus DMV Caulimovirus
Dandelion latent virus DLV Carlavirus
Daphne virus X DVX Potexvirus
Dasheen mosaic virus DaMV Potyvirus
Dendrobium vein necrosis virus DVNV Closterovirus I?
Desmodium yellow mottle virus DYMV Tymovirus
Digitaria striate virus DSV Rhabdoviridae
Dulcamara mottle virus DuMV Tymovirus
Eggplant mosaic virus EMV Tymovirus
Eggplant mottled crinkle virus EMCV Tombusvirus
Eggplant mottled dwarf virus EMDV Rhabdoviridae

Elderberry carlavirus ECV Carlavirus
Elderberry latent virus EBLV Unclassified
Elm mottle virus EMoV Ilarvirus
Erysimum latent virus ErLV Tymovirus
Euphorbia mosaic virus EuMV Geminivirus
Festuca cryptic virus FCV Unclassified
Festuca necrosis virus FNV Closterovirus I?
Fiji disease virus FDV Reoviridae (Fijivirus)
Figwort mosaic virus FiMV Caulimovirus
Finger millet mosaic virus FMMV Rhabdoviridae
Foxtail mosaic virus FoMV Potexvirus
Frangipani mosaic virus FMV Tobamovirus
Galinsoga mosaic virus GaMV Tombusvirus?
Garland chrysanthemum temperate virus GCTeV Unclassified
Glycine mosaic virus GMV Comovirus
Glycine mottle virus GMoV Tombusvirus
Gomphrena virus GV Rhabdoviridae
Grapevine Bulgarian latent virus GBLV Nepovirus
Grapevine chrome mosaic virus GCMV Nepovirus
Grapevine fan leaf virus GFLV Nepovirus
Grapevine leafroll-associated virus GLR-AV Closterovirus II
Grapevine virus A GVA Closterovirus II
Grapevine virus B GVB Closterovirus II?
Helenium virus S HeVS Carlavirus
Henbane mosaic virus HMV Potyvirus
Heracleum latent virus HLV Closterovirus II
Heracleum virus 6 HV6 Closterovirus I?
Hibiscus chlorotic ringspot virus HCRSV Tombusvirus?
Hibiscus latent ringspot virus HLRV Nepovirus
Hippeastrum mosaic virus HiMV Potyvirus
Hop latent virus HoLV Carlavirus
Hop mosaic virus HoMV Carlavirus
Hop trefoil cryptic virus HTCV Unclassified
Hordeum mosaic virus HorMV Potyvirus?
Horseradish latent virus HLaV Caulimovirus
Hydrangea ringspot virus HRSV Potexvirus
Hypochoeris mosaic virus HyMV Tobamovirus?
Impatiens latent virus ILV Potexvirus?
Indian peanut clump virus IPCV Tobamovirus?
Iris mild mosaic virus IMMV Potyvirus
Ivy vein clearing virus IVCV Rhabdoviridae
Kalanchoe virus 1 KV1 Carlavirus
Kennedya yellow mosaic virus KYMV Tymovirus
Lamium mild mosaic virus LMMV Comovirus?
Legume yellows virus LYV Luteovirus
Leek yellow stripe virus LYSV Potyvirus
Lettuce mosaic virus LMV Potyvirus
Lettuce necrotic yellows virus LNYV Rhabdoviridae
Lettuce speckles mottle virus LSMV Unclassified
Lilac chlorotic leafspot virus LCLV Closterovirus III
Lilac mottle virus LiMV Carlavirus
Volume I 203
Lilac ring mottle virus LRMV Ilarvirus

Lily symptomless virus LSV Carlavirus
Lily virus X LVX Potexvirus
Lolium enation virus LEV Reoviridae (Fijivirus)
Lolium mottle virus LoMV Unclassified
Lonicera latent virus LLV Carlavirus
Lucerne Australian latent virus LALV Nepovirus
Lucerne enation virus LuEV Rhabdoviridae
Lucerne transient streak virus LTSV Sobemovirus?
Lychnis ringspot virus LRSV Hordeivirus
Maclura mosaic virus MaMV Potyvirus?
Maize chlorotic dwarf virus MCDV Maize chlorotic dwarf virus
group
Maize chlorotic mottle virus MCMoV Unclassified
Maize mosaic virus MMV Rhabdoviridae
Maize rayado fino virus MRFV Unclassified
Maize rough dwarf virus MRDV Reoviridae (Fijivirus)
Maize sterile stunt virus MSSV Rhabdoviridae
Maize streak virus MSV Geminivirus
Maize stripe virus MStV Unclassified
Maize white line mosaic virus MWLMV Unclassified
Malva yellows virus MYV Luteo virus
Melandrium yellow fleck virus MYFV Bromovirus
Melilotus latent virus MeLV Rhabdoviridae
Mirabilis mosaic virus MiMV Caulimovirus
Mulberry latent virus MLV Carlavirus
Mulberry ringspot virus MRV Nepovirus
Mungbean yellow mosaic virus MYMV Geminivirus
Muskmelon vein necrosis virus MkVNV Carlavirus
Myrobalan latent ringspot virus MLRV Nepovirus
Nandina mosaic virus NanMV Potexvirus
Nandina stem-pitting virus NSPV Closterovirus II?
Narcissus latent virus NLV Carlavirus
Narcissus mosaic virus NaMV Potexvirus
Narcissus tip necrosis virus NTNV Unclassified
Narcissus yellow stripe virus NYSV Potyvirus
Nerine latent virus NeLV Carlavirus
Nerine virus X NeVX Potexvirus
Nicotiana velutina mosaic virus NVMV Tobamovirus?
Northern cereal mosaic virus NCMV Rhabdoviridae
Oat blue dwarf virus OBDV Unclassified
Oat mosaic virus OMV Potyvirus?
Oat sterile dwarf virus OSDV Reoviridae (Fijivirus)
Oat striate mosaic virus OSMV Rhabdoviridae
Odontoglossum ringspot virus ORSV Tobamovirus
Okra mosaic virus OkMV Tymovirus
Olive latent ringspot virus OLRV Nepovirus
Onion yellow dwarf virus OYDV Potyvirus
Ononis yellow mosaic virus OYMV Tymovirus
Orchid fleck virus OFV Rhabdoviridae?
Pangola stunt virus PaSV Reoviridae (Fijivirus)
Panicum mosaic virus PanMV Unclassified
204 Adas of Plant Viruses
Papaya mosaic virus PapMV Potexvirus

Papaya ringspot virus PRSV Potyvirus
Parsley latent virus ParLV Unclassified
Parsley rhabdovirus PRV Rhabdoviridae
Parsley virus 5 PalVS Potexvirus?
Parsnip mosaic virus ParMV Potyvirus
Parsnip virus 3 ParV3 Potexvirus?
Parsnip yellow fleck virus PYFV Unclassified
Paspalum striate mosaic virus PSMV Geminivirus
Passiflora latent virus PaLV Carlavirus
Passionfruit woodiness virus PWV Potyvirus
Peach rosette mosaic virus PRMV Nepovirus
Pea early-browning virus PEBV Tobravirus
Pea enation mosaic virus PEMV Pea enation mosaic virus
group
Pea green mottle virus PGMV Comovirus
Pea leafroll virus PeLRV Luteovirus
Pea mild mosaic virus PMMV Comovirus
Pea seed-borne mosaic virus PSMV Potyvirus
Pea streak virus PeSV Carlavirus
Peanut clump virus PCV Tobamovirus?
Peanut mottle virus PeMoV Potyvirus
Peanut stunt virus PSV Cucumovirus
Peanut yellow mottle virus PYMV Tymovirus
Pelargonium flower break virus PFBV Unclassified
Pelargonium leaf curl virus PLCV Tombusvirus
Pelargonium line pattern virus PLPV Unclassified
Pelargonium vein clearing virus PVCV Rhabdoviridae
Pelargonium zonate spot virus PZSV Ilarvirus?
Pepino latent virus PeLV Carlavirus
Pepino mosaic virus PepMV Potexvirus
Pepper mottle virus PepMoV Potyvirus
Pepper veinal mottle virus PVMV Potyvirus
Petunia asteroid mosaic virus PAMV Tombusvirus
Petunia vein clearing virus PeVCV Caulimovirus
Phleum mottle virus PhMoV Unclassified
Physalis mosaic virus PhMV Tymovirus
Pisum virus PV Rhabdoviridae
Pittosporum vein yellowing virus PVYV Rhabdoviridae
Plantago mottle virus P1MV Tymovirus
Plantago severe mottle virus P1SMV Potexvirus
Plantago virus 4 P1V4 Caulimovirus
Plantain virus X P1VX Potexvirus
Plum pox virus PPV Potyvirus
Poa semilatent virus PSLV Hordeivirus
Poinsettia cryptic virus PCrV Unclassified
Pokeweed mosaic virus PokMV Potyvirus
Poplar mosaic virus PopMV Carlavirus
Potato aucuba mosaic virus PAuMV Potexvirus?
Potato black ringspot virus PBRV Nepovirus
Potato leafroll virus PLRV Luteovirus
Potato mop-top virus PMTV Tobamovirus?
Volume I 205
Potato virus A PVA Potyvirus

Potato virus M PVM Carlavirus
Potato virus S PVS Carlavirus
Potato virus T PVT Closterovirus III
Potato virus X PVX Potexvirus
Potato virus Y PVY Potyvirus
Potato yellow dwarf virus PYDV Rhabdoviridae
Prune dwarf virus PDV Ilarvirus
Prunus necrotic ringspot virus PNRSV Ilarvirus
Quail pea mosaic virus QPMV Comovirus
Radish mosaic virus RaMV Comovirus
Radish yellow edge virus RYEV Unclassified
Raphanus virus RV Rhabdoviridae
Raspberry bushy dwarf virus RBDV Ilarvirus?
Raspberry ringspot virus RRV Nepovirus
Raspberry vein chlorosis virus RVCV Rhabdoviridae
Red clover cryptic virus RCCV Unclassified
Red clover mottle virus RCMV Comovirus
Red clover necrotic mosaic virus RCNMV Dianthovirus
Red clover vein mosaic virus RCVMV Carlavirus
Ribgrass mosaic virus RMV Tobamovirus
Rice black-streaked dwarf virus RBSDV Reoviridae (Fijivirus)
Rice dwarf virus RDV Reoviridae (Phytoreovirus)
Rice gall dwarf virus RGDV Reoviridae (Phytoreovirus)
Rice grassy stunt virus RGStV Unclassified
Rice hoja blanca virus RHBV Unclassified
Rice necrosis mosaic virus RNMV Potyvirus?
Rice ragged stunt virus RRSV Reoviridae?
Rice stripe virus RStV Unclassified
Rice transitory yellowing virus RTYV Rhabdoviridae
Rice tungro isometric virus RTIV Unclassified
Rice tungro bacilliform virus RTBV Unclassified
Rice yellow mottle virus RYMV Sobemovirus?
Robinia mosaic virus RobMV Cucumovirus
Rubus yellow net virus RYNV Unclassified
Ryegrass mosaic virus RGMV Potyvirus?
Ryegrass cryptic virus RCV Unclassified
Saguaro cactus virus SCV Unclassified
Sammons' opuntia virus SOV Tobamovirus
Satsuma dwarf virus SDV Nepovirus
Scrophularia mottle virus ScrMV Tymovirus
Shallot latent virus SLV Carlavirus
Soilborne wheat mosaic virus SBWMV Tobamovirus?
Solanum nodiflorum mottle virus SNMV Sobemovirus?
Solanum yellows virus SYV Luteo virus
Sonchus virus SonV Rhabdoviridae
Sonchus yellow net virus SYNV Rhabdoviridae
Southern bean mosaic virus SBMV Sobemovirus
Sowbane mosaic virus SoMV Sobemovirus?
Sowthistle yellow vein virus SYVV Rhabdoviridae
Soybean dwarf virus SoyDV Luteovirus
Soybean mosaic virus SoyMV Potyvirus
Spinach latent virus SPLV Ilarvirus

Spinach temperate virus STeV Unclassified
Squash mosaic virus SqMV Comovirus
Strawberry crinkle virus StCV Rhabdoviridae
Strawberry latent ringspot virus SLRV Nepovirus?
Strawberry vein banding virus SVBV Caulimovirus
Subterranean clover mottle virus SCMoV Sobemovirus?
Subterranean clover red leaf virus SCRLV Luteovirus
Sugarcane mosaic virus SCMV Potyvirus
Sunn-hemp mosaic virus SHMV Tobamovirus
Sweet clover necrotic mosaic virus SCNMV Dianthovirus
Sweet potato mild mottle virus SPMMV Potyvirus?
Tephrosia symptomless virus TeSV Tombusvirus?
Thistle mottle virus ThMoV Caulimovirus
Tobacco etch virus TEV Potyvirus
Tobacco leaf curl virus TLCV Geminivirus
Tobacco mosaic virus TMV Tobamovirus
Tobacco mosaic U2 virus T2MV Tobamovirus
Tobacco necrosis virus TNV Tobacco necrosis virus
group
Tobacco necrotic dwarf virus TNDV Luteovirus
Tobacco rattle virus TRV Tobravirus
Tobacco ringspot virus TRSV Nepovirus
Tobacco streak virus TSV Ilarvirus
Tobacco vein mottling virus TVMV Potyvirus
Tobacco yellow dwarf virus TYDV Geminivirus
Tomato aspermy virus TAV Cucumovirus
Tomato black ring virus TBRV Nepovirus
Tomato bushy stunt virus TBSV Tombusvirus
Tomato golden mosaic virus ToGMV Geminivirus
Tomato mosaic virus ToMV Tobamovirus
Tomato ringspot virus ToRSV Nepovirus
Tomato spotted wilt virus TSWV Tomato spotted wilt virus
group
Tomato top necrosis virus TTNV Nepovirus?
Tomato yellow dwarf virus ToYDV Geminivirus
Tomato yellow leaf curl virus ToYLV Geminivirus
Tomato yellow mosaic virus ToYMV Geminivirus
Tomato yellow top virus TYTV Luteovirus
Tulare apple mosaic virus TAMV Ilarvirus
Tulip breaking virus TBV Potyvirus
Tulip virus X TuVX Potexvirus
Turnip crinkle virus TCV Tombusvirus?
Turnip mosaic virus TuMV Potyvirus
Turnip rosette virus TRoV Sobemovirus
Turnip yellow mosaic virus TYMV Tymovirus
Turnip yellows virus TYV Luteovirus
Ullucus virus C UVC Comovirus
Velvet tobacco mottle virus VTMoV Sobemovirus?
Vicia cryptic virus VCV Unclassified
Viola mottle virus VMV Potexvirus
Watermelon mosaic virus WMV Potyvirus
Volume 1 207
Wheat chlorotic streak virus WCSV Rhabdoviridae

Wheat dwarf virus WDV Geminivirus
Wheat rosette stunt virus WRSV Rhabdoviridae
Wheat spindle streak mosaic virus WSSMV Poty virus?
Wheat streak mosaic virus WSMV Poty virus?
Wheat yellow leaf virus WYLV Closterovirus I
Wheat yellow mosaic virus WYMV Potyvirus?
White clover large cryptic virus WCLCV Unclassified
White clover mosaic virus WC1MV Potex virus
White clover small cryptic virus WCSCV Unclassified
White clover temperate virus WCTeV Unclassified
Wild cucumber mosaic virus WCMV Ty mo virus
Winter wheat (Russian) mosaic virus WWMV Rhabdoviridae
Wound tumor virus WTV Reoviridae (Phytoreovirus)
Yam internal brown spot virus YIBSV Unclassified
Youcai mosaic virus YMV Tobamovirus
Zucchini yellow fleck virus ZYFV Potyvirus
Zygocactus virus X ZVX Potexvirus
Volume I 209
INDEX
A Bean golden mosaic virus, 4, 33—38, 41—44

DNA, 36—37
Abbreviations, 199—207 Bean stipple streak strain, tobacco necrosis virus,
173
Abutilon mosaic virus, 33
Acerarogallia sanguinolenta, 74 Bean summer death virus, 33—35
Beet curly top virus, 33—34, 37—38
Atyrthosiphon pelargonii, 139
Adansonian approach, classification, 8 Beet leaf curl virus, 74
African cassava mosaic disease, 33 Beet leafhopper, 154
Beetle transmission
Agallia constricta, 74
Sobemovirus, 153—154
Aldehyde fixation, general discussion, 6
Alfalfa mosaic virus, 8—9, 11 Tymovirus, 117
American wheat striate mosaic virus, 75, 85—86 Beet mild yellowing virus, 138
Beet western ye.lows virus, 137—141, 147—148
Amorphous material, electron-dense, 173, 176
barley yellow dwarf virus compared with, 137—
Ananas comosus, 76
139, 141
Andean potato latent virus, 117—119
properties of, 138
Animal rhabdoviruses, plant rhabdoviruses com-
Beet yellow net virus, 139
pared with, 81, 85
Anligenic relationships, 17, 23, 33, 48, 75, 119, Beet yellows virus, 11
137—139, 181 Belladonna mottle virus, 118—119
Antiserum, 4, 48, 52, 63, 101, 137—138 Bemisia tabaci, 33—34
Beta virus 3, 74
Aphid transmission
persistent, Luteovirus, 137—139, 141 Beta vulgaris, 147—149
Blebs, 93
Rhabdoviridae, 73—75
semipersistent, Caulimovirus, 17—18, 22 Blisters, perinuclear, 90, 92
Sobemovirus, 154 Blueberry aphid, 154
Aphis Blueberry red ringspot virus, 18
coreopsidis, 75 Blueberry shoestring virus, 153—154, 156—157
craccivora, 74, 139 Bottom (B) particle component, 118—120, 122
gossypii, 139 Brassica
idaei, 15 oleracea, 24—25, 28—29
rubicola, 139 pekinensis, 124—125, 129—132
Arrhenanthcrum blue dwarf virus, 48, 50, 52, 57 Brevicoryne brassicae, 74
Artichoke, 190 Brevipalpus phoenicis, 74
Artichoke mottled crinkle virus, 182, 190 Broccoli necrotic yellows virus, 73—74, 77, 83,
Atropa belladonna, 76 85—86, 88
Aulacorthum solani, 137 properties of, 74, 85
Autoradiographic studies, general discussion, 6 Brome mosaic virus, 11
Bromovirus, taxonomic groups, 9, 11
Bubble-like structures, tonoplast, 187—190
B Bullet-shaped nucleocapsid, 81, 84
Bullet-shaped Rhabdoviridae particle, 73, 76, 81,
Bacitliform particles, 73, 76, 81—82, 111, 115 84
Bacteria, 1, 5 Bunyaviridae, tomato spotted wilt virus compared
Banana bunchy top virus, 139 with, 101, 103
Barley, 91 Buoyant densities, 81, 85, 111, 120, 154, 171,
Barley stripe mosaic virus, 11 182—183
Barley yellow dwarf virus, 10, 137—142
Groups I and II, classification into, 137—139,
141 C
MAY isolate, 137—138, 141
PAV isolate, 138, 141 Cabbage, 24—25, 28—29, 124—125, 129—132
properties of, 138 Cabb-B-S cauliflower mosaic virus strain, 20—21
RMV isolate, 138 Cacao mottle leaf virus, 111
RPV isolate, 138, 141 Cacao swollen shoot virus, 111
SGV isolate, 138 Cacao yellow mosaic virus, 118—119, 122
Barley yellow striate mosaic virus, 73—75, 80—81, Cajanus cajan, 76
90—91 Calcium ion, 155
properties of, 74 Callisiephus chinensis, 76
210 At lax of Plant Viruses
Capsid. see also Nucleocapsid. 17. 47—48. 120, Cell-free protein synthesis systems, general discus-
137. 155. 187 sion. 5
formation of, 21 Cell inclusions, see Inclusions
polypeptides. 55—56. 1 1 1 Cell wall proliferations. 60
Capsomere. 36. 120 Cereal chlorotic mottle virus, 74
Cap structure, lacking, mRNAs. 173 Cereal tillering disease virus. 48. 52
Carlavirus. laxonomic groups. 9. 11 Chaeliixiphon fragaefolii. 75. 139
Carnation etched ring virus. 17—19. 27 Clienopoiliuin
Carnation Italian ringspot virus. 182 ainaraniii-olor, 102, 162
Carnation latent virus. 11 quinmi. 162. 164. 190
Carnation mottle virus. 182 Chinese cabbage. 124—125. 129—132
Carnation ringspot virus, 11 Cliloris inn-ana. 38—39
Carnation yellow stripe virus. 171 Chloris striate mosaic virus. 33—39
Carrot. 145. 150 Chloroplast. 27. 102, 157. 187
Carrol latent virus, 74 involvement with Tymoviruses. 12, 117. 123—
Carrot red leaf virus. 141. 145. 150 131
properties of. 138 peripheral vesicles. 12. 124—128. 130—131. 195
Cassava latent virus, 33—34. 37—38 rounded. 124, 126, 128. 131
Cassava vein mosaic virus. 18 swollen. 124, 128
Cation removal, at high pH. 185 ultrastructure, alterations in. 190
Cauliflower mosaic virus. 4—5, 17—25. 27—28. Cliiindrilla juncca, 76
56 Chromatin. 38—39. 90
cytopathology. 10. 23—25, 27 Chrysanthemum sp.. 76
DNA, 5. 21—23 Chrysomelid beetles. 154
deletion in. 27 Cicadulina mbila, 34
discontinuities (gaps) in. 21—22 Circular single-stranded DNA. 33. 36
nuclear transcription. 23 Circulifer lenellus. 34. 154
gene product. 21—23 Cisternae, cytoplasm, inclusions in. 102—103. 105.
genome. 21—23. 27 108—109
replication of, 23 Cistron, 122, 156, 172—173, 187
strains Citrus. 75
Cabb-B-S. 21—22 Classification, viruses, into groups, see also Taxon-
CM4-I84, 21—22, 27 omic group, 7 — I I
CMI 1841, 21 Clitoria yellow vein virus, 118—119, 130
D/H"1. 21 Cloning. 5, 21
Cauliflower mosaic virus-specific RNA, 23 Clostcrovirus, taxonomic groups, 9, 11
Caulimovirus. see also specific viruses by name. CM4-I84 cauliflower mosaic virus strain, 21—22.
8—10. 17—29 27
coal protein. 17—23, 27—28, 56 CMI 1841 cauliflower mosaic virus strain, 21
cytopathology, 8. 23—29 Coat protein
general discussion. 8—10. 17 amino acid composition data. 117
genome, 10, 17—18. 21—23, 27 amino-acid replacements. 7
physical map of, 22 Caulimovirus studies, 17—23. 27—28, 56
host range. 17 cistron. 122, 156
mechanical transmission, 17 Geminivirus studies. 33—34, 37
panicle, 10, 17—21.23—27 gene product, 21—22
morphology. 8, 10. 17 general discussion. 7. 10—11
nucleus of infected plant cells, presence in, Luteovirus studies. 138, 140
27—28 mRNA, 119. 122—123, 156
size (diameter). 10. 17—20 size, 10—11
structure. 10. 17—21 Sobemovirus studies, 154—156
properties of, 10, 18 subunits, 183—187
replication of, 23 tobacco necrosis virus satellite virus studies. 172
seed transmission, 17 tobacco necrosis virus studies, 171—172
serological relationships, 17 Tombusvirus studies. 181—187
slructure and composition. 10, 17—23 Tymovirus studies. 117—120, 128—130
taxonomic groups, 8—10. 17—18 Cocksfoot mottle virus. 153—154, 156—157
thin section illustrations. 24—26, 28—29 Codons. 22—23. 27, 37
vectors. 17—18. 22 Coffee ringspot virus, 74
viroplasm. 8. 22—29 Colocasia bobone disease virus, 74
Celery yellow spot virus, 139 Colocasia virus 2, 111
Volume I 211
Comovirus. 8—10 Deittlnibiiiin pluiltienopsix. 94

Computer analysis, general discussion. 3—I Density-gradient ccnirifugation. general discussion.
Concanavalin A. 21 5^
Coniagium vivum JhtiJum. I Deoxyribonucleic acid, see also spixific types by vi-
Convoluted membranes. 141. 144 rus name
Cotton anthocyanosis virus. 139 Caulimovirus studies. 17—18. 21—23. 27
Cow parsnip mosaic virus. 74 double-stranded, see Double-stranded DNA
Cowpea mosaic virus, 10 Ceminivirus studies. 33—34. 36—38
Crystalline aggregates. 6. 37. 40 general discussion, 4—5
Crystalline monolayers. 4 replication, 27. 37
Cucumber. 171 single-stranded, 4
Cucumber green mottle mosaic virus. 12 viral, see Viral DNA
Cucumber mosaic virus, 6. 11 Deoxyribonucleic acid viruses. 21
Cucumber necrosis virus. 171. 182—183 Dcoxyribonucleoprotein. 38
Cucumis DE panicle, see Double-enveloped panicle
melo. 76 Dcproteinization. 5
satims. 102 Dcsmodium yellow mottle virus. I IS—119. 133
Cucumovirus Detergents, dissociation with. 81—83. 86. 101
cytopathology, 173, 190 D/H5" cauliflower mosaic virus strain. 21
laxonomic groups, 9, 11 Diamond knife, general discussion, d
tonoplast vesicles. 173 Dianthovirus, 8—9, 11
Cylindrical inclusions. 8 Dictyosome, 190
Cymbidium ringspot virus, 181—182, 190 Digitaria striate virus, 74
Cynara virus, 74 DiscontinuUies, DNA. 21—22
Cynosurus mottle virus. 153—154, 156—157 Discovery, viruses. 1
Cypovirus, 54 Dissociation techniques, virus particles. 81—83. 86.
Cytidine, 117 155
Cytochemical studies, general discussion, 6—7 Divalent metal ions. 155
Cytopathic effects, 8—12 DNA. see Deoxyribonucleic acid
Cytopathology. see also as subheading beneath spe- Double-enveloped particles. 102. 106. 108—109
cific virus names Double-membraned vesicles, 124—125, 127, 130,
panicle, 8—12 141
virus. 5—7 Double-stranded DNA
Cytoplasm, virus-infected cells, virus panicles in Caulimovirus studies. 17—18. 21—22
cistemae, 102—103. 105, 108—109 Geminivirus studies, 38
Geminivirus studies, 37 general discussion, 4
Luteovirus studies, 140—141, 143—147 Luteovirus studies, 140
maize chlorotic dwarf virus studies. I I I , 113 Double-stranded RNA
Reoviridae studies, 57—63 general discussion, 4. 7. 9—10
Rhabdoviridae studies, 73—75, 86, 90, 93, 95 Luteovirus studies. 141
Sobemovirus studies, J57. 160—163. 165—166 Reoviridae studies, 47—18. 50. 55—56. 63
tobacco necrolic virus studies, 173, 176 Sobemovirus studies. 157
tomato spotted wilt virus studies, 102—103, 105, tobacco necrosis virus studies, 172
108—109 Tombusvirus studies. 186, 190
Tombusvirus studies, 187—189. 191. 193. 195 Tymovirus studies. 128
Tymovirus studies. 128, 130—131. 133 virus-specific, 186, 190
Cytoplasmic multivesicular bodies. 181. 190—191, Dulcamara mottle virus. 118—119
193—195
Cytoplasmic polyhedrosis virus, 48, 63
Cytoplasmic tubules, 63—64. 66. 68 E
Early history, I
D EDTA, see Ethylenediaminetetraacetie acid
Eggplant mosaic virus. 117—119, 122, 127
Dahlia mosaic virus, 17—18, 25 Eggplant mottled crinkle virus, 182. 190
Dahlia variabilis, 25 Eggplant mottled dwarf virus. 73—75. 86—87
Datura stramonium, 105—107, 191—192 structural proteins, 86
Daucus carota, 145, 150 Electron beam damage, 3
Defective isolates, tomato spoiled will virus, 103 Electron-dense amorphous material, 173. 176
Deletions, RNA, 57 Electron-lucent inclusions, 27, 29
Dendrobium hybrid, 76 Electron-lucent regions, 128
Electron-lucent viroplasm, 93 French bean. 38. 41—44. 133. 160—161. 171.

Electron microscopy, general discussion. 1—6 173. 178, 194—195
grids for, 4 Fungus transmission
minimal exposure technique. 3 cucumber necrotic virus. 183
ELISA, see Enzyme-linked immunosorbem assay tobacco necrotic virus. 171. 183
Embedding resins, general discussion, 6
Empty protein shells. 119—120, 140
accumulation in nucleus. 128—130. 133 G
Enations, 47, 60, 76
Endonuclease Sal I, 21 Galinsoga mosaic virus, 12, 182—184
Endoplasmic reticulum, 86, 88—90, 93, 128, 130, cytopathology, 190, 194—195
141. 157. 173. 176, 190 particles, 184. 194—195
Endria inimica. 75 properties of, 182—183
Envelope, nuclear, 90. 93. 128 thin section illustrations, 194—195
Enveloped panicles Gall cells. 61—62, 64. 66
double envelopes. 102, 106. 108—109 Galls, 47, 60
envelope structure, 76—81 Gaps, DNA, 21—22
general discussion, 3, 6, 9—10 Garden pea, 126
Rhabdoviridae studies. 73. 75—85, 90, 93 Gel elcctrophorcsis, general discussion, 5
tomato spotted wilt virus studies. 101—103, 106. Germinate particles, 33—36
108—109 Geminivirus, see also specific viruses by name. 8—
Enzyme activities, 56—57, 83 10. 33^44
Enzyme-linked immunosorbent assay, general dis- coat protein, 33—34, 37
cussion, 4 cytopathology, 34, 37—44
Erysimum latent virus, 117—118 genome, 10, 33, 36—37
Ethylenediamine tetraacetic acid, 155, 185 host range, 33
Euonymus japonica. 76 mechanical transmission, 33
Euphorbia mosaic virus, 34. 37 particle, 10, 33—44
Evolution, general discussion, 6 cytoplasmic, 37
electron microscope detection, 37
germinate, 33—36
F morphology. 8, 10, 33—36
nucleus of infected plant cells, presence in, 34,
Festuca gigantea, 76 37—40
Fibrillar material. I l l , 124, 127—128, 130, 141. phloem cells, presence in, 37—38
144—146, 150, 194 size. 10, 33
Fibrillar regions, 37—38, 41—44 structure, 10, 33—36
Fibrillar rings, 37, 42—43 replication, 38
Fibrils. 147. 157. 160—164, 193 seed transmission, 33
virus-specific, 161—162, 164 structure and composition, 10, 33—37
Figwort mosaic virus, 17—18, 21 taxonomic groups, 8—10, 33
Fiji disease virus. I. 10—11, 48, 50, 52. 55—58, vectors, 33—34, 38
60—63 Gene products, 21—23, 55
subviral particle, 55—57, 62, 67 General discussion, I—12
Fijivirtu, 8, 50—54, 56—57 Genes, protein, 187
cytopathology, 57, 60—68 Genetic engineering, plants, 81
genome, 50, 57, 63 Genome, see also as subheading beneath specific vi-
morphology, 47—54 rus names, 4—5, 9—11
particle, 51—54, 56—58, 60—68 size, 10—11
model of. 55 Genome-linked viral protein, 140. 156
structure. 51—54, 56—57 Gerbera sp., 76
structure, dimensions, 54 German strain, tobacco necrosis virus, 171
serological relationships, 47—48, 52 Glycine mottle virus, 181—182
taxonomic group. 10—II. 47—48. 50—52 Glycosylated protein, 83, 86, 101, 103
viroplasms, 60—66 Glycosylation, 21
Filaments, 105. 107 Gomphrena globosa, 190
kinky. 60. 62—63. 66—67 Gomphrena virus, 74
Filaree red leaf virus, 139 G protein, see Glycosylated protein
Finger millet mosaic virus. 74 Graminella nigrifrons, 74
Fixatives, general discussion. 6, 76 Granular inclusions, see also Viroplasm. 90—91.
Flowerbreaking, I 102, 111
Volume I 213
Granular regions, 38, 41, 44 tomato spotted wilt virus studies. 102—103, 105.
Grids, electron microscope, general discussion, 4 108—109
Ground nul rosette assistor virus, 139 Tombusvirus studies, 181—182. 190
Groups, see Taxonomic groups Tymovirus studies. 128—133
virus-specific, 115
Infected plant cells, sec Virus-infected plant cells
H Insect vectors, see Vectors: specific vectors by
name
Heavy metal salts. 3 Insect viruses, 47—48, 57, 63, 90
Hepatitis B virus, 23 International Committee on Taxonomy of Viruses.
Hibiscus chlorotic ring spot virus, 182—184 7—9
Histopathology, general discussion, 57, 60 Ionic detergents, 81—83, 86
History, I Ions, metal, 155
Holcus lanatus, 76 Ms germanica. 76
Hordeivirus, taxonomic groups, 9, 11 Isometric particles, 1—2,4, 10—12.33—34. 117.
Horseradish latent virus. 18 181
Host, hypersensitive, local lesion formation in, 190 recognition of, in thin sections. 6
Host range Ivy vein clearing virus, 74
Caulimovirus. 17
Geminivirus, 33
Luteovirus, 137—139 K
maize chlorotic dwarf virus, 111
maize rayado fino virus, 117 25K, 36K proteins. 187
Reoviridae, 47—18 42K. 49K, 55K proteins, 18—21
Rhabdoviridae, 73 150K, I95K. 2IOK proteins, 122—123
Sobemovirus, 153 Kcnnedya yellow mosaic virus, 118—119, 126
tobacco necrosis virus, 171 Kinky filaments, 60, 62—63, 66—67
tomato spotted will virus, 101
Tombusvirus, 181
Tymovirus, 117 L
Hyadaphis foeniculi. 139
Hypcromyzus laciucae, 74—75 Laburnum anagyroides, 76
Hyperplasia, 57, 63 Lacebug transmission, Rhabdoviridae, 73—74
Hypersensitive hosts, local lesion formation in, 190 Laelia sp., 76
Laminate inclusions, 111
Laodelphax siriatellus, 74—75
I Leaf hopper assay, rice tungro virus studies, 111
Leafhopper transmission
Icosahedral particles, small, 8 persistent
Identification, viruses, 8—12 Geminiviruses, 33—34
Ilarvirus. taxonomic groups, 9, 11 maize rayado fino virus, 117
Immunochemtcal studies, general discussion, 6—7 Reoviridae, 47
Inclusion bodies, 5, 23, 57 Rhabdoviridae, 73—75
Inclusions scmipersistent, maize chlorotic dwarf virus, 111
Caulimovirus studies, 17—18, 23, 27—29 Sobemovirus, 154
cytoplasmic, see Cytoplasm, virus-infected cells, Leafminer transmission, Sobemovirus. 154
virus particles in Legume yellow virus, 138
electron-lucent, 27, 29 Lesions, local, formation in hypersensitive hosts,
Geminivirus studies, 34, 37—40 190
general discussion, 8 Lettuce nccrotic yellows virus, 3, 73—76, 79—81,
granular, see also Viroplasm, 90—91 83, 85—86, 88—90. 92—93
laminate, 111 aberrant, possible origin of, 81
maize chlorotic dwarf virus studies. I I I , 113 disassembly of, diagram, 86
nucleic, see Nucleus, virus-infected cells, virus properties and characteristics of. 10—II, 74, 85
particles in structural proteins, 86
perinuclear. see Perinuclear space synthesis and distribution, diagram, 93
Reoviridae studies, 57—63 Linear integration, 3
Rhabdoviridae studies. 73, 82, 84, 86—87, 90— Liriomyza langei. 154
93 Local lesion formation, in hypersensitive hosts. 190
rice tungro disease studies, I I I , 115 Lolium cnation virus, 48, 52, 57
Sobemovirus studies. 157 Lolium sp., 76
214 AI lax of Plain Viriisea
I. protein. 83. 86 Maniltoi esrulenta. 76

Lucerne cnution virus. 74 Maxonapliix pepperi. 154
Lucerne transient streak virus. 153—157. 159. 165 Matrix protein. 73. 83. 86. 101
cytopathology. 157. 165 Maturation, tomato spotted will virus particles.
negatively stained example. 159 102—103. 108—109
properties of. 153—154 Mature particle. 102
KNA. 156 MAY isolate, barley yellow dwarf virus. 137—138,
thin section illustration. 165 141
Lupinus pal\'pli\ilu\. 76 Mechanical transmission
Luteovirus. 8—10. 137—150 Caulimovirus. 17
coat protein. 138. 140 Geminivirus, 33
cytopathology. 138—141. 143—150 Luteovirus. 137. 141
differences in. correlation with serological dif- maize chlorolic dwarf virus. 111
ferences. 141 maize rayado fino virus. 117
sequence of cytological events, 140—141 Reoviridae. 47
genome. 10. 137. 140 Rhabdoviridae, 74—75
host range. 137—139 Sobemovirus. 153
mechanical transmission. 137. 141 tobacco necrosis virus, 171
negatively stained illustrations. 142 tomato spotted will virus. 101. 103
panicle. 'lO. 137—147. 149 Tombusvirus. 181
in cytoplasm. 140—141. 143—147 Tymovirus. 117
morphology. 8. 10. 137—140 Melilotus latent virus. 74
in nucleus. 141, 149 Melon variegation virus. 90
polyhedral. 137—138 Membrane-bound structures. 190
polypeptide. 137. 140 Membrane-bound tonoplast vesicles. 173
shape. 137—140 Membranes, convoluted. 141. 144
size. 1.17—139 Membranous cistcrnae. cytoplasm. 102—103. 105.
structure. 10. 137—140. 142 108
phloem cells and tissues, confinement to. 137— Membranous inclusions. 190
138. 140—141, 143—150 Membranous tubules. 105. 107
purification, improved methods for. 137 Mesophyll and mesophyll cells. I I I . 124—127.
sedimentation coefficients. 137—138. 140 131—133
seed transmission. 137 Messenger RNA, 5. 56—57
serological relationships. 137—139. 141 coat protein. 119. 122—123. 156
structure and composition, 10. 137. 139—140. 3' end termination, lack of Poly A. 173
142 5' end termination, lack of cap structure. 173
symptoms, 137. 139 I9S Messenger RNA. 22—23
laxonomic groups. 8—10. 137—139 Metal ions, divalent. 155
thin section illustrations. 143—150 Metal salts, heavy. 3
vectors. 137—139. 141 Metal shadowing, general discussion. I
vesicles. 141. 144—148. 150 Methodologies, general discussion. I —12
Lysosomc-like bodies, 60 Microbodies. 190
/,v.v.«/r//-».v, 73 Microtome, general discussion, 6
Milk-vetch dwarf vims, 139
Minimal exposure technique, general discussion. 3
M Mirabilis jalapa. 26
Mirabilis mosaic virus, 17—18. 26
Macrosiphum avenue. 137—138 Mirid transmission. Sobemovirus. 154
Magnesium ion. 155 Mite transmission. Rhabdoviridae, 73—74
Maize, 65—66. 113 Mitochondria, 12. 130. 141. 145. 150. 157. 181—
Mai/.e chlorotic dwarf virus. 9—10. 111—115 183, 187. 195
Maize mosaic virus, 74—76, 90, 93 ultrastructure. alterations in, 190
Maize rayado fino virus. 117. 120 Monolaycrs. particle, preparation of. 4
Mai/.e rough dwarf virus. 48. 50—53, 60. 65—66, Morphology, virus particles, see Particle,
68 morphology
capsid polypcptides. 56 M particle, see Mature particle
panicle structure, 53 M protein, see Matrix protein
subviral particles. 63 mRNA, sec Messenger RNA
Maize sterile stunt virus. 74 Multivesicular bodies. 171. 181—182, 190—195
Maize streak virus. 10. 33—38 in cytoplasm. 181. 190—191. 193—195
Malva yellows virus. 138 virus-induced. 191—195
Volume I 215
Mungbean yellow mosaic virus. 34. 37 Oal sterile dwarf virus. 48. 50. 52. 57. 59—60.
Mutants. 57 63—64. 66
My:us perxicne. 139 Oat striate mosaic virus. 7-1—75
Okra mosaic virus. 118—119. 122. 128
Olpitliiint
N hnixxitw. 183
sp.. 171
Negative-sense RNA. 9. 73. 83 Ononis yellow mosaic virus. 118—119
Negative staining technique, general discussion, sec Optical diffraction, general discussion. 3—'
also subheading "negative staining illustra- Orhivirux. 48. 50—52
tions" beneath specific virus names. 2—3. Orchid. 75. 95
8. 10—11. 76 Orchid fleck virus. 75. 90. 95
Negative-stranded RNA. 101. 103 Orosius tirgentalux. 34
Neoplasia. 47. 57. 60
Nephotenix (ipiatlis. 75
Nepovirus. 8—10 p
Nesoclutha patliila. 34. 74
Nicotiana Pangola stunt virus. 48. 50. 52
benlhamiana. 82 Paracrystalline monolayers. 4
Cleveland!!. 88—89. 92. 102. 163—164. 188— Parenchyma cells. 141. 143—144. 146. 150
189. 193 Parsley rhabdovirus. 74
x edwardsonii. 173. 176—177 Particle, see also as subheading beneath specific vi-
glutinosa. 102. 127 rus names
labaaim. 87. 102 baciltiform, see Bacilliform particles
Nomenclature. 7—8 bullet-shaped. 73. 76. 81. 84
Nonionic detergents. 81. 86. 101 cytopathology. see also Cytopathology. 8—12
Northern cereal mosaic virus. 73—74 enveloped, see Enveloped panicles
N protein. 83. 86 germinate. 33—36
NS protein. 83. 86 isometric, see Isometric particles
Nuclear envelope. 90. 93. 128 monolayers. preparation of. 4
Nuclear membrane. 90. 92—93. 95—96 morphology, see also as subheading beneath spe-
Nuclear replication, viral DNA. 27 cific virus names. 3. 8. 10—11
Nuclear transcription, viral DNA. 23 polyhedral, see Polyhedral panicles
Nuclear ultrastructure. alterations in. 190 shape, see also as subheading beneath specific vi-
Nucleic acid, see also Deoxyribonucleic acid: Ri- rus names. 1—3. 8—9
bonucleic acid. 1—5. 33. 103. 117. 128. size, see also as subheading beneath specific virus
130. 140. 144. 157. 190. 193 names, I—2. 8—11
cloning. 5 structure, see also as subheading beneath specific
molecules, delivery of. 36 virus names. 1—4
Nuclcocapsid. see also Capsid. 76. 81—85, 93— swelling. 155, 185
94. 96. 102—103 B Particle, see Bottom panicle component
bullet-shaped. 81.84 DE Panicle, see Double-enveloped panicle
envelopes. 90. 93 M Panicle, see Mature particle
protein. 103
T Particle, see Top particle component
Nucleoplasm, 38. 4,2—44. 90. 93. 141
Paspalum dilatation. 38, 40
Nuclcoprotein. I. 5. 76. 81. 101. 120. 122. 140
Paspalum striate mosaic virus. 34. 37—38. 40
Nuclcotide. 36—37. 120. 122—123
PAY isolate, barley yellow dwarf virus. 138. 141
Nucleotide sequences and sequencing. 5, 7. 37,
pBR322 plasmid. 21
117. 172. 186
Pea enation mosaic virus, taxonomic groups. 9. 11
Nucleus, virus-infected cells, virus particles in
Pea (bean) leafroll virus. 138
Caulimovirus studies, 27—28
Peanut yellow mottle virus. 117—118
Gcminivirus studies. 34. 37—40
Luteovirus studies. 141. 149 Pelargonium, 190
Rhabdoviridae studies. 74—75. 87. 90—96 Pelargonium leaf curl virus. 182. 190
Sobemovirus studies. 157. 160. 163. 165—166 Pelargonium vein clearing virus. 74. 82
tomato spotted wilt virus studies. 102 Pentalonia nigronfrvostt. 139
Tombusvirus studies. 187. 189—190 Peregrinus inaidis, 74
Perinudear blisters. 90. 92
Perinuclear space. 73. 82. 84. 86—87. 90. 92—93.
o 141. 147—148
Peripheral vesicles. 124—128. 130—131. 150. 190.
Oats, 64 193—195
Peroxisome. 181—182. 191 small, ribosomes differentiated from. 6. 173

Persistent aphid transmission. 137—139. 141 Polymerase. 56. 83. 183
Persistent Icaflioppcr transmission. 33—34. 117 Polypeptide
Persistent thrip transmission. 101 capsid. 55—56. 1 1 1
Persistent whitefly transmission. 33. 34 Geminivirus studies. 10. 33—35
Petunia asteroid mosaic virus. 182 general discussion. 10—II
Petunia vein clearing virus. 18 Luteovirus studies. 137. 140
pi! inai/e chlorotic dwarf virus studies. 10. I l l
high, cation removal at. 185 Reoviridae studies. 10. 48. 55—57
staining and. 2—3 Sobemovirus studies. 155—156
Plialacnopsis sp.. 76 tobacco necrosis virus studies. 171—172
Phaseolus vulgaris. 41—44. 133, 160—161, 194— Tombusvirus studies. 183
195 Tymovirus studies, 10. 117. 120. 122
Phleum mottle virus. 153 Polyribonuclcotide. 48
Phloem tissue and cells Polyribosome. 90
Gcminivirus studies. 37—38 Positive-sense RNA. 9. 117. 137. 140. 153. 171—
Luteovinis studies, 137—138, 140—141, 143— 172, 181
150 Posttranslational cleavage. 122—123
maize chlorotic dwarf virus studies. 111, 115 Potassium phosphotungstate. see Phosphotungstic
parenchyma cells. 141. 143—144. 146. 150 acid
Reoviridae studies. 57. 60 Potato, 139
Phosphorylation, 21 Potato leafroll virus. 137—141. 144. 146. 148
Phosphotungstic acid staining, general discussion. properties of. 138
2—3~. 76. 80 RNA, 140
Photographic rotation technique, general discussion, Potato virus X virus. 11
3 Potato virus Y virus. 11
Phycomycete fungus transmission, cucumber necro- Potato yellow dwarf virus. 73—74. 83. 85—86
sis and tobacco necrosis viruses, 183 properties and characteristics of. 10—11. 74. 85
Physalis fltiridana. 144. 146 structural proteins. 86
Physalis mild chlorosis virus. 139 Potexvirus. 9. I I . 173
Physalis mosaic virus. 117—118 Potyvirus. 8—9. 11
Physalis vein blotch virus. 139 Protein
Phytorem-irus. 8. 10—II. 47—60 Caulimovirus studies. 18—23. 27—28
cytopathology. 57—60 coat, see Coat protein
genome. 50. 57 Geminivirus studies. 33—34. 36—37
morphology, 47—54 general discussion. 1—2. 4—5. 7. 10—11
particle. 51—58 glycosylated. see Glycosylated protein
structure. 51—56 Luteovirus studies. 138, 140
structure, measurement of. 54 matrix, see Matrix protein
taxonomic group. 10—II. 47—50 nucleocapsid. 103
viroplasms, 57—60 posttranslational cleavage of. 122—123
Piesma quadraium. 74 Reoviridae studies. 48. 50. 55—56. 60. 63
Pinwhecl inclusions. 8 Rhabdoviridae studies. 73. 76. 81. 83. 86
Pisum sativum. 126 shells, see Protein shells
Pisum virus. 74 Sobemovirus studies, 154—156
Pittosporum vein yellowing virus, 74 structural, Rhabdoviridae. 83. 86
Planlago lanceolata, 76 synthesis, 5. 122
Plamago mottle virus. 118—119 tobacco necrosis virus studies. 171—172
Plantago virus 4, 18 tomato spotted wilt virus studies. 101. 103
Plant cells, infected, see Virus-infected plant cells Tombusvirus studies. 181—187
Plamhopper transmission Tymovirus studies. 10. 117—120. 122—123.
Reoviridae, 47. 63 128—130. 133
Rhabdoviridae. 74—75 viral, see Viral protein
Plasmids. 21—22 25K. 36K Protein. 187
Plasmodesmata. 23, 25. 27. 140—141. 144—145, 42K Protein. 18—21
190 49K.55K Protein. 20—21
Pogostfmon patchouli. 76 I50K, 195K. 2IOK Protein. 122—123
Poly A. 173 G Protein, see Glycosylated protein
Polyamines. 120 L Protein. 83. 86
Polyhedral particles, I I I . 115. 137—138, 153. M Protein, see Matrix protein
171—173 N Protein. 83, 86
Volume I 217
NS Protein. 83. 86 vertebrate viruses, 23
Protein genes. 187 viral DNA, 27. 38
Protein-RNA reactions. 120 viral RNA. 122. 128. 172—173. 186
Proteins I-1V. tomato spotted will virus panicle. 101 Restriction endonuclcase. 17. 22. 36
Protein shells. 117. 155. 1S5 Retrovirus, 23
empty. 119—120. 128—130, 133. 140 Reverse transcription. 23
accumulation in nucleus. 128—130, 133 Rhabdoviridae. 8—11. 73—96
Proteolysis, 17 animal (vertebrate) rhabdoviruscs compared with.
Protoplast. 6. 37. 140—141 81.85
Pseimmoteitix buoyant densities. 81. 85
alienus. 34 cytopathology. 85—86
striatus. 75 genome, 10. 83—85
PTA, see Phosphotungstic acid host range. 73
Pythiaceous fungus transmission, tobacco necrotic infection by. mechanism of. 83—85
virus, 171 mechanical transmission. 74—75
negative staining illustrations. 76—81, 85, 94
nuclcocapsid. 76. 81—85. 90. 93—94, 96
R particle. 10. 73—96
bacilliform. 73. 76. 81—82
Rabies. 73 bullet-shaped, 73. 76, 81, 84
Random aggregates, 37—38 in cytoplasm, 73—75. 86. 90, 93. 95
Ranunculus repens, 76 dissociation techniques. 81—83. 86
Raphamis virus. 74 in endoplasmic reticulum. 86, 88—90. 93
Raspberry leaf curl virus. 139 envelopes, 73. 75—85. 90, 93
Raspberry vein chlorosis. 75 envelope structure. 76—81
Reoviridae. see also specific viruses by name, 8— model of, 83
11.47—68 morphology, 8, 10. 76—86
capsid polypcptides, 55—56 in nucleus. 74—75. 87. 90—96
cytopathology. 8, 57—68 in perinuclear space. 73. 82. 84, 86—87, 90,
genome. 10, 47—48, 50. 56—57. 63 92—93
host range. 47^8 shape, 73. 76—84
mechanical transmission. 47 size. 73. 76. 81, 85
particle. 10, 47—68 structure, 10, 76—86
morphology. 8. 10. 47—54 in vesicles. 86. 88—90. 92—93
shell. 47—56. 60, 63 properties of. 74—75. 85
structure, 10, 48—57, 63 sedimentation coefficients, 81, 85
subviral, 48. 52—57. 62. 66. 68 seed transmission, 73
properties of, 48 serological relationships. 73. 75
serological relationships. 47^48, 52 site of assembly, in vivo. 74—75
structure and composition. 10—11, 48—57. 63 spokewhecl structures, 93, 95—96
symptoms. 47 structural proteins. 83. 86
taxonomic groups. 8—11. 47—52 structure and composition, 10. 76—85
thin section illustrations, 58—59, 61—62. 64— subgroups I and II. 73—75
67 symptoms. 76
tubular structures, 57, 63—64. 66. 68 synthesis and distribution, diagram. 93
vectors, 47. 57. 63 taxonomic groups. 8—II. 73—76
viroplasm. 8, 57—66 thin section illustrations, 82, 84. 87—89. 91—
Reovirus. 8. 47—48. 57, 60 92. 95—96
Rcovirus-like panicles. 47 thread-like structures. 89—90
Replication vectors, 73—75. 90
Caulimovirus, 23 vesicles. 86. 88—90. 92—93
DNA, 27. 37—38 viroplasm, 90—91, 93
Geminivirus. 38 Rhabdorirus. 3, 8, 73—96
maize rayado fino virus. 117 Rhabdovirus-like particles. 73. 75—76. 84. 90—94
RNA. 122. 128, 141. 157. 172—173. 186 devoid of envelope, 75
tobacco necrosis virus. 173—174 Rhopalosiphum
lobacco necrosis virus satellite virus. 172—174 maidis, 138
tobacco necrotic dwarf virus. 141 padi. 138. 154
Tombusvirus. 190 Ribonuclease, 6. 57, 172—173. 176
turnip yellow mosaic virus. 120 ribosome removal by. 140, 143—146, 150. 160.
Tymovirus. 120 162—166. 173. 176. 191—192. 195
Rihonucleic acid, see also specific types by virus Rice (ungro virus. I l l —12. 114—115
name Rice yellow mottle virus. 153—154. 156—157
capping at 5' end, 122—123 RMV isolate, barley yellow dwarf virus. 138
Caulimovirus studies. 22—23 RNA, see Ribonucleic acid
deletions in. 57 mRNA. see Messenger RNA
double-stranded, see Double-stranded RNA rRNA. see Ribosomal RNA
3'end. + RNA-like sequence at, 122—123 tRNA. see Transfer RNA
Geminivirus studies. 36 vRNA. see Viral RNA
general discussion. 4—7. 9—11 RNAse, see Ribonuclease
Luteovirus studies, 137—138, 140—141 Root cell, 65
mai/.e chlorotic dwarf virus studies. 111 Rotation technique, photographic, general discus-
messenger, see Messenger RNA sion. 3
negative-sense. 9. 73. 83 Kotavirus. 48
negative-stranded. 101. 103 Rounded chloroplast. 124. 126. 128. 131
positive-sense. 9. 117, 137. 140. 153. 171—172. RPV isolate, barley yellow dwarf virus, 138. 141
181 rRNA. see Ribosomal RNA
protein-RNA reactions, 120
Reoviridae studies. 47—48. 50, 55—57, 60. 63
replication and replicative form. 122, 128. 141, s
157. 172—173. 186
Rhabdoviridae studies. 73. 76. 81. 83. 85. 90 Saguaro cactus vims, 182
ribosomal. see Ribosomal RNA Saintpaulia sp.. 76
satellite. 5, 154. 156. 185. 187 Sal 1 endonuclease, 21
single-stranded, see Single-stranded RNA Sambucta sp., 76
Sobemovirus studies. 153—158 Sandfly fever virus, 103
subgenomic. 119—120. 122—123. 156. IBS- Sarracenia purpun'a, 76
IS? Smtelite RNA. 5, 154. 156. 185. 187
tobacco necrosis virus studies. 171—173 Satellite viruses, 5
tomato spotted wilt virus studies. 101. 103 tobacco necrosis virus, see Tobacco necrosis vi-
Tombusvirus studies. 181—183. 185—187. 190 rus, satellite virus
transfer, see Transfer RNA Schiiaphis graminuin, 138
Tymovirus studies. 117—120. 122—123. 128 Scrophularia mottle virus. 118—119
viral, see Viral RNA SDI. see Serological differentiation indices
viroid-like. 153—154. 156 Sedimentation coefficients
virus-like, 153—154 carnation yellow stripe virus, 171
Ribonucleic acid I. 2. 3. 156. 158 Luteoviruses, 137—138. 140
8S Ribonucleic acid. 23 maize chlorotic dwarf virus, 111
35S Ribonucleic acid. 23 Rhabdoviridae, 81. 85
Ribonucleic acid polymerase (transcriptase). 56, 83 Sobemoviruses. 153—154
Ribonucleic acid-ribonucleic acid hybridization. 117 tobacco necrosis virus, 171—172
Ribonueleoprotcin. 38 Tombusviruses. 181—183
Ribonucleotide sequences, 21 Tymoviruses. 118—120
Ribosomal RNA, 6 Seed transmission
Ribosome. 23. 26. 62. 90. 128. 130 Caulimovirus. 17
RNAse removal of, 140. 143—146. 150. 160. Geminivirus, 33
162—166. 173. 176. 191—192. 195 Luteovirus. 137
small polyhedral virus particles differenliated maize chlorotic dwarf virus. 111
from, 6, 173 Rhabdoviridae, 73
Rice. 115 Sobemovirus. 153—154
Rice black-streaked dwarf virus. 48. 50. 52 tomato spotted wilt virus, 101
Rice dwarf virus. 47. 49—50. 52. 54. 56—57 Tombusvirus. 181
capsid polypeptides, 56 Tymovirus. 117
Rice gall dwarf virus. 47. 50. 52. 54. 57 Semiaphis heraclei. 74
Rice giallume virus. 138 Semipersistcnt aphid transmission. 17—18, 22
Rice ragged stunt virus. 47. 53—54. 67 Scmipersistenl leafhoppcr transmission, 111
eytopathology. 57. 59. 63 Serological differentiation indices. 117. 119. 171
genome. 50. 57. 63 Serology and serological relationships
morphology. 54 Caulimovirus studies. 17
particle. 53—54. 59. 63 general discussion. 8
taxonomic group. 47 Luteovirus studies. 137—139, 141
Rice transitory yellowing virus. 75. 86 Reoviridae studies. 47—48. 52
Volume 1 219
Rhabdoviridae studies. 73. 75 tubules. 157—158. 164

Sobcinovirus studies. 153 vectors. 153—154
Tombus virus studies. IS I vesicles. 157. 160—161. 164
Tymovirus studies. 117. 119 viroid-like RNA. 153—154. 156
SGV isolate, barley yellow dwarf vim.s. 138 Sogiitfllu longifurciferu. 74
Shadow casting, general discussion, I Solanum nodiflorum mottle virus. 153—158. 164
Shape, particle. 1—3. 8—9 cylopathology. 157. 164
Shell, particle, see also Protein shells. 47—56. 60. properties of. 153—154
63 RNA. 156
Single-stranded DNA thin section illustration. 164
circular. 33. 36 Solanum yellows virus, 138
Geminivirus studies. 33—34. 36. 38 Sonchus olfraccuus. 84
general discussion. 4 Sonchus virus. 73. 75. 86
Single-stranded RNA Sonchus yellow net virus. 73. 83. 85—86. 90
Geminivirus studies. 36 properties of. 75. 85
general discussion. 4—7. 9—11 structural proteins, 86
Luteovirus studies. 137. 140 Soubane mosaic virus. 153—157. 159, 162
maize chlorotic dwarf virus studies, 1 1 1 cytopathology. 157. 162
Reoviridae studies. 56—57 negatively stained illustration. 159
Rhabdoviridae studies. 73. 83. 85 properties of. 153—154
Sobemovirus studies. 153. 156 RNA. 156
tomato spotted wilt virus studies. 101. 103 thin section illustrations. 162
Tombusvirus studies. 181 South African grapevine strain, tobacco necrosis vi-
Tymovirus studies. 117. 120 rus. 171
Size Southern bean mosaic virus. 10. 140. 153—157.
coat protein. 10—11 159—160. 183
genome. 10—! 1 cytopathology. 157. 160
particle, see also subheading "panicle, size" be- genome. 156
neath specific virus names. I—2. 8—11 negatively stained illustration. 159
Small-angle neutron scattering, general discussion. 4 particle structure. 155. 159. 183
Small icosaderal particles. 8 properties of. 153—154
Small polyhedral virus particles, recognition of, 6 thin section illustrations. 160—161
Sobemovirus. 8—10. 140. 153—166. 183 Sowthistle yellow vein virus. 73. 78, 81. 83—84
buoyant densities. 154 properties of. 75
coal protein. 154—156 structural proteins, 86
cytopathology. 156—158. 160—166 Soybean dwarf virus, 137—138
fibrils. 157. 160—164 Spcrmine and spcrmidine. 120
genome. 10. 153. 156 Spokewheel structures. Rhabdoviridae. 93. 95—96
host range, 153 Staining, see Negative staining technique
mechanical transmission, 153 Strawberry crinkle virus. 75
negatively stained illustrations, 159 Strawberry mild yellow edge virus. 139
particle. 10. 153—166. 183 Strawberry vein banding virus. 17—18
in cytoplasm. 157. 160—163. 165—166 Structural proteins. Rhabdoviridae. 83. 86
divalent metal cations, dependence on. 155 Subgenomic RNA. 119—120. 122—123, 156,
morphology. 8, 10, 153. 155 185—187
in nucleus. 157. 160. 163. 165—166 Subterranean clover, 143, 166
polyhedral. 153 Subterranean clover mottle virus. 153—157. 166
polypcplides, 155 Subterranean clover red leaf virus. 137—142
Subterranean clover stunt virus. 139
shape. 10. 153. 155
Sugar beet. 141. 147—149
size. 10. 153. 155
Sugarcane gall cells. 58. 61—62
structure. 10. 153. 155. 159. 183
Svcdberg's equation, 18
swelling. 155
SVP, see Subviral particle
in vacuoles, 151. 163
Swelling, panicles, 155, 185
properties, 153—154 Swollen chloroplast. 124. 128
sedimentation coefficients, 153—154 Synthesis, virus, 60, 90. 93
seed transmission. 153—154
serological relationships. 153
structure and composition. 10. 153, 155—156, T
159
taxonomic groups. 8—10. 140. 153—154 Taxonomic group, virus, see also as subhead be-
thin section illustrations. 160—166 neath specific virus names. 7—12. 199—207
characteristics, main. 10—11 structure, 181. 183—187

classification into. 7—11 properties of, 181—182
general discussion. 7—12 protein analyses of, 183
list of. by virus names. 199—207 RNA. 185—186
type members. 10—II thin section illutrations. 188—189. 191
Taxonomy, viruses. 7—11 Tomato golden mosaic virus. 34, 37—38
Thin section illustrations, see as subheading beneath Tomato spotted will virus, 8—10, 101—109
specific virus names Bunyaviridae compared with, 101, 103
Thistle mottle virus. 18 cytopathology. 102—103. 106—109
Thread-like structures. Rhabdoviridae. 89 defective isolates. 103
Thrip persistent transmission, tomato spotted wilt genome, 10, 101
virus. 101 host range, 101
Thymus x cilriodorus. 76 mechanical transmission. 101. 103
Tipula iridescent virus, 2 negative staining illustrations. 104
Tobacco leaf curl virus, 33—34. 37 particle. 8. 10, 101—109
Tobacco mosaic virus, I, 4—6. I I . 118 properties, 103
Tobacco necrosis virus. 8. 10, 171—178, 183, 190 RNA. 101
coat protein, 171—172 seed transmission, 101
cytopathology. 173—174. 176—178. 190 structure and composition. 10. 101, 104—105
genome. 172 taxonomic group. 8—10. 101
host range. 171 thin section illustrations. 105—109
mechanical transmission, 171 vectors. 101, 103
negatively stained illustration. 175 viroplasm. 102. 106. 108
particle. 8. 10. 171—178. 183 Tomato yellow dwarf virus, 33—34, 37
in cytoplasm. 173. 176 Tomato yellow leaf curl virus. 34. 37
morphology. 8. 10. 171 — 172 Tomato yellow mosaic virus. 34. 37
polypcptide. 171—172 Tomato yellow net virus. 139
shape. 10. 171—172 Tomato yellow top virus. 137—138
size. 10,.171—172 Tombusvirus, 8—10. 181—195
structure. 171—172 buoyant densities. 182—183
properties of, 171—172 coat protein. 181—187
replication. 173—174 subunits. 183—187
RNA, 172—173 cytopathology, 181—183. 187—195
satellite virus. 171—176. 178 genome, 181, 185—187
sedimentation coefficients. 171—172 host range, 181
serological differentiation indices. 171 local lesion formation, in hypersensitive hosts.
strains. 171—173 190
structure and composition. 10. 171—172 mechanical transmission. 181
taxonomic groups, 8—10. 171 negatively stained illustration. 184
thin section illustrations. 176—178 particle. 181—193. 195
tonoplast vesicles, 173. 177 in chloroplast, 187, 190
vectors. 171. 183 in cytoplasm, 187—189. 191. 193. 195
Tobacco necrotic dwarf virus. 138. 140—141 in mitochondria, 187. 190
Tobacco rattle virus. 11 morphology. 8, 10, 181
Tobacco ringspot virus. 10 in nucleus, 187, 189—190
Tobacco streak virus. 11 polypeptide, 183
Tobacco vein distorting virus. 139 shape. 10, 181
Tobacco yellow dwarf virus. 33—35 size. 10, 181. 183
Tobacco yellow net virus. 139 structure, 10, 181, 183—187
Tobacco yellow vein assistor virus. 139 structure, models, 183—187
Tobamovirus swelling, 185
cytopathology. 8—12. 181. 190 in vacuoles, 187—191
taxonomic groups. 9. 11 properties of. 181—183
tonoplast vesicles. 173 replication of. 190
Tobravirus. taxonomic groups, 9, 11 sedimentation coefficients, 181—181
Tomato, 102, 108—109 seed transmission, 181
Tomato bushy stunt virus. 2. 5. 10. 181—193 serological relationships, 181
cytopathology, 189—190 structure and composition. 10, 181. 183—187
negatively stained illustration. 184 taxonomic groups. 8—10. 181—184. 190
particle. 181—189. 191—192 thin section illustrations. 188—189. 191—195
model, structure, 183—187 ultrastructural alterations caused by, 190
Volume I 221
vectors. 181, 183 cytopathology. 117. 123—133. 190

Tonoplast genome. I0.'ll7. 120. 122—123
bubble-like structures, 187—190 model (physical map). 123
vesicles, 173 host range, 117
Top (T) panicle componenl. 118—119 maize rayado fino virus and. 117. 120
Torophagus proserpine. 74 mechanical transmission. 117
T particle, see Top particle component negatively stained illustration, 121
Transcriptase activity, 56—57. 73, 83 particle, 125, 127—128. 130—133
Transfer RNA. 119. 122—123 bottom (B) component. 118—120. 122
Transfer RNA-like sequence, 3' end of RNA, 122— in cytoplasm, 128. 130—131, 133
123 isometric, 117
Translation products, 122—123 model, structure, 119, 122
Transmission, see Mechanical transmission; Seed morphology, 8, 10, 117—122
transmission; Vectors polypeptide, 10, 117. 120
Trifoliuw shape. 10. 117
incarnatum, 76, 84 size, 10. 117. 119—120
repens, 76 structure, 10, 117—122
subterraneum, 143, 166 three-dimensional model, 119, 122
Triticum aesiivum. 76 top (T) component. 118—119
tRNA. sec Transfer RNA properties of, 117—118. 120
Tropaeohtm majus. 102 replication of, 120
Tubular membranes. 105, 107 sedimentation coefficients, 118—120
Tubular structures, 57. 63—64, 66, 68 seed transmission, 117
virus-specific, 164 serological differentiation indices. 117. 119
Tubules, I I I . 157—158. 164 structure and composition. 10, 117—123
Tulip. 1. 171 taxonomic groups. 8—10. 117—120
Tumors, 47. 57, 60 thin section illustrations. 124—127, 129—133
Tungro disease, see also Rice tungro virus. 114— vectors, 117
115 vesicles, 124—128. 130—131
Turnip crinkle virus, 181—183. 185—187 Type members, taxonomic groups. 10—II
cytopathology, 190
particle structure, 181, 185
properties of, 181—183 u
RNA. 186—187
Turnip rosette virus, 153—156 infrastructure, alterations in. 190
Turnip yellow mosaic virus, 10, 117—126, 128— Uranyl acetate staining, general discussion. 2—3.
132 11,76
cytopathology. 123—125. 128—132
stages of post-infection cytologica) events,
128—132
V
genome. 10, 120, 122—123
model (physical map), 123 Vacuoles, I I I . 115, 126. 128. 137. 141. 146. ISO.
negatively stained illustration, 121 157, 163. 187—191
particle. 125. 127—128, 130—132 Vectors, see also specific types by name
model, structure, 119, 122 caulimovirus. 17—18, 22
morphology, 10, 117—122 cucumber necrosis virus, 183
shape, 10. 117 geminivirus, 33—34. 38
size, 10, 117. 119—120 Luteovirus, 137—139. 141
structure, 10. 118—122 maize chlorotic dwarf virus studies. 111
three-dimensional image. 119. 122 Reoviridae. 47. 57. 63
properties of, 118, 120 Rhabdoviridae. 73—75. 90
replication of. 120 Sobemovirus. 153—154
RNA. 120. 122—123. 128 tobacco necrosis virus. 171. 183
structure and composition. 10, 118—122 tomato spotted wilt virus, 101, 103
thin section illustrations. 124—126. 129—132 Tombusvirus. 181, 183
Turnip yellow virus, 138 Tymovirus. 117
Tymovirus. 8—10, 12, 117—133. 190 Velvet tobacco mottle virus. 153—157. 159. 163
antigenic relationships. 119 cytopathology, 157, 163
buoyant densities. 120 negatively stained illustration, 159
chloroplasl involvement, 117, 123—133 properties of. 153—154
coat protein, 117—120, 128—130 RNA. 156
thin section illustration. 163 general discussion, 5—7

Vernacular nomenclature system. 7 Luteovirus studies, 140—141, 143—150
Vertebrate rhabdoviruses. plan! rhubdoviruses com- maize chorotic dwarf virus studies. I I I . 113
pared with. 81. 85 mechanism of cell infection. 83—85
Vertebrate viruses. 23. 101. 103 nucleus, see Nucleus virus-infected cells
Vesicles Reoviridae studies, 57—68
double-membraned. 124—125. 127. 130. 141 Rhabdoviridae studies, 76, 82, 84—96
galinsoga mosaic virus studies, 12, 194—195 rice lungro virus studies. 111—112. 114—115
general discussion. 12 Sobemovirus studies. 157—158. 160—166
Luteovirus studies. 141. 144—148. 150 tobacco necrosis virus studies. 173—174. 176—
maize chlorotic dwarf virus studies. 1 1 1 178
multivesicular bodies, see Multivesicular bodies tomato spotted wilt virus studies, 102—103,
peripheral, see Peripheral vesicles 106—109
Rhabdoviridae studies. 86. 88—90. 92—93 Tombusvirus studies. 187—195
sites of viral RNA replication. 128 Tymovirus studies. 123—133
Sobemovirus studies. 157. 160—161. 164 Virus-like panicles. 47. 73, 75—76. 84. 90—94.
tomato spotted wilt virus studies. 102 114—115. 190
Tombusvirus studies. 190, 193—195 Virus-like RNA. 153—154
lonoplast, 173 Virus panicle, see Particle
Tymovirus studies. 12. 124—128. 130—131 Virus-specific amorphous electron-dense material.
virus-induced. 144. 146—148 173. 176
virus-specific. 160 Virus-specific antisera. see also Antiserum, 4
Vesicular stomatitis virus. 73. 85 Virus-specific double-stranded RNA, 186. 190
Vesiculation. 182—183 Virus-specific fibrils. 161—162. 164
Vesiculovirus. 73 Virus-specific inclusions. 115
Vi/tna sinonsix. 76. 102 Virus-specific tubules, 164
Viral DNA. see also Dcoxyribonucleic acid Virus-specific vesicles, 160
replication of. 27. 38 Virus synthesis. 60, 90, 93
synthesis. 27 VPg, see Genome-linked viral protein
transcription of. 23 vRNA, see Viral RNA
Viral genome, see Genome
Viral protein. 120, 171 w
genome-linked. 140. 156
Viral RNA. see also Ribonucleic acid. 5. 87. 90,
Wheat chlorotic streak virus. 75
103. 120. 128—130. 183. 185—186. 190
Wheat dwarf virus, 34, 37
replication and replicative form, 122, 128. 172—
Wheat rosette stunt virus, 75
173. 186
Whitefly transmission. Geminiviruscs. 33—34
tRNA-like sequence at 3' end. 122—123
Wild cucumber mosaic virus, 118—119, 122
Viroid-like RNA. 153—154. 156
Viroplasm Winter wheat (Russian) mosaic virus. 75
Wound tumor virus, 4. 10—II. 47. 49—52. 54—
Caulimovirus studies. 8. 22—29
57.60
electron-lucent, 93
capsid polypeptides. 55—56
general discussion. 8
ex-vectorial isolates. 57
protein. 22. 27
Reoviridae studies. 8. 57—66
Rhabdoviridae studies. 90—91. 93 X
tomato spotted will virus studies, 102, 106. 108
Virus-induced inclusion body protein, 23 X-bodies. 5
Virus-induced multivesicular bodies. 191—195 X-ray diffraction, general discussion. 1. 4
Virus-induced vesicles. 144. 146—148
Virus-infected plant cells
Caulimovirus studies. 27—28 z
cytoplasm, see Cytoplasm, virus-infected cells
Geminivirus studies. 34. 37—40 Zca mays. 76

Atlas of Plant Viruses

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Atlas of Plant Viruses

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Atlas

R. I. B. Francki, Ph.D. Robert G. Milne, Ph.D.

Reissued 2018 by CRC Press

© 1985 by CRC Press, Inc.

No claim to original U.S. Government works

Library of Congress Cataloging in Publication Data

Francki, R. I. B., 1930-

Includes bibliographies and indexes.

A Library of Congress record exists under LC control number: 84006999

ISBN 13: 978-1-315-89085-2 (hbk)

R. I. B. Francki is a Reader in Plant Pathology at the Waite Agricultural Research

T. Hatta, Ph.D., is a head of the Plant Pathology Division, Aburahi Laboratories, of

II. The Viral Genome 4

III. Virus Cytopathology 5

IV. Virus Taxonomy 7

V. Particle Morphology, Cytopathology, and Virus Identification 8

II. Virus Structure and Composition 17

II. Virus Structure and Composition 33

II. Virus Structure and Composition 48

II. Virus Structure and Composition 76

II. Cytopathology 102

II. Cytopathology 111

III. Rice Tungro Virus — a Possible Member of the Group 111

II. Virus Structure and Composition 118

III. Cytopathology 123

II. Virus Structure and Composition 139

III. Cytopathology 140

II. Virus Structure and Composition 155

III. Cytopathology 156

II. Virus Structure and Composition 171

III. Cytopathology 173

II. Virus Structure and Composition 183

III. Cytopathology 187

II. Virus Structure and Composition 2

II. Virus Structure and Composition 27

II. Virus Structure and Composition 47

II. Virus Structure and Composition 54

II. Virus Structure and Composition 69

II. Virus Structure and Composition 84

II. Virus Structure and Composition 110

III. Cytopathology 116

II. Virus Structure and Composition 135

III. Cytopathology 140

II. Virus Structure and Composition 149

III. Cytopathology 153

II. Virus Structure and Composition 161

II. Virus Structure and Composition 175

III. Cytopathology 177

II. Virus Structure and Composition 189

III. Cytopathology 191

II. Particle Structure and Composition 222

III. Cytopathology 227

II. Phleum Mottle and Similar Viruses 235

III. Carnation Mottle and Some Similar Viruses 236

V. Cacao Swollen Shoot and Structurally Similar Viruses 237

VI. Rice Stripe and Similar Viruses 237

VII. Carrot Mottle and Similar Viruses 243

VIII. Cryptic Viruses 244

IX. Concluding Remarks 248

List of Virus Abbreviations and Taxonomic Groups 257

Who first "discovered" viruses or developed the concept "virus" as we understand it

I. VIRUS PARTICLE STRUCTURE

II. THE VIRAL GENOME

advance was the development of deproteinization techniques yielding undegraded nucleic