TRANSLOCATION, DISTRIBUTION AND

METABOLISM OF TRIFORINE IN PLANTS

A. FUCHS
Laboratory of Phytopathology, Agricultural University, Wageningen, The Netherlands,
and

W. OST
Celamerck GmbH & Co, KG, lngelheim, Germany

After short-term root-treatment of plants with SH-triforine (labelled in the

piperazine ring) or x4C-triforine (labelled in the side chain), the label was translocated
primarily to the leaves present at the time of treatment, without any redistribution of
the label taking place after termination of the treatment. Autoradiography showed the
radioactivity to be very evenly distributed over the leaf blades, without any appreciable
accumulation in leaf margins. In infected leaves, however, there was a tendency for
increased accumulation at the infection sites.
In dilute aqueous solution triforine was decomposed rather rapidly, the chemical
half-life being three days only. From these solutions several non-fungitoxic degrada-
tion products have been isolated and their chemical structures determined. In plants,
decomposition of triforine was considerably slower than in aqueous solutions, the
half-life of the compound varying depending on plant species. In plants, at least four
conversion products could be demonstrated, one of which is piperazine. Whether the
other three compounds are identical with or structurally related to the breakdown
products found in aqueous solution is not yet known.
Its low persistence and low animal toxicity suggest that triforine does not offer
serious hazards to the environment.

Introduction
Several reports on uptake, translocation and m e t a b o l i s m o f triforine in plants
have been published recently (Fuchs 1971; Fuchs e t al. 1971, 1972; Ost e t al. 1971;
Ebenebe 1972; Eichler 1972; Von Bruchhausen and Stiasni 1973; E b e n e b e et al.
1974). F r o m these reports, it has become evident that triforine is r e a d i l y taken up
via plant roots, and translocated to stems and leaves. A n apparent lack o f a c c u m u l a -
tion o f triforine in the roots (Fuchs 1971; E b e n e b e 1972; E b e n e b e e t al. 1974) and a
rather high turnover rate o f the fungicide in aerial plant parts (Fuchs e t a l . 1972)
seem to account for an " e a r l y " m a x i m u m o f triforine concentrations in the shoots,
which, d e p e n d i n g on the e x p e r i m e n t a l conditions used, was found 2-8 d a y s after
administration o f the fungicide.

Presented at the Third International Congress of Pesticide Chemistry (IUPAC), Helsinki, Finland,
1974.

Archives of Environmental Contamination 30

and Toxicology Vol. 4, 30-43 (1976)
~1976 by Springer-Verlag New York Inc.
 Triforine in Plants 31

In aerial plant parts triforine was found to be metabolized to at least four

metabolites (Fuchs et al. 1972; Von Bruchhausen and Stiasni 1973), one of which
was identified as piperazine.

In the experiments to be briefly described here, translocation and distribution as

well as metabolism of triforine in plants were examined more closely by using both
3H- and l~C-labelled triforine, whereas analysis of each single plant part provided a
detailed picture of these phenomena. These processes were studied in relation to
such parameters as mode and duration of application, age of plants, etc. In addition,
metabolism of triforine was compared with non-enzymatic degradation of the fun-
gicide in aqueous solution, in order to further elucidate the chemical nature of the
breakdown products, thus permitting more exact conclusions regarding the fate of
triforine in plants. A more detailed description of the results will be given elsewhere
(Fuchs and De Vries 1974, Fuchs et al. 1974).

Materials and methods

3H-triforine (uniformly 3H-labelled in the piperazine ring) or ~4C-tdforine (un-

iformly 14C-labelled in the trichloroethyl moiety of side chains) was administered
via the roots of plants belonging to different species, either by immersing the plants
with their roots in aqueous solutions or by applying the fungicide as soil drenches.
After appropriate periods of " i n c u b a t i o n " , plants were harvested and divided into
roots, hypocotyls, cotyledons, epicotyls, internodes, leaves, leaflets, etc. After
fresh weight determinations had been made, all parts were extracted in 10 to 50 ml
of methanol, depending on fresh weight. Some of the ~4C-triforine-treated plants
were left intact and dried between filter papers as evenly and quickly as possible;
then, autoradiographs were prepared by exposing Kodak No-Screen Medical X-ray
film to the dried plants.

Radioactivity in the extracts of all separate plant parts was measured by liquid
scintillation counting (LSC) and expressed as dpm/mg fresh weight. The degree of
accumulation of radioactive compounds (triforine + possible breakdown products)
in each single part was calculated relative to average radioactivity (expressed as
dpm/mg fresh weight) of total aerial plant parts, this ratio providing the best
approximation of the distribution of the label over the various plant parts. In
addition, in some cases the amount of radioactivity present in specific plant parts
(e.g., primary leaves, first two leaves present at the time of application of radioac-
tive triforine) was also given relative to total amount of radioactivity in aerial plant
parts. In order to transform the data on distribution of radioactive compounds in
general into data on distribution of triforine, extracts were further subjected to
thin-layer chromatography (solvent butanol/acetic acid/water 4:1:1). TLC's were cut
into ten equal pieces, with Rr-values 0.00 to 0.10 to 0.90 to 1.00, and radio-activity
in them was measured by LSC; in the case of 14C-triforine, analysis of TLC's also
proceeded by radioscanning. The analytical procedures used separated triforine from
its degradation products and thus enabled calculation of total amounts of radioactiv-
ity present as 3H- or 14C-triforine in all single plant parts, also in relation to the
32 A. Fuchs and W. Ost

incubation period, and thus permitted conclusions on the rate of conversion of

triforine in plants.

Chromatographic behaviour of degradation products was compared with that of

non-metabolic, hydrolytic breakdown products of triforine. These products were
obtained by incubating non-buffered aqueous solutions of triforine (30 /~g/ml) at
room temperature in closed vials in darkness and in diffuse daylight. The decrease in
concentration of the parent compound and the formation of ionogenic chlorine were
followed quantitatively for four weeks and seven months, respectively. The appear-
ance of other breakdown products was examined periodically for up to one year,
using TLC as the method of analysis. The main hydrolysis products were isolated
after freeze-drying of the aqueous solution, and after chromatographic purification
identified by mass spectrometry and by comparison with authentic reference sub-
stances.
Results
From Tables I and II it is evident that, after short-term treatment o f bean and
tomato seedlings with 3H-triforine, radioactivity was rapidly accumulated in the
leaves present at the time of treatment, without subsequent redistribution into the
later expanding leaves. These conclusions are based on the following observations:
a) The degree of accumulation of radioactivity in the roots (relative to average
radioactivity in aerial plant parts) dropped rapidly after termination of the treatment.
b) In the primary leaves (bean) or the cotyledons and first leaves (tomato) the degree
of accumulation of radioactivity increased continuously, not, however, because of a
continued supply with radioactive substances, but because of a decrease of the
average radioactivity in the aerial parts due to the growth of plants; since all new
growth was virtually devoid of radioactivity, the first leaves became relatively
" r i c h e r " in radioactive label, c) The percentage of total radioactivity in the first
expanded leaves increased rapidly to almost 90% of the total radioactivity in aerial
plant parts.

An entirely different distribution pattern of radioactivity was found after short-

term treatment of " a d u l t " plants; here, radioactivity was much more evenly distri-
buted over all plant parts, in a concentration gradient from roots to youngest leaves
in barley and bean plants (Tables III and IV; see: application time: 6 hr).

Uptake of 3H-triforine by barley plants showed a rapid saturation at about 60% of

the administered label (Fig. 1). The total activity in the roots hardly increased after
six hr (Table III), the rate of translocation of label to stem and leaves exceeding the
rate of uptake from 12 hr onwards. Nevertheless, after 42 hr about 70% of the label
was still retained in the roots. Radioactivity was translocated to the leaves at the
expense of label in roots and stems, with preference for the first leaf up to 18 hr;
after 42 hr the highest degree of accumulation (also in absolute amounts o f radioac-
tivity) was found in the third leaf. Degrees of accumulation of radioactivity in the
tillers were almost time-independent over the entire application period, the highest
accumulation being found in the " s e c o n d " tiller.
 Table I. Distribution of radioactivity and metabolism of triforine after administration of 3H-triforine to bean plants,
cv. Dubbele Witte z.dr. (Age of plants at time of administration o f triforine: 9 days)

Age (days)

9a 22 33 43 54

,--t
Number of leaves (primary + trifoliate leaves) 2+0 2+1 2+2 2+3 2+5
--.

Degree of accumulation of radioactivity roots 13.17 0.42 0.59 0.70 0.76

in relation to average radioactivity in int. b 1 + 2 1.05 0.41 0.63 1.28 1.32 -~"
aerial plant parts primary 1. 1.12 2.40 3.69 5.95 14.58

Percentage of radioactivity in primary leaves (with 41.6 86.4 84.6 86.0 89.0
respect to total radioactivity in aerial plant parts)

Percentage of radioactivity in primary leaves present 85.5 c 64.6 52.3 41.5 40.2
as 3H-triforine

a Immediately after administration of 3 H-~riforine; hint. = internode; tin roots.

 Table II. Distribution of radioactivity and metabolism of triforine after administration of 3H-triforine to tomato plants,
cv. Bonnet Beste (Age of plants at time of administration of triforine: 26 days)

Age (Days)

26 a 33 49 58 77

Number of leaves (cotyledons + true leaves) >

2+2 2+3 2+5 2+6 0+9

Degree of accumulation of radioactivity roots 8.20 1.01 1.52 2.11 3.17 ~r

in relation to average radioactivity in cot. b 1 0.72 1.10 4.82 10.39 _e
aerial plant parts cot.b2 rn
0.56 1.13 3.79 10.06 _c e~
leaf 1 1.27 1.88 8.08 11.20 14.09
leaf 2 1.41 2.14 4.70 4.00 8.02

Percentage of radioactivity in cotyledons and 74.5 86.8 87.2 88.9 85.1 d

first and second leaf (with respect to total radio-
activity in aerial plant parts)

Percentage of radioactivity in second leaf present 87.3 53.6 16.0 10.6 3.4
as 3H-triforine

almmediately after administration of 3H-triforine; b c o t . = cotyledon; edropped; donly in leaf 1 and 2

 Triforine in Plants 35

In bean plants, uptake of ~H-triforine took place much more slowly than in barley
plants, after 18 hr only a third of the total amount administered being taken up. As
can be derived from the low degree of accumulation in the roots after 42 hr,
aH-triforine, however, was readily translocated to the stems and leaves, only 50%
being still retained in the roots at that time (Table IV). In the stems, radioactivity
was initially present in a concentration gradient from roots to tips; after prolonged
" i n c u b a t i o n " , a maximum in the degree of accumulation was found in the second
internodes. With regard to the leaves, the label was preferentially translocated to the
youngest leaf present.

Since in the latter experiments, even after 42 hr of "incubation", from 75 to 90%

of the radioactivity was present as 3H-triforine, the data given can be assumed to
reflect the actual distribution of the parent compound. However, in experiments as
described in Table I and II, the amounts of radioactivity present do not represent the

Table IlI. Distribution of radioactivity after administration of 3H-triforine to "adult"

barley plants, cv. Cambrinus, in relation to duration of uptake;administered 13 830 600
dpm per plant (Age of plants at time of administration: 33 days)

Degree of accumulation of radio-

activity in relation to average Amounts of radioactivity (in dpm/mg
radioactivity in aerial plant parts fresh weight) per plant part

Applicationtime(hr) AppHcationtime(hr)

6 12 18 42 6 12 18 42

roots 28.03 20.02 11.25 5.41 1972 1999 1896 1162

"stem" 2.53 2.19 1.86 1.33 182 219 311 292
leaf 1 3.24 3.57 2.92 1.60 213 347 497 347
leaf 2 1.74 1.66 1.44 2.35 130 161 244 512
leaf 3 1.47 1.57 1.24 2.48 106 157 208 532
leaf4 0.96 1.19 1.24 1.27 69 119 208 277
leaf 5 0.50 0.53 0.78 0.94 38 54 128 200
leaf 6 0.22 0.26 0.43 0.47 16 26 71 100
leaf 7 0.36 0.31 0.38 0.36 27 32 63 77
tiller 1 0.85 0.93 1.20 0.97 63 93 203 210
tiller 2 I. 19 1.12 1.37 1.18 89 111 233 254
tiller 3 0.88 0.85 0.94 0.80 65 86 159 171
total uptake (ha dpm) 6732550 7454100 7671200 7934950
total uptake (in % of amount administered) 48.7 53.9 55.5 57.4
radioactivity in roots, as % of total uptake 88.1 85.0 78.6 70.1
 36 A. Fuchs and W. Ost

actual amounts of 3H-triforine, and, hence, the degrees of a c c u m u l a t i o n in the

various plant parts might not reflect the distribution of triforine. Therefore, in these
experiments the rates of degradation of triforine were estimated from the available
data using methods as indicated under " M a t e r i a l s and m e t h o d s " ; for bean and
tomato plants, the biological half-lives of triforine proved to be ca. 40 and 10 days,
respectively (Fig. 2). Preliminary results showed that these values should not be
taken too absolutely. Although they are valid for the aerial parts indicated (Tables I
and II), in the roots the biological half-lives appeared to be dissimilar.

Table IV. Distribution of radioactivity after administration of 3H-triforine to "adult"

bean plants, cv. Dubbele ;r z.dr., in relation to duration of uptake; administered
13 830 600 dpm per plant (Age of plants at time of administration: 46 days)

Degree of accumulation of radioactivity in relation to

average radioactivity in aerial plant parts

Application time

6 hr 12 hr
pet- leaf pet- leaf
iole blade iole blade

roots 10.47 a - - roots 6.91 a w m

internode 1 6.44 - 0.75 b internode 1 4.07 0.56 b

internode 2 5.08 1.28 0.41 internode 2 4.67 1.16 0.32
internode 3 2.60 1.07 0.38 internode 3 2.85 1.94 0.26
internode 4 0.80 0.43 0.55 internode 4 0.95 0.85 1.58

Application time

18 hr 42 hr

pet- leaf pet- leaf

iole blade iole blade

roots 3.71 a _ _ roots 1.58 a - - w

internode 1 2.45 - 0.67 b internode I 2.20 _ 1.01 b

internode 2 2.53 1.00 0.47 internode 2 2.56 0.96 0.81

internode 3 2.22 0.90 0.41 internode 3 2.28 O.95 O.57
internode 4 1.34 0.91 1.81 internode 4 1.08 0.60 1.66

aradioactivity in roots, as % of total uptake: 87.1 (6 hr), 79.0 (12 hr), 55.7 (18 hr) and
50.5 (42 hr), respectively.
baverage degree of accumulation of radioactivity in primary leaves.
 Triforine in Plants 37

15-
A m o u n t o f radioactivity administered

10
Q

X ,-O
...._ O ......-.=------ O --
E
O.
"lD

0 I I I I
0 10 20 30 40
Duration o f application (hours)

Fig. 1. Uptake of 3H-triforine by barley plants, in relation to duration of application.

In tomato plants, for instance, after 58 days c a . 30% of the label in the roots
was still in the form of 3H-triforine. However, it should be realized, that (especially
for roots) such data do not merely reflect metabolic rates, but also rates of trans-
location of the parent compound and its degradation products into stem and leaves.

100 -

~_ 5 0

0 I i I I~ -
0 10 20 310 40 50

Time in days after administration o f triforine

Fig. 2. Rate of conversion of triforine in primary leaves of bean plants and in the second leaf
of tomato plants after short-term (ca. 6-hr) treatment, and in the 6th leaf of pea plants after
long-term (one-week) treatment.
 38 A. Fuchs and W. Ost

Experiments on the distribution of ~4C-triforine in pea plants provided results

comparable to those of the experiments with 3H-triforine. The biological half-life in
pea proved to be ca. 40 days (Fig. 2). Other results will be outlined elsewhere
(Fuchs and De Vries 1974).

The experiments discussed so far provided an approximate idea about uptake and
translocation of triforine in plants; however, they did not give any information about
the distribution of triforine within single leaves. Therefore, autoradiographs of
healthy as well as diseased plants, treated with ~4C-triforine, were made after one
and four days of "incubation". The diseased plants were employed to examine a
possible effect of fungal infection on the distribution of triforine. In the healthy
plants of all species chosen (viz. bean, pea, tomato and wheat), the radioactive label
proved to be very evenly distributed over the leaf blades, without any appreciable
accumulation in the leaf margins. In rust-infected bean leaves, on the other hand,
there was a slight tendency toward increased accumulation of label at the infection
sites (Fig. 3); in rust-infected wheat leaves label seemed to be preferentially trans-
located to the most heavily infected leaves. Perhaps, leaf damage, leading to
enhanced evaporation, conditioned this increased accumulation; this seems to hold
true at any rate for the healthy leaves of tomato plants, which upon prolonged
"incubation" reacted to high concentrations of triforine (50/J,g a.i./ml) with distinct
lesions in the leaves with concomitant accumulation of label at these sites.

i ~ %'`

~ o ~

~ /x" 9

.og..' ~ o

Fig. 3. Effect of rust (Uromyces phaseoli) infection on distribution of label in 14C-triforine-

treated bean leaves. Left: bean leaf, showing typical green islands around infection sites;
right: autoradiograph, showing slightly increased concentrations of radioactive label around
infection sites.
 Triforine in Plants 39

TLC-analysis of extracts of 3H-triforine-treated plants invariably revealed four

areas on the TLC-plates carrying substantial amounts of radioactivity, with Rf-
values of 0.03, 0.50, 0.75 and 0.901, respectively (Fuchs et al. 1972). TLC-
radioscans of t4C-triforine-treated plants also showed four maxima of radioactivity
(Fig. 4) with Rf-values of 0.12, 0.40, 0.67 and 0.90, respectively. One of the four
products of each of both radioactive triforines is obviously characteristic for the type
of labelling: the ZH-containing compound (Re-value 0.03) is almost certainly
piperazine (Eichler 1972, Fuchs et al. 1972, Von Bruchhausen and Stiasni 1973);
the 14C-containing compound (Re-value 0.12) must be a substance lacking the
piperazine ring, and is thus a substance arising from (part of) the side chain(s) of
triforine. One of the three other substances common to both types of labelled
triforine, with Re-value 0.90, is the parent compound itself; the identity of the two
remaining ones is not yet known. Probably, they are identical with or structurally
related to two of the hydrolysis products, which are formed upon sterilization of
triforine (cf. Fuchs et al. 1971) or arise more gradually when aqueous triforine
solutions are kept at room temperature. Even under such mild experimental condi-
tions, the parent compound proved to be hydrolyzed rather quickly, in darkness as
well as in diffuse daylight, no triforine being left after four weeks of incubation.

Fig. 4. TLC-radioscan of extract of t4C-triforine-treated pea leaf. The highest peak (at left) is
due to unchanged triforine. Solvent: butanol/acetic acid/water 4:1:1.

1Because of the method used, the last three are only approximate values; they vary from plate to plate
between 0.40-0.60; 0.70-0.80 and 0.80-1.00, respectively.
 40 A. Fuchs and W. Ost

Table V. Formation of CI- in aqueous solutions o f triforine

(30 lzg/ml) in relation to incubation time

After (time) % of theoretical amount

30 min < 1%
10 days 37%
28 days 71%
50 days 86%
7 months > 95%

The chemical half-life of the fungicide in triforine solutions containing 30/.xg a.i./ml
proved to be only three days.

Time-course measurements of chloride formed showed intermediates with intact

CC13-groups to be rapidly hydrolyzed further, with formation of inorganic chloride
(Table V). Since this was present at about 50% as free hydrogen chloride, the pH of
the solution decreased from 5.5 to 3.6 within three days, remaining constant
afterwards.

OHC--NH--CH--CCI 3
I
N
~ ," Unstable intermediate

I
OHC-NH-CH--CCl3 OHC_NH_CH_CCI3
Triforine I

/ Substance I
J . OH I

%0-0/
I ~ OH
1
H.HCI H.HCI
N N N

N N N
I /OH I /OH H.HCl
o~C-CH .C-CH
OH O// ~ OH Piperazine
Substance II Substance III
Fig. 5. Suggested pathway of hydrolytic breakdown of triforine in aqueous sotution (30
/~g/ml), kept at room temperature for one year.
 Triforine in Plants 41

Identification of the various degradation products revealed hydrolysis to proceed

as indicated in Fig. 5. The asymmetric piperazine derivative N-(1-formamido-
2,2,2-trichloroethyl)- N'-2,2-dihydroxyacetyl- piperazine (I) was the first product to
be isolated in substantial amounts after one to three weeks of incubation. It was
further degraded with formation of the water-soluble intermediate bis-glyoxylic
piperazine dihydrate (II) and an N-monosubstituted piperazine derivative (III). The
latter breakdown product seems to be only stable in solution, or as a salt, and is
probably N-glyoxylic piperazine. Finally, piperazine itself was produced (isolated as
a dihydrochloride) and, in addition, a number of other chlorine-free, water-soluble
products, arising from the side chains. At elevated temperatures, hydrolysis was
greatly accelerated, so that some of the above-mentioned hydrolysis products could
not even be demonstrated under such experimental conditions.

The terminal residues of triforine hydrolysis constitute, beside piperazine, very

probably a series of low molecular weight organic acids, which partly are common
in plants, such as glyoxylic, formic and glycolic acid, and CO2.

Discussion

A critical quantitative evaluation of the results obtained on uptake, translocation

and metabolism of triforine is hampered in various ways. For instance, in the roots
the amounts (and thus concentrations) of triforine are the resultant of, a) uptake of
triforine from the substrate (aqueous solution, soil), in which chemical breakdown
of the parent compound will proceed continuously, b) translocation to the stems and
leaves, and c) metabolic and/or non-metabolic conversion of triforine into other
products. Therefore, values on biological half-life in roots can hardly be assumed to
represent the intrinsic value for the biological half-life of triforine characteristic of
the plant species involved. In fact, only those plant parts in which any decrease in
triforine concentrations is entirely due to breakdown, can be considered to give
reliable results in the estimation of biological half-life. In the experiments described
in Tables I and II, the primary leaves of bean plants and the first two leaves of
tomato plants seem to fulfill these requirements. In these instances, after short-term
treatment, triforine attained a maximum level within a few days without further
uptake or redistribution of fungicide taking place after this period. Therefore, any
change in triforine concentrations can be ascribed to chemical and/or metabolic
conversion.

The estimated values for leaves of bean and tomato plants, which were found to be
ca.40 and 10 days, respectively, might be supposed at least to approximate the
intrinsic value of the biological half-life of triforine for these species. These values
are so dissimilar from the chemical half-life in aqueous solution (three days), that it
is difficult to ascribe the differences to factors other than environmental ones. The
conclusion seems inevitable that triforine, in some way or other, is protected against
breakdown within the plant. In fact, a similar conclusion was drawn by Eichler
(1972), who attributed the very slow degradation of triforine in the peel of apples to
" i n c l u s i o n " of the fungicide in the lipophylic material of the cuticle.
 42 A. Fuchs and W. Ost

Autoradiographs of 14C-triforine-treated plants evoke the impression of a

homogeneous distribution of label over the leaf blades; therefore, it seems an
attractive idea to suggest that triforine is primarily translocated to the lipophylic
outer layer of the epidermis. There, it could not only be protected against degrada-
tion, for instance, because of adsorption to lipid cell wall material, but could also
attain relatively high concentrations, high enough to protect the plant against dis-
eases caused by obligate parasites, like mildews and rusts, and other foliar diseases
caused by non-obligate pathogens. This hypothesis might also constitute an explana-
tion for the remarkable fact that, in vitro and in vivo, activities of triforine are by no
means positively correlated (Drandarevski and Fuchs 1973, Fuchs and Drandarevski
1973), and that it is found to be active against an increasing number of foliar
pathogens (Drandarevski and Fuchs 1973, Fuchs and Drandarevski 1973), as has
been once more shown recently (Szkolnik 1974).

Comparison of the TLC data on the four conversion products of triforine left three
compounds, with Rrvalues of 0.12, 0.40 to 0.50 and 0.67 to 0.75, unidentified.
The fourth one, with an Rrvalue 0.03, which does not contain a 14C-labelled side
chain is certainly piperazine (cf. Eichler 1972. Fuchs et al. 1972, Von Bruchhausen
and Stiasni 1973). The one with the Rr-value of 0.12 must be a substance arising
from the side chain, since it lacks the piperazine ring; it might be glyoxylic acid or
structurally related to it. The remaining ones, which contain both the piperazine ring
and at least one of the C-atoms of the side chains, could be identical with or
structurally related to two of the three substances I, II and III. However, their exact
chemical structures await final identification.

Its low persistence in aqueous solution and in plants as well as its low toxicity to
mammals, fish and bees (Schicke and Veen 1969) suggest that triforine does not
offer serious hazards to the environment.

References

Drandarevski, C. A., and A. Fuchs: The in vitro and in vivo antifungal activity of
triforine. Meded. Fak. Landb.wetensch. Gent 38, 1525 (1973).
Ebenebe, C: Wirkung des systemischen Fungizids Triforine gegen Weizenbraunrost
und Welkekrankheiten der Tomate. Dissertation, Giessen (1972).
Ebenebe, C., V. von Bruchhausen, and F. Grossmann: Dosage-response curve of
wheat brown rust to triforine supplied via root treatment. Pesticide Sci. 5, 17
(1974).
Eichler, D.: Ober das Abbauverhalten von Triforine dargestellt an einigen aus-
gew~ihlten Kulturpflanzen. Meded. Fac. Landb.wetensch. Gent 37, 831
( 1972).
Fuchs, A.: Opname, transport en omzetting van triforine in planten. Gewasbe-
scherming 2, 142 (1971).
 Triforine in Plants 43

Fuchs, A., M. Viets-Verweij, J. Vfrrs, and F. W. de Vries: Some observations on

activity and metabolism of a new systemic fungicide, N , N ' - b i s - ( l -
formamido-2,2,2-trichloroethyl)- piperazine (CELA W 524). Acta Phytopath.
Hung. 6, 347 (1971).
Fuchs, A., M. Viets-Verweij, and F. W. de Vries: Metabolic conversion in plants
of the systemic fungicide triforine(N,N'-bis-(1-formamido-2,2,2-
trichloroethyl)- piperazine; CELA W 524). Phytopath. Z. 75, 111 (1972).
Fuchs, A., and C. A. Drandarevski: Wirkungsbreite und Wirkungsgrad von
Triforine in vitro and in vivo. Z. Pfl. Krankh. Pfl. Sch. 80, 403 (1973).
Fuchs, A., and F. W. de Vries: Uptake, translocation and metabolic fate of t~C-
triforine in plants. In preparation. (1974)
Fuchs, A., F. W. de Vries, and A. M. J. Aalbers: Uptake, translocation and
metabolic fate of aH-triforine in plants. In preparation. (1974).
Ost, W., V. von Bruchhausen, and C. Drandarevski: Transport of the systemic
fungicide Cela W 524 in barley plants (Part I). Pesticide Sci. 2, 219(1971).
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Manuscript received September 28, 1974; accepted November 11, 1974