Turk J Biol

33 (2009) 283-290
TBTAK
doi:10.3906/biy-0807-23

Genotoxic effects of herbicide Illoxan

(Diclofop-Methyl) on Allium cepa L.

Deniz YZBAIOLU1, Fatma NAL1, Cengiz SANCAK2

1
Department of Biology, Faculty of Arts and Science, Gazi University, Ankara - TURKEY
2Department of Field Crops, Faculty of Agriculture, Ankara University, Ankara - TURKEY

Received: 21.07.2008

Abstract: The genotoxic potential of the commercial herbicide Illoxan (containing 284 g/L diclofop-methyl) was
determined by using chromosome aberrations in Allium cepa root tip cells. The EC50 value was determined as 150.00 mg/L
using a root growth inhibition test and then the roots were treated with 37.50, 75.00, and 150.00 mg/L concentrations
for 12, 24, and 48 h. The results indicated that Illoxan significantly increased the abnormal cell frequency at all
concentrations and treatment periods when compared with their controls, and this increase was dose-dependent for the
24 and 48 h treatments. On the other hand, Illoxan significantly decreased the mitotic index (MI) in all treatments when
compared with their controls. The decrease in the mitotic index was slightly dose-dependent for the 24 and 48 h
treatments. Illoxan did not affect the percentage of mitotic stages. For the pretreated root tips, Illoxan (except 37.50 mg/L
at 12 and 24 h) significantly increased the frequency of abnormal cells in a dose-dependent manner. This study indicates
that Illoxan decreased the mitotic index and produced clastogenic and aneugenic types of abnormalities in Allium cepa
root tip cells. The data obtained in this study showed that plant bioassays can be used as an important test battery to
detect possible genotoxicity of chemicals.

Key words: Pesticide, herbicide, Illoxan, diclofop-methyl, genotoxic effects, Allium cepa

lloksan herbisitinin Allium cepa L.da genotoksik etkileri

zet: Bu almada, illoksan herbisitinin ticari formunun (284 g/L diclofop-methyl iermektedir) genotoksik potansiyeli,
Allium cepa kk ucu hcrelerinde kromozomal anormallikler kullanlarak belirlenmitir. EC50 deeri, byme inhibisyon
testi kullanlarak 150,00 mg/L olarak belirlenmi ve daha sonra kkler 37,50, 75,00 ve 150,00 mg/L konsantrasyonlar ile
12, 24 ve 48 saat muamele edilmitir. Sonular, illoksann tm muamele sreleri ve konsantrasyonlarda anormal hcre
frekansn kontrole gre nemli dzeyde artrdn gstermektedir, bu art 24 ve 48 saatlik uygulamalarda doza baldr.
Dier taraftan illoksan mitotik indeksi (MI) tm uygulamalarda kontrole gre anlaml oranda drmtr. Mitotik
indeksteki d 24 ve 48 saatlik uygulamalarda dk oranda doza baldr. lloksan mitotik safhalarn frekansn
etkilememitir. n muameleli kk ularnda, illoksan anormal hcre frekansn (12 ve 24 saatlik uygulamalarda 37,50
mg/L hari) doza bal olarak nemli oranda artrmtr. Bu almada illoksann Allium cepa kk ucu hcrelerinde
mitotik indeksi drd, klastojenik ve anjenik tip anormalliklere neden olduu belirlenmitir. Bu almann verileri,
kimyasal maddelerin muhtemel genotiksik etkilerinin belirlenmesinde bitki deneme sistemlerinin nemli bir deneme
sistemi olarak kullanlabileceini gstermektedir.

Anahtar szckler: Pestisit, herbisit, illoksan, diclofop-methyl, genotoksik etki, Allium cepa

283
Genotoxic effects of herbicide Illoxan (Diclofop-Methyl) on Allium cepa L.
Introduction
Due to their widespread use in agriculture,
pesticides are some of the compounds most
frequently released into the environment. Despite the
beneficial effects associated with the use of pesticides,
many of these chemicals may pose potential hazards
to humans and to nature. The genotoxic effects of
some pesticides have been evaluated by several
researchers through several test systems (1-5). Plant
bioassays, which are considerably more sensitive and
simple in comparison with animal bioassays, have
been validated in international collaborative studies
under the United Nations Environment Program
(UNEP), World Health Organization (WHO), and US
Environmental Protection Agency (US-EPA), and
proven to be efficient tests for the genotoxicity
monitoring of environmental pollutants (6-8).
In this study, an Allium test system was used in the
evaluation of genotoxic activity of the herbicide
Illoxan with the active ingredient diclofop-methyl.
Diclofop-methyl is a selective systemic herbicide,
absorbed primarily through leaves, with some
absorption through roots in moist soil. Diclofop-
methyl undergoes rapid transformation, which is
translocated within the plant. This herbicide is a fatty
acid synthesis inhibitor, and inhibits acetyl CoA
carboxylase (ACCase). It destroys the cell membrane,
prevents the translocation of assimilates to the roots,
reduces the chlorophyll content, and inhibits
photosynthesis and meristem activity. It is registered
for use on the post-emergence control of wild oats,
wild millets, and other annual grass weeds in wheat,
barley, rye, and broad-leaved crops such as soybeans,
sugar beet, legumes, sunflowers, clover, brassicas,
carrots, lettuce, spinach, potatoes, cucumbers, peas,
beans, tomatoes, alliums, and herbs (9). Diclofop-
methyl has been classified as likely to be carcinogenic
to humans by the laboratory studies based on the rats
and mice (10).
Diclofop-methyl tested negative for genotoxic
effects in a bacterial reverse mutation test in
Salmonella typhimurim, an in vitro mammalian cell
gene mutation test with Chinese hamster V79 cells,
an in vitro mammalian chromosomal aberration test
in primary human lymphocytes, an in vivo
cytogenetic test in bone marrow cells of the Chinese
hamster, unscheduled DNA synthesis in primary rat
hepatocytes and in A549 human lung carcinoma in
284
vitro, a dominant lethal test in male NMRI mice, a
mouse bone marrow micronucleus test, and
mutagenic activity with Saccharomyces cerevisiae (11).
Although there are studies showing the negative
effects of diclofop-methyl, there has been no study on
the effects of this pesticide in plant test systems used
for scoring the effects of chemicals.
The present investigation was conducted using
chromosome aberrations, detecting clastogenic
activity qualitatively and quantitatively, in an Allium
cepa test system in order to assess the genotoxic
potential of the commercial herbicide Illoxan
(containing 284 g/L of diclofop-methyl). We have
chosen Allium cepa as a test organism and a
commercial formulation as a test compound for the
following reasons: 1) Some pesticides have negative
effects on some organisms as indicated above while
showing positive effects in other organisms (12). 2)
Plant and animal assays are differentially responsive to
pesticides. Unlike animals, some chemicals, including
pesticides, are metabolically activated by plant
peroxidases and may give different responses than
those of mammalian cytochromes P-450 (5). 3) Many
studies report the importance of evaluating the
potential toxicities of complete formulations, rather
than just evaluating the toxicities of the active
components, because it is the commercial formulation
that has its application in commercial agriculture (5).
4) There has been no study, to our knowledge, on the
genetic effects of Illoxan or diclofop-methyl in plant
test systems.
Materials and methods
Illoxan belongs to the diphenylether/
chlorophenoxy group of herbicides. Its chemical
abstract name is methyl 2-[4-(2,4 dichlorophenoxy)
phenoxy] propanoate or 2-[- 4 (2,4
dichlorophenoxy) phenoxy] methyl propanoate. Its
molecular weight is 341.2. Its chemical formula is
C16H14Cl2O4.
Cl O O CH COOCH3
CH 3
Cl
Figure 1. The structural formula of Illoxan.
 D. YZBAIOLU, F. NAL, C. SANCAK

Approximately equal-sized onion bulbs (Allium
cepa L., 2n = 16) were used as the test material. The
outer scales of the bulbs were carefully removed and
the dry plates were scraped away without destroying
the root primordia. The growth inhibition test was
carried out as described by Fiskesj (13), with minor
modifications. Five onion bulbs were set up in 10, 25,
50, 75, 100, 125, 150, 200, 250, and 300 mg/L
concentrations. A set of 5 bulbs was also set up in
distilled water. After 4 days, the mean value of 10
measurements of each onion, i.e. 50 roots for each
concentration, was expressed as a percent of the
control value. This measurement was used to calculate
the EC50 value, which is the concentration where root
growth is reduced by 50% compared with the control.
The EC50 value of Illoxan was determined as 150.00
mg/L and then the root tips were treated with 37.50
(EC50/4), 75.00 (EC50/2), and 150.00 (EC50) mg/L
concentrations.
Bulbs were placed over the test tubes filled with
distilled water at room temperature (20 2 C). When
the roots reached 1.5-2 cm, they were treated with
each concentration of Illoxan for 12, 24, and 48 h.
Twelve bulbs were used for each concentration,
treatment period, and control group. Following all the
treatments, the root tips were fixed, macerated,
stained, and squashed as described earlier by
Yzbaolu et al. (3). The mitotic index (MI) was
found by observing 1000 cells and counting the stages
of mitotic cells and mitotic abnormalities. Slides were
prepared from different onions; 1000 cells were
screened from each of 10 onions yielding 10,000 cells
for each concentration and duration of time. In
addition, in pretreated roots, 100 metaphase cells were
scored and aberrations were recorded for each of the
concentrations and treatment times (14). Using the z-
distribution test method, the data obtained for the
mitotic index were statistically analyzed for the
frequency of mitotic phases and mitotic disturbance.
Metaphase aberrations were analyzed using a 2 test.
Results
Table 1 shows the mitotic index in Allium cepa root
tip cells. The mitotic index significantly decreased in
all treatments when compared with their controls.

Table 1. Mitotic index (MI), type and percentage of mitotic abnormalities in the root tips of A. cepa exposed to Illoxan.

Conc. No. of MISE % Abnormalities % % Abnormal

(mg/L) dividing Total interphase
cells Stickiness Laggards C-mitosis Bridges Multipolarity Fragments abnormalities cells

12 h
Control 707 7.070.25 0 0 0.28 0 0 0 0.28 0
37.5 427 4.270.20a 2.34 6.56 4.68 0.94 0.94 0 15.46** 0.02
75 270 2.700.16a 12.22 2.96 3.33 0.74 0 1.11 20.36** 0.03
150 475 4.750.21a 4.84 2.74 1.47 0.42 0.21 0.21 9.89* 0

24 h
Control 668 6.680.25 0 0 0 0.15 0 0 0.15 0
37.5 401 4.010.19a 2.99 3.24 1 0.25 0.75 0 8.23* 0
75 336 3.360.18a 5.36 3.27 0 1.78 0 0.89 11.30** 0.04
150 431 4.310.20a 3.48 3.71 3.02 0.70 0.46 0.7 12.07** 0.02

48 h
Control 607 6.070.24 0 0.16 0 0.16 0 0 0.32 0
37.5 383 3.830.19a 4.18 2.35 1.04 0.52 0.52 0 8.61* 0.02
75 321 3.210.18a 2.49 0 7.48 1.87 0 0 11.84** 0.01
150 362 3.620.18a 14.92 4.97 5.52 3.59 0.55 0 29.55** 0.04

Frequency of abnormalities (%) 40.30 24.73 21.53 8.32 2.99 2.13

a Significantly different from the control P < 0.001 (z test)

2
* Significantly different from the control P < 0.01 ( test)
** Significantly different from the control P < 0.001 (2test)

285
Genotoxic effects of herbicide Illoxan (Diclofop-Methyl) on Allium cepa L.
This decrease was slightly dose dependent at 24 h (r =
-0.56) and 48 h (-0.69). However, Illoxan had no
significant effect on the percentage of the mitotic
stages. Illoxan significantly increased the frequency
of abnormal cells in all concentrations and in all
treatment times when compared with their controls
(Table 1). This increase was dose dependent in the 24
h and 48 h treatments (r = 0.84, r = 0.99 respectively).
Six types of aberrations were recorded: stickiness,
laggards, C-mitosis, bridges, multipolarity, and
fragments (Figure 2). Stickiness was the most
common abnormality. Illoxan also induced micro-
nucleated and bi-nucleated cells at interphase.
However, these aberrations were not significant when
compared to the controls.
The results of metaphases obtained from -
monobromonaphthalene pretreated root tip cells are
summarized in Table 2. This herbicide induced four
types of structural aberrations in metaphase cells:
fragments, sister union, chromosome, and chromatid
breaks (Figure 3). This herbicide also induced
polyploidy, a numerical aberration (Figure 3). Illoxan
significantly increased the frequency of abnormal cells
(except 37.50 mg/L at 12 and 24 h) in a dose-
dependent manner (r = 0.76 at 12 h, r = 0.87 at 24 h,
r = 0.87 at 48 h).
Discussion
This study investigated genotoxicity of the
pesticide Illoxan in Allium cepa root tip cells. The
commercial form of the pesticide was tested because
this is the form that is utilized in agriculture and
introduced into the environment. Allium cepa was
used as the test system because pesticides are applied
on plants in agriculture and plants may produce
unique genotoxic metabolites.
In this study, Illoxan decreased the mitotic index in
Allium cepa root tip cells. This decrease was
significant in all concentrations when compared to
the control. The decreasing of the MI was slightly dose
dependent at 24 h and 48 h. These results show that
concentrations of Illoxan used were cytotoxic in
Allium. A similar result in mitosis has been observed
from the treatment of the herbicides racer (3),
atrazine (12), and arsenal (15). There are some
possible mechanisms for chemically decreased mitotic
286
index in plant cells. The first is that a decrease in MI
could be due to blocking of G1 suppressing DNA
synthesis (16). The second possible mechanism is a
blocking of G2 preventing the cell from entering
mitosis (17). The lowering of the mitotic index might
have been achieved by the inhibition of DNA
synthesis at the S-phase (18). Another possible
mechanism is explained by Chand and Roy (19).
These authors proposed that 2, 4-dinitrophenol
inhibits DNA and RNA synthesis by reducing the
oxidative phosphorylation in plants, resulting in lower
levels of ATP.
Illoxan increased the percentage of abnormal cells
in Allium cepa. This increase was significant in all
concentrations applied when compared to the control
and was dose dependent. Common abnormalities
were stickiness, laggards, and C-mitosis. Fiskesj (13)
reported that sticky chromosomes indicate highly
toxic chemical effects that are usually not reversible
and will probably lead to cell death. These results were
reported by many investigators following treatment
with some pesticides (2, 20). Generally, a mitotic
poison causes disturbance of the spindle apparatus,
resulting in a C-mitosis effect, which means the
complete absence of a spindle. A weak C-mitotic
effect produces lagging chromosomes that do not
attach to the spindle apparatus. The formation of C-
mitosis, lagging chromosomes and multipolarity may
be due to the disturbance in the spindle formation
which was effected by the herbicide (1, 21).
Pentachlorophenol (PCP), 2,4-dichlorophenoxyacetic
acid (2,4-D), and 2-chloro-2,6-diethyl-N-
(butoxymethyl) acetanilide (butachlor) induced
chromosome aberrations occur at a statistically
significant level. These herbicides induce breaks,
bridges, stickiness, and laggards in Allium cepa root
tips (20). Illoxan induced abnormalities in interphase
cells. However, the frequency was not high or
significant when compared to the control.
Illoxan also induced a significant increase of
chromosomal aberrations at the metaphase stage in
pretreated root tip cells in all concentrations and
treatment periods when compared with their
respective controls. While this increase was significant
in only 75 and 150 mg/L concentrations at 12 h and 24
h treatments, the increase was significant in all
concentrations at 48 h. This herbicide induced
 D. YZBAIOLU, F. NAL, C. SANCAK

a
b

d
c

e f

Figure 2. Different types of aberrations induced by Illoxan in Allium cepa root tips. a) Stickiness b) C-mitosis c) Chromosome fragment
d) Chromosome bridge e) Lagging chromosome f) Multipolarity.

287
Genotoxic effects of herbicide Illoxan (Diclofop-Methyl) on Allium cepa L.

Table 2. The types and numbers of chromosome aberrations in metaphase cells in -monobromonaphthalene pretreated root tips of A.
cepa exposed to Illoxan.

Aberrations
Total
Conc. cells Fragments Sister Chromosome Polyploidy Chromatid % Total
(mg/L) scored union breaks breaks aberrations

12 h
Control 100 - - - - - -
37.5 100 1 - - - - 1
75 100 9 - - - - 9**
150 100 2 4 1 - - 7**

24 h
Control 100 - - - - - -
37.5 100 1 - - - - 1
75 100 2 4 1 - - 7**
150 100 - 3 3 - 1 7**

48 h
Control 100 - - - - - -
37.5 100 2 1 1 - - 4*
75 100 2 5 1 1 - 9**
150 100 - 2 6 1 - 9**

*Significantly different from the control P < 0.05 (2 test)

**Significantly different from the control P < 0.01 (2 test)

fragments, sister union, chromosome and chromatid
breaks as structural aberrations and poliploidy as a
numerical aberration in metaphase cells in this study.
Sister union is the breakage followed by reunion of
both sister chromatids at an identical site (22). This
abnormality resulted from terminal deletion.
Terminal deletions cause sticky chromatid tips and
then these sticky tips may join to each other (23).The
herbicide flurochloridone induced a significant
increase in chromosomal aberrations at the
metaphase stage in all concentrations and treatment
periods when compared to the control groups. These
abnormalities manifested as chromosome breaks,
fragments and sister union (3). elik et al. (4)
reported that dinocap induced a significant increase
in chromosomal aberrations in pretreated root tips
when compared to control groups. The four types of
abnormalities recorded were chromosome breaks,
fragments, tetraploidy, and sister union at the
metaphase stage.
Although diclofop-methyl has been classified as
"likely to be carcinogenic to humans" based on
laboratory studies in rats and mice (10), the diclofop-
methyl tested negative for genotoxic effects in a
bacterial reverse mutation test, an in vitro mammalian
cell gene mutation test, an in vitro mammalian
chromosomal aberration test, an in vivo cytogenetic
test, and a mouse bone marrow micronucleus test as
mentioned in the introduction. However, the results
of this study indicate that Illoxan decreased the
mitotic index and produced clastogenic and
aneugenic types of abnormalities in Allium cepa root
tip cells.
Some compounds, such as triazines, maleic
hydrazide, and sodium azide (24), appear to be plant
specific, i.e. they are activated by genotoxic
metabolites that are not formed in mammals (25, 26).
More recently, the group of Gomez-Arroyo conducted
experiments with extracts from Vicia faba roots and

288
 D. YZBAIOLU, F. NAL, C. SANCAK

kr
k

a b

Figure 3. The chromosome aberrations in metaphase cells in -monobromonaphthalene pretreated root tips of Allium cepa exposed to
Illoxan. a) chromatid (kr) and chromosome (k) breaks b) Polyploidy c) Sister union (arrow).

cultured human lymphocytes (27). They found that
sister chromatid exchange (SCE) induction occurred
with 2 herbicides (molinate and butylate), which per
se did not cause positive effects in human
lymphocytes. These results show that plants can
metabolize chemicals that are not genotoxic per se to
mammalian cells. These findings are of crucial
importance in the case of pesticides that give negative
results in routine genotoxicity testing. If these
compounds are utilized on crop plants and
consequently convert to DNA-reactive metabolites,
ingestion of the plants could pose a threat to human
health. These kinds of metabolites may also be
mutagens in bacteria as well (28).
In conclusion, plant models such as micronucleus
(MN), chromosomal aberration (CA), and sister
chromatid exchange (SCE) assays have been found to
be highly sensitive in the detection of hazards arising
from pesticides, industrial contamination, and heavy
metals. The data obtained here support these findings
and indicate that plant bioassays can be used as an
important and integral part of test batteries for the
detection of genotoxic effects of chemicals used in the
environment.

289
Genotoxic effects of herbicide Illoxan (Diclofop-Methyl) on Allium cepa L.
Acknowledgements
The authors would like to thank the Gazi
University Research Fund for financial support under
grant no. FEF 05/99-07.
References
1. Badr A, Hamoud MA, Haroun SA. Effect of the herbicide
Gespax on mitosis, mitotic chromosomes and nucleic acids in
Vicia faba L. root meristems. Proc Saudi Biol Soc 8: 359-370,
1985.
2. Yzbaolu D. Cytogenetic effects of fungicide Afugan on the
meristematic cells of Allium cepa L. Cytologia 68: 237-243, 2003.
3. Yzbaolu D, nal F, Sancak C et al. Cytological effects of the 18.
herbicide racer flurochloridone on Allium cepa. Caryologia
56: 97-105, 2003.
4. elik M, Yzbaolu D, nal F et al. Effects of Dinocap on the
mitosis of Allium cepa L. Cytologia 70: 13-22, 2005.
5. Dimitrov BD, Gadeva PG, Benova DK et al. Comparative
genotoxicity of the herbicides Roundup, Stomp and Reglone in
plant and mammalian test systems. Mutagenesis 21: 375-382,
2006.
6. Grant WF. Higher plant assays for the detection of
chromosomal aberrations and gene mutations - a brief historical
background on their use for screening and monitoring
environmental chemicals. Mutat Res 426: 107-112, 1999.
7. Ma TH. The international program on plant bioassays and the
report of the follow-up study after the hands-on workshop in
China. Mutat Res 426: 103-106, 1999.
8. Yi HL, Meng ZQ. Genotoxicity of hydrated sulfur dioxide on
root tips of Allium sativum and Vicia faba. Mutat Res 537: 109-
114, 2003.
9. Tomlin CDS (ed.). The Pesticide Manual 13th Ed. BCPC, 2003.
10. United States Environmental Protection Agency (USEPA).
Prevention, Pesticides and Toxic Substances (7508C) R.E.D.
FACTS Diclofop-Methyl. EPA-738-F-00-007, 2000.
11. Cancer Assessment Review Committee (CARC). Evaluation of
the Carcinogenic Potential of Diclofop-methyl. P.C. Code:
110902, 2000.
12. Bolle P, Mastrangelo S, Tucci P et al. Clastogenicity of atrazine
assessed with the Allium cepa test. Environ Mol Mutagen 43:
137-141, 2004.
13. Fiskesj G. The Allium test as a standart in environmental
monitoring. Hereditas 102: 99-112, 1985.
14. Nicoloff H, Kappas A. Benomyl induced mitotic disturbances in
Hordeum vulgare. Mutat Res 189: 271-275, 1987.
15. Grisolia CK, Bilich MR, Formigli LM. A comparative
toxicologic and genotoxic study of the herbicide arsenal, its
active ingredient imazapyr, and the surfactant nonylphenol
ethoxylate. Ecotox Environ Safety 59: 123-126, 2004.
290
Corresponding Author:
Deniz YZBAIOLU
Department of Biology, Faculty of Art and Science,
Gazi University, Ankara - TURKEY
deniz@gazi.edu.tr
16. Shcneiderman MH, Dewey WC, Highfeld DP. Inhibition of
DNA synthesis in synchronized Chinese hamster cell treated in
G1 with cycloheximide. Exp Cell Res 67: 147-155, 1971.
17. Vant Hof J. The action of IAA and kinetin on the mitotic cycle
of proliferative and stationary phase excised root meristem. Exp
Cell Res 51: 167-176, 1968.
Sudhakar R, Ninge Gowda KN, Venu G. Mitotic abnormalities
induced by silk dyeing industry effluents in the cell of Allium
cepa. Cytologia 66: 235-239, 2001.
19. Chand S, Roy SC. Effects of herbicide 2,4-dinitrophenol on
mitosis, DNA, RNA and protein synthesis in Nigella sativa L.
Biol Plantarum 23: 198-202, 1981.
20. Ateeq B, Abdul FM, Niamat AM et al. Clastogenicity of
Pentachlorophenol, 2,4-D and Butachlor evaluated by Allium
root tip test. Mutat Res 514: 105-113, 2002.
21. Haliem AS. Cytological effects of the herbicide sencor on
mitosis of Allium cepa. Egypt J Bot 33: 93-104, 1990.
22. Murli H. Screening assay for chromosomal aberrations in
Chinese hamster ovary (CHO) cells with argentyn 23, Final
Report, 2003.
23. Renczoullar E, la HB, Kayraldz A et al. The genotoxic effect
of the new acaricide etoxazole. Russ J Genet 40(11): 1300-1304,
2004.
24. Valeminsky J, Gicher T. Mutagenic activity of promutagens in
plants: indirect evidence of their activation. Mutat Res 197: 221-
242, 1988.
25. Gentile JM, Gentile GJ, Bultman J et al. An evaluation of the
genotoxic properties of insecticides following plant and animal
activation. Mutat Res 101: 19-29, 1982.
26. Plewa MJ, Wagner ED, Gentile GJ et al. An evaluation of the
genotoxic properties of herbicides following plant and animal
activation. Mutat Res 136: 233-245, 1984.
27. Calderon-Segura ME, Gomez-Arroyo S, Villalobos-Pietrini R
et al. In vivo and in vitro promutagen activation by Vicia faba of
thiocarbamate herbicides molinate and butylate to products
inducing sister chromatid exchanges in human lymphocyte
cultures. Mutat Res 438: 81-88, 1999.
28. Majer BJ, Grummt T, Uhl M et al. Use of plant bioassays for the
detection of genotoxins in aquatic environment. Acta
Hydrochim Hydrobiol 33: 45-55, 2005.