RESEARCH LETTER

Fusarium response to oxidative stress by H2O2 is trichothecene

chemotype-dependent
Nadia Ponts1, Leslie Couedelo2, Laetitia Pinson-Gadais2, Marie-Noelle Verdal-Bonnin2, Christian
Barreau2 & Florence Richard-Forget2
1
Department of Cell Biology and Neuroscience, University of California at Riverside, Riverside, CA, USA; and 2UPR 1264, MYCSA, INRA, Villenave
dOrnon, France

Correspondence: Florence Richard-Forget, Abstract

UPR 1264, MYCSA, INRA, 71 Av. Edouard
Bourleaux, BP81, 33883 Villenave dOrnon-
cedex, France. Tel.: 133 557 122 483; fax:
133 557 122 500; e-mail:
fforget@bordeaux.inra.fr
Received 21 October 2008; accepted 27
January 2009.
First published online 23 February 2009.
DOI:10.1111/j.1574-6968.2009.01521.x
Editor: Bernard Prior
Keywords
Fusarium; type B trichothecenes; H2O2;
oxidative stress; catalase.
The present study aims at clarifying the impact of oxidative stress on type B
trichothecene production. The responses to hydrogen peroxide (H2O2) of an array
of Fusarium graminearum and Fusarium culmorum strains were compared, both
species carrying either the chemotype deoxynivalenol (DON) or nivalenol (NIV).
In both cases, levels of in vitro toxin production are greatly influenced by the
oxidative parameters of the medium. A 0.5 mM H2O2 stress induces a two- to 50-
fold enhancement of DON and acetyldeoxynivalenol production, whereas the
same treatment results in a 2.4- to sevenfold decrease in NIV and fusarenone X
accumulation. Different effects of oxidative stress on toxin production are the
result of a variation in Fusariums antioxidant defence responses according to the
chemotype of the isolate. Compared with DON strains, NIV isolates have a higher
H2O2-destroying capacity, which partially results from a significant enhancement
of catalase activity induced by peroxide stress. A 0.5 mM H2O2 treatment leads to a
1.3- to 1.7-fold increase in the catalase activity of NIV isolates. Our data, which
show the higher adaptation to oxidative stress developed by NIV isolates, are
consistent with the higher virulence of these Fusarium strains on maize compared
with DON isolates.

isolates, and (2) the DON chemotype, which includes DON-

Introduction
Fusarium head blight (FHB), also known as scab, is one of
the most damaging diseases of small grain cereals (Windels,
2000). FHB may be caused by an array of ear blight
pathogens, including Fusarium species from the discolour
section (mainly Fusarium culmorum, Fusarium graminear-
um, and Fusarium avenaceum in Europe) and Microdochium
nivale (Bottalico & Perrone, 2002). The main concern
regarding FHB arises from the ability of some Fusarium
species to produce mycotoxins in kernels. These mycotoxins
are harmful to both human and animal consumers. The
most frequent mycotoxins encountered in blighted grains
throughout the European countries are type B trichothe-
cenes (Logrieco & Visconti, 2004). They include deoxyniva-
lenol (DON), acetyldeoxynivalenol (ADON), nivalenol
(NIV), and fusarenone X (FX). Based on the production of
type B trichothecenes, isolates of F. graminearum and F.
culmorum have been divided into two chemotypes: (1) the
NIV chemotype, which includes NIV- and FX-producing
and ADON-producing isolates (Ichinoe et al., 1983). DON
can also be produced by NIV isolates, but in very low
amounts that are usually o 1% of NIV or FX (Desjardin &
Plattner, 2003). DON isolates cannot produce NIV (Llorens
et al., 2004). According to Lee et al. (2002), these differences
in type B trichothecene production by the two chemotypes
are determined by differences in Tri7 and Tri13 gene
structures. Both Tri7 and Tri13 are nonfunctional in isolates
of the DON chemotype.
Type B trichothecenes are chemically stable (Lauren &
Smith, 2001). They survive food and feed processing, lead-
ing to a potential risk to human and animal health.
Furthermore, because these mycotoxins are produced within
the growing crop, the most efficient way to minimize their
presence in grains is to develop prevention strategies. Today,
controlling the accumulation of type B trichothecenes in
kernels remains a challenge (Yuen & Schoneweis, 2007).
Identifying and understanding the mechanisms that regulate
the biosynthesis of type B trichothecenes can provide

FEMS Microbiol Lett 293 (2009) 255262 Journal compilation c 2009 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. No claim to original French government works
256
valuable insights into how to limit their occurrence in
kernels.
Previous data support the hypothesis that plant com-
pounds involved in plantfungi interactions are able to
interfere with mycotoxin production in host tissues (Bou-
tigny et al., 2008). In particular, neo-produced reactive
oxygen species (the generation of which is one of the earliest
responses of plant cells under biotic stresses) were shown to
trigger aflatoxin production by Aspergillus parasiticus (Re-
verberi et al., 2007). With regard to the fungus Fusarium, the
presence of hydrogen peroxide (H2O2) in the culture
medium was demonstrated to be a prerequisite to the
accumulation of DON and 15-ADON (Ponts et al., 2006).
Finally, oxidative stress induced by H2O2 leads to increased
expressions of Tri genes (Ochiai et al., 2007; Ponts et al.,
2007). The conclusions from these previous reports were
that DON production by Fusarium could be an element of a
global response against stress, and/or an element of an
adaptation response against oxidative stress. With regard to
NIV production, data are scarce and apparently contra-
dictory. According to preliminary trials performed by Ponts
et al. (2006), NIV and FX production seem to be restrained
by H2O2 oxidative stress. On the contrary, Ochiai et al.
(2007) have reported an upregulation of Tri5 gene expres-
sion in a F. culmorum strain belonging to the NIV chemo-
type. These observations remain to be confirmed and
clarified, while the mechanisms involved in differential
response to oxidative stress need to be elucidated.
Fungal cells possess both enzymatic and nonenzymatic
components that can protect cells from oxidative damage
(Belozerskaya & Gessler, 2007). Among these, catalases
(H2O2: H2O2 oxidoreductase, EC 1.11.1.6) are central
components, and prevent the formation of the highly
reactive hydroxyl radical by decomposing H2O2 into water
and oxygen. Most research works that investigated the role
of catalase in response to oxidative stress of filamentous
fungi were devoted to Neurospora crassa (Chary & Natvig,
1989; Michan et al., 2002), Aspergillus fumigatus, Aspergillus
niger, and Aspergillus nidulans (Kawasaki et al., 1997; Taka-
suka et al., 1999; Bai et al., 2003). Information regarding the
Fusarium genus remains fragmentary. Only a few species
have been investigated. The most extensive studies focused
on Fusarium oxysporum (Kono et al., 1995; Angelova et al.,
2005), Fusarium accuminatum (Ayar-Kayali et al., 2002),
and Fusarium equiseti (Ayar-Kayali & Tarhan, 2004). They
all led to the same conclusion, i.e. catalases play a pivotal
role in the antioxidant defence network of Fusarium. In
addition, H2O2-induced increase of catalase activity was
clearly demonstrated by Angelova et al. (2005).
In order to learn more about a potential link between
oxidative stress and type B trichothecene production, the
responses to H2O2 of an array of F. graminearum and F.
culmorum strains were compared. Both these species possess
Journal compilation 
c 2009 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. No claim to original French government works
N. Ponts et al.
either the chemotype DON or NIV. Furthermore, the
mechanisms by which Fusarium strains manage oxidative
stress by involving catalase enzyme were investigated.
Materials and methods
Fungal strains, media, and culture conditions
Ten different F. graminearum and F. culmorum strains were
used for this study: five F. graminearum (three with the DON
chemotype INRA 349, 202, and 155 and two with the NIV
chemotype INRA 91 and 127) and five F. culmorum (two
with the DON chemotype INRA 117 and 127 and three
with the NIV chemotype INRA 337, 132, and 319). All
strains belong to the INRA-MycSA laboratory collection,
with the exception of F. graminearum INRA 349, alias
CBS185.32 (Centraal Bureau voor Shimmelcultures, the
Netherlands). Fungal strains were maintained on PDA slants
for 8 days at 25 1C. Spore suspensions were prepared by
adding sterile-distilled water to the slants with gentle shaking.
To support type B trichothecene biosynthesis, liquid
cultures were routinely performed in GYEP (glucose, yeast
extract, peptone) medium (Miller et al., 1983), and per-
formed in triplicate. GYEP medium (100 mL) in 500-mL
Erlenmeyer flasks were inoculated with 106 spores and
incubated at 25 1C and 150 r.p.m. in the dark.
Effect of H2O2 on type B trichothecene
production
H2O2 supplementation
GYEP liquid cultures were supplemented with H2O (con-
trol) or H2O2 (0.5 mM) following two different methods:
synchronized with inoculation (H2O2t0) or daily (H2O2d).
In this last case, 0.5 mM H2O2 was added to each culture
flask every 24 h for 27 days. Following incubation, broths
were filtered as described by Ponts et al. (2006). Filtrates
were analysed for type B trichothecene production and
H2O2 consumption. Fungal biomass was measured by
weighing the mycelia after 48 h of freeze drying. For each
trial, it was ensured that supplementation did not modify
pH values of the treated batches compared with the control.
It was also ensured that fungal biomass accumulation was
not affected by the treatment compared with the control
(data not shown).
Type B trichothecenes analysis
An aliquot (15 mL) of the filtrate was extracted with 30 mL
of ethyl acetate; 20 mL of the organic phase was evaporated
to dryness at 70 1C under a stream of nitrogen. Dried
samples were resuspended in 200 mL of methanol/water
(50%, v/v) before analysis by HPLC-DAD (Bily et al.,
FEMS Microbiol Lett 293 (2009) 255262
Fusarium response to a peroxide stress
2004). Quantification was performed using an external
calibration with DON, ADON, NIV, and FX standard
solutions prepared from commercial pure powders (Sigma-
Aldrich, St. Louis).
Time course of H2O2 levels in culture media
H2O2 was assayed in filtered broth following the procedure
described by Ponts et al. (2006), adapted from Richard-
Forget & Gauillard (1997). Briefly, 500 mL of 50 mM guaia-
col in McIlvaine buffer (pH 5.5) and 1.25 U of horseradish
peroxidase (HRP) were added to 1 mL of filtered medium.
Absorbance was monitored at 470 nm for 1 min. Quantifica-
tion was performed by external calibration with H2O2
standard solutions. The absence of fungal peroxidase activity
in filtrates was always verified. All reagents were purchased
from Sigma-Aldrich.
Catalase activity during cultivation of Fusarium
spp.
Preparation of cell-free extracts
Harvested mycelia, grown in GYEP liquid cultures, were
washed twice with distilled water, frozen at  20 1C, pow-
dered in liquid nitrogen with a Dangoumo blender (Prola-
bo, Fontenay-sous-Bois, France), and freeze dried. Freeze-
dried mycelia (250 mg) were suspended in 1.75 mL of cold
Tris-HCl 0.2 M, pH 7.6, containing 1% PVP, 0.5 mM
dithiothreitol, and 0.1 mL mL1 of Pefablocs SC-protease
inhibitor (Roche, Manheim, Germany). The homogenate
was held for 30 min at 40 1C. Cell debris was removed by
centrifugation (21 000 g, 30 min, and 4 1C).
Catalase activity
Catalase activity in cell-free extracts was determined using
the method of Aebi (1994). The reaction mixture contained
10 mM H2O2 in a 100 mM phosphate buffer at pH 7.0 and
50 mL of cell-free extracts in a total volume of 3 mL. The
decomposition of H2O2 was followed directly by a decrease
in A240 nm at 25 1C, and quantified using an external calibra-
tion curve obtained with H2O2 standard solutions.
Protein assay
Protein content was determined according to Bradford
(1976), using bovine serum albumin as a standard.
Electrophoretic experiments
Isoelectrofocusing in polyacrylamide gels was performed
with the PhastSystem (Pharmacia) using Phastgelss, pH
FEMS Microbiol Lett 293 (2009) 255262
Published by Blackwell Publishing Ltd. No claim to original French government works
257
39, following the manufacturers instructions. Catalase
activity was visualized on gels following a procedure mod-
ified from Clare et al. (1984). Gels were soaked in 50 mM
phosphate buffer, pH 7.0, supplemented with 50 mg mL1 of
HRP at room temperature for 45 min, followed by two
washes with the phosphate buffer. Gels were then treated
with 5 mM H2O2 for 10 min. After rinsing twice with
distilled water, gels were stained with 0.5 mg mL1 of diami-
nobenzidine in phosphate buffer.
Expression of results and statistical analysis
All results (toxin and H2O2 contents, catalase activity) were
computed from experiments performed in triplicate. Data
were expressed as their arithmetic mean values  SD. Cata-
lase activity was expressed as micromoles of H2O2 con-
sumed per minute and per gram of dry fungal biomass.
Toxin levels were expressed in micrograms of toxin per gram
of dry fungal biomass. Because all of the observed effects on
DON and ADON or NIV and FX accumulation were
consistent for all conditions, DON and ADON or NIV and
FX yields were summed without distinguishing the chemical
species (DON1ADON, NIV1FX). Values were compared
by performing the standard Students t-test. The P value 0.05
was chosen as the point of statistical significance.
Results and discussion
Impact of H2O2 on trichothecene B production
by Fusarium spp.
Previous studies showed that 0.5 mM H2O2 led to a sig-
nificant increase of DON and ADON accumulation in
submerged cultures of F. graminearum (Ponts et al., 2006).
These results also suggested that the effect of H2O2 might
depend on the species and/or the chemotype of the con-
sidered Fusarium strain. Here, we investigate these hypoth-
eses by comparing the effect of 0.5 mM H2O2 on type B
trichothecene production by four different strains of Fusar-
ium: two of them belong to the species graminearum (INRA
349 with the DON chemotype, and INRA 91 with the NIV
chemotype), and the other two belong to the culmorum
species (INRA 155 with the DON chemotype, and INRA 337
with the NIV chemotype). Two procedures of H2O2 supple-
mentation were used, i.e. synchronized with inoculation
(H2O2t0) or daily (H2O2d), and accumulation of DON1A-
DON or NIV1FX was measured after 21 days of culture.
Dry mycelial mass accumulation was not affected by the
H2O2 treatments after 21 days of incubation (data not
shown). The results are summarized in Table 1. Independent
of the considered species or the supplementation procedure,
a significant increase in the DON1ADON levels was ob-
served in cultures treated with H2O2. This observation is
consistent with previously published results (Ponts et al.,
Journal compilation c 2009 Federation of European Microbiological Societies
258 N. Ponts et al.

Table 1. Type B trichothecene accumulation in liquid cultures of Fusarium graminearum and Fusarium culmorum isolates
F. graminearum F. culmorum

DON115-ADON NIV1FX DON115-ADON NIV1FX

Isolates INRA 349 INRA 91 INRA 155 INRA 337

Control 6296 (  173) 617 (  22.5) 43.7 (  3.1) 7360 (  228)
H2O2t0w 13208 (  377) 245 (  22.2) 476.6 (  18.5) 2150 (  41)
H2O2dz 44242 (  1004) 88.7 (  5.3) 2202 (  181) 3097 (  96)
Values are given as the average toxin yield (mg g1 dry mycelium)  SD in three independent experiments.
w
Twenty-one-day-old GYEP cultures supplemented by H2O2 at the beginning of the culture.
z
Twenty-one-day-old GYEP cultures supplemented by H2O2 daily.

2006). Significantly higher production of DON1ADON 12

was observed in cultures subjected to a continuous H2O2

Toxin yields (mg of toxins g1

treatment than cultures treated only at inoculation. Con- 10
versely, levels of NIV1FX were significantly reduced in

H2O2-treated cultures, irrespective of the species. The in-
tensities of the diminution ranged from 58% to 86%,
depending on the considered strain and the H2O2 supple-
mentation procedure. These data strongly suggest that type
B trichothecene production is differentially modulated by
oxidative stress by H2O2 according to the chemotype rather
than the species of the Fusarium strain.
The effect of 0.5 mM H2O2 on NIV1FX production was
further characterized: toxin accumulation was monitored in
the culture media for up to 27 days (Fig. 1). Cultures were
supplemented with H2O2 following the two procedures used
in this study, i.e. synchronized with inoculation (H2O2t0) or
daily (H2O2d). NIV1FX was rapidly produced in the
control samples and reached 10 mg g1 after 27 days of
incubation. However, after both H2O2 treatments, NIV1FX
reached detectable levels only after 5 days of incubation. The
rate of NIV1FX accumulation was significantly decreased in
H2O2-treated samples throughout the incubation period
and reached 5 mg g1 after 27 days. This result supports a
previous hypothesis, i.e. regulation of DON1ADON accu-
mulation is conditioned by events occurring during spore
germination (Beyer et al., 2005; Ponts et al., 2006), and
extends it to NIV1FX.
The mechanisms by which oxidative stress by H2O2 can
differentially modulate toxin accumulation, that is to say
increasing DON1ADON production while reducing
NIV1FX yields, are unknown. These differences may reflect
the uneven abilities of Fusarium strains to protect them-
selves from H2O2 oxidative damage, according to their
chemotype. This question was addressed by assaying the
ability of various Fusarium strains to metabolize H2O2.
Time course of H2O2 levels in culture media
The concentrations of H2O2 in Fusarium culture media were
monitored over 27 days. Typical curves obtained for Fusar-
dry mycelium)
8
6
4
2
0
0 5 10 15 20 25 30
Time (days)
Fig. 1. NIV1FX biosynthesis by Fusarium culmorum (INRA 337) in GYEP
medium supplemented with 0.5 mM H2O2 at different times in the
culture course, or not supplemented. The solid line represents the
control; the dashed lines represent the kinetics of toxin accumulation in
H2O2 media. ,, Control, , daily H2O2, , H2O2t0.  
ium strains cultures in 0.5 mM H2O2t0-supplemented GYEP
media, or not supplemented, are shown in Fig. 2. Both DON
and NIV chemotype strains were used. H2O2 assays in
control (H2O supplemented) media indicated the occur-
rence of a constitutive production of H2O2 by the fungus.
H2O2 accumulation was perceptible after 3 days and in-
creased between 3 and 5 days for the DON1ADON-produ-
cing strain (INRA 349). The time-course curve of this H2O2
accumulation follows the pattern of DON and ADON
production by INRA 349 (Ponts et al., 2006). With regard
to the NIV1FX-producing Fusarium strain INRA 337,
detected amounts of H2O2 were quantifiable after 5 days in
the nonsupplemented culture, and increased continuously
to reach nearly 30 mM after 27 days. This pattern of H2O2
accumulation also parallels the time course of NIV and FX
accumulation (see Fig. 1).
In GYEP media supplied with 0.5 mM H2O2, introduced
H2O2 was significantly consumed during the first days of
culture; the NIV Fusarium strain seemed more efficient in
decomposing H2O2, at a consumption rate two times higher
than that observed in media inoculated with the

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Published by Blackwell Publishing Ltd. No claim to original French government works
Fusarium response to a peroxide stress
600
500
400
[H2O2] (M)
300
200
100
0 2
0 5 10 15 20 25 30
Time (days)
Fig. 2. Time course of H2O2 amounts in liquid culture media inoculated
with Fusarium graminearum INRA 349 (DON/ADON producer) and
Fusarium culmorum INRA 337 (NIV/FX producer). Dashed lines corre-
spond to GYEP media supplemented with 0.5 mM H2O2 (H2O2t0), and
solid lines represent nonsupplemented GYEP media. n, 0.5 mM H2O2-

supplied GYEP medium, , 0.5 mM H2O2-supplied culture of INRA 349,
0.5 mM H2O2-supplied culture of INRA 337, &, control culture of INRA
349, and , control culture of INRA 337.
DON1ADON-producing strain (Fig. 2). While 70% of the
introduced H2O2 was metabolized after 3 days in the DON/
ADON Fusarium strain culture media, the total initial H2O2
amount was destroyed by the NIV Fusarium strain during
the same period of time. Similar differences were observed
for the other two strains: INRA 91 and 155 (data not
shown).
Our data suggest that Fusarium strains of the NIV
chemotype are more efficient at metabolizing H2O2 and
therefore alleviate oxidative stress. The different H2O2-
destroying capacities of the two Fusarium chemotypes,
leading to different levels of oxidative stress encountered by
the strain when toxin biosynthesis is induced, could partially
explain the fact that exogenous H2O2 supplementation led
to opposite effects on DON1ADON and NIV1FX bio-
synthesis.
To further investigate the response of Fusarium strains to
oxidative stress, catalases that have been shown to play a key
role in H2O2 detoxification by microorganisms were char-
acterized in Fusarium cells of strains belonging to the two
chemotypes.
Catalases in DON and NIV Fusarium cell extracts:
total activity and isoforms
Catalase activity was first followed during the asexual life
cycle of F. graminearum (strain INRA 349). Catalase activity
was higher during the lag phase, i.e. during spores germina-
tion and initiation of mycelial growth (Fig. 3). Similar data
were reported for N. crassa (Michan et al., 2002) and A.
nidulans (Navarro et al., 1996). In the two former reports,
FEMS Microbiol Lett 293 (2009) 255262
259
(a) 20 0.7
Catalase activity (M of H2O2
0.6
16
Fungal biomass (g)
0.5
min1 g1)
12
0.4
8 0.3
0.2
4
0.1
0 0.0
3 8 11 15 21
Time (days)
(b)
Fig. 3. Catalase activity during the asexual life cycle of Fusarium
graminearum (strain INRA 349) grown in GYEP medium. (a) Catalase
activities in the lag, exponential, and stationary phases expressed in
micromoles of H2O2 consumed per minute per gram of dry fungal
biomass. (b) Isoelectrophoretic patterns of catalase using extracts from
cells indicated in (a). Each lane was loaded with 10 mg of proteins and
duplicated. Band-staining procedure is described in Materials and meth-
ods.
three catalases were identified and the high level of catalase
activity in the lag phase was ascribed to the specific
accumulation of the Cat-1 (CatA for A. nidulans) isoform
in spores. As it clearly appeared on the isoelectrophoretic
profiles reported in Fig. 3, two catalase isoforms (isoelectric
points close to 5.3 and 6.8) are present in F. graminearum
cell extracts during the lag phase and one of them, the most
basic one, disappears during the exponential growth phase.
Catalase activities were compared in cell extracts of 3-
day-old cultures, supplemented or not with H2O2 (0.5 mM,
at the beginning of the culture) for five DON-producing
strains and five NIV isolates (Fig. 4). Constitutive levels of
catalase were not significantly different between the two
types of isolates. For DON and NIV strains, minimal values
were close to 40 mmol min1 g1 and maximal values were
nearing 130 mmol min1 g1. However, our results clearly
demonstrated that a peroxide stress induced an opposite
modulation of catalase activity, depending on the chemo-
type of the considered strain (Fig. 4). In H2O2-treated fungal
cells of NIV isolates, the activity of catalase was significantly
increased, with a level of induction ranging between 1.3 and
1.7. In contrast, peroxide stress caused a decrease (about
1.22.2-fold) in the catalase activity of DON isolates,
compared with the control. To our knowledge, no data
concerning the induction of oxidative enzymes in F. grami-
nearum and F. culmorum upon H2O2 treatment have been
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Published by Blackwell Publishing Ltd. No claim to original French government works
260
Fig. 4. Ratios of catalase activities of cell-free extracts from a 3-day-old
fungal culture grown in GYEP supplemented with 0.5 mM H2O2 and in
control. Ten strains were tested: five Fusarium graminearum (three with
the DON chemotype INRA 349, 202, and 155 and two with the NIV
chemotype INRA 91 and 127) and five Fusarium culmorum (two with
the DON chemotype INRA 117 and 127 and three with the NIV
chemotype INRA 337, 132, and 319). The data are presented as Box
and Whisker plots. A box represents the range in which 50% of the data
lie. The horizontal line within the box represents the median; the dashed
line represents the mean. The other 50% is reported as a vertical line,
extended upwards and downwards of the box.
published before. In H2O2-treated fungal cells of F. oxyspor-
um, an induction of catalase was reported by Angelova et al.
(2005). The H2O2 concentration used in this report was,
however, very high (30 mM), considerably higher than the
physiological concentrations that Fusarium could encoun-
ter. With H2O2 concentrations close to 0.5 mM, such as that
used in our work, an increase in the catalase activities of A.
nidulans (Kawasaki et al., 1997) and Streptomyces coelicor
(Lee et al., 1993) has already been described. The significant
enhancement of catalase activities by peroxide stress ob-
served for NIV isolates is in accordance with the higher
H2O2-destroying capacity that was previously reported for
these strains. These data suggest that Fusarium strains of the
NIV chemotype are able to develop a better adaptation to
oxidative stress compared with DON isolates.
Conclusion
For several years, mechanisms responsible for the onset of
mycotoxins in fungi have been the subject of considerable
speculation and research. Actually, greater knowledge of the
factors controlling their synthesis would ultimately allow
repressing their production in kernels. Oxidative stress
appears to be a common factor related to different mycotox-
ins biosynthesis. Oxidative stress was shown to trigger
aflatoxin production by A. parasiticus (Reverberi et al.,
2007) and also supposed to be a prerequisite for DON1A-
DON accumulation by F. graminearum (Ponts et al., 2006).
Our results show the strong effect of H2O2 stress on
NIV1FX accumulation by F. graminearum and F. culmor-
um. However, while DON1ADON production was en-
Journal compilation 
c 2009 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. No claim to original French government works
N. Ponts et al.
hanced by a peroxide stress (Ponts et al., 2006), our data
demonstrated a significant inhibition of NIV1FX produc-
tion by H2O2. This observation indicates that type B
trichothecene production is differentially modulated by
H2O2 oxidative stress, according to the chemotype of the
toxigenic strain. This differential modulation cannot result
from a differential transcriptional regulation of Tri genes.
Indeed, whatever the considered isolates (DON or NIV
strains), treatment with H2O2 activates Tri5 transcription
(Ochiai et al., 2007; Ponts et al., 2007). This differential
modulation might result from differences in antioxidant
defence responses between isolates of the two chemotypes
and therefore in the efficiency to detoxify H2O2. According
to our results, isolates of the NIV chemotype have a higher
H2O2-destroying capacity compared with DON isolates.
This higher ability to cope with H2O2 was partially ascribed
to a significant enhancement of catalase activities by per-
oxide stress, resulting in a better adaptation of NIV isolates.
Further studies involving other antioxidant enzymes, such
as glutathione peroxidases as well as Fusarium strains
capable of producing zearalenone, type A trichothecenes
and fumonisins, will provide a further insight into this
matter.
The higher adaptation to oxidative stress developed by
Fusarium isolates with the NIV chemotype is consistent with
the higher virulence of these Fusarium strains on maize
compared with DON isolates (Carter et al., 2002). However,
the manner in which trichothecenes influence the pathogen-
esis of F. graminearum remains highly complex and is
chemotype as well as host specific (Maier et al., 2006).
Actually, isolates of the DON and NIV chemotypes were
reported to be equally pathogenic on wheat (Carter et al.,
2002). According to our results, the host/virulence interac-
tions could be ascribed to different host H2O2-production
patterns in response to F. graminearum and F. culmorum
infection.
Acknowledgements
This work was partially supported by a grant from the
Conseil Regional de la Region Aquitaine (France) and the
INRA. The authors would like to thank Carol Park for her
proof reading.
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Journal compilation 
c 2009 Federation of European Microbiological Societies FEMS Microbiol Lett 293 (2009) 255262
Published by Blackwell Publishing Ltd. No claim to original French government works