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Received: 21 April 2020    Accepted: 11 September 2020

DOI: 10.1111/ppa.13289

ORIGINAL ARTICLE

Five-year survey uncovers extensive diversity and temporal

fluctuations among fusarium head blight pathogens of wheat
and barley in Brazil

Carolina B. Pereira1 | Todd J. Ward2 | Emerson M. Del Ponte3  | Gláucia Mara

Moreira3 | Mark Busman2 | Susan P. McCormick2 | Heraldo R. Feksa1,4 |
Juliano L. De Almeida4 | Dauri J. Tessmann1

1
Departamento de Agronomia, Universidade
Estadual de Maringá, Av. Colombo 5790, Abstract
Maringá, Paraná 87020-900, Brazil We conducted a five-year survey (2011–2015) of barley and wheat fields in Paraná
2
Mycotoxin Prevention and Applied
state, Brazil, obtaining 754 Fusarium isolates from spikes with fusarium head blight
Microbiology Research Unit, National
Center for Agricultural Utilization (FHB)-symptoms. Multilocus genotyping and TEF-1α gene sequence analyses con-
Research, Agricultural Research Service, US
firmed the dominance of the F. graminearum species complex (FGSC, 75.7%), but F.
Department of Agriculture, Peoria, IL, USA
3
Departamento de Fitopatologia,
poae (11.5%), as well as F. avenaceum and related members of the F. tricinctum species
Universidade Federal de Viçosa, Viçosa, complex (FTSC, 8.1%) appeared as substantial contributors to FHB. Within the FGSC,
Brazil
4
F. graminearum of the 15-ADON genotype was dominant (63%), followed by F. meridi-
Fundação Agrária de Pesquisa
Agropecuária – FAPA, Vitória, Entre Rios, onale of the NIV genotype (23.1%), F. cortaderiae of the NIV (7%) or 3-ADON (2.6%)
Guarapuava, Paraná 85139-400, Brazil genotypes, and F. austroamericanum (3.8%) of the 3-ADON genotype. Substantial
Correspondence variation in pathogen composition was observed across years, with F. poae and F.
Dauri J. Tessmann, Departamento de meridionale frequencies significantly elevated in some years. Most F. poae strains
Agronomia, Universidade Estadual de
Maringá, Av. Colombo 5790, Maringá, produced DAS, diANIV, and butenolide, but not neosolaniol, T-2, or HT-2. All FTSC
Paraná, 87020-900, Brazil. species produced moniliformin. Enniatin production was widespread among FTSC
Email: [email protected]
species, with the single F. acuminatum strain found to be the strongest producer of
Funding information enniatins. Our findings confirm FGSC as a major contributor to FHB and expand con-
Coordenação de Aperfeiçoamento de
Pessoal de Nível Superior, Grant/Award siderably our knowledge of the presence, frequency, and conditions under which
Number: Finance Code 001; Conselho other pathogens may emerge, altering the spectrum of toxins that may accumulate
Nacional de Desenvolvimento Científico e
Tecnológico, Grant/Award Number: Project in grain.
310719/2016-6; Fundação Agrária de
Pesquisa Agropecuária; USDA-ARS National KEYWORDS
Program for Food Safety
Fusarium avenaceum, fusarium head blight, Fusarium poae, Hordeum vulgare, mycotoxin,
Triticum aestivum

a result of infections by airborne spores of Fusarium spp. that land

1 |  I NTRO D U C TI O N on the plant during flowering and early grain development stages
(Del Ponte et al., 2007). Among several species of Fusarium that are
Fusarium head blight (FHB) is a disease of major economic concern pathogenic to these cereals, species of the F. graminearum species
to wheat and barley growers due to the potential to reduce yield complex (FGSC), such as F. graminearum and F. asiaticum, are re-
and contaminate grain with mycotoxins at levels that lead to grain garded as the main FHB pathogens worldwide, including in Brazil
downgrade or rejection by food and brewing industries (McMullen (Del Ponte et al., 2015). The FGSC is a subset of the broader F. sam-
et al., 2012). The disease symptoms manifest on the wheat heads as bucinum species complex (FSAMSC), which includes species such as

© 2020 British Society for Plant Pathology     1

Plant Pathology. 2020;00:1–10. wileyonlinelibrary.com/journal/ppa |
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2       PEREIRA et al.

F. poae that are also commonly associated with FHB in some regions. FHB in this region, the results were used to test the hypotheses that
In addition, F. avenaceum and other members of the F. tricinctum host or year of sampling drive pathogen composition (Ward et al.,
species complex (FTSC) are known to contribute to FHB epidemics, 2008; Del Ponte et al., 2015).
mainly in temperate regions (O’Donnell et al., 2013; Bec et al., 2015).
The main concern with the presence of multiple toxigenic spe-
cies relates to the wider range of mycotoxins that they may pro- 2 | M ATE R I A L A N D M E TH O DS
duce in grain. For instance, FGSC strains are known to produce
B-trichothecenes, but subtypes have been determined for each 2.1 | Survey area, seasons, and sampling
strain depending on their ability to produce deoxynivalenol (DON)
or nivalenol (NIV), and their respective acetylated forms 3-acetyl(A) During five consecutive growing seasons (2011 to 2015), commer-
DON (3-ADON chemotype), 15-ADON (15-ADON chemotype), cial wheat and barley fields from the central-southern region of
and 4,15di-ANIV (NIV chemotype) (Savard and Blackwell, 1994). Paraná state, Brazil, were visited during the grain filling period, and
Recently, a subpopulation of F. graminearum has been found to pro- heads exhibiting FHB symptoms were manually harvested. In the re-
duce the type A trichothecene NX-2 (NX-2 chemotype) instead of gion, virtually all fields are cultivated under the no-till system with
DON or NIV (Varga et al., 2015). The genes responsible for the dif- two growing seasons per year: wheat, barley, or oats grown during
ferential production of these toxins have been identified and assays the winter, and soybean or maize during the summer. Wheat and
have been developed to reliably predict toxin type based on DNA barley are sprayed with fungicides three to five times each crop sea-
sequence differences (Garmendia et al., 2018). FGSC are also known son to control foliar diseases and FHB. The minimum and maximum
to produce zearalenone, but differences among the species of the distance between two fields was 26.2 and 237 km, respectively. The
complex are still unknown. FTSC strains can produce moniliformin fields were located within the latitudes of  −  24.45 to  −  26.05 and
(MON) and a series of cyclic depsipeptides such as enniatin ana- longitudes of  −  51.45 to  −  52.16, encompassing 17 municipalities.
logues (ENs) and beauvericin (BEA) (Jestoi, 2008; Luz et al., 2017). The surveyed wheat and barley fields were cultivated under no-till
On the other hand, F. poae can produce NIV and type A trichoth- following soybean as a summer crop.
ecenes in addition to BEA (Vanheule et al., 2017).
In Brazil, molecular surveys since the mid-2000s have contrib-
uted to understanding FGSC diversity associated with FHB in wheat 2.2 | Fungal isolation, purification, and
(Scoz et al., 2009; Astolfi et al., 2012; Del Ponte et al., 2015) and DNA extraction
barley (Astolfi et al., 2011; Castañares et al., 2016; Machado et al.,
2017). For those crops, F. graminearum is the dominant species from All kernels were excised from each head and the Fusarium-damaged
the FGSC (>80% frequency) along with four other DON- or NIV- kernels selected for isolations. These were surface sterilized in 70%
producing species within the complex. Expanding these molecular ethanol for 1 min, immersed in 1% sodium hypochlorite (1% available
surveys to other major cereal crops documented host-specific dif- chlorine) for 1 min, and rinsed three times with autoclaved distilled
ferences in the relative prevalence of FGSC species. For instance, water. After drying, kernels were placed on top of wet filter paper
F. meridionale dominates in maize stalk and ear rots (Kuhnem et al., inside a germination box and incubated for 7 days at 20 ± 2°C with a
2015) and F. asiaticum dominates in rice kernels (Gomes et al., 2015). 12 hr light/dark cycle. Fragments of mycelia from a typical Fusarium
Collectively, these studies suggest that FGSC composition and toxi- spp. colony grown on the kernels were transferred to water agar
genic profile are strongly shaped by host and regional factors. For (WA) medium and subcultured onto a synthetic low-nutrient agar
example, F. meridionale was found to be a more significant contribu- (SNA) using a single spore (Leslie and Summerell, 2006). Isolates were
tor to FHB of wheat in Paraná state, where maize production is more frozen at  −80°C in 20% glycerol for long-term storage at the U. S.
common, than in Rio Grande do Sul (RS) state (Del Ponte et al., 2015). Department of Agriculture, Peoria, IL, USA and at the Universidade
In addition, there is evidence of a possible cultivar effect shaping Estadual de Maringá, Brazil. A mycelial plug was transferred from the
the composition of FGSC species, as was demonstrated recently for purified colonies to 6-cm Petri plates containing V8 juice agar and in-
barley grown in RS state (Machado et al., 2017). cubated at 26°C for 7–15 days. Mycelia were harvested with scalpel
Previous studies on FHB pathogens in Brazil have largely focused blades and stored at −20°C. Total genomic DNA was extracted using
on the diversity within the FGSC. Little is known about the presence the Quick-DNA Fungal/Bacterial 96 Kit (Zymo Research) according
and relative contributions of other species to FHB of wheat and bar- to the manufacturer's instructions.
ley in Brazil, despite the potential significance of this variation for
disease and mycotoxin control programmes. In this study, we con-
ducted a five-year survey of FHB diversity in the central-southern 2.3 | Molecular identification
wheat-producing area of Paraná state, Brazil. This is one of the most
important wheat and barley production regions in South America Species and trichothecene genotypes were identified for FGSC
and has not been well represented in previous studies. In addition isolates using a multilocus genotyping (MLGT) assay (Ward et al.,
to documenting the species and mycotoxin diversity associated with 2008). The MLGT is based on allele-specific primer extension
PEREIRA et al. |
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(ASPE) reactions targeting single nucleotide polymorphisms via bootstrap analysis employing 1,000 pseudoreplicates of the
(SNPs) for the discrimination of 16 species within the FGSC, five data.
related species, and their associated trichothecene types (Ward
et al., 2008; Garmendia et al., 2018). Sequences of the partial
translation elongation factor (TEF-1α) gene were obtained using 2.6 | Mycotoxin analysis
previously published primers (O’Donnell et al., 1998) for all iso-
lates that failed identification in the MLGT assay. PCR was carried Isolates were screened for production of mycotoxins by inoculation
out in a final volume of 25 µl, containing 50 ng genomic DNA on cracked maize kernels (25 g + 11 ml water) as previously described
and 10 × PCR buffer, 50 mM MgSO 4 , 10 mM deoxynucleoside (Aoki et al., 2015). After 14 days’ incubation in the dark, 10 g of each
triphosphate, 0.6 mM each primer, and 5 U Platinum Taq high fi- culture was extracted with 20 ml 86:14 (vol/vol) acetonitrile:water
delity polymerase (Invitrogen). PCR conditions were initial dena- for 30 min with shaking. After the extracts were clarified via fil-
turation for 2 min at 96°C; followed by 35 cycles of 30 s at 94°C, tration, they were analysed by high-performance liquid chroma-
15 s at 53°C, and 45 s at 68°C. PCR products were analysed on a tography–mass spectrometry (HPLC-MS) using a ThermoDionex
1% agarose gel and were purified with MultiScreen-PCR96 filter Ultimate U3000 liquid chromatograph (ThermoScientific) cou-
plates (Millipore). Sequencing reactions were performed using Big pled to a Thermo Exactive high resolution mass spectrometer
Dye Terminator Sequencing kit v. 3.1 (Applied Biosystems) and (ThermoScientific). A 0.6 ml/min reverse-phase gradient (40%–95%)
analysed with an ABI 3730 DNA Analyzer (Applied Biosystems). flow between acetonitrile and water over 5 min was used to elute
Sequences were edited and aligned manually using SeqAssem fungal metabolites from a 50 × 2 mm C18 column (Kinetex XB-
(Hepperle, 2004). Isolates were identified to the species level C18, 2.6 µm particle size, 10 nm pore size; Phenomenex). HPLC-MS
using sequence similarity searches of the Fusarium MLST (https:// analyses were conducted in positive electrospray ionization mode to
fusar​
i um.mycob​
a nk.org/) and Fusarium ID (http://isola​
te.fusar​ detect mycotoxins. HPLC-MS comparisons of ion mass and elution
iumdb.org/blast.php) databases (Geiser et al., 2004; O’Donnell time with purified standards were used to identify the metabolites
et al., 2010). Species identifications were applied when query (Busman et al., 2012; Busman, 2017). Limits of quantitation for au-
sequences had at least 99% sequence similarity with reference rofusarin, BEA, enniatin B (ENNB), enniatin B1 (ENNB1), enniatin A
sequences. (ENNA), and enniatin A1 (ENNA1) were 1 ng/µl.

2.4 | Analysis of species composition and frequency 2.7 | Toxin production in liquid culture

Descriptive statistics summarized species composition and fre- F. poae strains were screened for trichothecenes, culmorin, and
quency between hosts and among years. The χ2 independence test butenolide production in a liquid medium that induces trichothecene
or Fisher's exact test (for small sample size) were used to evaluate production. Each strain was initially grown on V8 juice agar plates
differences in species composition in specific comparisons. All data (20% V8 juice, 0.3% CaCO3, 2% agar; Stevens, 1974). Twenty millili-
processing and analyses, as well as graphical work, were performed tres of agmatine media (30 g sucrose, 1.14 g agmatine, 1 g KH2PO 4,
running R v. 3.6.0 (2019-04-26; R Core Team, 2019). To encourage 0.5 g MgSO 4.7H2O, 0.5 g KCl, 10 mg FeSO 4.7H2O) and 200 μl of
and facilitate reproducibility, a website (https://emdel​ponte.github. trace element solution (per 100 ml: 5 g citric acid, 5 g ZnSO 4.7H2O,
io/paper​
-Fusar​
ium-PR/) was generated to navigate through the 0.25 g CuSO 4.5H2O, 50 mg MnSO 4.H2O, 50 mg H3BO3, 50 mg
documented code and all files are freely available and permanently NaMoO 4.2H2O) per 1 L distilled water (Gardiner et al., 2009) in
stored as an Open Science Framework project available at https:// 50 ml Erlenmeyer flasks were inoculated with two 0.5 cm2 plugs cut
osf.io/dzc9h/. from V8 plates. The cultures were grown at 28°C on a rotary shaker
at 200 rpm in the dark. After 7 days, each culture was transferred
to a 50 ml conical tube and extracted with 8 ml ethyl acetate with
2.5 | Phylogenetic analysis of FTSC isolates shaking for 30 min. The mixture was separated with centrifugation
(1,000  × g) and the top ethyl acetate layer was transferred to a 1
Phylogenetic analysis was conducted using TEF-1α sequences dram vial and dried under a nitrogen stream. The residue was then
from FTSC isolates identified in this study and 50 reference iso- resuspended in 1 ml ethyl acetate for gas chromatography-mass
lates (O’Donnell et al., 2012; Cerón-Bustamante et al., 2018; spectrometry (GC-MS).
Moreira et al., 2020). DNA sequences were edited with SeqAssem GC-MS analyses were performed with an Agilent 5873 fitted
(Hepperle, 2004) and aligned using the MUSCLE algorithm im- with an HP-5MS column (30 m, 0.25 mm, 0.25 µm), and detected
plemented in MEGA X (Kumar et al., 2018). Phylogenetic analysis with a mass spectrometer with an electron impact source. Helium
was also conducted in MEGA using maximum likelihood with the was used as the carrier gas with a 20:1 split ratio and a 20 ml/
K2  + G model of molecular evolution as determined by Bayesian min split flow. Samples were injected at 150°C, the temperature
information criterion (BIC) scores. Branch support was assessed held for 1 min, and then the column was heated at 30°C/min to
 |
4       PEREIRA et al.

280°C and held for 7.7 min. Individual peaks in chromatograms frequencies of the nine species/complexes and host (wheat or bar-
were examined for trichothecenes. Under these conditions, T-2 ley) (Fisher's exact test, p = .071).
toxin is detected at 10.6 min, neosolaniol at 8.5 min, 4,15-diANIV
at 8.1 min, 4,15-diacetoxyscirpenol (DAS) at 7.4 min, 15-ADON
at 7.1 min, and butenolide at 3.1 min. All chromatographic peaks 3.2 | Yearly frequency of Fusarium spp.
were examined and identified based on comparison of retention
time and ion fragmentation patterns with a commercial NIST li- The frequencies of the species/complexes of the two hosts in com-
brary and a library prepared with purified standards. Extracted bination were dependent on the year (Fisher's exact test, p < .001).
ion chromatograms were generated to screen for any components FGSC accounted for the majority of isolates in each of the five
with a prominent m/z 121 ion, characteristic of T-2 toxin, neoso- years; their frequencies ranged from 51.9% to 92.4% regardless of
laniol, and related compounds. The limit of detection was 0.25 µg the host. F. poae was present in all collections except wheat from
toxin/ml culture. 2011. Three distinct peaks in frequency of F. poae were observed
in isolates from wheat in 2013 (Figure 2a) and from barley in 2012
and 2013 (Figure 2b). A slight increase in FFSC was also observed in
3 |   R E S U LT S 2012 (Figure 2a,b). While FTSC was present in all years, FIESC oc-
curred more sporadically, but also peaked in 2012 in isolates from
3.1 | Species diversity both hosts, similar to FFSC (Figure 2). The rare species (others) were
collectively observed in four out of the five years.
A total of 754 isolates of Fusarium spp. were obtained from spikes
with symptoms. The number of isolates was similar between the
hosts (Figure 1a), but variable across years; the lowest sample 3.3 | FGSC composition and toxin genotypes
size was obtained from 2011 (n  = 54) and the highest from 2013
(n  = 295) (Figure 1b). Isolates were identified as members of nine Four species were identified in the subcollection of 571 FGSC iso-
species or species complexes (Figure 1c), among which FGSC ac- lates from both hosts. Among them, F. graminearum dominated the
counted for the majority of isolates and occurred at similar frequen- collection (63.3%) followed by F. meridionale (23.1%), F. cortaderiae
cies between the hosts (79.9% and 72.4% of the wheat and barley (9.6%), and F. austroamericanum (3.8%) (data not shown). Differences
strains, respectively). The second most prevalent species overall in their relative frequencies in wheat and barley were not statisti-
was F. poae, recovered more frequently from barley (14.5%) than cally significant (χ2 = 7.40, p = .06). For example, frequencies of F.
wheat (7.8%), followed by FTSC isolates found in similar frequency graminearum were 58% and 69% in barley and wheat, and of F. me-
in wheat (7.5%) and barley (8.5%) (Figure 1c). Members of the FIESC ridionale were 26% and 19% in barley and wheat, respectively.
and FFSC were each represented by around 2% of all isolates from The four species were found in all years regardless of the differ-
both hosts. The isolates of F. cerealis were obtained from wheat, and ence in sample size. Their frequencies, wheat and barley combined,
the single-isolate representatives of the F. oxysporum species com- varied among the years (Fisher's exact test, p  < .001). F. gramin-
plex (FOSC), F. subtropicale, and F. armeniacum were obtained from earum was dominant in most year × host combinations, but its rela-
barley (Figure 1c). No significant relationship was found between the tive frequency was slightly reduced in 2012 and 2013, years when

F I G U R E 1   Number of Fusarium spp.

isolates in relation to host of origin (a) and
cropping season (b) obtained from heads
showing symptoms of fusarium head
blight in wheat and barley fields sampled
from several fields in the central-southern
region of Paraná state, Brazil, over five
years. Isolates were identified as members
of nine species or species complexes using
DNA sequence-based methods (c)
PEREIRA et al. |
      5

F I G U R E 2   Relative yearly frequencies

of Fusarium species complex/species
in a collection of 754 isolates obtained
from heads of wheat (a) and barley (b)
conditioned host of origin. FFSC = F.
fujikuroi species complex; FGSC = F.
graminearum species complex; FIESC = F.
incarnatum-equiseti species complex;
FTSC = F. tricinctum species complex;
Others = F. subtropicale, F. armeniacum,
and FOSC (F. oxysporum species
complex)

F I G U R E 3   Number of isolates per species

of the Fusarium graminearum species complex
(FGSC) conditioned to year of sampling in
wheat (a) and barley (b)

F. meridionale peaked in both hosts but was more evident in barley 3.5 | Mycotoxin production by F. poae
(Figure 3a,b). A slight increase of F. meridionale was also noted in
2015 in barley (Figure 3b). Three toxins were found in isolates of F. poae (20 from barley and
A single toxin type was observed for all species except for F. cor- 12 from wheat): 29/32 produced DAS, 24/32 produced diANIV, and
taderiae isolates, which were either NIV (73%) or DON (27%). All F. 20/32 produced butenolide. Neosolaniol, T-2, or HT-2 were not de-
graminearum isolates were of the 15-ADON type, and all F. meridio- tected in cultures of any of the isolates.
nale and F. austroamericanum isolates were of the NIV type.

3.6 | Mycotoxin production by FTSC

3.4 | FTSC diversity
ENs, MON, and BEA were detected, but only the first two myco-
Comparison of TEF-1α sequences with reference sequences in the toxins could be quantified. The frequency and concentration levels
Fusarium MLST database suggested three FTSC species in the col- varied across the species.
lection of 61 isolates. The results of phylogenetic analysis were con-
sistent with the species identifications derived from Fusarium MLST.
Three well-supported clades (bootstrap  ≥  87%) were formed. The 3.6.1 | F. avenaceum
largest one included 51 F. avenaceum, 30 in wheat (Figure 4a) and 21
in barley (Figure 4b). The second in frequency was FTSC11 with nine All strains produced MON, but not all produced the different
isolates (five from wheat and four from barley). F. acuminatum was forms of ENs. Five out of 30 strains did not produce any form, and
represented by a single isolate collected in wheat in 2013. F. avena- 60% to 80% of the strains produced ENNA1, ENNB, and ENNB1
ceum was found in all but one season at a frequency ranging from 2 at median levels ranging from 0.94 to 15.28 ng/µl; ENNA1 was
to 10 isolates and FTSC11 was found in all but two years in barley detected at much lower concentration than ENNB and ENNB1
and all but one in wheat (Figure 4). (Table 1).
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6       PEREIRA et al.

F I G U R E 4   Number of isolates
belonging to two clades of the Fusarium
tricinctum species complex (FTSC)
conditioned to year of sampling in wheat
(a) and barley (b). A single F. acuminatum
isolate was reported in wheat in 2013

TA B L E 1   Summary for the mycotoxin

Mycotoxin concentration (ng/µl)
analysis data for 42 strains of the Fusarium
Speciesa  Mycotoxin Frequency Mean Median Min. Max. tricinctum species complex (FTSC) isolated
from heads with symptoms of fusarium
FTSC11 Enniatin A1 3 0.40 0.27 0.10 0.84
head blight collected from wheat and
Enniatin B 6 9.78 8.26 3.99 19.43 barley fields in central-southern Brazil
Enniatin B1 4 1.25 1.02 0.60 2.38 (Guarapuava, Paraná state) over five
growing seasons (2011 to 2015)
Moniliformin 9 3.46 0.23 0.00 9.10
F. acuminatum Enniatin A 1 7.76 — — —
Enniatin A1 1 76.38 — — —
Enniatin B 1 103.54 — — —
Enniatin B1 1 116.30 — — —
Moniliformin 1 3.05 — — —
F. avenaceum Enniatin A 1 0.02 — — —
Enniatin A1 20 3.57 0.94 0.15 22.46
Enniatin B 24 28.59 15.28 0.16 106.81
Enniatin B1 22 15.51 3.57 0.36 146.84
Moniliformin 30 4.21 2.87 0.00 17.50
a
FTSC11, nine strains; F. acuminatum, one strain; F. avenaceum, 30 strains.

3.6.2 | FTSC11 4 | D I S CU S S I O N

Similar to the other species, all produced MON. The frequency Results of this study considerably expand our knowledge of the di-
of enniatin-producing strains was similar to F. avenaceum, but versity of Fusarium species associated with FHB in wheat and barley
the concentration levels were much lower, especially for ENNB in Brazil, in particular for a major small grain production region in
and ENNB1 (Table 1). The maximum ENNB produced by one South America. The central-southern region of Paraná state has not
strain was similar to the median levels produced by F. avenaceum been well represented in previous surveys, that identified FGSC iso-
strains. lates collected mainly from northern and western Paraná state (Del
The only F. acuminatum strain produced MON at a concentration Ponte et al., 2015). The analysis of the total FHB diversity obtained
similar to the mean concentration levels of the other two species. from a multiseason collection in a new region confirmed FGSC as
All forms of enniatin were produced by this strain, including ENNA, the main FHB pathogen, but also revealed the presence of other
which was produced only by this strain and one F. avenaceum strain non-FGSC species commonly known to cause FHB in other wheat-
(trace levels). Finally, this F. acuminatum strain produced very high growing regions, such as F. avenaceum and related FTSC members,
amounts of ENNA compared to the other two species (>70-fold as well as F. poae.
higher than the median) and amounts of ENNB and ENNB1 similar to FTSC species are known to prevail in temperate, higher latitude
only two out of 30 F. avenaceum strains that produced these toxins climates such as northern Europe and Canada (Bottalico and Perrone,
in large quantities. 2002; Uhlig et al., 2007). Studies conducted at lower, still cooler,
PEREIRA et al. |
      7

latitudes have shown the presence of FTSC in wheat. In Kentucky, malting and brewing parameters (Nielsen et al., 2014). Evidence of
five of 68 strains from wheat belonged to two FTSC species (Bec the large diversity of species in this region may also be explained
et al., 2015). In Mexico, 75% of 80 toxigenic isolates from wheat by the occurrence of opportunistic pathogens at a relatively lower
in the central highlands (cooler climate) belonged to F. avenaceum frequency.
and a sister species (Cerón-Bustamante et al., 2018). In Brazil, two Our data suggest yearly shifts in species frequencies, especially
previous studies have reported FTSC in wheat and barley. A small a decrease of the dominant F. graminearum population in favour of
collection of FTSC strains was isolated from wheat with symptoms other contributors. In 2012, F. poae prevalence peaked in barley and
and from air above the canopy reproduced FHB symptoms in wheat F. meridionale peaked in both wheat and barley. The 2012 season
at severity levels similar to FGSC (Moreira et al., 2020). A recent sur- was relatively drier and the FHB incidence in the untreated plots of
vey also showed the presence of FTSC in commercial barley grain experiments was generally low (Feksa et al., 2019). We hypothesize
from southern Brazil (Piacentini et al., 2019). The concern with an that less-dominant pathogens may be more important in regions or
apparent emergence of FTSC among FHB pathogens relates to their years where weather is not favourable to F. graminearum outbreaks
toxigenic potential. In the Mexican study, most strains produced of FHB. In that context, understanding the overall spectrum of FHB
ENNB or chlamydosporol. These species may potentially produce pathogens and their mycotoxin potential may help inform toxin
MON, which is known to cause muscular weakness, cardiovascular monitoring programmes when conditions are less favourable for F.
problems, and reduced immune function, and also a reduced weight graminearum. The ecological or seasonal factors driving these fluc-
grain (Uhlig et al., 2007). Our study showed that most of the FTSC tuations should be further investigated, as this will have important
strains, regardless of the species, were able to produce MON. This implications for food safety. The influence of climate changes on the
is an important finding as data on grain contamination with this my- composition of species associated with FHB has been reported in
cotoxin in Brazil is only available for maize as a result of F. verticillioi- the literature (Parikka et al., 2012).
des infection (Oliveira et al., 2017). The in planta inoculation studies When restricting the analysis to FGSC strains, our data con-
conducted in cooler regions where F. avenaceum is more common, firmed (a) the dominance of F. graminearum of the 15-ADON type,
such as Canada and Poland, have shown the ability of this group to followed by F. meridionale, of the NIV type at a frequency similar
produce mycotoxin in barley and wheat grains (Goliński et al., 1997; to the other regions of Paraná state; and (b) the minor role of NIV-
Abramson et al., 2002). producing isolates of F. austroamericanum and F. cortaderiae in FHB
Another known FHB-causing species reported for the first epidemics. We confirmed the role of pathogen species in shaping
time at relatively high frequency in Brazil was F. poae, which rep- the composition of the trichothecene genotypes in Brazil (Astolfi
resented 11% of the collection. F. poae is known to be a typical et al., 2012; Del Ponte et al., 2015; Gomes et al., 2015; Kuhnem
type B trichothecenes producer (NIV), but has also been known to et al., 2016). Despite analysing close to 2,000 isolates from dif-
produce type A trichothecene such as DAS (Vanheule et al., 2017; ferent hosts, no chemotype polymorphism within F. graminearum
O’Donnell et al., 2018). In the Americas, F. poae isolates have been from Brazil (exclusively 15-ADON) has been found thus far. Also,
found in barley and wheat from Argentina and Canada (Bourdages the study did not find the F. graminearum NX-2 genotype. A pre-
et al., 2006; Stenglein et al., 2012), wheat in Paraguay (Arrúa et al., vious study that included isolates from Brazil (n = 165) as well as
2019), and barley from Uruguay (Garmendia et al., 2018). In Europe, Argentina and Uruguay also did not find the NX-2 genotype (Kelly
reports exist in Germany, Italy, and other countries (Birzele et al., et al., 2016).
2002; Xu et al., 2005; Covarelli et al., 2015). Identification of F. The 3-ADON type in F. graminearum was reported for sev-
poae strains based only on morphological characteristics is unre- eral strains in Argentina and for one isolate from Uruguay
liable and may lead to inaccurate conclusions regarding the toxin (Alvarez et al., 2009; Umpiérrez-Failache et al., 2013). In Brazil,
production potential for this species. The species F. langsethiae the 3-ADON chemotype is unique to F. cortaderiae and F. aus-
is similar morphologically to F. poae; however, it is more closely troamericanum strains from wheat (Del Ponte et al., 2015) and
related to F. sporotrichioides than F. poae (Torp and Nirenberg, barley (Astolfi et al., 2011; Castañares et al., 2016). This pattern
2004), and synthesizes the trichothecenes T-2 and HT-2, toxins seems unique compared with other intensively sampled wheat
not usually reported for F. poae. Therefore, DNA sequence-based regions of North America, Europe, and China. There, the domi-
identifications are required, and PCR assays may provide a rapid nant species, either F. graminearum or F. asiaticum, possess more
and reliable alternative to distinguish between these two species than one chemotype that alternate dominance depending on
(Stenglein, 2009). the region, previous crop, or year. For example, 15-ADON and
DON and NIV have multiple adverse effects at the molecu- 3-ADON strains of F. graminearum have been found in Europe
lar level, such as inhibition of protein, DNA, and RNA synthesis, where the former is more common in southern and central (Tóth
with NIV representing an added concern for food safety, because et al., 2005; Boutigny et al., 2014) and the latter in north-west-
several reports have shown its toxicity is higher than DON (Cheat ern Europe (Yli-Mattila et al., 2009; Fredlund et al., 2013). In
et al., 2015). The presence of NIV-producing F. poae, in addition to China, F. asiaticum of the 3-ADON type has been replacing
F. meridionale, as FHB pathogens in Brazil may represent an addi- the NIV-producing populations of this species (Zhang et al.,
tional risk for wheat and barley, and could have negative effects on 2012). In Japan, 3-ADON and NIV chemotypes from F. asiaticum
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8       PEREIRA et al.

predominate in wheat (Karugia et al., 2009). In North America, REFERENCES

the 15-ADON chemotype is predominant in F. graminearum, but Abramson, D., McCallum, B., Smith, D.M. and Tekauz, A. (2002)
a high frequency of the 3-ADON chemotype isolates has been Moniliformin in barley inoculated with Fusarium avenaceum. Food
Additives & Contaminants, 19, 765–769.
found in some parts of USA and Canada (Gale et al., 2007; Ward
Alvarez, C.L., Azcarate, M.P. and Pinto, V.F. (2009) Toxigenic potential of
et al., 2008) and NIV and NX-2 strains of F. graminearum have Fusarium graminearum sensu stricto isolates from wheat in Argentina.
been reported at relatively low levels (Gale et al., 2011; Kelly International Journal of Food Microbiology, 135, 131–135.
et al., 2016). Finally, we found that NIV-producing FGSC spe- Aoki, T., Vaughan, M.M., McCormick, S.P., Busman, M., Ward, T.J., Kelly,
A. et al. (2015) Fusarium dactylidis sp. nov., a novel nivalenol tox-
cies were more prevalent in barley than in wheat, which is in
in-producing species sister to F. pseudograminearum isolated from
agreement with previous studies of isolates from these crops in orchard grass (Dactylis glomerata) in Oregon and New Zealand.
RS state (Astolfi et al., 2012; Del Ponte et al., 2015; Machado Mycologia, 107, 409–418.
et al., 2017). Arrúa, A.A.A., Arrúa, J.M.M., Caza, M.C.C., Iehisa, J., Caballero, Y.M.R.,
Ríos, D.F. et al. (2019) First report of Fusarium poae associated with
In summary, using state of the art methods of Fusarium spe-
fusarium head blight in wheat in Paraguay. Plant Disease, 103, 580.
cies identification, we report a large diversity of toxigenic species Astolfi, P., dos Santos, J., Schneider, L., Gomes, L.B., Silva, C.N.,
associated with FHB in wheat and barley in a major production Tessmann, D.J. et al. (2011) Molecular survey of trichothecene
region of southern Brazil. F. graminearum remains the most domi- genotypes of Fusarium graminearum species complex from barley
nant in wheat and barley, but FTSC and F. poae are reported here in southern Brazil. International Journal of Food Microbiology, 148,
197–201.
for the first time as significant contributors to FHB in Brazil and
Astolfi, P., Reynoso, M.M., Ramirez, M.L., Chulze, S.N., Alves, T.C.A.,
may be considered emergent pathogens. In addition, but at much Tessmann, D.J. et al. (2012) Genetic population structure and tricho-
lower levels, we report FFSC, FOSC, FIESC, F. cerealis, and a new thecene genotypes of Fusarium graminearum isolated from wheat in
species within the FSAMSC, namely F. subtropicale (Pereira et al., southern Brazil. Plant Pathology, 61, 289–295.
Bec, S., Ward, T., Farman, M., O’Donnell, K., Hershman, D., Van Sanford,
2018). The significant temporal changes in FHB species compo-
D. et al. (2015) Characterization of Fusarium strains recovered from
sition could have direct impacts on the type of mycotoxins that wheat with symptoms of head blight in Kentucky. Plant Disease, 99,
accumulate in grain. Future surveys should focus on mycotoxins 1622–1632.
potentially produced by the entire spectrum of Fusarium species Birzele, B., Meier, A., Hindorf, H., Kramer, J. and Dehne, H.W. (2002)
Epidemiology of Fusarium infection and deoxynivalenol content in
diversity reported here. The continuous monitoring of the popu-
winter wheat in the Rhineland, Germany. European Journal of Plant
lation may be informative to clarify the reasons for the temporal Pathology, 108, 667–673.
variation of the population in a certain year. Bottalico, A. and Perrone, G. (2002) Toxigenic Fusarium species and my-
cotoxins associated with head blight in small-grain cereals in Europe.
European Journal of Plant Pathology, 108, 611–624.
AC K N OW L E D G E M E N T S
Bourdages, J.V., Marchand, S., Rioux, S. and Belzile, F.J. (2006) Diversity
This study was financed in part by the Coordenação de and prevalence of Fusarium species from Quebec barley fields.
Aperfeiçoamento de Pessoal de Nível Superior – Brazil (CAPES) – Canadian Journal of Plant Pathology, 28, 419–425.
finance code 001, for a scholarship for the first author, and also Boutigny, A.L., Ward, T.J., Ballois, N., Iancu, G. and Ioos, R. (2014)
Diversity of the Fusarium graminearum species complex on French
by the Fundação Agrária de Pesquisa Agropecuária (FAPA),
cereals. European Journal of Plant Pathology, 138, 133–148.
Cooperativa Agrária Agroindustrial, and the Brazilian National Busman, M. (2017) Utilization of high performance liquid chromatog-
Council for Scientific and Technological Development (CNPQ pro- raphy coupled to tandem mass spectrometry for characterization
ject 310719/2016-6). Funding was also provided by the USDA-ARS of 8-O-methylbostrycoidin production by species of the fungus
National Program for Food Safety. We also thank Nathan Orwig, Fusarium. Journal of Fungi, 3, E43.
Busman, M., Butchko, R.A. and Proctor, R.H. (2012) LC-MS/MS method
Thomas Usgaard, Amy Kelly, and Amy McGovern for excellent assis-
for the determination of the fungal pigment bikaverin in maize ker-
tance. Mention of trade names or commercial products in this article nels as an indicator of ear rot. Food Additives and Contaminants Part
is solely for the purpose of providing specific information and does A, 29, 1736–1742.
not imply recommendation or endorsement by the United States Castañares, E., Dinolfo, M.I., Del Ponte, E.M., Pan, D. and Stenglein,
S.A. (2016) Species composition and genetic structure of Fusarium
Department of Agriculture (USDA). USDA is an equal opportunity
graminearum species complex populations affecting the main
provider and employer. We declare that we have no conflicts of in- barley growing regions of South America. Plant Pathology, 65,
terest to this work. 930–939.
Cerón-Bustamante, M., Ward, T.J., Kelly, A., Vaughan, M.M., McCormick,
S.P., Cowger, C. et al. (2018) Regional differences in the composi-
DATA AVA I L A B I L I T Y S TAT E M E N T
tion of fusarium head blight pathogens and mycotoxins associated
The data that support the findings of this study are freely available with wheat in Mexico. International Journal of Food Microbiology, 273,
and permanently stored as an Open Science Framework project 11–19.
available at https://osf.io/dzc9h/. Cheat, S., Gerez, J.R., Cognie, J., Alassane-Kpembi, I., Bracarense, A.P.,
Raymond-Letron, I. et al. (2015) Nivalenol has a greater impact than
deoxynivalenol on pig jejunum mucosa in vitro on explants and in
ORCID vivo on intestinal loops. Toxins, 7, 1945–1961.
Emerson M. Del Ponte  https://orcid.org/0000-0003-4398-409X Covarelli, L., Beccari, G., Prodi, A., Generotti, S., Etruschi, F., Juan, C.
Dauri J. Tessmann  https://orcid.org/0000-0001-7193-1783 et al. (2015) Fusarium species, chemotype characterization and
PEREIRA et al. |
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trichothecene contamination of durum and soft wheat in an area Kuhnem, P.R., Ward, T.J., Silva, C.N., Spolti, P., Ciliato, M.L., Tessmann,
of central Italy. Journal of the Science of Food and Agriculture, 95, D.J. et al. (2016) Composition and toxigenic potential of the Fusarium
540–551. graminearum species complex from maize ears, stalks and stubble in
Del Ponte, E.M., Fernandes, J.M.C. and Bergstrom, G.C. (2007) Influence Brazil. Plant Pathology, 65, 1185–1191.
of growth stage on Fusarium head blight and deoxynivalenol produc- Kumar, S., Stecher, G., Li, M., Knyaz, C. and Tamura, K. (2018) MEGA
tion in wheat. Journal of Phytopathology, 155, 577–581. X: molecular evolutionary genetics analysis across computing plat-
Del Ponte, E.M., Spolti, P., Ward, T.J., Gomes, L.B., Nicolli, C.P., Kuhnem, forms. Molecular Biology and Evolution, 35, 1547–1549.
P.R. et al. (2015) Regional and field-specific factors affect the compo- Leslie, J.F. and Summerell, B.A. (2006) The Fusarium Laboratory Manual.
sition of Fusarium head blight pathogens in subtropical no-till wheat Ames, IA: Blackwell Publishing.
agroecosystem of Brazil. Phytopathology, 105, 246–254. Luz, C., Saladino, F., Luciano, F.B., Mañes, J. and Meca, G. (2017)
Feksa, H.R., Do Couto, H.T.Z., Garozi, R., De Almeida, J.L., Gardiano, Occurrence, toxicity, bioaccessibility and mitigation strategies
C.G. and Tessmann, D.J. (2019) Pre- and post-infection application of beauvericin, a minor Fusarium mycotoxin. Food and Chemical
of strobilurin-triazole premixes and single fungicides for control of Toxicology, 107, 430–439.
fusarium head blight and deoxynivalenol mycotoxin in wheat. Crop Machado, F.J., Nicolli, C.P., Möller, P.A., Arruda, R., Ward, T.J. and Del
Protection, 117, 128–134. Ponte, E.M. (2017) Differential triazole sensitivity among members
Fredlund, E., Gidlund, A., Sulyok, M., Börjesson, T., Krska, R., Olsen, M. of the Fusarium graminearum species complex infecting barley grains
et al. (2013) Deoxynivalenol and other selected Fusarium toxins in in Brazil. Tropical Plant Pathology, 42, 197–202.
Swedish oats—occurrence and correlation to specific Fusarium spe- McMullen, M., Bergstrom, G., De Wolf, E., Dill-Macky, R., Hershman,
cies. International Journal of Food Microbiology, 167, 276–283. D., Shaner, G. et al. (2012) A unified effort to fight an enemy
Gale, L.R., Harrison, S.A., Ward, T.J., O’Donnell, K., Milus, E.A., Gale, of wheat and barley: Fusarium head blight. Plant Disease, 96,
S.W. et al. (2011) Nivalenol-type populations of Fusarium gramin- 1712–1728.
earum and F. asiaticum are prevalent on wheat in southern Louisiana. Moreira, G.M., Machado, F.J., Pereira, C.B., Neves, D.L., Tessmann, D.J.
Phytopathology, 101, 124–134. et al. (2020) First report of the Fusarium tricinctum species complex
Gale, L.R., Ward, T.J., Balmas, V. and Kistler, H.C. (2007) Population causing fusarium head blight of wheat in Brazil. Plant Disease, 104, 586.
subdivision of Fusarium graminearum sensu stricto in the upper Nielsen, L.K., Cook, D.J., Edwards, S.G. and Ray, R.V. (2014) The prev-
Midwestern United States. Phytopathology, 97, 1434–1439. alence and impact of Fusarium head blight pathogens and myco-
Gardiner, D.M., Kazan, K. and Manners, J.M. (2009) Nutrient profiling toxins on malting barley quality in UK. International Journal of Food
reveals potent inducers of trichothecene biosynthesis in Fusarium Microbiology, 179, 38–49.
graminearum. Fungal Genetics and Biology, 46, 604–613. O’Donnell, K., Cigelnik, E. and Nirenberg, H.I. (1998) Molecular sys-
Garmendia, G., Pattarino, L., Negrín, C., Martínez-Silveira, A., Pereyra, S., tematics and phylogeography of the Gibberella fujikuroi species com-
Ward, T.J. et al. (2018) Species composition, toxigenic potential and plex. Mycologia, 90, 465–493.
aggressiveness of Fusarium isolates causing head blight of barley in O’Donnell, K., Humber, R.A., Geiser, D.M., Kang, S., Park, B., Robert,
Uruguay. Food Microbiology, 76, 426–433. V.A.R.G. et al. (2012) Phylogenetic diversity of insecticolous fusaria
Geiser, D.M., del Mar Jiménez-Gasco, M., Kang, S., Makalowska, I., inferred from multilocus DNA sequence data and their molecular
Veeraraghavan, N., Ward, T.J. et al. (2004) FUSARIUM-ID v. 1.0: a identification via FUSARIUM-ID and Fusarium MLST. Mycologia, 104,
DNA sequence database for identifying Fusarium. European Journal 427–445.
of Plant Pathology, 110, 473–479. O’Donnell, K., McCormick, S.P., Busman, M., Proctor, R.H., Ward, T.J.,
Goliński, P., Kostecki, M., Kaptur, P., Wojciechowski, S., Kaczmarek, Z., Doehring, G. et al. (2018). Marasas et al. 1984 “Toxigenic Fusarium
Wiśniewska, H. et al. (1997) Fusarium head blight and moniliformin species: identity and mycotoxicology” revisited. Mycologia, 110, 1–23.
accumulation in kernels of 18 winter wheat cultivars inoculated with O’Donnell, K., Rooney, A.P., Proctor, R.H., Brown, D.W., McCormick,
Fusarium avenaceum (3 years study). Cereal Research Communications, S.P., Ward, T.J. et al. (2013) Phylogenetic analyses of RPB1 and RPB2
25, 673–675. support a middle Cretaceous origin for a clade comprising all agricul-
Gomes, L., Ward, T., Badiale-Furlong, E. and Del Ponte, E. (2015) Species turally and medically important fusaria. Fungal Genetics and Biology,
composition, toxigenic potential and pathogenicity of Fusarium gram- 52, 20–31.
inearum species complex isolates from southern Brazilian rice. Plant O’Donnell, K., Sutton, D.A., Rinaldi, M.G., Sarver, B.A.J., Balaje, S.A.,
Pathology, 64, 980–987. Summerell, R.C. et al. (2010) Internet-accessible DNA sequence
Hepperle, D. (2004) SeqAssem. A sequence analysis tool, contig as- database for identifying fusaria from human and animal infections.
sembler and trace data visualization tool for molecular sequences. Journal of Clinical Microbiology, 48, 3708–3718.
Win32-Version. Distributed by the author via: http://www.seque​ Oliveira, M.S., Rocha, A., Sulyok, M., Krska, R. and Mallmann, C.A. (2017)
ntix.de [Accessed 15 September 2020] Natural mycotoxin contamination of maize (Zea mays L.) in the South
Jestoi, M. (2008) Emerging Fusarium-mycotoxins fusaproliferin, beau- region of Brazil. Food Control, 73, 127–132.
vericin, enniatins, and moniliformin – a review. Critical Reviews in Parikka, P., Hakala, K. and Tiilikkala, K. (2012) Expected shifts in Fusarium
Food Science and Nutrition, 48, 21–49. species’ composition on cereal grain in Northern Europe due to cli-
Karugia, G.W., Suga, H., Gale, L.R., Nakajima, T., Ueda, A. and matic change. Food Additives & Contaminants: Part A, 29, 1543–1555.
Hyakumachi, M. (2009) Population structure of Fusarium asiaticum Pereira, C.B., Ward, T.J., Tessmann, D.J., Del Ponte, E.M., Laraba, I.,
from two Japanese regions and eastern China. Journal of General Vaughan, M.M. et al. (2018) Fusarium subtropicale, sp. nov., a novel ni-
Plant Pathology, 75, 110–118. valenol mycotoxin-producing species isolated from barley (Hordeum
Kelly, A.C., Proctor, R.H., Belzile, F., Chulze, S.N., Clear, R.M., Cowger, C. vulgare) in Brazil and sister to F. praegraminearum. Mycologia, 10, 1–12.
et al. (2016) The geographic distribution and complex evolutionary Piacentini, K.C., Rocha, L.O., Savi, G.D., Carnielli-Queiroz, L., Fontes, L.C.
history of the NX-2 trichothecene chemotype from Fusarium gramin- and Correa, B. (2019) Assessment of toxigenic Fusarium species and
earum. Fungal Genetics and Biology, 95, 39–48. their mycotoxins in brewing barley grains. Toxins, 11, 31.
Kuhnem, P.R., Del Ponte, E.M., Dong, Y. and Bergstrom, G.C. (2015) R Core Team (2019) R: A language and environment for statistical com-
Fusarium graminearum isolates from wheat and maize in New York puting. R Foundation for Statistical Computing, Vienna, Austria.
show similar range of aggressiveness and toxigenicity in cross-spe- Available at: https://www.R-proje​c t.org/ [Accessed 15 September
cies pathogenicity tests. Phytopathology, 105, 441–448. 2020]
 |
10       PEREIRA et al.

Savard, M.E. & Blackwell, B.A. (1994) Spectral characteristic of second- Varga, E., Wiesenberger, G., Hametner, C., Ward, T.J., Dong, Y.,
ary metabolites from Fusarium fungi. In: Miller, J.D. and Trenholm, Schöfbeck, D. et al. (2015) New tricks of an old enemy: isolates of
H.L. (Eds.) Mycotoxins in Grain: Compounds Other Than Aflatoxin. St Fusarium graminearum produce a type A trichothecene mycotoxin.
Paul, MN: Eagan Press, pp. 59–260. Environmental Microbiology, 17, 2588–2600.
Scoz, L.B., Astolfi, P., Reartes, D.S., Schmale, D.G.I.I.I., Moraes, M.G. Ward, T.J., Clear, R.M., Rooney, A.P., O'Donnell, K., Gaba, D., Patrick, S.
and Del Ponte, E.M. (2009) Trichothecene mycotoxin genotypes of et al. (2008) An adaptive evolutionary shift in Fusarium head blight
Fusarium graminearum sensu stricto and Fusarium meridionale in wheat pathogen populations is driving the rapid spread of more toxigenic
from southern Brazil. Plant Pathology, 58, 344–351. Fusarium graminearum in North America. Fungal Genetics and Biology,
Stenglein, S.A. (2009) Fusarium poae: a pathogen that needs more atten- 45, 473–484.
tion. Journal of Plant Pathology, 91, 25–36. Xu, X., Parry, D., Nicholson, P., Thomsett, M., Simpson, D., Edwards, S.
Stenglein, S.A., Dinolfo, M.I., Bongiorno, F. and Moreno, M.V. (2012) et al. (2005) Predominance and association of pathogenic fungi caus-
Response of wheat (Triticum spp.) and barley (Hordeum vulgare) to ing Fusarium ear blight in wheat in four European countries. European
Fusarium poae. Agrociencia, 46, 299–306. Journal of Plant Pathology, 112, 143–154.
Stevens, R.R. (1974) Mycology Guidebook. Seattle, WA: University of Yli-Mattila, T., Gagkaeva, T., Ward, T.J., Aoki, T., Kistler, H.C. and
Washington Press. O’Donnell, K. (2009) A novel Asian clade within the Fusarium
Torp, M. and Nirenberg, H.I. (2004) Fusarium langsethiae sp. nov. on cere- graminearum species complex includes a newly discovered cereal
als in Europe. International Journal of Food Microbiology, 95, 247–256. head blight pathogen from the Russian Far East. Mycologia, 101,
Tóth, B., Mesterházy, Á., Horváth, Z., Bartók, T., Varga, M. and Varga, 841–852.
J. (2005) Genetic variability of central European isolates of the Zhang, H., Van der Lee, T., Waalwijk, C., Chen, W.Q., Xu, J., Xu, J.S. et al.
Fusarium graminearum species complex. European Journal of Plant (2012) Population analysis of the Fusarium graminearum species com-
Pathology, 113, 35–45. plex from wheat in China show a shift to more aggressive isolates.
Uhlig, S., Jestoi, M. and Parikka, P. (2007) Fusarium avenaceum—the North PLoS One, 7, e31722.
European situation. International Journal of Food Microbiology, 119, 17–24.
Umpiérrez-Failache, M., Garmendia, G., Pereyra, S., Rodríguez
Haralambides, A., Ward, T.J. and Vero, S. (2013) Regional differences
How to cite this article: Pereira CB, Ward TJ, Del Ponte EM,
in species composition and toxigenic potential among Fusarium head
blight isolates from Uruguay indicate a risk of nivalenol contami-
et al. Five-year survey uncovers extensive diversity and
nation in new wheat production areas. International Journal of Food temporal fluctuations among fusarium head blight pathogens
Microbiology, 166, 135–140. of wheat and barley in Brazil. Plant Pathol. 2020;00:1–10.
Vanheule, A., De Boevre, M., Moretti, A., Scauflaire, J., Munaut, F., De https://doi.org/10.1111/ppa.13289
Saeger, S. et al. (2017) Genetic divergence and chemotype diversity
in the Fusarium head blight pathogen Fusarium poae. Toxins, 9, 255.