Zearalenone-producing species of Fusarium on barley seed

Isolates of Fusarium were obtained from samples of barley grain at harvest in 1971, 1973 and 1974. The ability of these isolates to produce the myco-toxin zearalenone was examined, the grain also being tested for its presence in 1974. The most common isolate was F. culmorum, 60% of which in 1974 produced zearalenone. Other species isolated which produced this myco-toxin were F. monilijorme, F. graminearum, F. avenaceum, F. nivale and F. sambucinum var coeruleum, the last three species have not previously been recorded as producing zearalenone.     

Occurrence of toxigenic strains of Fusarium in maize and barley in Germany

Summary A survey was made of maize and barley in Germany for the occurrence of toxigenic strains of Fusarium and of the mycotoxins produced in culture by these strains.The following 6 species of Fusarium were found: F. avenaceum, F. culmorum, F. equiseti, F.oxysporum, F. poae, and F. tricinctum. The species most commonly isolated from bird-damaged maize ears was F. avenaceum while F. culmorum was consistently isolated from maize stem rot. The predominant species in barley grain was F. poae while F. avenaceum, F. culmorum, and F. tricinctum were also isolated frequently.Cultures on autoclaved maize of all the Fusarium strains were assayed for toxicity by feeding to 1-day-old chickens for 14 days. Some strains of F. avenaceum, F. culmorum, F. equiseti, and F. oxysporum proved to be acutely toxic to chickens and caused mortality as well as marked reductions in weight gain and feed consumption. All the strains of F. poae and F. tricinctum had a low degree of toxicity.Culture material of all the strains were analyzed for the presence of 11 known Fusarium mycotoxins. The following 4 mycotoxins were detected in the strains examined: moniliformin in 9 out of 9 F. avenaceum strains (2 to 760 ppm) and in the single strain of F. oxysporum (1150 ppm); zearalenone in 4 out of 5 F. culmorum strains (320 to 1400 ppm); deoxynivalenol in 3 out of 5 F. culmorum strains.(1 to 15 ppm); and acetyldeoxynivalenol (1 to 2 ppm) in 3 out of 5 F. culmorum strains. This is the first report of moniliformin production by F. avenaceum and F. oxysporum and also the first report of the occurrence of moniliformin-producing Fusarium strains in Europe.     

Zearalenone formation by<Emphasis Type="Underline">Fusarium crookwellense</Emphasis> /Burgess,Nelson and Toussoun/ isolates from Poland and their pathogenicity towards cereals

Fusarium crookwellense /B.N. and T./ isolated from affected cereals in Poland formed zearalenone on wheat grain up to 602 mg/kg. Tested isolates have been found strong to severe pathogens of wheat, rye, triticale and barley seedlings and corn ears with pathogenicity similar to that ofF. culmorum andF. graminearum.     

Toxicity and Occurrence of Fusarium species and Mycotoxins in Late Harvested and Overwintered Grain from Norway, 1993

Samples of grain, harvested in October/November of 1993 or in the spring of 1994 from fields in Norway with overwintered grain, were collected. The presence of Fusarium species and the amount of mycotoxins produced by this genus were determined. The cytotoxic properties of the grain samples were examined with an in vitro methylthiazoltetrazolium (MTT)-cell culture assay using swine kidney and VERO cells as target cells. The total count of colony forming units of Fusarium species was about the same in the grain harvested in late autumn and in the spring, but the dominant species seemed to vary somewhat between the two groups. F. culmorum was found more in the samples from October/November while F. avenaceum was isolated more in the grain harvested in the spring. Only small amounts of F. sporotrichioides were present in the grain harvested in the autumn, and none was found in the overwintered grain. The deoxynivalenol content was significantly higher in the grain harvested in autumn than in the overwintered grain. Although very small amounts of toxins were detected in the overwintered grain, it was more cytotoxic than the grain harvested in October/November. A significant correlation between the cytotoxicity and the amount of F. avenaceum in the samples was found.     

Pathogenicity of Fusarium Isolates from Wheat, Rye and Triticale Towards Seedlings and their Ability to Produce Trichothecenes and Zearalenone

37 Fusarium isolates (F. culmorum 20, F. graminearum 4, F. avenaceum 3, F. solani 4, and F. equiseti 6 from wheat, rye and triticale) were examined for pathogenicity and tested for zearalenone (F-2) and trichothecenes production in vitro. Strong pathogens (F. culmorum and F. graminearum) produced deoxynivalenol and 3-acetyl-deoxynivalenol and zearalenone in considerable quantities.     

Toxin production by Fusarium culmorum IMI 309344 and F. graminearum NRRL 5883 on grain substrates

The production of deoxynivalenol, acetyl deoxynivalenol and zearalenone by Fusarium culmorum and F. graminearum on autoclave-sterilized grain (maize, rice, wheat and barley) was investigated. Fusarium culmorum produced significantly greater levels of toxins than F. graminearum. The four substrates examined differed in their ability to support toxin production. Toxin production on maize and rice was significantly greater than toxin production on barley or wheat.     

Effects of infection time and moisture on development of ear blight and deoxynivalenol production by Fusarium spp. in wheat

Wheat ears were inoculated with conidia of Fusarium spp. at different growth stages between ear emergence and harvest and moist conditions were maintained for up to 7 days subsequently by mist irrigation. Of the fungi tested (Fusarium culmorum, F. avenaceum, F. tricinctum, F. sporotrichioides and Microdochium nivale), only F. culmorum produced ear blight symptoms and grain samples were found subsequently to contain deoxynivalenol. Most ear infection and deoxynivalenol formation occurred following inoculation at about mid-anthesis. Small amounts of deoxynivalenol were formed and some F. culmorum was isolated even in the absence of ear blight symptoms. An overnight wet period was sufficient to initiate infection and deoxynivalenol formation but both were increased by extending the wet period up to at least 3 days. Recovery of Fusarium spp. from harvested grain was usually possible whether or not symptoms developed. F. culmorum usually persisted and often increased to moderately high levels after storage for 7 wk in a range of moisture conditions.     

Susceptibility of Barley Cultivars and Lines to Fusarium Infection and Mycotoxin Accumulation in Kernels

Heads of 12 barley genotypes (8 cultivars and 4 lines) were inoculated with conidial suspension of the following single isolates: F. culmorum no. 3, F. graminearum no. 122 and F. sporotrichioides no. ATCC 62 360. The number of kernels per head. 1000 Kernel weight and yield have been calculated for each genotype. Seed samples collected at harvest were analysed for each genotype. Seed samples collected at harvest were analysed for several trichothecene mycotoxins and zearalenone.The mycotoxin concentrations (mg/kg) in barley kernels inoculated with F. graminearum were as follows. deoxynivalenol (DON) 0.1 to 5.4 (av. 2.3). 3-acetyldeoxy-nivalenol (3-AcDON) 0.0–0.2 (av. 0.1), 15-acetyldeoxynivalenol (15-AcDON) 0.0–0.7 (av.0.2), nivalenol (NIV) 0.0–0.8 (av. 0.3). zearalenone (ZEA) 0.0–0.1 (av. 0.0); F. culmorum: DON 0.6 to 12.0 (av. 5.3), 3-AcDON 0.1 to 1.0 (av. 0.6). 15-AcDON nd. NIV 0.1–0.7 (av. 0.3). ZEA 0.1–0.5 (av. 0.2). F. sporotrichioides T-2 toxin 2.4–13.9 (av. 6.0), HT-2-toxin 0.1–0.8 (av.0.3) and neosolaniol 0.2–1.5 (av.0.7).     

The occurrence of culmorin and hydroxy-culmorins in cereals

Forty-five samples from 1988–1995 of naturally contaminated grain, barley, wheat and oats, three samples of mixed feed, and 16 samples of grain artificially inoculated with Fusarium culmorum during the flowering stage were analysed for deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-acetyl-DON), culmorin and hydroxy-culmorins. These compounds are secondary metabolites produced by the fungal species F. culmorum and F. graminearum. Acetonitrile-water extract of the samples was purified on a Mycosep^TM#225 column, derivetized using pentafluoropropionic anhydride (PFPA) and analysed by gas chromatography-mass spectrometry (GC-MS). The amount of each of culmorin, 5-, 12-, 14 and 15-hydroxy-culmorin and one unknown hydroxy-culmorin were determined relative to the amount of DON plus 3-acetyl DON for each sample. The ratio between the total amount of culmorin compounds and the DON compounds ranged from 0.14 to 1.07 in the samples. This study shows that there is a strong correlation between the amount of DON present in the grain and the amount of culmorin and hydroxy-culmorins present. The ratio of each of the culmorin compounds relative to the amount of DON compounds were in the same range in the grain artificially inoculated by F. culmorum as found in an earlier study for F. culmorum strains cultivated on rice, while the hydroxy-culmorin profile in the naturally contaminated grain was more similar to what was found for the F. graminearum cultures in the same study [1]. These results indicate that F. graminearum may be a relatively important source for DON in grain also in relatively cold areas. This revised version was published online in June 2006 with corrections to the Cover Date.     

Screening of zearalenone-producing Fusarium species in Egypt and chemically defined medium for production of the toxin

296 isolates of Fusarium spp. from 100 samples of cereale grains were examined for their ability to produce zearalenone on liquid culture medium. Thin layer Chromatographic analysis revealed that the mycotoxin was detected from 45 isolates, (F. oxysporum, 36; F. moniliforme, 8; and F. equiseti, one isolate). A suitable liquid medium and some optimal conditions for the biosynthesis of zearalenone were reported. Glutamine and riboflavin stimulated the production of the toxin. The maximum amount of zearalenone appeared at pH 7, after 12 days of incubation at 20 °C.     

Fusarium crookwellense,a newly isolated fungus from wheat in Japan: Its mycotoxin production and pathogenicity to wheat and barley

Fusarium crookwellense was isolated for the first time in Japan from scabby wheat harvested in Hokkaido in 1991. Four isolates were obtained and examined for their mycological features on culture media, mycotoxin production, and pathogenicity to wheat and barley. The texture of fungal colonies and the morphology of macroconidia on a potato dextrose agar medium were similar to those ofFusarium graminearum. AllF. crookwellense isolates examined produced nivalenol, 4-acetylnivalenol, and zearalenone on rice media at levels ranging from 0.9 to 22.5 µg/g, 0.5 to 25.0 µg/g, and 1.4 to 162.5 µg/g, respectively. All were pathogenic toward the wheat and barley strains tested, and scab symptoms were found on the heads and leaves of plants. This is the first report on domestic isolates ofF. crookwellense from the crop field in Japan.     

Use of Rep‐PCR for Genetic Diversity Analyses in Fusarium culmorum

Fusarium culmorum is a pathogen of economically important grain crops. In this work, Rep‐PCR was used to identify genetic diversity in F. culmorum isolates which have been collected from wheat fields in Turkey. Reproducible genomic fingerprints were amplified in each strain by PCRs of prokaryotic repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC) and BOX sequences. Totally 104 molecular markers were evaluated and similarity comparisons were shown as a dendrogram. The average genetic diversity was 52.3% ranging from 15.8% to 88.7% according to the Rep‐PCR data. Cluster analysis showed agreement with the distance of sampling locations. The highest genetic similarity (84.2%) was determined between two F. culmorum isolates (F1 and F2) originated from the same agro‐ecological region. Our results showed that Rep‐PCR is convenient and rapid for genetic diversity analyses and strain differentiation in F. culmorum.     

Colonization of barley roots by Fusarium culmorum and influence of Pseudomonas fluorescens on the process

The stages of barley root colonization by Fusarium culmorum were studied in sterile vermiculite by the method of fluorescent antibodies. The influence of the antagonistic bacterium Pseudomonas fluorescens on the process of root colonization by F. culmorum was demonstrated. In vermiculite inoculated with F. culmorum, the fungus density on the roots increased gradually. In the case of joint inoculation of vermiculite with the fungus and the bacterium, the F. culmorum density on the roots changed abruptly. It was shown that the site of primary colonization of the roots by the fungus was mainly the zone of root hairs. When Pseudomonas fluorescens was present on the roots, F. culmorum colonized not only root hairs, but also the elongation zone, during the first two days. Introduction of Pseudomonas fluorescens into vermiculite resulted in lower intensity of barley root rot.     

The impact of Fusarium culmorum infection on the protein fractions of raw barley and malted grains

Contaminating fungi, such as Fusarium species, produce metabolites that may interfere with normal barley grain proteolysis pattern and consequently, affect malt and beer quality. Protein compositional changes of an initial mixture of 20 % Fusarium culmorum infected and 80 % noninfected mature barley grains and respective malt are reported here. Proteolytic activity of infected barley grains (IBG) and respective malt, with controls (uninfected grains), were characterized using protease inhibitors from each class of this enzyme, including metallo-, cysteine, serine, and aspartic proteases, as well as uninhibited protease fractions. The proteins were extracted according to the Osborne fractionation and separated by size exclusion chromatography. Additionally, two-dimensional (2D) gel electrophoresis (GE) was used to analyze hydrophobic storage proteins isolated from the control and IBG. Analyses revealed that F. culmorum IBG had a twofold increase of proteolytic activity compared to the control sample, which showed an increase in all protease classes with aspartic proteases dominating. Infected and control malt grains were comparable with cysteine proteases representing almost 50 % of all proteolytic enzymes detected. Protein extractability was 31 % higher in IBG compared to the control barley. The albumin fraction showed that several metabolic proteins decreased and increased at different rates during infection and malting, thus showing a complex F. culmorum infection interdependence. Prolamin storage proteins were more hydrophobic during barley fungal infection. F. culmorum interfered with the grain hydrolytic protein profile, thereby altering the grain's protein content and quality.     

Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis

Background  

Fusarium head blight (FHB) is a disease of cereal crops, which has a severe impact on wheat and barley production worldwide. Apart from reducing the yield and impairing grain quality, FHB leads to contamination of grain with toxic secondary metabolites (mycotoxins), which pose a health risk to humans and livestock. The Fusarium species primarily involved in FHB are F. graminearum and F. culmorum. A key prerequisite for a reduction in the incidence of FHB is an understanding of its epidemiology.     

Phytopathogenic fungus Fusarium cerealis in Russia

The fungus Fusarium cerealis is described, which had not been previously identified in Russia. F. cerealis was isolated from wheat and barley roots and grains, as well as from the leaves of thistle collected in the Far Eastern, North Caucasus, and Central regions. F. cerealis strains may be misidentified as F. culmorum or F. graminearum, since the morphological characteristics of these fungi are similar.     

Occurrence of moniliformin,deoxynivalenol, and zearalenone in durum wheat (Triticum durum Desf.)

Forty-eight durum wheat samples from 5 locations in Austria were examined forFusarium infection andFusarium toxin content.F.gramlnearum andF.avenaceum were by far the prevailingFusarium species In durum wheat kernels, followed byEpoae, F.culmorum, andF.equlsetl. Ion-paired HPLC analyses of the samples showed moniliformin contents of kernels up to 0.88 mg/kg. All moniliformin contaminated samples also contained high levels of deoxynivalenol (up to 8.2 mg/kg) and lower levels of zearalenone (<0.33 mg/kg). The levels of zearalenone in naturally contaminated durum wheat samples did not correspond to the high yields of zearalenone found in cultures of the fusaria isolated from the durum wheat kernels. These conflicting results as well as some toxicologlcal aspects of the carry over ofFusarium toxins from durum wheat kernels into pasta are discussed.     

Production of trichothecene mycotoxins byFusarium graminearum andFusarium culmorum on barley and wheat

Wheat cultivars (Stoa, MN87150, SuMai-3, YMI-6, Wheaton) and barley cultivars (Robust, Excel, Chevron, M69) were inoculated in the field with isolates ofFusarium graminearum andF. culmorum. The diseased (Fusarium head blight) kernels were analyzed for deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON) and nivalenol (NIV).F. culmorum produced all three trichothecenes on all cultivars tested whereasF. graminearum only produced DON and 15-ADON. There was no well defined correlation between DON production in the host and resistance although the data tended to favor SuMai-3 as having definitive resistance to bothF. graminearum andF. culmorum.Minnesota Agricultural Experiment Station, Paper No. 20 279.     

Formation of [13C]- and [14C]zearalenone by Fusarium graminearum DSM 4529

Summary To raise the yields for the production of ¹⁴C-labelled zearalenone in Fusarium cultures the influence of growth conditions and known effectors or precursors of toxin biosynthesis was studied. Benzoic acid and 2,4-dihydroxybenzoic acid used as precursors decreased toxin formation; in the presence of different pesticides such as 2,4-dichlorophenoxyacetic acid, however, toxin production increased up to 140%. The known pathway of zearalenone biosynthesis could be confirmed from the relative extents of ¹³C-incorporation into the zearalenone molecule by incubating Fusarium graminearum DSM 4529 with d-(+)-[1-¹³C]glucose as carbon source. When grown in the presence of d-[U-¹⁴C]glucose or [2-¹⁴C]malonic acid the strain produced [¹⁴C]zearalenone with specific activities of 0.07 and 0.09 Ci/mg, the ¹⁴C-incorporation rates being 0.34% and 0.48%, respectively.     

Toxigenic Fusarium species infecting wheat heads in Poland

Toxigenic Fusarium species are common pathogens of wheat and other cereals worldwide. In total, 449 wheat heads from six localities in Poland, heavily infected with Fusarium during 2009 season, were examined for Fusarium species identification. F. culmorum was the most common species (72.1% on average) with F. graminearum and F. avenaceum the next most commonly observed, but much less frequent (13.4 and 12.5% respectively). F. cerealis was found in 1.8% of all samples, and F. tricinctum was found only in one sample (0.2%). Subsequent quantification of the three major mycotoxins (deoxynivalenol, zearalenone and moniliformin) in grain and chaff fractions with respect to associated prevailing pathogen species uncovered the following patterns. Moniliformin (MON) was found in low amounts in all samples with F. avenaceum present. In contrast, deoxynivalenol (DON) and zearalenone (ZEA) were the contaminants of F. culmorum- and F. graminearum-infected heads. The highest concentration of DON was recorded in grain sample collected in Radzików (77 µg g^?1). High temperatures in Central Poland during July and August accompanied with high rainfall in July were responsible for this high DON accumulation. Trichothecene, zearalenone, enniatin and beauvericin chemotypes were identified among 21 purified isolates using gene-specific PCR markers.