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Fusarium graminearum (Fg) is the primary causal agent of Fusarium head blight (FHB) in wheat, barley, and other small grains in North America and worldwide. FHB results in yield reduction and contaminates grain with mycotoxins that pose threats to human and livestock health. Three genetically distinct North American (NA) populations of Fg have been characterized, which are generally associated with differences in their predominant trichothecene chemotype: NA1/15-acetyl-deoxynivalenol (15-ADON), NA2/3-acetyl-deoxynivalenol (3-ADON), and NA3/3α-acetoxy, 7,15-dihydroxy-12,13-epoxytrichothec-9-ene (NX-2). Recent studies found that the NA3 population had significantly less spread on point-inoculated wheat spikes than the NA1 and NA2 populations, and NX toxins are important for Fg spread and initial infection in wheat. In this follow-up study, to compare the effect of the three populations on initial infection and mycotoxin production on different hosts, we dip-inoculated spikes of the moderately resistant wheat cultivar Alsen and the susceptible barley cultivar Voyager using five strains from each population to evaluate disease, trichothecene mycotoxin accumulation, and trichothecene production per unit of fungal biomass. In dip-inoculated wheat spikes, the NA3 population produced significantly more trichothecene per unit of fungal biomass and accumulated higher levels of trichothecene per plant biomass than the NA1 and NA2 populations, regardless of the disease levels caused by the three populations. In contrast to its critical role during wheat infection, NX toxins had no effect on barley infection. In dip-inoculated barley, the NA1 population was more infectious and caused more severe FHB symptoms than the NA2 and NA3 populations; however, the NA3 population produced significantly higher toxin per unit of fungal biomass in infected barley tissues than the NA1 population. This study provides critical information on the emerging NA3 population, which produces high levels of NX toxin and poses a potential food safety concern. [ABSTRACT FROM AUTHOR]

Fusarium graminearum, the causal agent of Fusarium head blight (FHB), produces various mycotoxins that contaminate wheat grains and cause profound health problems in humans and animals. Deoxynivalenol (DON) is the most common trichothecene found in contaminated grains. Our previous study showed that Arabidopsis-expressing F. graminearum trichothecene 3-O-acetyltransferase (FgTRI101) converted DON to 3-acetyldeoxynivalenol (3-ADON) and excreted it outside of Arabidopsis cells. To determine ifwheat can convert and excrete 3-ADON and reduce FHB and DON contamination, FgTRI101 was cloned and introduced into wheat cv Bobwhite. Four independent transgenic lines containing FgTRI101 were identified. Gene expression studies showed that FgTRI101 was highly expressed in wheat leaf and spike tissues in the transgenic line FgTri101-1606. The seedlings of two FgTri101 transgenic wheat lines (FgTri101-1606 and 1651) grew significantly longer roots than the controls on media containing 5 µg/mL DON; however, the 3-ADON conversion and excretion was detected inconsistently in the seedlings of FgTri101-1606. Further analyses did not detect 3-ADON or other possible DONrelated products in FgTri101-1606 seedlings after adding deuterium-labeled DON into the growth media. FgTri101-transgenic wheat plants showed significantly enhanced FHB resistance and lower DON content after they were infected with F. graminearum, but 3-ADON was not detected. Our study suggests that it is promising to utilize FgTRI101, a gene that the fungus uses for self-protection, for managing FHB and mycotoxin in wheat production. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) is one of the most devastating diseases of wheat and can lead to significant yield losses as well as a contamination of the crop with mycotoxins that are a major concern in cereal‐based food and feed products. The aims of this study were thus to investigate the relationship between resistance against multiple Fusarium species and to assess the potential of an indirect phenotypic and genomic selection for the resistance against the accumulation of several mycotoxins in wheat. Strong phenotypic and genetic correlations between the mycotoxin contents, FHB severity and FHB‐associated traits were observed, irrespective of if traits were assessed in trials inoculated with a DON or HT‐2/T‐2 producing Fusarium species. A multi‐stage phenotypic or genomic selection with low anther retention being used for an indirect selection among early generation selection candidates, followed by an evaluation of the pre‐selected set in disease nurseries, and lastly by assessing the mycotoxin content of the most promising genotypes is suggested as a suitable strategy to breed for wheat cultivars with reduced risk of mycotoxin accumulation. [ABSTRACT FROM AUTHOR]

The aim of this study was to compare the susceptibility of oats to Fusarium langsethiae infection, as measured by combined HT‐2 and T‐2 mycotoxin concentration (HT2 + T2) in harvested oat grain samples. Over 10 years (2004–2013), samples from single replicates of each UK Recommended List oat trial were analyzed for HT‐2 and T‐2. For spring oats, there were small but statistically significant differences between varieties, whereas for winter oats, they had a broader range and higher mean of HT2 + T2 concentration compared with spring oats. For winter oats, the short‐strawed varieties had consistently high HT2 + T2 levels compared with other varieties, whereas naked varieties were at the lower end of the range, and short, naked varieties had intermediate levels. A separate set of harvested oat grain samples of eight common varieties from 17 field experiments were analyzed by modified joint regression analysis. Results showed that environment had the strongest impact on HT‐2 and T‐2 concentrations but that the varietal susceptibility to HT‐2 and T‐2 contamination was highly stable across environments. This methodology can be used to calculate a Fusarium (HT2 + T2) resistance score for oats to aid grower selection of suitable varieties, as is available for Fusarium (DON) resistance for wheat varieties in many countries. [ABSTRACT FROM AUTHOR]

Fusarium fungi produce a diverse array of mycotoxic metabolites during the pathogenesis of cereals. Some, such as the trichothecenes and fumonisins, are phytotoxic, acting as non-proteinaceous effectors that facilitate disease development in cereals. Over the last few decades, we have gained some depth of understanding as to how trichothecenes and fumonisins interact with plant cells and how plants deploy mycotoxin detoxification and resistance strategies to defend themselves against the producer fungi. The cereal-mycotoxin interaction is part of a co-evolutionary dance between Fusarium and cereals, as evidenced by a trichothecene-responsive, taxonomically restricted, cereal gene competing with a fungal effector protein and enhancing tolerance to the trichothecene and resistance to DON-producing F. graminearum. But the binary fungal–plant interaction is part of a bigger ecosystem wherein other microbes and insects have been shown to interact with fungal mycotoxins, directly or indirectly through host plants. We are only beginning to unravel the extent to which trichothecenes, fumonisins and other mycotoxins play a role in fungal-ecosystem interactions. We now have tools to determine how, when and where mycotoxins impact and are impacted by the microbiome and microfauna. As more mycotoxins are described, research into their individual and synergistic toxicity and their interactions with the crop ecosystem will give insights into how we can holistically breed for and cultivate healthy crops. [ABSTRACT FROM AUTHOR]

Fusarium graminearum (Fg) is the primary causal agent of Fusarium head blight (FHB) in wheat, barley, and other small grains in North America and worldwide. FHB results in yield reduction and contaminates grain with mycotoxins that pose threats to human and livestock health. Three genetically distinct North American (NA) populations of Fg have been characterized, which are generally associated with differences in their predominant trichothecene chemotype: NA1/15-acetyl-deoxynivalenol (15-ADON), NA2/3-acetyl-deoxynivalenol (3-ADON), and NA3/3α-acetoxy, 7,15-dihydroxy-12,13-epoxytrichothec-9-ene (NX-2). Recent studies found that the NA3 population had significantly less spread on point-inoculated wheat spikes than the NA1 and NA2 populations, and NX toxins are important for Fg spread and initial infection in wheat. In this follow-up study, to compare the effect of the three populations on initial infection and mycotoxin production on different hosts, we dip-inoculated spikes of the moderately resistant wheat cultivar Alsen and the susceptible barley cultivar Voyager using five strains from each population to evaluate disease, trichothecene mycotoxin accumulation, and trichothecene production per unit of fungal biomass. In dip-inoculated wheat spikes, the NA3 population produced significantly more trichothecene per unit of fungal biomass and accumulated higher levels of trichothecene per plant biomass than the NA1 and NA2 populations, regardless of the disease levels caused by the three populations. In contrast to its critical role during wheat infection, NX toxins had no effect on barley infection. In dip-inoculated barley, the NA1 population was more infectious and caused more severe FHB symptoms than the NA2 and NA3 populations; however, the NA3 population produced significantly higher toxin per unit of fungal biomass in infected barley tissues than the NA1 population. This study provides critical information on the emerging NA3 population, which produces high levels of NX toxin and poses a potential food safety concern.

Fusarium graminearum, the causal agent of Fusarium head blight (FHB), produces various mycotoxins that contaminate wheat grains and cause profound health problems in humans and animals. Deoxynivalenol (DON) is the most common trichothecene found in contaminated grains. Our previous study showed that Arabidopsis-expressing F. graminearum trichothecene 3-O-acetyltransferase (FgTRI101) converted DON to 3-acetyldeoxynivalenol (3-ADON) and excreted it outside of Arabidopsis cells. To determine if wheat can convert and excrete 3-ADON and reduce FHB and DON contamination, FgTRI101 was cloned and introduced into wheat cv Bobwhite. Four independent transgenic lines containing FgTRI101 were identified. Gene expression studies showed that FgTRI101 was highly expressed in wheat leaf and spike tissues in the transgenic line FgTri101-1606. The seedlings of two FgTri101 transgenic wheat lines (FgTri101-1606 and 1651) grew significantly longer roots than the controls on media containing 5 µg/mL DON; however, the 3-ADON conversion and excretion was detected inconsistently in the seedlings of FgTri101-1606. Further analyses did not detect 3-ADON or other possible DON-related products in FgTri101-1606 seedlings after adding deuterium-labeled DON into the growth media. FgTri101-transgenic wheat plants showed significantly enhanced FHB resistance and lower DON content after they were infected with F. graminearum, but 3-ADON was not detected. Our study suggests that it is promising to utilize FgTRI101, a gene that the fungus uses for self-protection, for managing FHB and mycotoxin in wheat production.

ABSTRACT: Contamination of feed with mycotoxins has become a severe issue worldwide. Among the most prevalent trichothecene mycotoxins, T-2 toxin is of particular importance for livestock production, including poultry posing a significant threat to animal health and productivity. This review article aims to comprehensively analyze the pathological consequences, metabolism, and toxic effects of T-2 toxin in poultry. Trichothecene mycotoxins, primarily produced by Fusarium species, are notorious for their potent toxicity. T-2 toxin exhibits a broad spectrum of negative effects on poultry species, leading to substantial economic losses as well as concerns about animal welfare and food safety in modern agriculture. T-2 toxin exposure easily results in negative pathological consequences in the gastrointestinal tract, as well as in parenchymal tissues like the liver (as the key organ for its metabolism), kidneys, or reproductive organs. In addition, it also intensely damages immune system-related tissues such as the spleen, the bursa of Fabricius, or the thymus causing immunosuppression and increasing the susceptibility of the animals to infectious diseases, as well as making immunization programs less effective. The toxin also damages cellular processes on the transcriptional and translational levels and induces apoptosis through the activation of numerous cellular signaling cascades. Furthermore, according to recent studies, besides the direct effects on the abovementioned processes, T-2 toxin induces the production of reactive molecules and free radicals resulting in oxidative distress and concomitantly occurring cellular damage. In conclusion, this review article provides a complex and detailed overview of the metabolism, pathological consequences, mechanism of action as well as the immunomodulatory and oxidative stress-related effects of T-2 toxin. Understanding these effects in poultry is crucial for developing strategies to mitigate the impact of the T-2 toxin on avian health and food safety in the future.

Grain mold and stalk rot are among the fungal diseases that cause significant losses in sorghum worldwide and are caused by different Fusarium spp. The presence of Fusarium species in sorghum grains causes yield losses and mycotoxin contamination, which represents a risk to consumers. In this study, Fusarium graminearum species complex (FGSC) had a high incidence, followed by Fusarium fujikuroi species complex (FFSC) and F. incarnatum-equiseti species complex. Within FFSC, F. proliferatum, F. andiyazi, F. fujikuroi, F. thapsinum, F. verticillioides and F. subglutinans were identified, and this was the first report of F. fujikuroi in sorghum. The most frequent toxins found in sorghum samples were deoxynivalenol (DON) and zearalenone (ZEN). The presence of fumonisins and nivalenol (NIV) was detected at low levels. This study adds new knowledge about the occurrence of Fusarium species and mycotoxins in sorghum grains. Furthermore, this is the first report in Uruguay on fungicide sensitivity for Fusarium isolates from sorghum, which constitutes an important starting point for defining management practices to minimize fungal infection and mycotoxin contamination. [ABSTRACT FROM AUTHOR]

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T-2 toxin is a mycotoxin routinely found as a contaminant of cereal grains worldwide. A portable mass spectrometer was adapted to enable the detection of T-2 toxin in wheat and maize by APCI-MS. In order to facilitate rapid testing, a rapid cleanup was used. The method was able to detect T-2 toxin in soft white wheat, hard red wheat, and yellow dent maize and could be used to screen for T-2 at levels above 0.2 mg/kg. The HT-2 toxin was only detectable at very high levels (>0.9 mg/kg). Based on these results, the sensitivity was not sufficient to allow the application of the screening method to these commodities at levels recommended by the European Commission. With a cut-off level of 0.107 mg/kg, the method correctly classified nine of ten reference samples of wheat and maize. The results suggest that portable MS detection of T-2 toxin is feasible. However, additional research will be needed to develop an application sensitive enough to meet regulatory requirements. [ABSTRACT FROM AUTHOR]

Mycotoxin contamination in food poses health hazards to humans. Current methods of controlling mycotoxins still have limitations and more effective approaches are needed. During the past decades of years, variable environmental factors have been tested for their influence on mycotoxin production leading to elucidation of a complex regulatory network involved in mycotoxin biosynthesis. These regulators are putative targets for screening molecules that could inhibit mycotoxin synthesis. Here, we summarize the regulatory mechanisms of hierarchical regulators, including pathway-specific regulators, global regulators and epigenetic regulators, on the production of the most critical mycotoxins (aflatoxins, patulin, citrinin, trichothecenes and fumonisins). Future studies on regulation of mycotoxins will provide valuable knowledge for exploring novel methods to inhibit mycotoxin biosynthesis in a more efficient way. [ABSTRACT FROM AUTHOR]

Fusarium trichothecenes are among the mycotoxins of most concern to food and feed safety. Production of these mycotoxins and presence of the trichothecene biosynthetic gene (TRI) cluster have been confirmed in only two multispecies lineages of Fusarium: the Fusarium incarnatum-equiseti (Incarnatum) and F. sambucinum (Sambucinum) species complexes. Here, we identified and characterized a TRI cluster in a species that has not been formally described and is represented by Fusarium sp. NRRL 66739. This fungus is reported to be a member of a third Fusarium lineage: the F. buharicum species complex. Cultures of NRRL 66739 accumulated only two trichothecenes, 7-hydroxyisotrichodermin and 7-hydroxyisotrichodermol. Although these are not novel trichothecenes, the production profile of NRRL 66739 is novel, because in previous reports 7-hydroxyisotrichodermin and 7-hydroxyisotrichodermol were components of mixtures of 6–8 trichothecenes produced by several Fusarium species in Sambucinum. Heterologous expression analysis indicated that the TRI13 gene in NRRL 66739 confers trichothecene 7-hydroxylation. This contrasts the trichothecene 4-hydroxylation function of TRI13 in other Fusarium species. Phylogenetic analyses suggest that NRRL 66739 acquired the TRI cluster via horizontal gene transfer from a close relative of Incarnatum and Sambucinum. These findings provide insights into evolutionary processes that have shaped the distribution of trichothecene production among Fusarium species and the structural diversity of the toxins. [ABSTRACT FROM AUTHOR]

Type B trichothecenes commonly contaminate cereal grains and include five structurally related congeners: deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), fusarenon X (FX), and nivalenol (NIV). These toxins are known to have negative effects on human and animal health, particularly affecting food intake. However, the pathophysiological basis for anorexic effect is not fully clarified. The purpose of this study is to explore the potential roles of the brain-gut peptides substance P (SP) and glucagon-like peptide-17-36 amide (GLP-1) in anorexic responses induced by type B trichothecenes following both intraperitoneal (IP) and oral administration. SP and GLP-1 were elevated at 1 or 2 h and returned to basal levels at 6 h following exposure to DON and both ADONs. FX induced the production of both brain gut peptides with initial time at 1 or 2 h and duration > 6 h. Similar to FX, exposing IP to NIV caused elevations of SP and GLP-1 at 1 h and lasted more than 6 h, whereas oral exposure to NIV only increased both brain gut peptides at 2 h. The neurokinin-1 receptor (NK-1R) antagonist Emend® dose-dependently attenuated both SP- and DON-induced anorexic responses. Pretreatment with the GLP-1 receptor (GLP-1R) antagonist Exending9-39 induced a dose-dependent attenuation of both GLP-1- and DON-induced anorexic responses. To summarize, the results suggest that both SP and GLP-1 play important roles in anorexia induction by type B trichothecenes. [ABSTRACT FROM AUTHOR]

• Populations of Fusarium graminearum undergo frequent sexual recombination in the field. • 15ADON isolates from Uruguay and 3ADON isolates from North America are highly genetically similar. • The NA1, NA2 and SLA populations separated from a common ancestral population at approximately the same time. • Genome scans of natural selection identified regions that may play a role in population adaptation. Fusarium head blight (FHB) is a major disease of wheat and barley worldwide and is caused by different species in the genus Fusarium , Fusarium graminearum being the most important. We conducted population genomics analyses using SNPs obtained through genotyping by sequencing of over 500 isolates of F. graminearum from the US Upper Midwest, New York, Louisiana, and Uruguay. PCA and STRUCTURE analyses group our isolates into four previously described populations: NA1, NA2, Southern Louisiana (SLA) and Gulf Coast (GC). Some isolates were not assigned to populations because of mixed ancestry. Population structure was associated with toxin genotype and geographic origin. The NA1, NA2, and SLA populations are differentiated (F ST 0.385 – 0.551) but the presence of admixed isolates indicates that the populations are not reproductively isolated. Patterns of linkage disequilibrium (LD) decay suggest frequent recombination within populations. Fusarium graminearum populations from the US have great evolutionary potential given the high recombination rate and a large proportion of admixed isolates. The NA1, NA2, and Southern Louisiana (SLA) populations separated from their common ancestral population roughly at the same time in the past and are evolving with moderate levels of subsequent gene flow between them. Genome-wide selection scans in all three populations revealed outlier regions with the strongest signatures of recent positive natural selection. These outlier regions include many genes with unknown function and some genes with known roles in plant–microbe interaction, fungicide/drug resistance, cellular transport and genes that are related to cellular organelles. Only a very small proportion of outlier regions are shared as outliers among the three populations, suggesting unique host-pathogen interactions and environmental adaptation. [ABSTRACT FROM AUTHOR]

Mycotoxins such as deoxynivalenol introduce a health risk to the food supply and are costly to manage or avoid. Technologies for reducing or eliminating the toxicity of deoxynivalenol could be useful in a variety of processes, such as in preserving the value as animal feed of byproducts of ethanol production. We characterized transformation products of deoxynivalenol that were formed by the combination of a fungal laccase paired with the chemical mediator 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO), using chromatography, mass spectrometry, and nuclear magnetic resonance spectroscopy. Alcohol groups at the C3 and C15 positions of deoxynivalenol were oxidized to ketones, and the chemical mediator became covalently linked to the C4 position. Conditions experienced during gas chromatography led to the dissociation of TEMPO, forming 3,15-diketodeoxynivalenol. Understanding the range of possible modifications to deoxynivalenol and other trichothecenes is a necessary step toward effective remediation of contaminated grain. [ABSTRACT FROM AUTHOR]

The Fusarium graminearum species complex is the main causal agent of wheat head blight worldwide. Trichothecenes produced by the pathogen in infected grains have important food safety implications. Previously reported studies on trichothecene production have all focused on the conditions conducive to mycotoxin production, while the molecular mechanisms of trichothecene biosynthesis in Fusarium strains under normal or non-inducing conditions are still unclear. Here, a global analysis of the fungal gene expression of three strains using the Affymetrix Fusarium GeneChip under non-inducing conditions is reported. Differentially expressed genes were identified among strains with different trichothecene-production ability, and some novel genes associated with trichothecene biosynthesis were found by bioinformatics analysis. To verify the transcriptome results, proteomic analyses of the three strains were conducted under the same culture conditions. In total, 69 unique fungal proteins were identified in 77 protein spots. Combined with transcriptome and proteome analysis, 27 novel genes were predicted to be associated with trichothecene mycotoxin production. A protein, encoded by FGSG_01403, was found to be associated with trichothecene production via proteome analysis. Gene knock-out mutations of FGSG_01403 resulted in mutants with increased production of trichothecenes. Future functional analysis of the candidate genes identified in this study may reveal new insights into the negative regulation of trichothecene production in the Fusarium graminearum species complex. [ABSTRACT FROM AUTHOR]

Grain mold and stalk rot are among the fungal diseases that cause significant losses in sorghum worldwide and are caused by different Fusarium spp. The presence of Fusarium species in sorghum grains causes yield losses and mycotoxin contamination, which represents a risk to consumers. In this study, Fusarium graminearum species complex (FGSC) had a high incidence, followed by Fusarium fujikuroi species complex (FFSC) and F. incarnatum-equiseti species complex. Within FFSC, F. proliferatum, F. andiyazi, F. fujikuroi, F. thapsinum, F. verticillioides and F. subglutinans were identified, and this was the first report of F. fujikuroi in sorghum. The most frequent toxins found in sorghum samples were deoxynivalenol (DON) and zearalenone (ZEN). The presence of fumonisins and nivalenol (NIV) was detected at low levels. This study adds new knowledge about the occurrence of Fusarium species and mycotoxins in sorghum grains. Furthermore, this is the first report in Uruguay on fungicide sensitivity for Fusarium isolates from sorghum, which constitutes an important starting point for defining management practices to minimize fungal infection and mycotoxin contamination.

The secondary metabolites of Fusarium sporotrichioides, an endophytic fungus with anti-tumor activity isolated from Rauvolfia yunnanensis Tsiang, were investigated. Five trichothecenes, including one previously undescribed metabolite, were isolated and identified. Their structures were elucidated by means of extensive spectroscopic methods; the absolute configuration of compound 1 was determined by the ECD method. Surprisingly, 8-n-butyrylneosolaniol (3) exhibited stronger anti-tumor activity than T-2 toxin against Huh-7 cell line, with an IC50 value of 265.9 nM. 8-n-butyrylneosolaniol (3) promoted apoptosis induction in Huh-7 cells. Moreover, cell cycle analysis showed that cell cycle arrest caused by 8-n-butyrylneosolaniol (3) at the G2/M phase resulted in cell proliferation inhibition and pro-apoptotic activity. Further studies showed a significant decrease in mitochondrial membrane permeabilization and a significant increase in ROS generation, which led to the activation of caspase cascades and subsequent cleavage of PARP fragments. In conclusion, 8-n-butyrylneosolaniol (3) induced cell apoptosis in Huh-7 cells via the mitochondria-mediated apoptotic signaling pathway, which could be a leading compound for anti-tumor agents. [ABSTRACT FROM AUTHOR]

T-2 toxin is a mycotoxin routinely found as a contaminant of cereal grains worldwide. A portable mass spectrometer was adapted to enable the detection of T-2 toxin in wheat and maize by APCI-MS. In order to facilitate rapid testing, a rapid cleanup was used. The method was able to detect T-2 toxin in soft white wheat, hard red wheat, and yellow dent maize and could be used to screen for T-2 at levels above 0.2 mg/kg. The HT-2 toxin was only detectable at very high levels (>0.9 mg/kg). Based on these results, the sensitivity was not sufficient to allow the application of the screening method to these commodities at levels recommended by the European Commission. With a cut-off level of 0.107 mg/kg, the method correctly classified nine of ten reference samples of wheat and maize. The results suggest that portable MS detection of T-2 toxin is feasible. However, additional research will be needed to develop an application sensitive enough to meet regulatory requirements.

The influence of previous crops (oat, maize, or rape) and six variety of hulled oats (Avena sativa L.) were assessed qualitatively and quantitatively as to yield, thousand grain weight, and grain size in observations from 2009 to 2012. Harvested grain samples were analysed for the presence of mycotoxins (T-2 and HT-2 toxins, deoxynivalenol, and nivalenol) using LC/MS–MS. Fusarium poae was the species most often found on oats. Flamingsprofi and Salo were the most sensitive varieties of oat. The sensitivity of the variety to lodging as well as the earliness of the variety was correlated with the occurrence of fusariosis on oats. The highest average occurrence of Fusaria on oat grains was after use of maize as the pre-crop. The oat samples were most often contaminated with nivalenol. Nivalenol positively correlated with the weight of a thousand grains of oat and negatively correlated with the oats grain falling through the sieve. [ABSTRACT FROM AUTHOR]

This study aimed to assess the incidence and distribution of toxigenic fungi in Korean oat. Toxigenic fungi were isolated from oat samples collected from 12 oat fields from heading to harvest in 2017 and 2018. A total of 745 fungal colonies were isolated based on morphology and identified using marker genes. About 92% of the fungal isolates were Fusarium spp. and others were Penicillium (5.9%) and Aspergillus (2.1%). Fusarium isolates comprised mostly of F. asiaticum (83.1%), followed by F. incarnatum (5.4%), F. proliferatum (3.5%), F. fujikuroi (2.8%), F. tricinctum species complex (FTSC) 11 (1.5%) and F. graminearum (1.0%). About 97% of F. asiaticum was nivalenol type, and 3-acetyl deoxynivalenol (3.2%) and 15-acetyl deoxynivalenol (0.4%) types also were found. Pathogenicity test of the selected Fusarium isolates revealed that F. asiaticum isolates have a wide range of virulence depending on the tested plants. F. graminearum and FTSC 11 isolates from blighted spikelets were the most virulent in naked oat. All Fusarium isolates (n=18) except one (FTSC 11) produced nivalenol (0.2–7.6 μg/g), deoxynivalenol (0.03–6.1 μg/g), and zearalenone (0.1–27.0 μg/g) on rice medium. This study is first report that F. asiaticum causes Fusarium head blight disease of oat in Korea. These findings demonstrate the dominance of F. asiaticum in oat agroecosystems as in rice, wheat and barley in Korea.

A total of 1,159 Fusarium strains were isolated from sorghum grown in Danyang and Youngwol in 2017 and 2018. The isolates were analyzed to reveal genetic, toxigenic and pathogenic characteristics. Phylogenetic analysis using TEF-1α and RPB2 genes showed that the samples were contaminated with at least 17 Fusarium species. Among them, F. graminearum, F. proliferatum, F. thapsinum, F. incarnatum, and F. asiaticum were dominant species. In F. graminearum and F. asiaticum, F. graminearum-15-acetyl deoxynivalenol chemotype and F. asiaticum-nivalenol chemotype were frequent. Six Fusarium species tested produced one or more mycotoxins, except F. thapsinum and FTSC 11. F. proliferatum and F. fujikuroi had FUM1 gene (76.0% and 81.6%, respectively) and some isolates produced high level of fumonisin (over 1,000 μg/g). F. proliferatum and F. thapsinum were more virulent than other species on sorghum. These results indicate that Fusarium species in sorghum might produce multiple mycotoxins.

Type B trichothecenes commonly contaminate cereal grains and include five structurally related congeners: deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), fusarenon X (FX), and nivalenol (NIV). These toxins are known to have negative effects on human and animal health, particularly affecting food intake. However, the pathophysiological basis for anorexic effect is not fully clarified. The purpose of this study is to explore the potential roles of the brain-gut peptides substance P (SP) and glucagon-like peptide-17-36 amide (GLP-1) in anorexic responses induced by type B trichothecenes following both intraperitoneal (IP) and oral administration. SP and GLP-1 were elevated at 1 or 2 h and returned to basal levels at 6 h following exposure to DON and both ADONs. FX induced the production of both brain gut peptides with initial time at 1 or 2 h and duration > 6 h. Similar to FX, exposing IP to NIV caused elevations of SP and GLP-1 at 1 h and lasted more than 6 h, whereas oral exposure to NIV only increased both brain gut peptides at 2 h. The neurokinin-1 receptor (NK-1R) antagonist Emend® dose-dependently attenuated both SP- and DON-induced anorexic responses. Pretreatment with the GLP-1 receptor (GLP-1R) antagonist Exending9-39 induced a dose-dependent attenuation of both GLP-1- and DON-induced anorexic responses. To summarize, the results suggest that both SP and GLP-1 play important roles in anorexia induction by type B trichothecenes.

Mycotoxin contamination in food poses health hazards to humans. Current methods of controlling mycotoxins still have limitations and more effective approaches are needed. During the past decades of years, variable environmental factors have been tested for their influence on mycotoxin production leading to elucidation of a complex regulatory network involved in mycotoxin biosynthesis. These regulators are putative targets for screening molecules that could inhibit mycotoxin synthesis. Here, we summarize the regulatory mechanisms of hierarchical regulators, including pathway-specific regulators, global regulators and epigenetic regulators, on the production of the most critical mycotoxins (aflatoxins, patulin, citrinin, trichothecenes and fumonisins). Future studies on regulation of mycotoxins will provide valuable knowledge for exploring novel methods to inhibit mycotoxin biosynthesis in a more efficient way.

Fusarium trichothecenes are among the mycotoxins of most concern to food and feed safety. Production of these mycotoxins and presence of the trichothecene biosynthetic gene (TRI) cluster have been confirmed in only two multispecies lineages of Fusarium: the Fusarium incarnatum-equiseti (Incarnatum) and F. sambucinum (Sambucinum) species complexes. Here, we identified and characterized a TRI cluster in a species that has not been formally described and is represented by Fusarium sp. NRRL 66739. This fungus is reported to be a member of a third Fusarium lineage: the F. buharicum species complex. Cultures of NRRL 66739 accumulated only two trichothecenes, 7-hydroxyisotrichodermin and 7-hydroxyisotrichodermol. Although these are not novel trichothecenes, the production profile of NRRL 66739 is novel, because in previous reports 7-hydroxyisotrichodermin and 7-hydroxyisotrichodermol were components of mixtures of 6–8 trichothecenes produced by several Fusarium species in Sambucinum. Heterologous expression analysis indicated that the TRI13 gene in NRRL 66739 confers trichothecene 7-hydroxylation. This contrasts the trichothecene 4-hydroxylation function of TRI13 in other Fusarium species. Phylogenetic analyses suggest that NRRL 66739 acquired the TRI cluster via horizontal gene transfer from a close relative of Incarnatum and Sambucinum. These findings provide insights into evolutionary processes that have shaped the distribution of trichothecene production among Fusarium species and the structural diversity of the toxins.

Trichodermin, a trichothecene first isolated in Trichoderma species, is a sesquiterpenoid antibiotic that exhibits significant inhibitory activity to the growth of many pathogenic fungi such as Candida albicans , Rhizoctonia solani , and Botrytis cinerea by inhibiting the peptidyl transferase involved in eukaryotic protein synthesis. Trichodermin has also been shown to selectively induce cell apoptosis in several cancer cell lines and thus can act as a potential lead compound for developing anticancer therapeutics. The biosynthetic pathway of trichodermin in Trichoderma has been identified, and most of the involved genes have been functionally characterized. An exception is TRI3 , which encodes a putative acetyltransferase. Here, we report the identification of a gene cluster that contains seven genes expectedly involved in trichodermin biosynthesis (TRI3 , TRI4 , TRI6 , TRI10 , TRI11 , TRI12 , and TRI14) in the trichodermin-producing endophytic fungus Trichoderma taxi. As in Trichoderma brevicompactum , TRI5 is not included in the cluster. Functional analysis provides evidence that TRI3 acetylates trichodermol, the immediate precursor, to trichodermin. Disruption of TRI3 gene eliminated the inhibition to R. solani by T. taxi culture filtrates and significantly reduced the production of trichodermin but not of trichodermol. Both the inhibitory activity and the trichodermin production were restored when native TRI3 gene was reintroduced into the disruption mutant. Furthermore, a His-tag-purified TRI3 protein, expressed in Escherichia coli , was able to convert trichodermol to trichodermin in the presence of acetyl-CoA. The disruption of TRI3 also resulted in lowered expression of both the upstream biosynthesis TRI genes and the regulator genes. Our data demonstrate that T. taxi TRI3 encodes an acetyltransferase that catalyzes the esterification of the C-4 oxygen atom on trichodermol and thus plays an essential role in trichodermin biosynthesis in this fungus. [ABSTRACT FROM AUTHOR]

Trichodermin, a trichothecene first isolated in Trichoderma species, is a sesquiterpenoid antibiotic that exhibits significant inhibitory activity to the growth of many pathogenic fungi such as Candida albicans, Rhizoctonia solani, and Botrytis cinerea by inhibiting the peptidyl transferase involved in eukaryotic protein synthesis. Trichodermin has also been shown to selectively induce cell apoptosis in several cancer cell lines and thus can act as a potential lead compound for developing anticancer therapeutics. The biosynthetic pathway of trichodermin in Trichoderma has been identified, and most of the involved genes have been functionally characterized. An exception is TRI3, which encodes a putative acetyltransferase. Here, we report the identification of a gene cluster that contains seven genes expectedly involved in trichodermin biosynthesis (TRI3, TRI4, TRI6, TRI10, TRI11, TRI12, and TRI14) in the trichodermin-producing endophytic fungus Trichoderma taxi. As in Trichoderma brevicompactum, TRI5 is not included in the cluster. Functional analysis provides evidence that TRI3 acetylates trichodermol, the immediate precursor, to trichodermin. Disruption of TRI3 gene eliminated the inhibition to R. solani by T. taxi culture filtrates and significantly reduced the production of trichodermin but not of trichodermol. Both the inhibitory activity and the trichodermin production were restored when native TRI3 gene was reintroduced into the disruption mutant. Furthermore, a His-tag-purified TRI3 protein, expressed in Escherichia coli, was able to convert trichodermol to trichodermin in the presence of acetyl-CoA. The disruption of TRI3 also resulted in lowered expression of both the upstream biosynthesis TRI genes and the regulator genes. Our data demonstrate that T. taxi TRI3 encodes an acetyltransferase that catalyzes the esterification of the C-4 oxygen atom on trichodermol and thus plays an essential role in trichodermin biosynthesis in this fungus.

Mycotoxins such as deoxynivalenol introduce a health risk to the food supply and are costly to manage or avoid. Technologies for reducing or eliminating the toxicity of deoxynivalenol could be useful in a variety of processes, such as in preserving the value as animal feed of byproducts of ethanol production. We characterized transformation products of deoxynivalenol that were formed by the combination of a fungal laccase paired with the chemical mediator 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO), using chromatography, mass spectrometry, and nuclear magnetic resonance spectroscopy. Alcohol groups at the C3 and C15 positions of deoxynivalenol were oxidized to ketones, and the chemical mediator became covalently linked to the C4 position. Conditions experienced during gas chromatography led to the dissociation of TEMPO, forming 3,15-diketodeoxynivalenol. Understanding the range of possible modifications to deoxynivalenol and other trichothecenes is a necessary step toward effective remediation of contaminated grain.

Nepenthesins are categorized under the subfamily of the nepenthesin-like plant aspartic proteases (PAPs) that form a distinct group of atypical PAPs. This study describes the effect of nepenthesin 1 (HvNEP-1) protease from barley (Hordeum vulgare L.) on fungal histidine acid phosphatase (HAP) phytase activity. Signal peptide lacking HvNEP-1 was expressed in Pichia pastoris and biochemically characterized. Recombinant HvNEP-1 (rHvNEP-1) strongly inhibited the activity of Aspergillus and Fusarium phytases, which are enzymes that release inorganic phosphorous from phytic acid. Moreover, rHvNEP-1 suppressed in vitro fungal growth and strongly reduced the production of mycotoxin, 15-acetyldeoxynivalenol (15-ADON), from Fusarium graminearum. The quantitative PCR analysis of trichothecene biosynthesis genes (TRI) confirmed that rHvNEP-1 strongly repressed the expression of TRI4 , TRI5 , TRI6 , and TRI12 in F. graminearum. The co-incubation of rHvNEP-1 with recombinant F. graminearum (rFgPHY1) and Fusarium culmorum (FcPHY1) phytases induced substantial degradation of both Fusarium phytases, indicating that HvNEP-1-mediated proteolysis of the fungal phytases contributes to the HvNEP-1-based suppression of Fusarium. [ABSTRACT FROM AUTHOR]

Fusarium equiseti is a plant pathogen with a wide range of hosts and diverse effects, including probiotic effects. However, the molecular mechanisms underlying these effects remain unclear, hindering its effective utilization. The final assembly included 16 scaffolds of contiguous sequence without gaps. The total sequence length was 40,776,005 bp, and the GC content of 48.01%. In total, we annotated the putative function of 13,134 genes, accounting for 94.97% of the candidate genes. We identified two and 23 candidate genes that are likely involved in the production of mycotoxins zearalenone and trichothecene, respectively. A comparative genomic analysis supported the high quality of the F. equiseti assembly. Our comprehensive analysis of whole-genome sequence will serve as a valuable resource for future studies of expression, regulation, function and evolution of the genes of F. equiseti as well as studies into disease prevention and control. • The Whole-Genome Sequence of Fusarium equiseti was assembled and annotated. • Multiple genes were related to metabolism and pathogenicity. • A comparative genomic analysis was performed. • The genomic data provide a valuable resource for future studies. [ABSTRACT FROM AUTHOR]

Contamination of feed with mycotoxins has become a severe issue worldwide. Among the most prevalent trichothecene mycotoxins, T-2 toxin is of particular importance for livestock production, including poultry posing a significant threat to animal health and productivity. This review article aims to comprehensively analyze the pathological consequences, metabolism, and toxic effects of T-2 toxin in poultry. Trichothecene mycotoxins, primarily produced by Fusarium species, are notorious for their potent toxicity. T-2 toxin exhibits a broad spectrum of negative effects on poultry species, leading to substantial economic losses as well as concerns about animal welfare and food safety in modern agriculture. T-2 toxin exposure easily results in negative pathological consequences in the gastrointestinal tract, as well as in parenchymal tissues like the liver (as the key organ for its metabolism), kidneys, or reproductive organs. In addition, it also intensely damages immune system-related tissues such as the spleen, the bursa of Fabricius, or the thymus causing immunosuppression and increasing the susceptibility of the animals to infectious diseases, as well as making immunization programs less effective. The toxin also damages cellular processes on the transcriptional and translational levels and induces apoptosis through the activation of numerous cellular signaling cascades. Furthermore, according to recent studies, besides the direct effects on the abovementioned processes, T-2 toxin induces the production of reactive molecules and free radicals resulting in oxidative distress and concomitantly occurring cellular damage. In conclusion, this review article provides a complex and detailed overview of the metabolism, pathological consequences, mechanism of action as well as the immunomodulatory and oxidative stress-related effects of T-2 toxin. Understanding these effects in poultry is crucial for developing strategies to mitigate the impact of the T-2 toxin on avian health and food safety in the future. [ABSTRACT FROM AUTHOR]

The Fusarium graminearum species complex is the main causal agent of wheat head blight worldwide. Trichothecenes produced by the pathogen in infected grains have important food safety implications. Previously reported studies on trichothecene production have all focused on the conditions conducive to mycotoxin production, while the molecular mechanisms of trichothecene biosynthesis in Fusarium strains under normal or non-inducing conditions are still unclear. Here, a global analysis of the fungal gene expression of three strains using the Affymetrix Fusarium GeneChip under non-inducing conditions is reported. Differentially expressed genes were identified among strains with different trichothecene-production ability, and some novel genes associated with trichothecene biosynthesis were found by bioinformatics analysis. To verify the transcriptome results, proteomic analyses of the three strains were conducted under the same culture conditions. In total, 69 unique fungal proteins were identified in 77 protein spots. Combined with transcriptome and proteome analysis, 27 novel genes were predicted to be associated with trichothecene mycotoxin production. A protein, encoded by FGSG_01403, was found to be associated with trichothecene production via proteome analysis. Gene knock-out mutations of FGSG_01403 resulted in mutants with increased production of trichothecenes. Future functional analysis of the candidate genes identified in this study may reveal new insights into the negative regulation of trichothecene production in the Fusarium graminearum species complex.

The secondary metabolites of Fusarium sporotrichioides, an endophytic fungus with anti-tumor activity isolated from Rauvolfia yunnanensis Tsiang, were investigated. Five trichothecenes, including one previously undescribed metabolite, were isolated and identified. Their structures were elucidated by means of extensive spectroscopic methods; the absolute configuration of compound 1 was determined by the ECD method. Surprisingly, 8-n-butyrylneosolaniol (3) exhibited stronger anti-tumor activity than T-2 toxin against Huh-7 cell line, with an IC50 value of 265.9 nM. 8-n-butyrylneosolaniol (3) promoted apoptosis induction in Huh-7 cells. Moreover, cell cycle analysis showed that cell cycle arrest caused by 8-n-butyrylneosolaniol (3) at the G2/M phase resulted in cell proliferation inhibition and pro-apoptotic activity. Further studies showed a significant decrease in mitochondrial membrane permeabilization and a significant increase in ROS generation, which led to the activation of caspase cascades and subsequent cleavage of PARP fragments. In conclusion, 8-n-butyrylneosolaniol (3) induced cell apoptosis in Huh-7 cells via the mitochondria-mediated apoptotic signaling pathway, which could be a leading compound for anti-tumor agents.

Trichothecene mycotoxins such as deoxynivalenol (DON) are virulence factors of Fusarium graminearum, which causes Fusarium head blight, one of the most important diseases of small grain cereals. We previously identified a nonspecific lipid transfer protein (nsLTP) gene, AtLTP4.4, which was overexpressed in an activation-tagged Arabidopsis line resistant to trichothecin, a type B trichothecene in the same class as DON. Here we show that overexpression of AtLTP4.4 in transgenic wheat significantly reduced F. graminearum growth in 'Bobwhite' and 'RB07' lines in the greenhouse and reduced fungal lesion size in detached leaf assays. Hydrogen peroxide accumulation was attenuated on exposure of transgenic wheat plants to DON, indicating that AtLTP4.4 may confer resistance by inhibiting oxidative stress. Field testing indicated that disease severity was significantly reduced in two transgenic 'Bobwhite' lines expressing AtLTP4.4. DON accumulation was significantly reduced in four different transgenic 'Bobwhite' lines expressing AtLTP4.4 or a wheat nsLTP, TaLTP3, which was previously shown to have antioxidant activity. Recombinant AtLTP4.4 purified from Pichia pastoris exhibited potent antifungal activity against F. graminearum. These results demonstrate that overexpression of AtLTP4.4 in transgenic wheat suppresses DON accumulation in the field. Suppression of DON-induced reactive oxygen species by AtLTP4.4 might be the mechanism by which fungal spread and mycotoxin accumulation are inhibited in transgenic wheat plants. [ABSTRACT FROM AUTHOR]

Five trichothecenes including a new one, together with two previously undescribed benzene derivatives were isolated from the solid culture of Trichothecium sp. Their structures were established by 1D and 2D NMR data in conjunction with HR-ESI-MS analysis. Compounds 1-5 exhibited cytotoxicity against MCF-7 cell lines at various levels ranging from IC 50 of 7.23 to 16.95  μ M. Compound 6 decreased the concentration of blood lipids in zebra fish at the concentration of 20  μ M.

A total of 45 cereal samples (35 paddy and 10 sorghum) were collected from Mandya and Mysore districts of Karnataka during March, 2019 to study the mycotoxigenic Fusarium strains present in the field and market samples. All samples were brought to DFRL, Mysore and screened for Fusarium presence by agar plating method on Selective Fusarium Agar (SFA) containing streptomycin (1 mg/ml) with incubation at 25 ± 2°C for 3–5 days. In the study, almost 10 different Fusarium species association was revealed with the collected cereal samples. Fusarium species were micromorphologically identified and further confirmed by PCR-based detection using ITS1 and ITS4 primers followed by sequencing. PCR studies confirmed that all the tested fungal isolates belonged to Fusarium spp. with the amplicon size of 590 bp. NCBI data base confirmed the sequence similarity of 99 % to the genus Fusarium and accession numbers were obtained by submitting the sequences to the GenBank. The study disclosed the diversity in phytopathogenic Fusarium spp. in paddy and sorghum growing in different agro-climatic regions of Mysore and Mandya districts of Karnataka, India.

Fungi produce mycotoxins in the presence of appropriate temperature, humidity, sufficient nutrients and if the density of the mushroom mass is favorable. Although all mycotoxins are of fungal origin, all toxic compounds produced by fungi are not called mycotoxins. The interest in mycotoxins first started in the 1960s, and today the interest in mycotoxin-induced diseases has increased. To date, 400 mycotoxins have been identified and the most important species producing mycotoxins belongs to Aspergillus, Penicillium, Alternaria and Fusarium genera. Mycotoxins are classified as hepatotoxins, nephrotoxins, neurotoxins, immunotoxins etc. In this review genotoxic and also other health effects of some major mycotoxin groups like Aflatoxins, Ochratoxins, Patulin, Fumonisins, Zearalenone, Trichothecenes and Ergot alkaloids were deeply analyzed. [ABSTRACT FROM AUTHOR]

There is limited data available concerning the effect of T-2/HT-2 toxin or deoxynivalenol (DON) on invertebrates such as springtails, and no data on their life history and oxidative stress. Control maize and DON or T-2 toxin contaminated maize were fed to Folsomia candida with a toxin content of 16324 mg DON kg-1 or 671 mg T-2 kg-1 maize. Ten to twelve days old animals were investigated in a life-history test and a stress protein test. T-2 toxin did not affect Folsomia candida in any measured parameters. The DON exposed group showed decreased growth and reproduction, and a higher survival rate. DON treatment resulted in lower protein content, while reduced glutathione content was higher than in control. It suggests that DON activated the glutathione-related detoxification pathway, which possibly causes a higher survival rate. The results also suggest that the oral toxicity of DON or T-2 is lower than through physical contact. For that reason, DON and T-2 toxin contaminated maize is not suggested to be used as green manure in the native state. Alternative solutions could be using mycotoxin contaminated maize for biogas production, or after decontamination by bacterial strains, it can be used as organic fertilizer. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON), also known as vomitoxin, is a mycotoxin that can cause antifeeding and vomiting in animals. However, the mechanism of DON inducing anorexia is complicated. Studies have shown that intestinal hormones play a significant part in the anorexia caused by DON. We adopted the “modeling of acute antifeeding in mice” as the basic experimental model, and used two methods of gavage and intraperitoneal injection to explore the effect of intestinal hormones on the antifeedant response induced by DON in mice. We found that 1 and 2.5 mg/kg·bw of DON can acutely induce anorexia and increase the plasma intestinal hormones CCK, PYY, GIP, and GLP-1 in mice within 3 h. Direct injection of exogenous intestinal hormones CCK, PYY, GIP, and GLP-1 can trigger anorexia behavior in mice. Furthermore, the PYY receptor antagonist JNJ-31020028, GLP-1 receptor antagonist Exendin(9-39), CCK receptor antagonist Proglumide, GIP receptor antagonist GIP(3-30)NH2 attenuated both intestinal hormone and DON-induced anorectic responses. These results indicate that intestinal hormones play a critical role in the anorexia response induced by DON.

A total of 52 samples of finished fish feeds and ingredients were collected from smallholder farmers in Kenya, Tanzania, Rwanda and Uganda, and analyzed. Culture and molecular techniques were used to identify fungal isolates from the feedstock, and mycotoxin profiles were determined using liquid chromatography–tandem mass spectrometry. The most prevalent fungal species recovered in the samples was Asperigillus flavus (54.5%). Other fungal species recovered from the samples were Aspergillus tamarii (9.1%), Mucor velutinosus (9%), Phoma sp. (6.1%), Aspergillus niger (6%), Eurotium rubrum (3%) and Penicillium chrysogenum (3%). Fourteen mycotoxins were identified: aflatoxins B1, B2, G1 and G2, fumonisin B1 and B3, deoxynivalenol (DON) and acetyldeoxynivalenol (sum of 3-ADONand 15-ADON), ochratoxin A, roquefortine C, alternariol, T-2 toxin, and nivalenol. DON (92.9%), aflatoxins (64.3%) and fumonisins (57.1%) were the most prevalent within locally manufactured feeds, while no contamination was found in imported feed. Samples from Kenya were the most contaminated with aflatoxin (maximum 806.9 μg·kg−1). The high levels of aflatoxin and trichothecene type A and B contamination found in this study point to potential risks to fish performance and to the health of consumers of the fish and derived products.

This study evaluated the ability of isolated or semisynthesized trichothecene sesquiterpenes to prevent cancer emergence and proliferation and inhibit signal transducer and activator of transcription-3 (STAT3) phosphorylation through in vitro assays. Trichothecinol A (TTC-A), which bears a hydroxy group at C3, exhibited greater cancer prevention, antiproliferation, and STAT3 phosphorylation inhibition effects than trichothecin (TTC), which lacks a hydroxy group at C3. Furthermore, trichothecinol B (TTC-B), which is a reduced derivative of TTC and has similar cytotoxic effect, showed substantially weaker chemoprotection and STAT3 phosphorylation inhibition effects than TTC. These results clearly indicate that the hydroxy group at C3 and carbonyl group at C8 are crucial for inducing both potent chemoprevention and STAT3 phosphorylation inhibition.

Fusarium graminearum, the causal agent of Fusarium head blight (FHB), produces trichothecenes including deoxynivalenol (DON), nivalenol (NIV), and 3,7,15-trihydroxy-12,13-epoxytrichothec-9-ene (NX-3). These toxins contaminate grains and cause profound health problems in humans and animals. To explore exploiting a fungal self-protection mechanism in plants, we examined the ability of F. graminearum trichothecene 3-O-acetyltransferase (FgTri101) to detoxify several key trichothecenes produced by F. graminearum: DON, 15-ADON, NX-3, and NIV. FgTri101 was cloned from F. graminearum and expressed in Arabidopsis plants. We compared the phytotoxic effects of purified DON, NIV, and NX-3 on the root growth of transgenic Arabidopsis expressing FgTri101. Compared to wild type and GUS controls, FgTri101 transgenic Arabidopsis plants displayed significantly longer root length on media containing DON and NX-3. Furthermore, we confirmed that the FgTri101 transgenic plants acetylated DON to 3-ADON, 15-ADON to 3,15-diADON, and NX-3 to NX-2, but did not acetylate NIV. Approximately 90% of the converted toxins were excreted into the media. Our study indicates that transgenic Arabidopsis expressing FgTri101 can provide plant protection by detoxifying trichothecenes and excreting the acetylated toxins out of plant cells. Characterization of plant transporters involved in trichothecene efflux will provide novel targets to reduce FHB and mycotoxin contamination in economically important plant crops.

Fusarium head blight (FHB) is a major disease in wheat causing severe economic losses globally by reducing yield and contaminating grain with mycotoxins. In Canada, Fusarium graminearum is the principal etiological agent of FHB in wheat, producing mainly the trichothecene mycotoxin, deoxynivalenol (DON) and its acetyl derivatives (15-acetyl deoxynivalenol (15ADON) and 3-acetyl deoxynivalenol (3ADON)). Understanding the population biology of F. graminearum such as the genetic variability, as well as mycotoxin chemotype diversity among isolates is important in developing sustainable disease management tools. In this study, 570 F. graminearum isolates collected from commercial wheat crops in five geographic regions in three provinces in Canada in 2018 and 2019 were analyzed for population diversity and structure using 10 variable number of tandem repeats (VNTR) markers. A subset of isolates collected from the north-eastern United States was also included for comparative analysis. About 75% of the isolates collected in the Canadian provinces of Saskatchewan and Manitoba were 3ADON indicating a 6-fold increase in Saskatchewan and a 2.5-fold increase in Manitoba within the past 15 years. All isolates from Ontario and those collected from the United States were 15ADON and isolates had a similar population structure. There was high gene diversity (H = 0.803–0.893) in the F. graminearum populations in all regions. Gene flow was high between Saskatchewan and Manitoba (Nm = 4.971–21.750), indicating no genetic differentiation between these regions. In contrast, less gene flow was observed among the western provinces and Ontario (Nm = 3.829–9.756) and USA isolates ((Nm = 2.803–6.150). However, Bayesian clustering model analyses of trichothecene chemotype subpopulations divided the populations into two clusters, which was correlated with trichothecene types. Additionally, population cluster analysis revealed there was more admixture of isolates among isolates of the 3ADON chemotypes than among the 15ADON chemotype, an observation that could play a role in the increased virulence of F. graminearum. Understanding the population genetic structure and mycotoxin chemotype variations of the pathogen will assist in developing FHB resistant wheat cultivars and in mycotoxin risk assessment in Canada.

Trichothecenes are the most prevalent mycotoxins contaminating cereal grains. Some of them are also considered as the virulence factors of Fusarium head blight disease. However, the mechanism behind the structure-activity relationship for trichothecenes remains unexplained. Filling this information gap is a crucial step for developing strategies to manage this large family of mycotoxins in food and feed. Here, we perform an in-depth re-examination of the existing structures of Saccharomyces cerevisiae ribosome complexed with three different trichothecenes. Multiple binding interactions between trichothecenes and 25S rRNA, including hydrogen bonds, nonpolar pi stacking interactions and metal ion coordination interactions, are identified as important binding determinants. These interactions are mainly contributed by the key structural elements to the toxicity of trichothecenes, including the oxygen in the 12,13-epoxide ring and a double bond between C9 and C10. In addition, the C3-OH group also participates in binding. The comparison of three trichothecenes binding to the ribosome, along with their binding pocket architecture, suggests that the substitutions at different positions impact trichothecenes binding in two different patterns. Moreover, the binding of trichothecenes induced conformation changes of several nucleotide bases in 25S rRNA. This then provides a structural framework for understanding the structure-activity relationships apparent in trichothecenes. This study will facilitate the development of strategies aimed at detoxifying mycotoxins in food and feed and at improving the resistance of cereal crops to Fusarium fungal diseases.

Sorghum has gained popularity with consumers as a grain source with its gluten-free and high protein dietary characteristics. Acreage has increased recently, in part due to the demand for an alternative feed source for poultry and swine. New information is needed about the level of mycotoxin contamination in sorghum accessions, and accurate and affordable analytical methods need to be developed and tested to quantify mycotoxins in sorghum. A traditional method (solid-phase extraction chromatography with C18, followed by GC-MS) to quantify the mycotoxin deoxynivalenol (DON) produced inconsistent DON values following controlled spiking and recovery experiments with different sorghum accessions. Consequently, we incorporated a stable isotope (d1-DON) as an internal standard into a traditional GC-MS method. This method, stable isotope dilution assay (SIDA), was used to accurately determine DON levels in 196 sorghum samples representing 98 different accessions. Of the 98 accessions tested (two samples per accession), 76 of the accessions had DON levels that were greater than the limit of detection for both methods (0.20 μg g−1). For a USA regulatory threshold of 1 μg g−1, about one third of all the accessions (26/76) had at least 20% more DON using the SIDA method. For a regulatory threshold of 5 μg g−1, about 7% of all the accessions (5/76) had at least 20% more DON using the SIDA method. Using SIDA, the amount of DON in a sorghum sample can be accurately and reliably quantitated by basing calculations on the recovery of d1-DON, and this method may have future applications for quantifying DON from samples with complex matrices. [ABSTRACT FROM AUTHOR]

• T-2 toxin produced by Fusarium spp., induced resistance against postharvest diseases on potato tubers. • Dry rot development of tubers was significantly reduced by T-2 toxin application. • The mechanism of induced resistance includes the activation of ROS and phenylpropanoid metabolism in potato tubers tissue. Potato (Solanum tubersosum L) is the most important non-grain source of human diet for the people living at poverty line. Potato suffers from many notorious biotic stresses, including dry rot caused by Fusarium spp. Treatments of potato tubers with 1 mg L−1 of T-2 toxin produced by Fusarium spp. induced resistance against Fusarium sulphureum by new elicitor-like activity. To investigate the mechanism of induced resistance by T-2 toxin against dry rot of potato, tuber slices were treated with T-2 toxin, followed by the inoculation with F. sulphureum spores. Results in this study showed that the reduction of disease area was linked with higher abundance of reactive oxygen species (ROS) and new phenylpropanoid metabolites. T-2 toxin treatment increased ROS accumulation and induced the activities of enzymes involved in ROS and phenylpropanoid metabolism. At the same time, T-2 toxin treatment also up-regulated proteins involved in ROS and phenylpropanoid metabolism determined by iTRAQ (isobaric tags for relative and absolute quantitation) assay, as well as genes encoding proteins involved ROS and phenylpropanoid metabolism. Taken together, this study provides substantial evidences that T-2 toxin induced resistance related proteins and genes in potato tuber slices to suppress the dry rot severity of F. sulphureum. [ABSTRACT FROM AUTHOR]