Determination of the trichothecene mycotoxin chemotypes and associated geographical distribution and phylogenetic species of the Fusarium graminearum clade from China
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Abstract BACKGROUND Fusarium head blight ( FHB ) of wheat is an important disease causing yield losses and mycotoxin contamination. The aim of the work was to detect and characterise trichothecene producing Fusarium species in durum and soft wheat cultivated in an area of central Italy in 2009 and 2010 and to determine trichothecene contamination by LC‐MS / MS in the grain. RESULTS F. graminearum s. str. was the most frequent species. In 2009, the occurrence of F. avenaceum and F. poae was higher than in 2010. Among F. graminearum strains, the 15‐acetyl deoxynivalenol (15‐ ADON ) chemotype could be found more frequently, followed by nivalenol ( NIV ) and 3‐ ADON chemotypes, while all F. culmorum isolates belonged to the 3‐ ADON chemotype. All F. poae strains were NIV chemotypes. In vitro trichothecene production confirmed molecular characterisation. Durum wheat was characterised by a higher average DON contamination with respect to soft wheat, NIV was always detected at appreciable levels while type‐A trichothecenes were mostly found in durum wheat samples in 2009 with 6% of samples exceeding the contamination level recently recommended by the European Union. CONCLUSION Climatic conditions were confirmed to be predominant factors influencing mycotoxigenic species composition and mycotoxin contaminations. However, NIV contamination was found to occur irrespective of climatic conditions, suggesting that it may often represent an under‐estimated risk to be further investigated. © 2014 Society of Chemical Industry
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Fusarium graminearum is one of the most important causes of FHB or wheat scab in different part of the world. This fungus is able to produce diffrent Trichothecene mycotoxins such as Nivalenol (NIV) and Deoxynivalenol (DON) which are harmful for both human and animals. To determine chemotypes of Trichothecene, a total of 100 isolates from different fields of Golestan province in Iran including Gorgan, Kordkuy, Bandaregaz, Gonbad, Minodasht, Kalaleh and Azadshahr were identified as F. graminearum using morphological features. The identity of 96 isolates was confirmed by polymerase chain reaction (PCR) assay using F. graminearum species-specific primers (Fg16F/Fg16R). Based on sequences of Tri13 gene involved in the mycotoxin biosynthetic pathway, PCR assays were used to detect Nivalenol (NIV) and Deoxynivalenol (DON) chemotypes. Of the 96 tested isolates in Tri13 PCR assays, 70 classified as NIV chemotype and the remaining 26 isolates as DON producers. These results indicated that NIV chemotype was the most dominant chemotype in studied region. A greater proportion of NIV chemotype was found in Gorgan fields (P < 0.05, P < 0.0001), whereas greater proportion of DON was detected in Gonbad fields (P < 0.05, P < 0.0001). PCR assay-based chemotyping was confirmed by HPLC method. These results demonstrated that PCR assay and HPLC could be used as rapid, reliable and cost-effective methods for the detection and identification of mycotoxin-producing Fusarium-species and may thus help to develop strategies to avoid or reduce mycotoxin contamination of cereals.
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The role of Fusarium graminearum trichothecene-chemotypes in disease outcomes was evaluated by point inoculation in a series of wheat lines with different levels of resistance to Fusarium head blight (FHB). Four inocula, each consisting of a composite of four strains with either 15-acetyldeoxynivalenol (ADON) chemotypes from "traditional" or emergent populations, a 3-ADON chemotype, or a nivalenol (NIV) chemotype, were compared. The evaluated wheat included Canadian lines with different levels of FHB resistance/susceptibility and double haploid lines developed from crosses of these lines. Highly resistant lines were resistant to infection by all of the F. graminearum chemotypes evaluated. In the moderately susceptible/resistant wheat lines, the 3-ADON producers and the emergent 15-ADON population were, in some instances, more aggressive and resulted in higher Fusarium damaged kernel scores and levels of trichothecene accumulation. The data presented in this study demonstrate the importance of growing highly resistant wheat cultivars in the current climate of an evolving F. graminearum population, and suggest that moderate levels of FHB resistance may not be sufficient to minimize trichothecene contamination of grain from F. graminearum-infected wheat.
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Fusarium graminearum is an important pathogen that causes Fusarium head blight (FHB) in several cereal crops worldwide. The potential of this pathogen to contaminate cereals with trichothecene mycotoxins presents a health risk for both humans and animals. This study aimed to evaluate the potential of different trichothecene genotypes of F. graminearum isolated from an alternative host plant to produce mycotoxins under different spring wheat grain incubation conditions. Fourteen F. graminearum strains were isolated from seven alternative host plants and identified as 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) genotypes. These strains were cultivated on spring wheat grains at 25 °C and 29 °C for 5 weeks. The mycotoxins produced were analysed with a high-performance liquid chromatograph (HPLC) coupled to a Thermo Scientific TSQ Quantiva MS/MS detector. The obtained results showed that the F. graminearum strains from alternative host plants could produce nivalenol (NIV), deoxynivalenol (DON), fusarenon-X (FUS-X), 3-ADON, deoxynivalenol-3-ß-d-glucoside (D3G), 15-ADON, and zearalenone (ZEA). F. graminearum strains produced DON and ZEA under both temperatures, with the mean concentrations varying from 363 to 112,379 µg kg-1 and from 1452 to 44,816 µg kg-1, respectively. Our results indicated the possible role of dicotyledonous plants, including weeds, as a reservoir of inoculum sources of F. graminearum-induced Fusarium head blight, associated with the risk of mycotoxin contamination in spring wheat.
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Fusarium graminearum (Schwabe) contaminates agricultural crops and commodities with trichothecenes, mostly deoxynivalenol and its acetyl-derivatives. Current techniques available to detect final mycotoxin contamination products usually require an extended time lag between sampling and the corresponding report, and include different clean-up steps and eventually derivatisation. This study was aimed to develop a methodology to detect toxigenic F. graminearum prior to mycotoxin production. Headspace solid-phase microextraction coupled to capillary gas chromatography is shown to be useful to predict the potential of trichothecene mycotoxin formation by detecting the presence of F. graminearum at early stages of fungal growth in wheat cultivars, based on the detection of trichodiene (TRI), the volatile intermediate of trichothecenes. We showed that TRI is a useful marker to detect toxigenic Fusarium in wheat spikes from live plants, regardless of the actual development of Fusarium head blight (FHB). This is the first predictive methodology for FHB and trichothecene occurrence in field-collected samples. It might be a useful tool to help to prevent the risk of mycotoxin contamination.
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