A method for reducing the concentrations of Fusarium graminearum trichothecenes in durum wheat grain with the use of Debaryomyces hansenii.

Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is one of the most dangerous diseases of durum wheat. This hemibiotrophic pathogen transitions from the biotrophic phase, during which it penetrates host tissues and secretes trichothecenes, to the necrotrophic phase which leads to the destruction of host tissues. Yeasts applied to spikes often reduce mycotoxin concentrations, but the underlying mechanisms have not been fully elucidated. Therefore, the aim of this study was to analyze the concentrations trichothecenes in durum wheat grain and changes in the F. graminearum transcriptome under the influence the Debaryomyces hansenii antagonistic yeast strain. Debaryomyces hansenii cells adhered to and formed cell aggregates/biofilm on the surface of spikes and pathogenic hyphae. Biological control suppressed the spread of F. graminearum by 90 % and decreased the content of deoxynivalenol (DON) in spikes by 31.2 %. Yeasts significantly reduced the expression of pathogen's genes encoding the rpaI subunit of RNA polymerase I and the activator of Hsp90 ATPase, but they had no effect on mRNA transcript levels of genes encoding the enzymes involved in the biosynthesis of trichothecenes. The yeast treatment reduced the number of F. graminearum operational taxonomic units (OTUs) nearly five-fold and increased the number of D. hansenii OTUs more than six-fold in the spike mycobiome. The mechanisms that suppress infections should be explored to develop effective biological methods for reducing the concentrations mycotoxins in wheat grain.

Comparison of Volatiles Profile and Contents of Trichothecenes Group B, Ergosterol, and ATP of Bread Wheat, Durum Wheat, and Triticale Grain Naturally Contaminated by Mycobiota.

In natural conditions cereals can be infested by pathogenic fungi. These can reduce the grain yield and quality by contamination with mycotoxins which are harmful for plants, animals, and humans. To date, performed studies of the compounds profile have allowed for the distinction of individual species of fungi. The aim of this study was to determine the profile of volatile compounds and trichothecenes of group B, ergosterol, adenosine triphosphate content carried out on a representative sample of 16 genotypes of related cereals: triticale, bread wheat, and durum wheat. Based on an analysis of volatile compounds by means of gas chromatography mass spectrometry and with the use of an electronic nose, volatile profiles for cereals were determined. Differentiation is presented at four levels through discriminant analysis, heatmaps, principal component analysis (PCA), and electronic nose maps. The statistical model was built by subsequent incorporation of chemical groups such as trichothecenes (GC/MS), fungal biomass indicators ergosterol (HPLC) and ATP (luminometric) and volatiles. The results of the discriminatory analyses showed that the volatile metabolites most markedly differentiated grain samples, among which were mainly: lilial, trichodiene, p-xylene. Electronic nose analysis made it possible to completely separate all the analyzed cereals based only on 100 ions from the 50-150 m/z range. The research carried out using chemometric analysis indicated significant differences in the volatile metabolites present in the grain of bread wheat, durum wheat and triticale. The end result of the performed analyses was a complete discrimination of the examined cereals based on the metabolites present in their grain.

A comparison of contents of group A and B trichothecenes and microbial counts in different cereal species.

Concentrations of trichothecenes and the amounts of microbial biomass were compared in grain of bread wheat, durum wheat, triticale, rye, oat and barley. Grain samples came from lines regionalised in Poland grown under identical climatic and agricultural conditions in 2007. Among the six analysed cereals, the highest mean concentration of toxic metabolites of 151.89 µg/kg was found for grain of Triticum durum, whereas the lowest was for barley grain (25.56 µg/kg). The highest contamination with microscopic fungi was recorded in case of barley and rice grain (mean concentrations of ergosterol were 12.53 mg/kg and 11.24 mg/kg, respectively). In case of the analysed cereals, the total microbial biomass expressed in the amount of ATP turned out to be the highest (4.7 × 105 relative light units [RLU]) for rye and oat (3.2 × 105 RLU). The results of the applied classical discrimination analysis indicate a significant diversification of species in terms of all the 11 analysed metabolites.

Concentration of fungal biomass and trichothecenes in different parts of einkorn.

Analyses of ergosterol (ERG), adenosine-5'-triphosphate (ATP) and groups A and B trichothecenes were performed in three Triticum monococcum cultivars registered in Germany (Albini, Terzino and Tifi), grown in the organic system. The experiment was carried out on two dates: the first - in the final phase of flowering (BBCH 69) and the second -- in the phase of full ripeness (BBCH 89). The analyses were performed on shanks, glumes, grain and awns. Concentrations of analyzed metabolites in different parts of T. monococcum plants varied significantly. Mean ERG concentration in the first term was more than 30 times higher than immediately before harvest, whereas for group B trichothecenes it was 4 times higher. Contents of ATP and concentrations of group A trichothecenes were similar at both times. When analyzing parts of the spike, the highest amount of metabolites was recorded in shanks (ERG - 114 mg/kg, ATP 900,000 RLU, group A and B trichothecenes - 0.07 and 0.20 mg/kg, respectively), while the lowest in grain (ERG - 5 mg/kg, ATP 55,000 RLU, group A and B trichothecenes - 0.03 and 0.08 mg/kg, respectively). A higher ERG concentration was found in awns (65 mg/kg) than in glumes (41 mg/kg), whereas for ATP and group A and B trichothecenes by contrast higher concentrations were recorded in glumes (160,000 RLU, 0.06 and 0.029 mg/kg, respectively) than in awns (77,000 RLU, 0.05 and 0.014 mg/kg, respectively). Recorded results indicate a potential occurrence of trichothecenes in shanks, awns and glumes already during flowering, when grain has not yet developed. In these parts of plants, after harvest, the highest amounts of microorganisms and Fusarium toxins were found, which pose a threat for farmers and workers employed in the cereal industry.

Volatile metabolites in various cereal grains.

To date, studies on volatile metabolites in cereal grain have focused mainly on a single species. In this paper, results are presented of the analysis of volatile compounds in five cereal grain species (spring wheat, durum wheat, triticale, rye, oats and barley) based on representative sampling of at least 15 cultivars of individual species. Profiles of volatile compounds were determined using solid phase microextraction (SPME) and GC-TOF (time of flight mass spectrometry). Many of the volatile compounds were only present in single samples; however, several dozen were found in over 50% of samples and 46 volatiles were found in all samples. Among them there were six alcohols, 10 aldehydes and ketones, six terpenes, seven hydrocarbons and 11 benzene derivatives. The highest concentrations of these compounds were found in durum wheat, while the lowest were observed in triticale and rye.

Contents of microscopic fungi in dusts coming from cereal analysis laboratories.

Microscopic fungi - components of bioaerosol found in the workplace environment of individuals employed in the agricultural sector - constitute a considerable hazard for their health. This study includes quantitative and qualitative analyses of mycobionta contained in 20 samples of dusts collected from laboratories conducting analyses of cereals. A total of 27 species of viable microscopic fungi were isolated. The most frequently isolated genera Penicillium and Aspergillus, accounting for 27 percent and 26 percent of analyzed isolates. The content of fungal biomass was determined quantitatively using a fungal marker, ergosterol (ERG). Concentrations of this metabolite for all samples ranged from 0.48 mg/kg-212.36 mg/kg. Based on the analyses, it may be stated that the concentration of microfungi in settled dust from laboratories conducting analyses of cereals was varied, and in several cases markedly exceeded admissible concentration levels.