Fungal toxins, such as those produced by members of the order Hypocreales, have widespread effects on cereal crops, resulting in yield losses and the potential for severe disease and mortality in humans and livestock. Among the most toxic are the trichothecenes. Trichothecenes have various detrimental effects on eukaryotic cells including an interference with protein production and the disruption of nucleic acid synthesis. However, these toxins can have a wide range of toxicity depending on the system. Major differences in the phytotoxicity and cytotoxicity of these mycotoxins are observed for individual members of the class, and variations in toxicity are observed among different species for each individual compound. Furthermore, while diverse toxicological effects are observed throughout the whole cellular system upon trichothecene exposure, the mechanism of toxicity is not well understood. In order to comprehend how these toxins interact with the cell, we must first have an advanced understanding of their structure and dynamics. The structural analysis of trichothecenes was a subject of major interest in the 1980s, and primarily focused on crystallographic and solution-state Nuclear Magnetic Resonance (NMR) spectroscopic studies. Recent advances in structural determination through solution- and solid-state NMR, as well as computation based molecular modeling is leading to a resurgent interest in the structure of these and other mycotoxins, with the focus shifting in the direction of structural dynamics. The purpose of this work is to first provide a brief overview of the structural data available on trichothecenes and a characterization of the methods commonly employed to obtain such information. A summary of the current understanding of the relationship between structure and known function of these compounds is also presented. Finally, a prospectus on the application of new emerging structural methods on these and other related systems is discussed.

Barley (Hordeum vulgare L.) is the fourth largest cereal crop in the world. One of the most devastating diseases in barley worldwide is Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe. Several mycotoxins are produced by FHB infection, and deoxynivalenol (DON) is one of them responsible for the deterioration of grain quality. The current limited number of reliable molecular markers makes the development of FHB-resistant cultivars rather difficult and laborious. Moreover, there is a limited number of designed specific biomarkers that could distinguish the FHB resistance and mycotoxin accumulation in barley cultivars. This study investigated the phenolic compounds of ten different Canadian barley cultivars, grown in artificially FHB-infected and non-infected field trials. The enzyme-linked immunosorbent assay (ELISA) was used to assess the presence of DON in the harvested infected grains of each tested variety. High-performance liquid chromatography (HPLC) analysis was performed using both infected and non-infected samples. We identified differences among cultivars tested in non-infected samples through quantitative analysis of free and bound phenolic compounds. The resistant cultivars showed higher amounts of major bound phenolic compounds compared to the susceptible check CDC Bold. Additionally, the FHB-infected cultivars produced significantly higher amounts of sinapic acid (SIN) () and catechin (CAT) in the soluble free form of phenolics in barley compared to the non-infected subjects. This study suggests that phenolic compounds in barley could allow barley breeders to precisely identify and develop FHB-resistant barley germplasm and cultivars.

Fusarium head blight (FHB), caused by Fusarium graminearum, is currently the most devastating disease for barley (Hordeum vulgare) in Canada. Associated mycotoxins can compromise grain quality, where deoxynivalenol (DON) is considered particularly damaging due to its frequency of detection. Breeding barley with a lower DON content is difficult, due to the poor adaptation and malt quality of resistance sources. A set of European-derived heritage varieties were screened in an FHB nursery in Charlottetown, PE, with selections tested at Brandon, MB, between 2018–2022. Genetic evaluation demonstrated a distinct clustering of Canadian varieties from the heritage set. At Brandon, 72% of the heritage varieties ranked lower for DON content than did the moderately resistant Canadian check ‘AAC Goldman’, but resistance was associated with later heading and taller stature. In contrast with Canadian modern malting variety ‘AAC Synergy’, general deficiencies were observed in yield, enzyme activity, and extract, along with higher protein content. Nonetheless, several resistant varieties were identified with reasonable a heading date and yield, including ‘Chevallier Chile’, ‘Domen’, ‘Djugay’, ‘Hannchen’, ‘Heils Franken’, ‘Moravian Barley’, ‘Loosdorfer’ with ‘Golden Melon’, ‘Nutans Moskva’, and ‘Vellavia’, these being some of the most promising varieties when malting quality characteristics were also considered. These heritage resources could be used as parents in breeding to develop FHB-resistant malting barley varieties.

Liquid chromatography–mass spectrometry analysis of Pseudomonas chlororaphis subsp. phenazini S1Bt23 extracts detected phenazine-1-carboxylic acid (PCA) and 2-hydroxyphenazine (2-OH-PHZ) as the main phenazine derivatives. We investigated their relative contributions to the antagonistic activity of strain S1Bt23 against Pythium arrhenomanes, a root rot pathogen of corn. CRISPR-Cas9 knockouts were carried out on the phzF gene, required for PCA synthesis, and the phzO gene, which is involved in converting PCA to 2-OH-PHZ. Deletion of the phzF gene abrogated the production of PCA and 2-OH-PHZ, and the ΔphzF mutant lost the antagonism against Pythium arrhenomanes. In contrast, deletion of the phzO gene created a 2-OH-PHZ-negative mutant with intact antagonistic ability. Concordantly, S1Bt23 wild type and the ΔphzO mutant, but not the ΔphzF mutant, significantly bioprotected corn seeds of a susceptible inbred variety, CO441, from P. arrhenomanes. At equimolar amounts of 75 nM, synthetic PCA inhibited Pythium growth, whereas 2-OH-PHZ did not. This highlights the critical contribution of PCA to the biocontrol activity of strain S1Bt23 against P. arrhenomanes. Unexpectedly, deletion of phzO did not result in additional PCA accumulation. This suggests that the conversion of PCA to 2-OH-PHZ by S1Bt23 is a potential protective mechanism against the overproduction of lethal cellular doses. This study paves the way for bioengineering strain S1Bt23 into a more effective biopesticide.

Fusarium graminearum and F. avenaceum often co-contaminate Canadian durum wheat grain, resulting in the co-deposition of species-specific mycotoxins, including trichothecenes produced by F. graminearum and enniatins produced by F. avenaceum. These mycotoxins pose significant risks to human and animal health. Although these fungi commonly co-occur in infected wheat, relatively little is known about how they interact during host infection. Interactions between the two species were examined using co-inoculation experiments on durum wheat spikes. In pathology trials, co-inoculations often reduced both disease severity and trichothecene accumulation compared with inoculations of F. graminearum alone, despite F. graminearum greatly out-competing F. avenaceum in total fungal biomass. Transcriptomic profiling identified strong induction of the F. graminearum ABC transporter gene FgABCB8 during co-inoculation with an enniatin-producing F. avenaceum strain. When F. graminearum was grown in vitro, FgABCB8 was induced upon exposure to F. avenaceum culture filtrate, or the related cyclohexadepsipeptides enniatin B1 or beauvericin. Heterologous expression of FgABCB8 in yeast provided partial protection against enniatin and beauvericin toxicity. Gene disruption of FgABCB8 increased F. graminearum sensitivity to enniatins. These findings demonstrate that FgABCB8 expression enhances the ability of F. graminearum to tolerate enniatin-producing fungi.