Fusarium head blight caused by Fusarium graminearum is an important disease of wheat and barley. Previous studies have showed that all three MAP kinase genes, MGV1, FgHOG1, and GPMK1, are involved in regulating hyphal growth, sexual reproduction, plant infection, and stress responses in this pathogen. To determine the relationship between the Mgv1 and FgHog1 pathways, in this study, we generated and characterized the mgv1 Fghog1 double mutant. Deletion of FgHOG1 partially rescued the defects of the mgv1 mutant in vegetative growth and cell wall integrity but had no effects on its defects in plant infection and DON production. The mgv1 Fghog1 mutant grew faster and was more tolerant to cell wall stressors than the mgv1 mutant. Swollen compartments and cell burst were observed frequently in the mgv1 mutant but rarely in the mgv1 Fghog1 mutant when treated with fungicide fludioxonil or cell wall stressor Congo red. Conversely, the deletion of MGV1 also alleviated the hyperosmotic sensitivity of the Fghog1 mutant in vegetative growth. TGY assays indicated increased phosphorylation of FgHog1 in the mgv1 mutant, and TEY assays further revealed elevated activation of Gpmk1 in the mgv1 Fghog1 double mutant, particularly under cell wall stress conditions. Overall, our data showed that deletion of FgHOG1 partially suppressed the defects of the mgv1 mutant, possibly by affecting genes related to cell wall integrity and osmoregulation via the over-activation of Gpmk1 MAP kinase and avoiding intracellular turgor elevation.

Serine/arginine (SR) proteins are essential pre-mRNA splicing factors in eukaryotic organisms. Our previous studies have shownthat the unique SR-specific protein kinase Srk1 is important for RNA splicing and gene transcription in Fusarium graminearum, and interacts with two SR proteins, FgSrp1 and FgSrp2. In this study, we have identified an SR-like protein called Sgh1 in F. graminearum, which is orthologous to budding yeast paralogous Gbp2 and Hrb1. Our data have shownthat the Sgh1 is involved in vegetative growth, conidiation, sexual reproduction, DON synthesis, and plant infection. Moreover, the Sgh1 is mainly localized to the nucleus. RNA-seq analysis has shownthat the expression of over 1100 genes and the splicing efficiency in over 300 introns were affected in the Δsgh1 mutant. Although the RS domain and all three of the RRM domains are important for the Sgh1 functions, only the RS domain is responsible for its nuclear localization. Finally, we verified that the Sgh1 interacts with the unique SR-specific kinase Srk1 in F. graminearum by the yeast-two hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. Taken together, our results have revealed that the Sgh1 regulates the fungal development, plant infection, and the pre-mRNA processing, and the RS domain regulates the function of the Sgh1 by modulating its nucleocytoplasmic shuttling.

Summary: In wheat head blight fungus Fusarium graminearum , A‐to‐I RNA editing occurs specifically during sexual reproduction. Among the genes with premature stop codons (PSCs) that require RNA editing to encode full‐length proteins, FgBUD14 also had alternative splicing events in perithecia. In this study, we characterized the functions of FgBUD14 and its post‐transcriptional modifications during sexual reproduction. The Fgbud14 deletion mutant was slightly reduced in growth, conidiation and virulence. Although deletion of FgBUD14 had no effect on perithecium morphology, the Fgbud14 mutant was defective in crozier formation and ascus development. The FgBud14‐GFP localized to the apex of ascogenous hyphae and croziers, which may be related to its functions during early sexual development. During vegetative growth and asexual reproduction, FgBud14‐GFP localized to hyphal tips and both ends of conidia. Furthermore, mutations blocking the splicing of intron 2 that has the PSC site had no effect on the function of FgBUD14 during sexual reproduction but caused a similar defect in growth with Fgbud14 mutant. Expression of the non‐editable FgBUD14 ^{Intron2‐TAA} mutant allele also failed to complement the Fgbud14 mutant. Taken together, FgBUD14 plays important roles in ascus development, and both alternative splicing and RNA editing occur specifically to its transcripts during sexual reproduction in F . graminearum .

The wheat head blight disease caused by Fusarium graminearum is a major concern for food security and the health of both humans and animals. As a pathogenic microorganism, F. graminearum produces virulence factors during infection to increase pathogenicity, including various macromolecular and small molecular compounds. Among these virulence factors, secreted proteins and deoxynivalenol (DON) are important weapons for the expansion and colonization of F. graminearum. Besides the presence of virulence factors, sexual reproduction is also crucial for the infection process of F. graminearum and is indispensable for the emergence and spread of wheat head blight. Over the last ten years, there have been notable breakthroughs in researching the virulence factors and sexual reproduction of F. graminearum. This review aims to analyze the research progress of sexual reproduction, secreted proteins, and DON of F. graminearum, emphasizing the regulation of sexual reproduction and DON synthesis. We also discuss the application of new gene engineering technologies in the prevention and control of wheat head blight.

Microtubules (MTs), as transport tracks, play important roles in hyphal-tip growth in filamentous fungi, but MT-associated proteins involved in polarized growth remain unknown. Here, we found that one novel zinc finger protein, FgChy1, is required for MT morphology and polarized growth in Fusarium graminearum. The Fgchy1 mutant presented curved and directionless growth of hyphae. Importantly, the conidia and germ tubes of the Fgchy1 mutant exhibited badly damaged and less-organized beta-tubulin cytoskeletons. Compared with the wild type, the Fgchy1 mutant lost the ability to maintain polarity and was also more sensitive to the anti-MT drugs carbendazim and nocodazole, likely due to the impaired MT cytoskeleton. Indeed, the hyphae of the wild type treated with nocodazole exhibited a morphology consistent with that of the Fgchy1 mutant. Interestingly, the disruption of FgChy1 resulted in the off-center localization of actin patches and the polarity-related polarisome protein FgSpa2 from the hyphal-tip axis. A similar defect in FgSpa2 localization was also observed in the nocodazole-treated wild-type strain. In addition, FgChy1 is also required for conidiogenesis, septation, sexual reproduction, pathogenicity, and deoxynivalenol production. Overall, this study provides the first demonstrations of the functions of the novel zinc finger protein FgChy1 in polarized growth, development, and virulence in filamentous fungi. [ ] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .