Fusarium head blight caused by Fusarium graminearum is a serious fungal disease on wheat and maize worldwide, resulting in a significant economic loss. DUG3-mediated glutathione utilization has been revealed to play important roles in fungal differentiation, metabolism, stress adaptation, and plant infection. However, functional roles of the DUG3 homolog in F. graminearum remain uncharacterized. In the present study, FgDUG3 was knocked-out via homologous recombination to investigate functions of this gene. The deletion mutant (ΔFgDUG3) was normal in mycelial growth, but showed impairments in conidiation, conidial germination, and plant infection, compared to the wild-type strain. The defects of ΔFgDUG3 were recovered in the complemented strain (ΔFgDUG3-C). Transcriptomic analysis revealed that deletion of FgDUG3 caused significantly differential expression of genes, mainly related to metabolism, catabolism, cellular structure organization, and signal transduction. Taken together, these results suggest that FgDUG3 plays important roles in the differentiation and pathogenicity of F. graminearum.

The obligate biotrophic fungus Puccinia striiformis f. sp. tritici (Pst) frequently overcomes host immunity through rapid evolution. The wheat landrace Paozimai exhibits high-temperature all-stage resistance (HTAS) to Pst, a type of resistance expressed at relatively high temperatures across all growth stages. In this study, we investigated host–pathogen interactions using integrated transcriptomic and data-independent acquisition proteomic analyses. Dual RNA-seq analysis at 192 and 216 h post-inoculation (hpi) revealed a shift in enriched Pst pathways during the late latent period, from endoplasmic reticulum protein processing and fatty acid metabolism at 192 hpi to carbohydrate and glutathione metabolism at 216 hpi. In the host, differentially expressed genes (DEGs) were enriched in defense-related signaling pathways at 192 hpi and in metabolic pathways at 216 hpi. Co-expression network analysis identified ribosomal and protein biosynthesis processes that may contribute to defense. Proteomic analysis showed greater overlap across time points, with oxidative phosphorylation enriched at 192 hpi and glycan biosynthesis at 216 hpi. Furthermore, integrated multi-omics and co-expression network analysis identified 16 candidate wheat genes, including genes encoding ribosomal proteins and stress-response proteins. These findings describe temperature-associated host and pathogen responses during HTAS and nominate candidates for functional validation.