Long non-coding RNA (lncRNA) plays important roles in animals and plants. In filamentous fungi, however, their biological function in infection stage has been poorly studied. Here, we investigated the landscape and regulation of lncRNA in the filamentous plant pathogenic fungus Botrytis cinerea by strand-specific RNA-seq of multiple infection stages. In total, 1837 lncRNAs have been identified in B. cinerea. A large number of lncRNAs were found to be antisense to mRNAs, forming 743 sense-antisense pairs, of which 55 antisense lncRNAs and their respective sense transcripts were induced in parallel as the infection stage. Although small RNAs were produced from these overlapping loci, antisense lncRNAs appeared not to be involved in gene silencing pathways. In addition, we found the alternative splicing events occurred in lncRNA. These results highlight the developmental stage-specific nature and functional potential of lncRNA expression in the infection stage and provide fundamental resources for studying infection stage-induced lncRNAs.

Fusarium heading blight (FHB) is a devastating disease in wheat, primarily caused by field invasion of Fusarium graminearum. Due to the scarcity of resistant wheat varieties, the agricultural sector resorts to chemical fungicides to control FHB incidence. On the other hand, biocontrol represents a promising, eco-friendly approach aligned with sustainable and green agriculture concepts. In the present study, a bacterial endophyte, Pseudescherichia sp. (GSE25), was isolated from wheat seeds and identified through complete genome sequencing and phylogenetic analysis. In vitro testing of this endophytic strain demonstrated strong antifungal activity against F. graminearum PH-1 by inhibiting spore germination, suppressing germ tube growth, and causing cell membrane damage. Under field conditions, the strain GSE25 significantly reduced the FHB incidence and the associated deoxynivalenol mycotoxin accumulation by over 60% and 80%, respectively. These findings highlight the potential of the isolated bacterial endophyte Pseudescherichia sp. GSE25 strain as a biocontrol agent in protecting wheat from FHB-caused F. graminearum. This is the first report showing a biocontrol effect of Pseudescherichia sp. a strain against phytopathogens.

Alternative splicing plays a crucial role in enhancing the protein diversity of eukaryotic genomes. However, alternative splicing has not been extensively studied in Botrytis cinerea. In this study, we examined the distribution and regulation of alternative splicing in the filamentous plant pathogenic fungus B. cinerea through strand-specific RNA sequencing at various stages of infection of Solanum lycopersicum. During infection (pre-penetration and biotrophic stage), most spliceosome genes had upregulated expression levels, indicating that splicing is altered at this stage. A total of 3308 genes underwent alternative splicing in B. cinerea, resulting in 7466 alternative splicing events, most of which were stage-specific. Transcripts generated through alternative splicing typically exhibit lower expression levels, coding potential, and functional domains, which are more prevalent during the hyphal phase compared to the infestation phase. To conclude, our research offers an extensive analysis of the genome-wide alternative splicing in B. cinerea throughout the infection process, acting as a significant resource for further clarifying the pathogenic mechanisms associated with B. cinerea.

The nonsense-mediated mRNA decay (NMD) is extensively involved in physiological, pathological, and stress response processes in humans and plants. However, the NMD in phytopathogenic fungi has not yet been thoroughly investigated. In this study, we identified and performed domain analysis on the core components of the NMD in ten globally widespread phytopathogenic fungi that cause significant economic losses. The core components of NMD in these fungi exhibit high similarity to their homologous genes in humans, while also possessing certain specificities. The core factors of the NMD, including the Up-frameshift factors (UPFs) and the exon junction complex (EJC), are generally conserved among phytopathogenic fungi. Notably, suppressors with morphological effects on genitalia (SMG) genes are absent in these fungi, which bears some similarity to the EJC-independent NMD degradation mechanism observed in Saccharomyces cerevisiae. Interestingly, plant pathogenic fungi contain highly homologous genes of the EJC complex, suggesting the presence of an EJC-dependent NMD degradation mechanism. In summary, our findings demonstrate that NMD are prevalent in plant pathogenic fungi, providing a research foundation for subsequent studies on NMD in their growth, development, and involvement in pathogenic processes.