Multilayered Transcriptomic Reprogramming and Spliceosomal Divergence Shape Bipolaris sorokiniana Pathogenicity

Bipolaris sorokiniana, the causative agent of spot blotch of wheat, significantly limits wheat productivity. Despite the pathogens widespread impact, the in planta molecular mechanisms underpinning its virulence remain poorly characterized. We performed a comprehensive RNA-sequencing analysis of B. sorokiniana during attack on spot-blotch-resistant and susceptible wheat genotypes, integrating differentially expressed genes (DEGs), alternative splicing (AS) events, and identification of long non-coding RNAs (lncRNAs). The pathogen exhibited extensive host-genotype-dependent transcriptomic reprogramming, with 128 pathogen genes upregulated in the susceptible host. These genes were associated with ribosome biogenesis, RNA processing, and primary metabolic functions, supporting aggressive colonization. In contrast, pathogen attack on the resistant genotype triggered the upregulation of 58 pathogen genes associated with stress-responsive pathways, including sphingolipid and ceramide metabolism. This suggests a shift toward defensive metabolic reprogramming in the resistant host that restricted its proliferation. Our investigation uncovered five classes of AS events and 14 differentially expressed lncRNAs, revealing substantial post-transcriptional complexity. Notably, a subunit of the H/ACA small nucleolar ribonucleoprotein (snoRNP) complex emerged as a rare "triple-hit" candidate simultaneously identified as a DEG, differentially alternatively spliced gene, and target of a differentially expressed lncRNA, highlighting its potential as a central regulatory node in host-responsive stress adaptation. This study reveals a multilayered regulatory landscape involving transcriptional plasticity, alternative splicing, and lncRNA-mediated control, enabling B. sorokiniana to fine-tune its infection strategy in response to host resistance. This work advances the understanding of fungal pathogenesis and identifies molecular vulnerabilities that could be exploited for targeted, host-specific disease control.