Comparative Transcriptomics Reveals Important Genes Underlying Heat-Tolerant Sterility in Photo-Thermo-Sensitive Male Sterile Wheat in Seed Production Environments

Maintaining stable male sterility is fundamental for ensuring the genetic purity and productivity of two-line hybrid wheat. However, unexpected heat events during the fertility-sensitive period can induce fertility restoration in photo-thermo-sensitive male sterile (PTMS) lines, posing a major threat to hybrid seed production. In this study, we identified two BS-type PTMS lines, BS166 and BS192, that consistently maintained sterility under heat stress in a seed-production environment, indicating strong heat-tolerant sterility. To uncover the molecular basis underlying this stability, we compared four BS-type PTMS lines exhibiting contrasting heat responses through field assessments, controlled heat treatments, transcriptome sequencing, and weighted gene co-expression network analysis (WGCNA). A total of 19,105 differentially expressed genes were identified, with the bisque4 module showing a significant correlation with seed setting rate. KEGG enrichment analysis revealed that starch and sucrose metabolism, cutin, suberin, and wax biosynthesis, fatty acid biosynthesis, and plant hormone signal transduction pathways were highly associated with heat-tolerant sterility. Core genes within these pathways displayed transcriptional stability in BS166 and BS192 but were strongly induced in heat-sensitive lines. In situ hybridization and RT-qPCR further confirmed tapetum-specific expression of <i>TaBGLU32</i> and <i>TaLACS1</i>. Based on these findings, we propose a regulatory model explaining how PTMS lines maintain sterility stability under heat stress.