Integrated cytological, physiological, and comparative transcriptome profiling reveals the regulatory network for male sterility in Welsh onion (Allium fistulosum L.)

Welsh onion (Allium fistulosum L.) is a specialty crop with value as a vegetable, condiment, and medicine. However, the molecular mechanisms related to pollen abortion and cytoplasmic male sterility are poorly understood, limiting the heterosis utilization and hybrid production of A. fistulosum. This study investigated the dynamic differences between male-sterile (MS) plants and male-fertile (MF) plants at different anther developmental stages based on integrated cytological, physiological, and transcriptomic analyses. Cytological observations revealed that pollen abortion in the MS plants began during the tetrad stage and was caused by vacuolated microspores, abnormal degradation of the tapetum, and inadequate nutrient supply. The physiological results showed that MS plants exhibited low total soluble sugar and starch contents and decreased antioxidant enzyme activities (peroxidase and catalase). Comparative transcriptome analysis identified 7005 differentially expressed genes (DEGs) and functional enrichment analysis indicated that significant transcriptomic differences were related to oxidative phosphorylation, starch and sucrose metabolism, and cutin, suberine, and wax biosynthesis. The relative expression levels of key DEGs in these three important metabolic pathways were determined using quantitative real-time polymerase chain reaction. On this basis, a core transcriptome-mediated MS network was proposed for MS plants with four functional components: inhibited sucrose synthesis and transport; blocked cutin, suberine, and wax synthesis; accelerated reactive oxygen species generation; and abnormal tapetal programmed cell death. These results provide theoretical guidance for further exploring the molecular mechanisms of male sterility in A. fistulosum.