Maturation of Eupyrene Sperm upon Ejaculation Is Influenced by a Male Accessory Gland-Derived Serine Protease in .

is a globally significant fruit pest. Females achieve maximal reproductive output through efficient sperm utilization following a single copulation. Post-mating maturation of eupyrene sperm is a critical step in reproductive success. Here, we report that a male accessory gland-derived serine protease (named GmAGSP1) is essential for this process. GmAGSP1 was only distantly related to other identified sperm-activating SPs, and its transcript was highly expressed in the AG at 48 h after emergence. RNAi-mediated knockdown of in males did not affect courtship rate, copulation duration, or mating frequency, whereas male fertility decreased significantly. Mating with -knockdown males markedly impaired eupyrene sperm maturation in the spermatophores, with phenotypes including failure of eupyrene sperm bundles to dissociate normally and marked reduction in viability of the dissociated eupyrene sperm. Finally, untargeted metabolomic analysis preliminarily demonstrated marked alterations in multiple metabolic pathways within the spermatophore following mating with -knockdown males. This study advances our understanding of the regulatory mechanism of "sperm activation in the spermatophore's metabolic microenvironment mediated by male AG-derived SP" while providing critical insights for the development of novel genetic control strategies targeting .