Trifluoromethanesulfonamide Induces Male Sterility Through Systemic Metabolic Reprogramming and Anther-Specific Proline Deficiency

Chemical hybridization agents (CHAs) enable efficient, large-scale hybrid seed production, yet their mechanisms remain poorly understood. Understanding how CHAs induce male sterility at the metabolic level is important for both basic pollen biology and crop breeding. Here, we performed integrated metabolomic analyses to investigate the metabolic basis of the action of trifluoromethanesulfonamide (TFMSA) across multiple species and tissues. TFMSA treatment induced systemic metabolic reprogramming across species, prominently affecting amino acid metabolism, central carbon metabolism, and one-carbon metabolism. Although individual metabolite responses varied among species, pathway-level analyses consistently revealed coordinated modulation of carbon–nitrogen metabolic networks. In reproductive tissues, TFMSA induced tissue-specific metabolic changes. In cowpea anthers, proline was the only metabolite significantly altered and was strongly depleted, whereas in floral tissues several amino acids, including phenylalanine and tyrosine, were accumulated. Pathway analysis revealed altered amino acid metabolism, suggesting that systemic metabolic responses accompanied the proline reduction in anthers. These findings indicate that TFMSA induces male sterility through coordinated metabolic reprogramming across tissues and species, leading to depletion of key metabolites required for pollen development. This study provides a metabolic framework for understanding CHA-induced male sterility and highlights TFMSA as a powerful tool for probing metabolic regulation of pollen development.