Comparative analysis of gene expression and metabolites in female and male Cannabis sativa flowers

Cannabis sativa L., a dioecious plant, exhibits sex-specific medicinal active ingredients. However, their causes remain unexplored. In this study, C. sativa flowers exhibited significant intersexual morphological differences, particularly in glandular trichomes. 1676 DEGs and 700 DAMs were identified. Among 149 genes related to sexual differentiation, 77 encode hormone-related products presumably involved in dioecy. Female flowers had cannabinoid levels 16.88 times those of males. Metabolomic sequencing identified 28 distinct terpenoids. We identified CsDXPS1 , CsLOX2 , CsLOX1.5 , CsAAE18 , CsAAE6-1 , CsAAE6-2 , CsGGR, and the metabolite mevalonate-5P; all except AAE6 were highly expressed in female flowers, regulating terpene and cannabinoid production. Seven glandular-trichome-related DEGs showed significant positive correlation with cannabinoid and terpene abundance; female-enriched CsMYC4 (TPS activator) and CsMYB49 (secretion enhancer) emerged as pivotal transcriptional regulators. These results provide a theoretical basis for the directional selection of female and male strains with high production and cultivation value.