The medicinal plant Stephania yunnanensis is rich in aporphine alkaloids, a type of benzylisoquinoline alkaloid (BIA), with aporphine being the representative and most abundant compound, but our understanding of the biosynthesis of BIAs in this plant has been relatively limited. Previous research reported the genome of S. yunnanensis and preliminarily identified the norcoclaurine synthase (NCS), which is involved in the early stages of the BIA biosynthetic pathways. However, the key genes promoting the formation of the aporphine skeleton have not yet been reported. In this study, based on the differences in the content of crebanine and several other BIAs in different tissues, we conducted transcriptome sequencing of roots, stems, and leaves. We then identified candidate genes through functional annotation and sequence alignment and further analyzed them in combination with the genome. Based on this analysis, we identified three CYP80 enzymes (SyCYP80Q5-1, SyCYP80Q5-3, and SyCYP80G6), which exhibited different activities toward (S)- and (R)-configured substrates in S. yunnanensis and demonstrated strict stereoselectivity enroute to aporphine. This study provides metabolomic and transcriptomic information on the biosynthesis of BIAs in S. yunnanensis, offers valuable insights into the elucidation of BIA biosynthesis, and lays the foundation for the complete analysis of pathways for more aporphine alkaloids.

The NBS-LRR genes constitute the largest class of resistance (R) proteins in plants, capable of recognizing pathogen-secreted effectors to trigger immune responses. However, systematic studies of NBS-LRR genes in medicinal plants have not yet been reported. In this study, we performed a comprehensive genome-wide identification and analysis of the NBS-LRR gene family in the medicinal plant Salvia miltiorrhiza. A total of 196 NBS-LRR genes were identified, among which 62 possessed complete N-terminal and LRR domains. Multiple NBS-LRR proteins extracted from other model plants can be classified and distinguished on the phylogenetic tree according to subfamilies CNL, TNL, and RNL. Comparative analysis revealed a marked reduction in the number of TNL and RNL subfamily members within the Salvia species. Analysis of the expression patterns of SmNBS-LRR genes with transcriptomes data revealed a close association between SmNBS-LRRs and secondary metabolism. Promoter analysis demonstrated an abundance of cis-acting elements in SmNBS genes related to plant hormones and abiotic stress. Our study enhances the understanding of the NBS-LRR gene family in medicinal plants and provides a foundation for future functional characterization of the NBS-LRR genes in S. miltiorrhiza and its potential application in disease-resistance breeding.