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Tanshinones found in  Salvia  species are the main active compounds for the treatment of cardiovascular and cerebrovascular diseases, but their contents are hugely different in different species. For example, tanshinone IIA content in  Salvia castanea  Diels f.  tomentosa  Stib. is about 49 times higher than that in  Salvia miltiorrhiza  Bunge. The molecular mechanism responsible for this phenomenon remains largely unknown. To address this, we performed comparative transcriptomic and proteomic analyses of  S. miltiorrhiza  and  S. castanea . A total of 296 genes in  S. castanea  and 125 genes in  S. miltiorrhiza  were highly expressed at both the transcriptional and proteome levels, including hormone signal regulation, fungus response genes, transcription factors, and CYP450. Among these differentially expressed genes, the expression of  SmWRKY61  was particularly high in  S. castanea . Overexpression of  SmWRKY61  in  S. miltiorrhiza  could significantly increase the content of tanshinone I and tanshinone IIA, which were 11.09 and 33.37 times of the control, respectively. Moreover,  SmWRKY61  had a strong regulatory effect, elevating the expression levels of tanshinone pathway genes such as  DXS2, CMK, HMGS2, 1, KSL1, KSL2, CYP76AH1 , and  CYP76AK3 . For the WRKY family, 79  SmWRKY s were originally obtained and classified into three main groups. Collinearity analysis indicated a more specific extension of  WRKY  gene family in  Salvia  genus. In 55  Salvia  species, only 37 species contained the  WRKY61  sequence, and high  SmWRKY61  expression in some  Salvia  L. species was often accompanied by high tanshinone accumulation. The above results suggest that  SmWRKY61  is a highly effective regulator of tanshinone accumulation and may be a key factor resulting in high tanshinone accumulation in  S. castanea .

frontiers in plant science

Integrated Transcriptomics and Proteomics to Reveal Regulation Mechanism and Evolution of SmWRKY61 on Tanshinone Biosynthesis in Salvia miltiorrhiza and Salvia castanea