AUTHOR=Chen Yue , Wang Yanting , Guo Juan , Yang Jian , Zhang Xiaodan , Wang Zixuan , Cheng Ying , Du Zewei , Qi Zhechen , Huang Yanbo , Dennis Mans , Wei Yukun , Yang Dongfeng , Huang Luqi , Liang Zongsuo 

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

JOURNAL=Frontiers in Plant Science

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.820582

DOI=10.3389/fpls.2021.820582

ISSN=1664-462X

ABSTRACT=<p>Tanshinones found in <italic>Salvia</italic> 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 <italic>Salvia castanea</italic> Diels f. <italic>tomentosa</italic> Stib. is about 49 times higher than that in <italic>Salvia miltiorrhiza</italic> Bunge. The molecular mechanism responsible for this phenomenon remains largely unknown. To address this, we performed comparative transcriptomic and proteomic analyses of <italic>S. miltiorrhiza</italic> and <italic>S. castanea</italic>. A total of 296 genes in <italic>S. castanea</italic> and 125 genes in <italic>S. miltiorrhiza</italic> 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 <italic>SmWRKY61</italic> was particularly high in <italic>S. castanea</italic>. Overexpression of <italic>SmWRKY61</italic> in <italic>S. miltiorrhiza</italic> could significantly increase the content of tanshinone I and tanshinone IIA, which were 11.09 and 33.37 times of the control, respectively. Moreover, <italic>SmWRKY61</italic> had a strong regulatory effect, elevating the expression levels of tanshinone pathway genes such as <italic>DXS2, CMK, HMGS2, 1, KSL1, KSL2, CYP76AH1</italic>, and <italic>CYP76AK3</italic>. For the WRKY family, 79 <italic>SmWRKY</italic>s were originally obtained and classified into three main groups. Collinearity analysis indicated a more specific extension of <italic>WRKY</italic> gene family in <italic>Salvia</italic> genus. In 55 <italic>Salvia</italic> species, only 37 species contained the <italic>WRKY61</italic> sequence, and high <italic>SmWRKY61</italic> expression in some <italic>Salvia</italic> L. species was often accompanied by high tanshinone accumulation. The above results suggest that <italic>SmWRKY61</italic> is a highly effective regulator of tanshinone accumulation and may be a key factor resulting in high tanshinone accumulation in <italic>S. castanea</italic>.</p>