Salvia miltiorrhiza is well known for its clinical practice in treating heart and cardiovascular diseases. Its roots, used for traditional Chinese medicine materials, are usually brick-red due to accumulation of red pigments, such as tanshinone IIA and tanshinone I. Here we report a S. miltiorrhiza line (shh) with orange roots. Compared with the red roots of normal S. miltiorrhiza plants, the contents of tanshinones with a single bond at C-15,16 were increased, whereas those with a double bond at C-15,16 were significantly decreased in shh. We assembled a high-quality chromosome-level genome of shh. Phylogenomic analysis showed that the relationship between two S. miltiorrhiza lines with red roots was closer than the relationship with shh. It indicates that shh could not be the mutant of an extant S. miltiorrhiza line with red roots. Comparative genomic and transcriptomic analyses showed that a 1.0 kb DNA fragment was deleted in shh Sm2OGD3m. Complementation assay showed that overexpression of intact Sm2OGD3 in shh hairy roots recovered furan D-ring tanshinone accumulation. Consistently, in vitro protein assay showed that Sm2OGD3 catalyzed the conversion of cyptotanshinone, 15,16-dihydrotanshinone I and 1,2,15,16-tetrahydrotanshinone I into tanshinone IIA, tanshinone I and 1,2-dihydrotanshinone I, respectively. Thus, Sm2OGD3 functions as tanshinone 15,16-dehydrogenase and is a key enzyme in tanshinone biosynthesis. The results provide novel insights into the metabolic network of medicinally important tanshinone compounds.
Acknowledgements
We are thankful for financial support from the CAMS Innovation Fund for Medical Sciences (CIFMS) (2021-I2M-1-029) and the National Natural Science Foundation of China (81773836).
Author contributions
S.L. conceived and designed the study. X.P. and Y.C. conducted experiments and acquired data. X.P. and S.L. analysed and interpreted data. C.L., X.Q., X.C., F.M., and S.Z. helped with experiments and data analysis. X.L. provided plant materials. X.P. and S.L. wrote and revised the manuscript. All authors approved the final version of the manuscript.
Data availability
The data that supports the findings of this study are available in the supplementary material of this article. The genome sequence of S. miltiorrhiza line shh is available at NCBI BioProject PRJNA903271. The annotation information has been uploaded to the National Genomics Data Center (https://ngdc.cncb.ac.cn/) under the accession number WGS038566.
Conflict of interest statement
The authors declare no conflicts of interest.
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