Increased accumulation of the cardio-cerebrovascular disease treatment drug tanshinone in Salvia miltiorrhiza hairy roots by the enzymes 3-hydroxy-3-methylglutaryl CoA reductase and 1-deoxy-d-xylulose 5-phosphate reductoisomerase

Abstract

Tanshinone is widely used for treatment of cardio-cerebrovascular diseases with increasing demand. Herein, key enzyme genes SmHMGR (3-hydroxy-3-methylglutaryl CoA reductase) and SmDXR (1-deoxy-D-xylulose 5-phosphate reductoisomerase) involved in the tanshinone biosynthetic pathway were introduced into Salvia miltiorrhiza (Sm) hairy roots to enhance tanshinone production. Over-expression of SmHMGR or SmDXR in hairy root lines can significantly enhance the yield of tanshinone. Transgenic hairy root lines co-expressing HMGR and DXR (HD lines) produced evidently higher levels of total tanshinone (TT) compared with the control and single gene transformed lines. The highest tanshinone production was observed in HD42 with the concentration of 3.25 mg g(-1) DW. Furthermore, the transgenic hairy roots showed higher antioxidant activity than control. In addition, transgenic hairy root harboring HMGR and DXR (HD42) exhibited higher tanshinone content after elicitation by yeast extract and/or Ag(+) than before. Tanshinone can be significantly enhanced to 5.858, 6.716, and 4.426 mg g(-1) DW by YE, Ag(+), and YE-Ag(+) treatment compared with non-induced HD42, respectively. The content of cryptotanshinone and dihydrotanshinone was effectively elevated upon elicitor treatments, whereas there was no obvious promotion effect for the other two compounds tanshinone I and tanshinone IIA. Our results provide a useful strategy to improve tanshinone content as well as other natural active products by combination of genetic engineering with elicitors.

Molecular cloning and transgenic characterization of the genes encoding chalcone synthase and chalcone isomerase from the Tibetan herbal plant Mirabilis himalaica

“…There are a lot of successful cases in this field. In A. belladonna and A. acutangulus , biosynthesis of scopolamine was dramatically enhanced by overexpressing the committed enzymes, H6H and TRI in A. annua , the production of artemisinin was greatly promoted by overexpressing CYP71AV1 and aldehyde 11Δ(13) reductase ; in Catharanthus roseus , overexpressing strictosidine synthase led to the increased production of ajmalicine ; in Ophiorrhiza pumila , the anti‐cancer camptothecin was significantly improved by overexpressing geraniol‐10‐hydroxylase and strictosidine synthase ; in Salvia miltiorrhiza , biosynthesis of tanshinone was enhanced by different combination of the key enzymes . These cases strongly suggest that overexpression of key enzymes is a very potent tool for genetic improvement of metabolite biosynthesis.…”

SmMYB36, a Novel R2R3-MYB Transcription Factor, Enhances Tanshinone Accumulation and Decreases Phenolic Acid Content in Salvia miltiorrhiza Hairy Roots

“…Photos of hairy roots and their extracts were recorded to display the correlation between colour difference and the total tanshinone content 74, 101 . The sum of dihydrotanshinone I, cryptotanshinone, tanshinone I and tanshinone II A was calculated as total tanshinones in this research.…”

Impact of Abiotic Elicitors on In vitro Production of Plant Secondary Metabolites: A Review

“…In an attempt to enhance betalaines production, the hairy roots were exposed to metal ions [14]. It was reported that Ca 2+ , Ag + and Cd 2+ could improve the production of tropane alkaloids, scopolamine and hyoscyamine, in hairy roots cultures of Brugmansia candida [15,16] [17][18][19][20]. Among them, Ag + was considered as an effective elicitor for phenolic compound and tanshinone production in S. miltiorrhiza hairy roots and could improve rosmarinic acid, salvianolic acid B and tanshinones production.…”