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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

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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.

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Abbreviations

Sm:

Salvia miltiorrhiza

MVA:

Mevalonate

MEP:

Methylerythritol phosphate

HMGR:

3-Hydroxy-3-methylglutaryl CoA reductase

DXR:

1-Deoxy-d-xylulose 5-phosphate reductoisomerase

HT:

Dihydrotanshinone

CT:

Cryptotanshinone

T1:

Tanshinone I

T2A:

Tanshinone IIA

TT:

Total tanshinone

HPLC:

High-performance liquid chromatography

LC/MS:

Liquid chromatography-mass spectrometry

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Acknowledgments

This work was supported by the National Natural Science Fund (31270007, 31201261, and 30900110), the Fok Ying-Tong Education Foundation for Young Teachers in the Higher Education Institutions of China (131041), the New Century Talent Project (NECT-13-0902), the Shanghai Talent Development Fund and the Open Funding Project of the State Key Laboratory of Bioreactor Engineering, Shanghai Science and Technology Committee Project (10JC1412000 and 09QH1401900), the Shanghai Education Committee Fund (13ZZ104, 09ZZ138, and J50401), and the Key Science and Technology Project of Yangtze River Delta (10140702018). Miss Pin Liu’s (Shanghai Jiaotong University, China) kind assistance with HPLC analysis is also acknowledged.

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Authors and Affiliations

  1. Development Center of Plant Germplasm Resources, College of Life and Environment Sciences, Shanghai Normal University, Shanghai, 200234, China

    Min Shi, Xiuqin Luo, Guanhua Ju, Xiaolong Hao, Qiang Huang, Jianbo Xiao, Lijie Cui & Guoyin Kai

  2. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China

    Guoyin Kai

  3. Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, 11794, USA

    Xiaohong Yu

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  1. Min Shi
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Correspondence to Guoyin Kai.

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Shi and Luo are co-first authors

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Shi, M., Luo, X., Ju, G. et al. 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. Funct Integr Genomics 14, 603–615 (2014). https://doi.org/10.1007/s10142-014-0385-0

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