Abstract
Tanshinones are a group of bioactive abietane-type norditerpenoid quinone compounds in Salvia miltiorrhiza. Copalyldiphosphate synthase of S. miltiorrhiza (SmCPS) is the first key enzyme in tanshinone biosynthesis from the universal diterpene precursor geranylgeranyl diphosphate. Hairy roots of S. miltiorrhiza were transformed with Agrobacterium rhizogenes carrying an RNA interference (RNAi) construct designed to silence SmCPS, and we examined the resulting SmCPS expression and tanshinone accumulation. In SmCPS–RNAi hairy roots, the transcript level of SmCPS was reduced to 26 % while the dihydrotanshinone I and cryptotanshinone levels were decreased by 53 and 38 % compared to those of the vector control hairy roots; tanshinone IIA was not detected. Therefore, the decreased expression of SmCPS caused a decrease in tanshinone levels which verifies that SmCPS is a key enzyme for tanshinone biosynthesis in S. miltiorrhiza.
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Asano T, Kobayashi K, Kashihara E, Sudo H, Sasaki R, Iijima Y, Aoki K, Shibata D, Saito K, Yamazaki M (2013) Suppression of camptothecin biosynthetic genes results in metabolic modification of secondary products in hairy roots of Ophiorrhiza pumila. Phytochemistry 91:128–139
Cheng QQ, He YF, Li G, Liu YJ, Gao W, Huang LQ (2013) Effects of combined elicitors on tanshinone metabolic profiling and SmCPS expression in Salvia miltiorrhiza hairy root cultures. Molecules 18:7473–7485
DeBoer KD, Lye JC, Aitken CD, Su AKK, Hamill JD (2009) The A622 gene in Nicotiana glauca (tree tobacco): evidence for a functional role in pyridine alkaloid synthesis. Plant Mol Biol 69:299–312
Dong Y, Morris-Natschke SL, Lee KH (2011) Biosynthesis, total syntheses, and antitumor activity of tanshinones and their analogs as potential therapeutic agents. Nat Pro Rep 28:529–542
Fire A, Xu SQ, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811
Gao W, Hillwig ML, Huang LQ, Cui GH, Wang X, Kong J, Yang B, Peters RJ (2009) A functional genomics approach to tanshinone biosynthesis provides stereochemical insights. Org Lett 11:5170–5173
Guo J, Zhou YJ, Hillwig ML, Shen Y, Yang L, Wang YJ, Zhang XN, Liu WJ, Peters RJ, Chen XY, Zhao ZB, Huang LQ (2013) CYP76AH1 catalyzes turnover of miltiradiene in tanshinones biosynthesis and enables heterologous production of ferruginol in yeasts. Proc Natl Acad Sci USA 110:12108–12113
Hall DE, Zerbe P, Jancsik S, Quesada AL, Dullat H, Madilao LL, Yuen M, Bohlmann J (2013) Evolution of conifer diterpene synthases: diterpene resin acid biosynthesis in lodgepole pine and jack pine involves monofunctional and bifunctional diterpene synthases. Plant Physiol 161:600–616
Hannon GJ (2002) RNA interference. Nature 418:244–251
Keeling CI, Dullat HK, Yuen M, Ralph SG, Jancsik S, Bohlmann J (2010) Identification and functional characterization of monofunctional ent-copalyl diphosphate and ent-kaurene synthases in white spruce reveal different patterns for diterpene synthase evolution for primary and secondary metabolism in gymnosperms. Plant Physiol 152:1197–1208
Pavli OI, Panopoulos NJ, Goldbach R, Skaracis GN (2010) BNYVV-derived dsRNA confers resistance to rhizomania disease of sugar beet as evidenced by a novel transgenic hairy root approach. Transgenic Res 19:915–922
Peters RJ (2010) Two rings in them all: the labdane-related diterpenoids. Nat Prod Rep 27:1521–1530
Subramanian S, Graham MY, Yu O, Graham TL (2005) RNA interference of soybean isoflavone synthase genes leads to silencing in tissues distal to the transformation site and to enhanced susceptibility to Phytophthora sojae. Plant Physiol 137:1345–1353
Wang X, Morris-Natschke SL, Lee KH (2007) New developments in the chemistry and biology of the bioactive constituents of Tanshen. Med Res Rev 27:133–148
Zhou LM, Zuo Z, Chow MSS (2005) Danshen: an overview of its chemistry, pharmacology, pharmacokinetics, and clinical use. J Clin Pharmacol 4:1345–1359
Zhou YJ, Gao W, Rong QX, Jin GJ, Chu HY, Liu WJ, Yang W, Zhu ZW, Li GH, Zhu GF, Huang LQ, Zhao ZB (2012) Modular pathway engineering of diterpenoid synthases and the mevalonic acid pathway for miltiradiene production. J Am Chem Soc 134:3234–3241
Acknowledgments
This work was supported by the National Natural Science Foundation of China (81072990 and 30901965), the Foundation for the Author of National Excellent Doctoral Dissertation of China (201188), Beijing Natural Science Foundation (5102009), and Beijing Outstanding Talent Plan (2011D005018000002).
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Cheng, Q., Su, P., Hu, Y. et al. RNA interference-mediated repression of SmCPS (copalyldiphosphate synthase) expression in hairy roots of Salvia miltiorrhiza causes a decrease of tanshinones and sheds light on the functional role of SmCPS. Biotechnol Lett 36, 363–369 (2014). https://doi.org/10.1007/s10529-013-1358-4
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DOI: https://doi.org/10.1007/s10529-013-1358-4