RNA interference targeting CYP76AH1 in hairy roots of Salvia miltiorrhiza reveals its key role in the biosynthetic pathway of tanshinones.
Plant cytochrome P450s (CYPs) are well known as the largest family of enzymes that contribute to both primary metabolism and the chemical diversity of plant secondary metabolites. It is important to elucidate the in vivo role of CYPs in secondary metabolism, in order to apply them in the production of valuable metabolites in medicinal plants via metabolic engineering. CYP76AH1 has been suggested to catalyze the conversion of the carbon skeleton miltiradiene into the intermediate ferruginol, which is involved in the biosynthesis of tanshinones, the chief bioactive ingredients of Salvia miltiorrhiza. However, its role in planta remains to be elucidated. In this work, we constructed a CYP76AH1 RNAi system for hairy roots. Metabolic analysis of RNAi-AH1 hairy root lines showed a significantly increased accumulation of miltiradiene compared to the control lines. At the same time, the concentration of ferruginol decreased revealing the in vivo catalytic activity of CYP76AH1. The content of tanshinones decreased significantly after silencing of CYP76AH1, which verified its key role in the biosynthesis of tanshinones, and indicated that it could be used as a target for metabolic engineering.
Area Major Laboratory of Traditional Chinese Medicine, Beijing Normal University, Beijing 100875, China; State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: xiaoma1110@126.com.
Ying
Y
Ma
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China. Electronic address: maxiaohui1988@126.com.
Xiao-Hui
XH
Ma
Area Major Laboratory of Traditional Chinese Medicine, Beijing Normal University, Beijing 100875, China. Electronic address: mfy@bnu.edu.cn.
Fan-Yun
FY
Meng
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: zzlzhongyi@163.com.
Zhi-Lai
ZL
Zhan
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: guojuanzy@163.com.
Juan
J
Guo
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: huangluqi01@126.com.
Lu-Qi
LQ
Huang
D045784:Abietanes / Q000096:biosynthesis* / Q000235:genetics; D053898:Biosynthetic Pathways / Q000502:physiology*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference / Q000502:physiology*; D027561:Salvia miltiorrhiza / Q000235:genetics / Q000378:metabolism*
D045784:Abietanes; C021751:tanshinone; D003577:Cytochrome P-450 Enzyme System
Biosynthetic pathway; Cytochrome P450; RNAi; Salvia miltiorrhiza; Tanshinones