RNA interference targeting CYP76AH1 in hairy roots of Salvia miltiorrhiza reveals its key role in the biosynthetic pathway of tanshinones.
RNA interference targeting CYP76AH1 in hairy roots of Salvia miltiorrhiza reveals its key role in the biosynthetic pathway of tanshinones.
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DOI：
10.1016/j.bbrc.2016.06.036
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发表时间：
2016-08
影响因子：
3.1
通讯作者：
Ying Ma;Xiao-hui Ma;F. Meng;Z. Zhan;Juan Guo;Luqi Huang
中科院分区：
文献类型：
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作者：
Ying Ma;Xiao-hui Ma;F. Meng;Z. Zhan;Juan Guo;Luqi Huang
植物细胞色素p450 （CYPs）被认为是参与植物初级代谢和次生代谢产物化学多样性的最大的酶家族。阐明CYPs在次生代谢中的作用，以便通过代谢工程将其应用于药用植物中有价值的代谢物的生产，具有重要意义。CYP76AH1催化碳骨架米地拉二烯转化为中间体铁二醇，参与丹参主要生物活性成分丹参酮的生物合成。然而，其在植物中的作用仍有待阐明。本研究构建了毛状根的aCYP76AH1RNAi系统。RNAi-AH1毛状根系代谢分析显示，与对照相比，米替拉地尼积累量显著增加。同时，铁二醇浓度降低，显示其CYP76AH1的内催化活性。cyp76ah1沉默后，丹参酮含量显著下降，证实了其在丹参酮生物合成中的关键作用，提示其可作为代谢工程靶点。
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 thein vivorole 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 ofSalvia miltiorrhiza. However, its rolein plantaremains to be elucidated. In this work, we constructed aCYP76AH1RNAi 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 thein vivocatalytic activity of CYP76AH1. The content of tanshinones decreased significantly after silencing ofCYP76AH1, which verified its key role in the biosynthesis of tanshinones, and indicated that it could be used as a target for metabolic engineering.