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The transcription factor OpWRKY2 positively regulates the biosynthesis of the anticancer drug camptothecin in Ophiorrhiza pumila

查看全文 作　　者：Xiaolong [1]Hao;Chenhong [1]Xie;Qingyan [1]Ruan;Xichen [2]Zhang;Chao [2]Wu;Bing [1]Han;Jun [1]Qian;Wei [1]Zhou;Hans-Wilhelm [3]Nutzmann;Guoyin [1]Kai 高影响力作者 机构地区：[1]Laboratory of Medicinal Plant Biotechnology,College of Pharmacy,Zhejiang Chinese Medical University,310053 Hangzhou,China;[2]Institute of Plant Biotechnology,School of Life Sciences,Shanghai Normal University,200234 Shanghai,China;[3]The Milner Centre for Evolution,Department of Biology and Biochemistry,University of Bath,Claverton Down,Bath BA27AY,UK高影响力机构 出　　处：《Horticulture Research》索引2021年第8卷第1期,共14页高影响力期刊 基　　金：the National Key Research and Development Program of China(2018YFC1706203);National Natural Science Foundation of China(31571735,82073963,81522049,82003889);Zhejiang Provincial Ten Thousands Program for Leading Talents of Science and Technology Innovation(2018R52050);Zhejiang Provincial Program for the Cultivation of High-level Innovative Health Talents,Shanghai Science and Technology Committee Project(17JC1404300);Zhejiang Provincial Natural Science Foundation of China(LY20H280008,LQ21H280004);Zhejiang Chinese Medical University Research Foundation(2020ZR15). 摘　　要：The limited bioavailability of plant-derived natural products with anticancer activity poses major challenges to the pharmaceutical industry.An example of this is camptothecin,a monoterpene indole alkaloid with potent anticancer activity that is extracted at very low concentrations from woody plants.Recently,camptothecin biosynthesis has been shown to become biotechnologically amenable in hairy-root systems of the natural producer Ophiorrhiza pumila.Here,time-course expression and metabolite analyses were performed to identify novel transcriptional regulators of camptothecin biosynthesis in O.pumila.It is shown here that camptothecin production increased over cultivation time and that the expression pattern of the WRKY transcription factor encoding gene OpWRKY2 is closely correlated with camptothecin accumulation.Overexpression of OpWRKY2 led to a more than three-fold increase in camptothecin levels.Accordingly,silencing of OpWRKY2 correlated with decreased camptothecin levels in the plant.Further detailed molecular characterization by electrophoretic mobility shift,yeast one-hybrid and dual-luciferase assays showed that OpWRKY2 directly binds and activates the central camptothecin pathway gene OpTDC.Taken together,the results of this study demonstrate that OpWRKY2 acts as a direct positive regulator of camptothecin biosynthesis.As such,a feasible strategy for the over-accumulation of camptothecin in a biotechnologically amenable system is presented. 关 键 词：CAMPTOTHECIN enable ALKALOID
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