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通过过表达 pmt 和 h6h 基因提高颠茄转基因植物中的莨菪碱产量。

Enhancing the scopolamine production in transgenic plants of Atropa belladonna by overexpressing pmt and h6h genes.

机构信息

Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Sweet potato Research Center, School of Life Sciences, Southwest University, Chongqing 400715, China.

出版信息

Physiol Plant. 2011 Dec;143(4):309-15. doi: 10.1111/j.1399-3054.2011.01506.x. Epub 2011 Sep 21.

Abstract

Atropa belladonna is officially deemed as the commercial plant to produce scopolamine in China. In this study we report the simultaneous overexpression of two functional genes involved in biosynthesis of scopolamine, which encode the upstream key enzyme putrescine N-methyltransferase (PMT) and the downstream key enzyme hyoscyamine 6β-hydroxylase (H6H), respectively, in transgenic herbal plants Atropa belladonna. Analysis of gene expression profile indicated that both pmt and h6h were expressed at a higher level in transgenic lines, which would be favorable for biosynthesis of scopolamine. High-performance liquid chromatography result suggested that transgenic lines could produce higher accumulation of scopolamine at different levels compared with wild-type lines. Scopolamine content increased to 7.3-fold in transgenic line D9 compared with control lines. This study not only confirms that co-overexpression of pmt and h6h is an ideal method to improve the biosynthetic capacity of scopolamine but also successfully cultivates the transgenic line D9, which significantly enhanced the scopolamine accumulation. Our research can serve as an alternative choice to provide scopolamine resources for relative industry, which is more competitive than conventional market.

摘要

颠茄被官方认定为在中国生产莨菪碱的商业植物。在本研究中,我们报告了同时过量表达参与莨菪碱生物合成的两个功能基因,它们分别编码上游关键酶腐胺 N-甲基转移酶(PMT)和下游关键酶莨菪碱 6β-羟化酶(H6H),在颠茄转基因草本植物中。基因表达谱分析表明,pmt 和 h6h 在转基因系中表达水平更高,有利于莨菪碱的生物合成。高效液相色谱结果表明,与野生型相比,转基因系在不同水平上可以产生更高水平的莨菪碱积累。与对照系相比,转基因系 D9 中的莨菪碱含量增加了 7.3 倍。本研究不仅证实了 pmt 和 h6h 的共过表达是提高莨菪碱生物合成能力的理想方法,而且还成功培育了转基因系 D9,显著提高了莨菪碱的积累。我们的研究可以作为提供莨菪碱资源的替代选择,为相关产业提供竞争优势,优于传统市场。

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