Promoting scopolamine biosynthesis in transgenic Atropa belladonna plants with pmt and h6h overexpression under field conditions
Promoting scopolamine biosynthesis in transgenic Atropa belladonna plants with pmt and h6h overexpression under field conditions
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在田间条件下通过 pmt 和 h6h 过表达促进转基因颠茄植物中东莨菪碱的生物合成
DOI：
10.1016/j.plaphy.2016.04.034
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发表时间：
2016-09-01
影响因子：
6.5
通讯作者：
Liao, Zhihua
中科院分区：
文献类型：
--
作者：
Xia, Ke;Liu, Xiaoqiang;Liao, Zhihua
颠茄（Atropa belladonna）是生产药用托烷类生物碱（tropane alkaloids，TA）的重要植物来源之一。转基因A.颠茄，其中腐胺N-甲基转移酶（EC. 2.1.1.53）和莨菪碱6 β-羟化酶（EC. 1.14.11.14）过量表达尼日尔（HnH 6 H），研究田间条件下TA在成熟果实、叶、茎、初生根和次生根中的生物合成和分布。NtPMT和HnH 6 H在转基因植物中在转录水平上被检测到，而在野生型植物中未检测到。转基因不影响A.颠茄所有4个内源TA生物合成基因（AbPMT、AbTRI、AbCY 1380 F1和AbH 6 H）在次生根中均具有最高/排他性表达水平，表明TA主要在次生根中合成。转基因A.颠茄表现出令人印象深刻的富含东莨菪碱的化学型，这大大提高了颠茄的药用价值。颠茄地上部分的莨菪碱转化效率高于地下部分。在转基因植株的地上部分，莨菪碱完全转化为下游生物碱，尤其是东莨菪碱。在地下部分检测到莨菪碱、山莨菪碱和东莨菪碱，但东莨菪碱和山莨菪碱的含量高于莨菪碱。山莨菪碱在根中的高水平表明它可能难以从根转移到地上器官。转基因A.颠茄在田间条件下产生非常高的东莨菪碱水平（2.94 - 5.13 mg g（-1）），可为东莨菪碱生产提供更有价值的植物材料。(C)2016 Elsevier Masson SAS。All rights reserved.
Atropa belladonna is one of the most important plant sources for producing pharmaceutical tropane alkaloids (TAs). T1 progeny of transgenic A. belladonna, in which putrescine N-methyltransferase (EC. 2.1.1.53) from Nicotiana tabacum (NtPMT) and hyoscyamine 6 beta-hydroxylase (EC. 1.14.11.14) from Hyoscyamus niger (HnH6H) were overexpressed, were established to investigate TA biosynthesis and distribution in ripe fruits, leaves, stems, primary roots and secondary roots under field conditions. Both NtPMT and HnH6H were detected at the transcriptional level in transgenic plants, whereas they were not detected in wild-type plants. The transgenes did not influence the root-specific expression patterns of endogenous TA biosynthetic genes in A. belladonna. All four endogenous TA biosynthetic genes (AbPMT, AbTRI, AbCY1380F1 and AbH6H) had the highest/exclusive expression levels in secondary roots, suggesting that TAs were mainly synthesized in secondary roots. T1 progeny of transgenic A. belladonna showed an impressive scopolamine-rich chemotype that greatly improved the pharmaceutical value of A. belladonna. The higher efficiency of hyoscyamine conversion was found in aerial than in underground parts. In aerial parts of transgenic plants, hyoscyamine was totally converted to downstream alkaloids, especially scopolamine. Hyoscyamine, anisodamine and scopolamine were detected in underground parts, but scopolamine and anisodamine were more abundant than hyoscyamine. The exclusively higher levels of anisodamine in roots suggested that it might be difficult for its translocation from root to aerial organs. T1 progeny of transgenic A. belladonna, which produces scopolamine at very high levels (2.94 -5.13 mg g(-1)) in field conditions, can provide more valuable plant materials for scopolamine production. (C) 2016 Elsevier Masson SAS. All rights reserved.