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紫花苜蓿GDP-甘露糖3,5-表异构酶基因的过表达通过增加抗坏血酸积累增强转基因拟南芥的耐酸性、耐旱性和耐盐性。

Overexpression of an alfalfa GDP-mannose 3, 5-epimerase gene enhances acid, drought and salt tolerance in transgenic Arabidopsis by increasing ascorbate accumulation.

作者信息

Ma Lichao, Wang Yanrong, Liu Wenxian, Liu Zhipeng

机构信息

State Key Laboratory of Grassland Agro-ecosystems, School of Pastoral Agricultural Science and Technology, Lanzhou University, Lanzhou, 730020, People's Republic of China,

出版信息

Biotechnol Lett. 2014 Nov;36(11):2331-41. doi: 10.1007/s10529-014-1598-y. Epub 2014 Jun 30.

Abstract

GDP-mannose 3', 5'-epimerase (GME) catalyses the conversion of GDP-D-mannose to GDP-L-galactose, an important step in the ascorbic acid (ascorbic acid) biosynthetic pathway in higher plants. In this study, a novel cDNA fragment (MsGME) encoding a GME protein was isolated and characterised from alfalfa (Medicago sativa). An expression analysis confirmed that MsGME expression was induced by salinity, PEG and acidity stresses. MsGME overexpression in Arabidopsis enhanced tolerance of the transgenic plants to salt, drought and acid. Real-time PCR analysis revealed that the transcript levels of GDP-D-mannose pyrophosphorylase (GMP), L-galactose-phosphate 1-P phosphatase (GP) and GDP-L-galactose phosphorylase (GGP) were increased in transgenic Arabidopsis (T3 generation). Moreover, the ascorbate content was increased in transgenic Arabidopsis. Our results suggest that MsGME can effectively enhance tolerance of transgenic Arabidopsis to acid, drought and salt by increasing ascorbate accumulation.

摘要

GDP-甘露糖3',5'-表异构酶(GME)催化GDP-D-甘露糖转化为GDP-L-半乳糖,这是高等植物中抗坏血酸生物合成途径中的重要一步。在本研究中,从紫花苜蓿(Medicago sativa)中分离并鉴定了一个编码GME蛋白的新cDNA片段(MsGME)。表达分析证实,MsGME的表达受盐度、聚乙二醇和酸度胁迫诱导。MsGME在拟南芥中的过表达增强了转基因植物对盐、干旱和酸的耐受性。实时PCR分析表明,转基因拟南芥(T3代)中GDP-D-甘露糖焦磷酸化酶(GMP)、L-半乳糖-1-磷酸磷酸酶(GP)和GDP-L-半乳糖磷酸化酶(GGP)的转录水平升高。此外,转基因拟南芥中的抗坏血酸含量增加。我们的结果表明,MsGME可以通过增加抗坏血酸积累有效地增强转基因拟南芥对酸、干旱和盐的耐受性。

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