GDP-D-mannose pyrophosphorylase （GMPase） catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced...GDP-D-mannose pyrophosphorylase （GMPase） catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced into potato by Agrobacterium-mediated transformation. Two transgenic lines with higher GMPase expression were selected using qPCR and protein blot analyses. The results showed that the content of L-ascorbic acid （AsA） and the ratio of AsA/ DHA （dehydroascorbate） significantly increased in both leaves and tubers of transgenic potato plants. Both pigment content and photosynthetic rate were much higher in transgenic plants than in wild-type plants. Transgenic plants showed a distinguishable change in phenotype from the wild-type plants. Furthermore, transgenic plants showed delayed senescence.展开更多
抗坏血酸(Vc)是重要的抗氧化剂,在茶树体内的各种生理代谢及抵御非生物胁迫中起重要作用,Vc含量关系到绿茶品质优劣。GDP-D-甘露糖焦磷酸化酶(GMP)是茶树Vc生物合成途径中的关键酶之一。本研究通过RACE-PCR方法从茶树中克隆了GMP c DNA...抗坏血酸(Vc)是重要的抗氧化剂,在茶树体内的各种生理代谢及抵御非生物胁迫中起重要作用,Vc含量关系到绿茶品质优劣。GDP-D-甘露糖焦磷酸化酶(GMP)是茶树Vc生物合成途径中的关键酶之一。本研究通过RACE-PCR方法从茶树中克隆了GMP c DNA全长序列,共计1510 bp,其中包含1 086 bp的开放阅读框(ORF),编码361个氨基酸,推测蛋白分子量为39.599 k Da。BLAST分析表明,茶树GMP基因氨基酸序列与猕猴桃的亲缘性最高,达到96%。实时定量PCR分析表明,茶树新梢芽下第三叶中GMP基因表达量最高,嫩茎中最低,不同品种茶树新梢叶片中表达量存在明显差异。高温胁迫初期,GMP基因表达量及抗坏血酸含量迅速升高,然后逐渐下降且均低于同期对照。展开更多
L-Ascorbic acid （AsA） plays an important role in plants and animals. In plants, GDP-D-mannose pyrophosphorylase （GMP） is essential in the AsA biosynthetic pathway. However, little is known about the genes encoding...L-Ascorbic acid （AsA） plays an important role in plants and animals. In plants, GDP-D-mannose pyrophosphorylase （GMP） is essential in the AsA biosynthetic pathway. However, little is known about the genes encoding GMP in soybean and here we report genetic and functional analysis of the GmGMP1 （Glycine max GDP-D-mannose pyrophosphorylase 1） gene in this species. GmGMP1 encoded a GDP-mannose pyrophosphorylase and exhibited higher transcript levels in the leaf than in the root, stem, flower, and seed. Transcript of this gene was ubiquitous in the vegetative and reproductive organs, and was induced by abiotic stress and light. Increasing expression of GmGMP1 in Arabidopsis and soybean through an overexpressing approach caused pronounced enhancement of AsA content, and was implicated in lowering the superoxide anion radical content and lipid peroxidation levels in Arabidopsis, and conferring tolerance to osmotic and high salt stresses during seed germination. The present study represents the first systematic determination of soybean genes encoding GDP-mannose pyrophosphorylase and provides useful evidence for the functional involvement of GmGMP1 in control of AsA content and conferring tolerance to osmotic and salt stress.展开更多
基金supported by the National Basic Re-search Program of China (2009CB119000)the National Natural Science Foundation of China (30771473)funded by Key Laboratory of Horticultural Crops Ge-netic Improvement, Ministry of Agriculture of China
文摘GDP-D-mannose pyrophosphorylase （GMPase） catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced into potato by Agrobacterium-mediated transformation. Two transgenic lines with higher GMPase expression were selected using qPCR and protein blot analyses. The results showed that the content of L-ascorbic acid （AsA） and the ratio of AsA/ DHA （dehydroascorbate） significantly increased in both leaves and tubers of transgenic potato plants. Both pigment content and photosynthetic rate were much higher in transgenic plants than in wild-type plants. Transgenic plants showed a distinguishable change in phenotype from the wild-type plants. Furthermore, transgenic plants showed delayed senescence.
文摘抗坏血酸(Vc)是重要的抗氧化剂,在茶树体内的各种生理代谢及抵御非生物胁迫中起重要作用,Vc含量关系到绿茶品质优劣。GDP-D-甘露糖焦磷酸化酶(GMP)是茶树Vc生物合成途径中的关键酶之一。本研究通过RACE-PCR方法从茶树中克隆了GMP c DNA全长序列,共计1510 bp,其中包含1 086 bp的开放阅读框(ORF),编码361个氨基酸,推测蛋白分子量为39.599 k Da。BLAST分析表明,茶树GMP基因氨基酸序列与猕猴桃的亲缘性最高,达到96%。实时定量PCR分析表明,茶树新梢芽下第三叶中GMP基因表达量最高,嫩茎中最低,不同品种茶树新梢叶片中表达量存在明显差异。高温胁迫初期,GMP基因表达量及抗坏血酸含量迅速升高,然后逐渐下降且均低于同期对照。
基金supported by the Genetically Modified Organisms Breeding Major Projects, China (2016ZX08004)the earmarked fund for China Agriculture Research System (CARS-004-PS10)the Program for Changjiang Scholars and Innovative Research Team in University, China (PCSIRT13073)
文摘L-Ascorbic acid （AsA） plays an important role in plants and animals. In plants, GDP-D-mannose pyrophosphorylase （GMP） is essential in the AsA biosynthetic pathway. However, little is known about the genes encoding GMP in soybean and here we report genetic and functional analysis of the GmGMP1 （Glycine max GDP-D-mannose pyrophosphorylase 1） gene in this species. GmGMP1 encoded a GDP-mannose pyrophosphorylase and exhibited higher transcript levels in the leaf than in the root, stem, flower, and seed. Transcript of this gene was ubiquitous in the vegetative and reproductive organs, and was induced by abiotic stress and light. Increasing expression of GmGMP1 in Arabidopsis and soybean through an overexpressing approach caused pronounced enhancement of AsA content, and was implicated in lowering the superoxide anion radical content and lipid peroxidation levels in Arabidopsis, and conferring tolerance to osmotic and high salt stresses during seed germination. The present study represents the first systematic determination of soybean genes encoding GDP-mannose pyrophosphorylase and provides useful evidence for the functional involvement of GmGMP1 in control of AsA content and conferring tolerance to osmotic and salt stress.
