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拟南芥中编码GDP-L-半乳糖磷酸化酶(一种参与抗坏血酸生物合成的酶)的VTC2和VTC5基因的表达分析。

Expression analysis of the VTC2 and VTC5 genes encoding GDP-L-galactose phosphorylase, an enzyme involved in ascorbate biosynthesis, in Arabidopsis thaliana.

作者信息

Gao Yongshun, Badejo Adebanjo Ayobamidele, Shibata Hitoshi, Sawa Yoshihiro, Maruta Takanori, Shigeoka Shigeru, Page Mike, Smirnoff Nicholas, Ishikawa Takahiro

机构信息

Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University.

出版信息

Biosci Biotechnol Biochem. 2011;75(9):1783-8. doi: 10.1271/bbb.110320. Epub 2011 Sep 7.

Abstract

Arabidopsis thaliana contains two GDP-L-galactose phosphorylase genes, VTC2 and VTC5, which are critical for ascorbate (AsA) biosynthesis. We investigated the expression levels of both VTC2 and VTC5 genes in wild-type A. thaliana and the AsA deficient mutants during early seedling growth. Ascorbate accumulated to an equal extent in all genotypes up to 5 d post-germination (DPG). The transcript level of VTC2 was dominant, and increased in parallel with AsA accumulation in the wild type. On the other hand, the expression of VTC5 compensated for the reduced VTC2 transcription levels in the AsA deficient mutant vtc2-1 in young seedlings. A luciferase activity assay indicated that the VTC5 promoter was more active in young (2 DPG) cotyledons and that the VTC2 and VTC5 promoters drove a day-to-night variation in expression. The present work provides clues to the precise roles of VTC2 and VTC5 in AsA biosynthesis in A. thaliana at the young seedling stage.

摘要

拟南芥含有两个GDP-L-半乳糖磷酸化酶基因VTC2和VTC5,它们在抗坏血酸(AsA)生物合成中起关键作用。我们研究了野生型拟南芥以及AsA缺陷型突变体在幼苗早期生长过程中VTC2和VTC5基因的表达水平。在萌发后5天(DPG)之前,所有基因型中的抗坏血酸积累程度相同。VTC2的转录水平占主导地位,并且在野生型中与AsA积累平行增加。另一方面,在幼苗中,VTC5的表达补偿了AsA缺陷型突变体vtc2-1中VTC2转录水平的降低。荧光素酶活性测定表明,VTC5启动子在幼嫩(2 DPG)子叶中更活跃,并且VTC2和VTC5启动子驱动表达的昼夜变化。目前的工作为VTC2和VTC5在拟南芥幼苗期AsA生物合成中的精确作用提供了线索。

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