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Analysis of Two l-Galactono-1,4-Lactone-Responsive Genes with Complementary Expression During the Development of Arabidopsisn thaliana

机译:拟南芥发育过程中两个具有互补表达的l-半乳糖-1,4-内酯响应基因的分析

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Unraveling the role of genes annotated as protein of unknown function is of importance in progression of plant science. l-Galactono-1,4-lactone (l-GalL) is the terminal precursor for ascorbic acid (AsA) biosynthesis in Arabidopsis thaliana, and a previous study showed two DUF (domains of unknown function) 642 family genes (At1g80240 and At5g25460, designated as DGR1 and DGR2, respectively) to be sensitive to it. In this work, leaves from wild-type Arabidopsis were fed with d-glucose, l-galactose, l-GalL and AsA, and the expression level of the At1g80240 and At5g25460 genes showed a specific response to l-GalL, but not to the other supplements despite the increases of the tissue AsA contents. Analysis of promoter–β-glucuronidase (GUS) transgenic plants showed the two genes to be complementarily expressed at the root apex and in the rest of the root excluding the apex, respectively, in both young and old seedlings, and to be expressed at the leaf primordia. The GUS activity under the control of the At5g25460 promoter was high in the cotyledon and leaf veins of young seedlings. These findings were consistent with the results of quantitative real-time PCR. Interestingly, the T-DNA insertion mutant of At5g25460 (SALK_125079) displayed shorter roots and smaller rosettes than Col-0; however, no phenotypic difference was observed between the T-DNA insertion mutant of At1g80240 and the wild type. This is the first report on the expression and functional analysis of these two DUF642 family genes, with the results revealing the contribution of DGR genes to the development of Arabidopsis.
机译:揭示被注释为功能未知的蛋白质的基因的作用在植物科学的发展中具有重要意义。 1-Galactono-1,4-lactone(1-GalL)是拟南芥中抗坏血酸(AsA)生物合成的末端前体,先前的研究显示了两个DUF(功能未知的域)642个家族基因(At1g80240和At5g25460,分别指定为DGR1和DGR2)对其敏感。在这项工作中,野生型拟南芥的叶片被喂以d-葡萄糖,l-半乳糖,l-GalL和AsA,并且At1g80240和At5g25460基因的表达水平显示出对l-GalL的特异性反应,但对其他补充剂,尽管组织中​​AsA含量增加。对启动子–β-葡萄糖醛酸苷酶(GUS)转基因植物的分析表明,这两个基因分别在年轻和老龄幼苗中在根尖和除根尖以外的其余根中互补表达,并在根尖表达。叶原基。在At5g25460启动子控制下,GUS活性在幼苗的子叶和叶脉中较高。这些发现与定量实时PCR的结果一致。有趣的是,与Col-0相比,At5g25460(SALK_125079)的T-DNA插入突变体显示出更短的根和更小的花环。但是,在At1g80240的T-DNA插入突变体与野生型之间没有观察到表型差异。这是关于这两个DUF642家族基因表达和功能分析的第一份报告,结果揭示了DGR基因对拟南芥发育的贡献。

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