茶树磷酸烯醇式丙酮酸转运子CsPPT2基因的克隆和分析

发布时间：2019-06-02 22:07

【摘要】：以茶树(Camellia sinensis)白叶1号为材料,应用RT-PCR和RACE技术克隆获得茶树磷酸烯醇式丙酮酸转运子家族一个基因CsPPT2的c DNA序列,并与茶树体内另一个磷酸烯醇式丙酮酸转运子家族的基因CsPPT1在不同时期和不同部位的表达进行比较。CsPPT2的c DNA全长1 469 bp,其中开放阅读框(ORF)为1 218 bp,编码406个氨基酸。通过生物学信息分析表明,CSPPT2蛋白分子量为44.6 k D,理论等电点为9.90,CsPPT2有6个跨膜区,属于疏水性叶绿体跨膜蛋白。聚类分析显示,茶树磷酸烯醇式丙酮酸转运子家族分为2个亚组,而CsPPT1与CsPPT2属于不同亚组。荧光定量结果分析表明,两个基因可能在茶树体内功能不同,CsPPT2在所考察的组织中均有表达,且在根和成熟叶片中表达量远大于CsPPT1;CsPPT1在茎和复绿前的幼嫩芽叶中表达量高于CsPPT2。在白化期起始时CsPPT2有短暂的升高,但伴随白化受到较大的抑制,表明CsPPT2的表达抑制可能是白叶1号低儿茶素高氨基酸品质形成的关键因素。
[Abstract]:A c-DNA sequence of a gene, CsPPT2, was obtained by using RT-PCR and RACE to obtain a gene CsPPT2 from the family of tea tree (Camellia sinensis). And compared with the expression of the gene CsPPT1 in the other phosphoenolpyruvate transporter family in the tea tree in different times and at different parts. The total length of c-DNA of CsPPT2 was 1,469bp, and the open reading frame (ORF) was 1,218 bp, encoding 406 amino acids. The molecular weight of the CSPPT2 protein is 44.6 kD, the theoretical isoelectric point is 9.90, and the CsPPT2 has 6 transmembrane domains, and belongs to the hydrophobic chloroplast transmembrane protein. The cluster analysis showed that the trienol-type pyruvate transport subfamily of the tea tree was divided into 2 subgroups, and CsPPT1 and CsPPT2 belong to different subgroups. The results of fluorescence quantitative analysis show that the two genes may be different in the tea tree, and CsPPT2 is expressed in the tissues examined, and the expression of CsPPT1 in the root and the mature leaves is much higher than that of CsPPT1; and the expression of CsPPT1 in the leaves of the young shoots before the stem and the compound green is higher than that of CsPPT2. At the beginning of the albino stage, the CsPPT2 has a short increase, but the concomitant whitening is greatly inhibited, indicating that the inhibition of the expression of CsPPT2 may be a key factor for the formation of the low-catechin high-amino-acid quality of the white-leaf No.1.
【作者单位】：南京农业大学茶叶科学研究所;福建省农业科学院茶叶研究所;
【基金】：现代农业产业技术体系建设专项资金(CARS-23) 江苏高校优势学科建设工程资助项目国家自然科学基金(31570691) 江苏省科技支撑计划(BE2009313-1)
【分类号】：S571.1

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