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cDNA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cDNA全长1β469βbp,其中开放阅读框（ORF）为1β218βbp,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44.6βkD,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 full length cDNA sequence of phosphoenolpyruvate transporter gene (CsPPT2) was obtained from tea plant(Camellia sinensis) cultivar ‘Baiye 1’ by polymerase chain reaction (PCR) and rapid amplification of cDNA ends PCR(RACE-PCR). The nucleotide sequence length of this gene was 1469 bp, containing a complete open reading frame (1218 bp) encoding 406 amino acids. Bioinformatic analysis showed that the predicted molecular weight of the protein was 44.6 kD, and the theoretic isoelectric point was 9.90. CsPPT2 has 6 trans-membrane regions, which may belong to the hydrophobic chloroplast trans-membrane protein. Phylogenetic tree analysis showed the phosphoenolpyruvate transporter family in tea plant could be divided into two subgroups, with CsPPT1 and CsPPT2 belonged to different subgroups. The expression of CsPPT1 and CsPPT2 were compared in different periods and plant organs. It showed that CsPPT2 was expressed in all tested tissues, with higher expression level in roots and mature leaves than those of CsPPT1. But the expression level of CsPPT1 in young shoots was higher than CsPPT2. CsPPT2 was transiently elevated at the beginning of the albino, but inhibited during the development of albino, indicates that the inhibition of the expression of CsPPT2 may act as a key factor for low-catechins and high-amino acids in ‘Baiye 1’.
关键词
CsPPT2 /
白叶1号 /
表达分析 /
茶树 /
磷酸烯醇式丙酮酸转运子
Key words
Baiye 1 /
CsPPT2 /
expression analysis /
phosphoenolpyruvate transporter /
Camellia sinensis
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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）
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