茶树谷胱甘肽过氧化物酶编码基因CsGPX1功能分析

doi:10.13305/j.cnki.jts.2019.04.003

摘要/Abstract

摘要： 为明确茶树[Camellia sinensis (L.) O. Kuntze.]谷胱甘肽过氧化物酶（Glutathione peroxidases,GPX）编码基因CsGPX1的功能,本文利用茶树转录组数据克隆获得CsGPX1的编码区全长序列,进行序列比对与分析,在此基础上,将CsGPX1在烟草中进行过表达获得转CsGPX1基因烟草,并比较野生型烟草与转基因烟草耐旱性的差异,验证CsGPX1功能。序列分析表明,CsGPX1编码序列（CDS）长723 bp,编码240个氨基酸。BLAST比对发现,该基因与桃树（Prunus persica）的GPX基因具有高度同源性（>85%）。CsGPX1编码的氨基酸序列具有GPX蛋白保守特征序列和区别于其他家族成员的特有的结构域——磷脂氢谷胱甘肽过氧化物酶（PHGPX）。干旱胁迫处理结果显示,过表达CsGPX1烟草株系抗旱性强于野生型。GPX酶活性测定结果表明,转基因植株的GPX酶活性高于野生型植株。以上研究结果表明,过表达CsGPX1可提高转基因烟草的耐旱能力,说明CsGPX1可能与茶树的耐旱性能相关。本研究为提高茶树的耐旱性能研究提供了新的理论途径,并对降低茶园管理的成本具有一定的积极意义。

关键词: 茶树, 谷胱甘肽, 过氧化物酶, 非生物胁迫, 抗旱机理

Abstract: To clarify function of glutathione peroxidase-encoding gene CsGPX1 in Camellia sinensis (L.) O. Kuntze, the full-length sequence of the coding region of CsGPX1 was obtained by transcriptome data of tea plant, and sequence analysis and function prediction were performed. Then, the CsGPX1 was overexpressed in tobacco. Subsequently, the difference in drought tolerance between wild-type and transgenic tobacco was compared to verify the function of CsGPX1. Sequence analysis indicates that CsGPX1 was 723 bp in length, encoding 240 amino acids. The Blast alignment reveals that the GPX showed more than 85% of identity in amino acid with GPX from Prunus persica. The amino acid sequence encoded by CsGPX1 has a conserved characteristic sequence of the GPX protein and a phospholipid hydrogen glutathione peroxidase (PHGPX) which is unique to other family members. The results of drought stress treatment show that the drought resistance of the over-expressing of CsGPX1 in tobacco lines was stronger than that of wild-type. The results of GPX enzyme activity assay show that the GPX enzyme activity of the transgenic plants was higher than that of the wild type plants. All of above results verify that the over-expression of CsGPX1 improved the drought tolerance of transgenic tobacco, suggesting that CsGPX1 might be related to the drought resistance in tea plants. This study could provide a new strategy to improve the drought resistance of tea plants, which would significantly reduce the management cost of tea plantation.

Key words: Camellia sinensis, glutathione, peroxidase, abiotic stress, drought resistance mechanism

中图分类号:

S571.1
Q52

刘赛, 刘硕谦, 龙金花, 吴敦超, 陈宇宏, 刘丽萍, 刘仲华, 田娜. 茶树谷胱甘肽过氧化物酶编码基因CsGPX1功能分析[J]. 茶叶科学, 2019, 39(4): 382-391. doi: 10.13305/j.cnki.jts.2019.04.003.

LIU Sai, LIU Shuoqian, LONG Jinhua, WU Dunchao, CHEN Yuhong, LIU Liping, LIU Zhonghua, TIAN Na. Functional Analysis of Glutathione Peroxidase Encoding Gene CsGPX1 in Camellia sinensis[J]. Journal of Tea Science, 2019, 39(4): 382-391. doi: 10.13305/j.cnki.jts.2019.04.003.

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