茶树Na+/H+逆向转运蛋白基因CsNHX1、CsNHX2的克隆及表达分析

陈江飞; 余津铭; 杨建坤; 余有本; 肖斌; 杨亚军; 王伟东

doi:10.13305/j.cnki.jts.2018.06.002

茶叶科学 >

2018 , Vol. 38 >Issue 6: 559 - 568

DOI: https://doi.org/10.13305/j.cnki.jts.2018.06.002

茶树Na⁺/H⁺逆向转运蛋白基因CsNHX1、CsNHX2的克隆及表达分析

陈江飞 ,
余津铭 ,
杨建坤 ,
余有本 ,
肖斌 ,
杨亚军 ,
王伟东

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1. 西北农林科技大学园艺学院,陕西杨凌 712100;
2. 中国农业科学院茶叶研究所,浙江杭州 310008

陈江飞,男,硕士研究生,主要从事茶树生理与分子生物学研究。

收稿日期: 2018-03-16

修回日期: 2018-07-25

网络出版日期: 2019-12-15

基金资助

中国博士后科学基金面上项目（2016M602873）、现代农业产业技术体系专项资金（CARS-19）、中央高校基本科研业务费专项资金（2452017074）

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Cloning and Expression Analysis of Na⁺/H⁺ Antiporter Gene CsNHX1 and CsNHX2 in Tea Plant (Camellia sinensis)

CHEN Jiangfei ,
YU Jinming ,
YANG Jiankun ,
YU Youben ,
XIAO Bin ,
YANG Yajun ,
WANG Weidong

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1. College of Horticulture, Northwest A&F University, Yangling 712100, China;
2. Tea Research Institute of the Chinese Academy of Agricultural Sciences, Hangzhou 310008, China

Received date: 2018-03-16

Revised date: 2018-07-25

Online published: 2019-12-15

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摘要

Na⁺/H⁺逆向转运蛋白（Na⁺/H⁺ antiporter,NHX）在植物生长发育与逆境响应过程中扮演着重要角色。本研究以龙井长叶茶树品种为材料,克隆获得了茶树CsNHX1和CsNHX2基因cDNA全长序列,GeneBank登录号分别为：MG722977和MG515211。生物信息学分析结果显示,CsNHX1和CsNHX2的cDNA全长分别为1β691βbp和1β757βbp,均包含1个1β626βbp的开放阅读框,编码541个氨基酸,预测分子量为59.5βkD和59.7βkD,理论等电点为7.07和8.79;蛋白序列分析结果显示,CsNHX1和CsNHX2属于典型的跨膜蛋白,均含有保守的Na⁺/H⁺ Exchanger结构域;进化树分析显示,CsNHX1和CsNHX2均为IC类中定位于液泡膜上的Class I成员。qRT-PCR结果显示,干旱、低温和盐胁迫能够显著诱导CsNHX1和CsNHX2上调表达;外源ABA处理下,CsNHX1和CsNHX2表达水平整体变化趋势不显著;高温胁迫处理下,茶树CsNHX1表达水平显著降低,而CsNHX2表达水平逐渐增加,表明茶树CsNHX1和CsNHX2参与了茶树对多种环境胁迫的响应过程,但对于不同逆境胁迫的应答模式存在一定差异。

关键词： 茶树; NHX; 基因克隆; 表达分析

本文引用格式

陈江飞 , 余津铭 , 杨建坤 , 余有本 , 肖斌 , 杨亚军 , 王伟东 . 茶树Na⁺/H⁺逆向转运蛋白基因CsNHX1、CsNHX2的克隆及表达分析[J]. 茶叶科学, 2018 , 38(6) : 559 -568 . DOI: 10.13305/j.cnki.jts.2018.06.002

Abstract

The Na⁺/H⁺ antiporter (NHX) plays an important role in plant growth, development and stress response. In this study, the full-length cDNA sequences of CsNHX1 and CsNHX2 (GenBank: MG722977 and MG515211) were cloned from tea cultivar ‘Longjingchangye’. Bioinformatics analysis showed that the full-length cDNA of CsNHX1 and CsNHX2 were 1β691βbp and 1β757βbp, all containing a 1β626βbp open reading frame and encoding 541 amino acids. The molecular weights of CsNHX1 and CsNHX2 were 59.5βkD and 59.7βkD and pI were 7.07 and 8.79, respectively.The results of protein sequence analysis showed that CsNHX1 and CsNHX2 contained the conserved Na⁺/H⁺ exchange domain, and belong to the typical transmembrane proteins. Phylogenetic analysis of plant NHX revealed that CsNHX1 and CsNHX2 are the member of Class I that localized to the vacuolar membrane in IC subfamily. In addition, qRT-PCR results showed that the expressions of CsNHX1 and CsNHX2 were induced by drought, low-temperature and salt stress. In contrast, exogenous ABA could not induce the expressions of CsNHX1 and CsNHX2. In addition, the expression level of CsNHX1 in tea plant decreased significantly, but that of CsNHX2 increased gradually under heat stress, indicating that CsNHX1 and CsNHX2 were differently involved in tea plant responding to environmental stress, and possibly through different responding modes.

Key words： tea plant; NHX; gene cloning; expression analysis

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