新疆无苞芥Na+/H+ 逆向转运蛋白基因OpNHX1 的克隆、表达分析与功能验证

生物技术通报 ›› 2014, Vol. 0 ›› Issue (7): 74-80.

新疆无苞芥Na⁺/H⁺ 逆向转运蛋白基因OpNHX1 的克隆、表达分析与功能验证

赵云霞, 郭丹丽, 魏艳玲, 黄先忠

石河子大学生命科学学院农业生物技术重点实验室, 石河子 832003

收稿日期:2014-01-09 出版日期:2014-07-15 发布日期:2014-07-16
作者简介:赵云霞, 女, 硕士研究生, 研究方向：植物分子遗传；E-mail：yunxiaz_shzu@163.com
基金资助:
国家自然科学基金项目（31060149）, 石河子大学高层次人才启动项目（RCZX200902）, 新世纪优秀人才支持计划（NCET-12-1072）

Cloning, Expressing and Functional Analysis of Na⁺/H⁺ Antiporter Gene OpNHX1 from Olimarabidopsis pumila in Xinjiang

Zhao Yunxia, Guo Danli, Wei Yanling, Huang Xianzhong

Key Laboratory of Agrobiotechnology, College of Life Sciences, Shihezi University, Shihezi 832003

Received:2014-01-09 Published:2014-07-15 Online:2014-07-16

摘要/Abstract

摘要： 从耐盐植物无苞芥中克隆获得了1个Na⁺/H⁺逆向转运蛋白基因NHX1, 命名为OpNHX1（GenBank登录号：KC200248）。OpNHX1基因cDNA全长2153 bp, 包含一个1 605 bp的开放阅读框, 编码534个氨基酸。系统进化树分析表明, OpNHX1编码产物与拟南芥、小盐芥亲缘关系较近, 属于同一进化分支。实时荧光定量PCR分析表明, 该蛋白基因在无苞芥根、茎、叶、花和荚果中均有表达, 其中茎中表达量最高。半定量RT-PCR分析表明, 该基因受高盐、干旱、低温及ABA的诱导上调表达。进一步将该基因在拟南芥中过量表达, 显著提高了转基因植株在盐胁迫下的存活率, 说明OpNHX1基因参与了植物的耐盐性。酵母功能互补试验结果显示, 该基因转化酵母Δnhx1后可以补充NHX1的缺失, 表明OpNHX1参与Na⁺/H⁺的转运。

关键词: Na⁺/H⁺逆向转运蛋白, 无苞芥, 抗逆, 酵母功能互补

Abstract: A NHX1 gene, designated as OpNHX1（GenBank Accession No. KC200248）was isolated from Olimarabidopsis pumila, a plant for stress tolerance research. The cDNA of OpNHX1 was 2 153 bp in full length, contained an open reading frame（ORF）of 1 605 bp encoding a peptide of 534 amino acids residues. Phylogenetic analysis revealed that OpNHX1 had high sequence similarities with AtNHX1 and EhNHX1 and they belong to the same clade. The expression of OpNHX1 is induced by salt, PEG, cold, and abscisic acid. OpNHX1 is detected in roots, stems, leaves, flowers and siliques of Olimarabidopsis pumila, with the highest expression in stems. Importantly, overexpression of OpNHX1 in Arabidopsis thaliana improved the survival rate of transgenic plants under salt stress. This study provides valuable information to understand more about the molecular mechanism of salt tolerance in plant. Heterologous expression of OpNHX1 in W303-1B Δnhx1 yeast Na⁺/H⁺ antiporter mutants showed functional complementation.

Key words: Na⁺/H⁺ antiporter, Olimarabidopsis pumila, Stress tolerance, Functional complementation in yeast

赵云霞, 郭丹丽, 魏艳玲, 黄先忠. 新疆无苞芥Na⁺/H⁺ 逆向转运蛋白基因OpNHX1 的克隆、表达分析与功能验证[J]. 生物技术通报, 2014, 0(7): 74-80.

Zhao Yunxia, Guo Danli, Wei Yanling, Huang Xianzhong. Cloning, Expressing and Functional Analysis of Na⁺/H⁺ Antiporter Gene OpNHX1 from Olimarabidopsis pumila in Xinjiang[J]. Biotechnology Bulletin, 2014, 0(7): 74-80.

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