桑树Na+/H+逆向转运蛋白基因的克隆及功能分析

doi:10.3969/j.issn.1004-1524.2017.06.03

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (6): 874-881.DOI: 10.3969/j.issn.1004-1524.2017.06.03

桑树Na⁺/H⁺逆向转运蛋白基因的克隆及功能分析

刘岩, 计东风, 沈国新, 魏佳, 林天宝, 朱燕, 吕志强^*

浙江省农业科学院蚕桑研究所,浙江杭州 310021

收稿日期:2016-08-23 出版日期:2017-06-20 发布日期:2017-09-07
通讯作者: 吕志强,E-mail: 13958131715@139.com
作者简介:刘岩(1976—),女,河南焦作人,博士,副研究员,主要从事桑蚕分子生物学研究。E-mail: mayanly @sina.com
基金资助:
家蚕基因组生物学国家重点实验室开放课题(sklsgb2013015); 现代农业产业技术体系建设专项(蚕桑)(CARS-22-ZJ0105); 浙江省农业(畜禽)新品种选育重大科技专项(2016C02054)

Cloning and functional characterization of a Na⁺/H⁺ antiporter gene from mulberry

LIU Yan, JI Dongfeng, SHEN Guoxin, WEI Jia, LIN Tianbao, ZHU Yan, LYU Zhiqiang^*

Institute of Sericulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2016-08-23 Online:2017-06-20 Published:2017-09-07

摘要/Abstract

摘要： 盐害是威胁作物生长的重要因素之一,培育和种植耐盐作物品系是进行盐地开发利用的有效途径。该研究通过RACE方法,从特优1#桑品种克隆获得了Na⁺/H⁺逆向转运蛋白基因(MaNHX)的全长cDNA(2 016 bp),编码蛋白预测分子量59.28 ku,等电点6.67,含保守位点(⁸²LFFIYLLPPI⁹¹)。用根癌农杆菌介导法,将MaNHX基因导入拟南芥中,结果表明,在拟南芥中异源表达MaNHX能明显提高拟南芥对盐胁迫的耐受性。转基因植株NaCl胁迫下叶绿素含量更高、抗氧化活性增强,MDA含量下降。结果证实,异源表达MaNHX转基因拟南芥可以通过维持细胞膜稳定性、减少膜损伤和透性伤害等机制应对盐胁迫。该研究可为桑树的抗盐品种培育提供候选参考基因。

关键词: Na⁺/H⁺逆向转运蛋白基因, 盐胁迫, 桑树, 转基因

Abstract: Salinity is a major factor that adversely affects the growth and yield of plants worldwide. Plants Na⁺/H⁺ antiporters (NHX) play significant role in adaption to salt stress by compartmentalizing excess cytosolic Na⁺ into vacuoles and maintaining ionic equilibrium. Here we cloned an orthologous of NHX1 gene from You1#, a popular mulberry variety used especially in Southwest China. The length of MaNHX cDNA was 2 016 bp with a deduced molecular mass of 59.28 ku and pI 6.67. The consensus amiloride binding motif (⁸²LFFIYLLPPI⁹¹) was observed in the third putative transmembrane domain. Heterologous expression of MaNHX in Arabidopsis enhanced plants salt tolerance, which was verified by retaining higher chlorophyll content, higher antioxidase activity, slower reudction of malondialdehyde (MDA) content in transgenic lines than those in WT plants when confronted with the NaCl stress. Consequently, our work suggested that MaNHX was a candidate salt tolerance determinant gene in future mulberry breeding.

Key words: Na⁺/H⁺ antiporter pump, salinity stress, mulberry, transgenic

中图分类号:

S718.46

刘岩, 计东风, 沈国新, 魏佳, 林天宝, 朱燕, 吕志强. 桑树Na⁺/H⁺逆向转运蛋白基因的克隆及功能分析[J]. 浙江农业学报, 2017, 29(6): 874-881.

LIU Yan, JI Dongfeng, SHEN Guoxin, WEI Jia, LIN Tianbao, ZHU Yan, LYU Zhiqiang. Cloning and functional characterization of a Na⁺/H⁺ antiporter gene from mulberry[J]. , 2017, 29(6): 874-881.

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桑树Na⁺/H⁺逆向转运蛋白基因的克隆及功能分析

Cloning and functional characterization of a Na⁺/H⁺ antiporter gene from mulberry

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桑树Na+/H+逆向转运蛋白基因的克隆及功能分析

Cloning and functional characterization of a Na+/H+ antiporter gene from mulberry

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桑树Na⁺/H⁺逆向转运蛋白基因的克隆及功能分析

Cloning and functional characterization of a Na⁺/H⁺ antiporter gene from mulberry