[1]李大红,蒋炳伸,邬海燕,等.苜蓿MsDREB1基因的诱导表达增强大豆的耐盐性[J].大豆科学,2017,36(01):17-23.[doi:10.11861/j.issn.1000-9841.2017.01.0017]
　LI Da-hong,JIANG Bing-shen,WU Hai-yan,et al.MsDREB1 Overexpression Improves Tolerance to Salt Stress in Transgenic Glycine max L.[J].Soybean Science,2017,36(01):17-23.[doi:10.11861/j.issn.1000-9841.2017.01.0017]
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苜蓿MsDREB1基因的诱导表达增强大豆的耐盐性

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第36卷 期数: 2017年01期 页码: 17-23 栏目: 出版日期: 2017-01-20
Title:
MsDREB1 Overexpression Improves Tolerance to Salt Stress in Transgenic Glycine max L.
作者:
李大红; 蒋炳伸; 邬海燕; 李鸿雁
（黄淮学院 生物与食品工程学院，河南 驻马店 463000）
Author(s):
LI Da-hong; JIANG Bing-shen; WU Hai-yan; LI Hong-yan
（College of Biology and Food Engineering,Huanghuai University，Zhumadian 463000，China）
关键词:
大豆; MsDREB1; rd29A启动子; 耐盐性
Keywords:
Soybean;  MsDREB1;  rd29A promoter;  Salt tolerance
DOI:
10.11861/j.issn.1000-9841.2017.01.0017
文献标志码:
A
摘要:
基因转录调节是植物对非生物胁迫适应机制的一个重要方面，转录调节因子在胁迫信号转导途径中调节下游基因的表达，在建立植物对胁迫适应性过程中起到重要作用。DREB是功能多样的转录调节因子蛋白家族，家族成员在植物响应非生物胁迫方面扮演着重要角色。本研究以苜蓿MsDREB1基因为目的基因，分别把MsDREB1克隆到35S启动子与rd29A启动子之后，并把两种载体用农杆菌介导转入大豆基因组中，通过Southern检测转基因植株。15 d龄的幼苗在200 mmol·L-1NaCl胁迫条件下，用RT-PCR分析基因不同时间的表达差异；并测定叶绿素、丙二醛、H2O2、SOD、相对根长及相对地上部分长度。结果表明：转MsDREB1基因在两种启动子驱动下均有一定耐盐能力，但存在差异。在非胁迫下35S启动子调控的MsDREB1为超量表达，而rd29A启动子调控MsDREB1表达量较低；在盐胁迫下，rd29A：MsDREB1表达量高于35S：MsDREB1的表达量；MsDREB1超量表达抑制植株正常生长。MsDREB1 诱导表达耐盐性效果更明显，其植株脯氨酸含量、SOD活性均显著高于MsDREB1超量表达，而H2O2和MDA含量则显著低于MsDREB1超量表达。结果说明MsDREB1作为转录调节因子参与了植物的渗透调节，对植物的耐盐性具有贡献。 该试验研究两种启动子调控的转MsDREB1基因大豆耐盐效果，为MsDREB1基因在大豆耐盐基因工程中的应用提供参考。
Abstract:
Transcriptional regulation is one of the important factors in plant adaption to abiotic stress. Transcription factors play a critical role in regulating downstream genes expression in stress signaling pathway and the plant acclimation to adverse environments. Among transcription factors involved in abiotic stress response, DREB represents a large protein family which displays diverse roles in various biological processes. In this study, MsDREB1 was cloned into the 35S promoter and the rd29A promoter. The two vectors were transferred into soybean genome by Agrobacterium mediated. The transgenic plants were detected by Southern blot. Difference in gene expression of 15 day-old seedlings under 200 mmol·L-1NaCl stress conditions was analyzed by RT-PCR in different time, and the content of chlorophyll, MDA content, SOD, H2O2, relative root length and relative length of the above-ground parts of plants were determined. The results showed that there were significant differences in the regulation of MsDREB1 expression between the two promoters. MsDREB1 gene was overexpressed under the regulation of CaMV-35S promoter, but the expression was lower under the regulation of rd29A promoter under non stress. Under salt stress, the expression level of rd29A:MsDREB1 was higher than that of 35S:MsDREB1. Overexpression of MsDREB1inhibited the normal growth of plant. Both of the two transgenic lines had a certain ability to salt tolerance, but there were some differences. The effect of salt resistance of rd29A: MsDREB1 was more obvious. The chlorophyll content, proline content, SOD activity of rd29A:MsDREB1 were significantly higher than those of 35S: MsDREB1, while H2O2 and MDA content were significantly lower than that of 35S:MsDREB1 overexpression. Therefore, MsDREB1 functioned as a transcript regulator contributing to osmotic regulation in soybean plant under NaCl stress.

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备注/Memo

基金项目：河南省科技发展计划（112300410042）。

第一作者简介：李大红（1969-），男，博士，副教授，主要从事植物逆境生理研究工作。E-mail:lidahong27@163.com。
通讯作者：蒋炳伸（1975-），女，博士，副教授，主要从事植物生态学研究工作。E-mail:574618863@qq.com。

更新日期/Last Update: 2017-03-14