逆境诱导基因GsSNARE1的耐逆功能分析.pdfVIP

中国农业科学 2013,46(11):2191-2200 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2013.11.002 逆境诱导基因GsSNARE1 的耐逆功能分析 孙明哲，孙晓丽，于清悦，纪 巍，才 华，朱延明 （东北农业大学/农业生物功能基因重点实验室，哈尔滨 150030） 摘要：【目的】分离GsSNARE1，并分析其耐逆功能，为深入研究GsSNARE1 功能及分子机制奠定基础。【方法】 以耐盐野生大豆G50109 为材料，利用酵母双杂交验证GsSNARE1 与GsCBRLK 的互作关系，通过real-time PCR 分 析干旱和盐处理后GsSNARE1 的表达特性，对GsSNARE1 进行原核表达，分析GsSNARE1 抗盐旱功能。【结果】获得 与GsCBRLK 互作的GsSNARE1，同源克隆其全长CDS，并在酵母体内验证了二者的相互作用；Real-time PCR 分析 表明 GsSNARE1 受盐、干旱胁迫诱导，经PLACE 分析，发现其启动子中含有多个逆境胁迫相关的顺式作用元件； GsSNARE1 在植物不同组织中均表达；GsSNARE1 的表达降低了重组菌对盐、干旱胁迫的耐性。【结论】验证了GsSNARE1 与GsCBRLK 的互作关系，GsSNARE1 在不同组织中均表达，且受盐、干旱胁迫诱导，GsSNARE1 的表达降低了重组菌 耐盐、耐旱能力。 关键词：野生大豆； SNARE 蛋白；功能分析；盐胁迫；干旱胁迫 Functional Analysis of a Stress-Induced SNARE Gene GsSNARE1 in Response to Salt and Drought Stresses SUN Ming-zhe, SUN Xiao-li, YU Qing-yue, JI Wei, CAI Hua, ZHU Yan-ming (Northeast Agricultural University/Key Laboratory of Agricultural Biological Functional Genes, Harbin 150030) Abstract: 【Objective 】The aim of this study is to isolate GsSNARE1, characterize its function under environmental stress, and provide an important basis for studying the precise function and molecular mechanism of GsSNARE1. 【Method 】Glycine soja 50109 was used as gene cloning material, and the interaction between GsSNARE1 and GsCBRLK was verified by yeast two hybrid analysis. Real-time PCR analysis was used to analyze the expression profile of GsSNARE1 under stress conditions and in different plant tissues. The GsSNARE1 protein was expressed in E. coli, and its function was analyzed under salt and drought stresses. 【Result 】In this study, the GsCBRLK interacting protein GsSNARE1 was isolated, the full length GsSNARE1 gene was cloned,