大豆GmWRKY35基因的克隆及其增强烟草耐旱能力研究

发布时间：2018-04-29 03:20

本文选题：大豆 + 烟草　；参考：《大豆科学》2017年05期

【摘要】：大豆易遭受干旱、盐、低温等非生物胁迫,严重导致产量下降。利用分子生物学技术提高作物抗性是作物育种的有效途径。锌指蛋白是植物中常见的重要转录因子,能够调控多种胁迫诱导基因的表达,有效提高综合抗性。在这项研究中,利用RT-PCR方法克隆大豆(Glycine max L.)GmWRKY35基因。其cDNA为1 500 bp,编码一个499个氨基酸的蛋白质,预测其分子量为54.89 kD,等电点(p I)为6.74。亚细胞定位分析表明,GmWRKY35定位于细胞核。实时荧光定量PCR分析显示,GmWRKY35转录能被干旱诱导。把GmWRKY35基因通过农杆菌介导转化烟草(Nicotiana tabacum L.)中。在干旱胁迫下,与野生型烟草相比,转基因烟草植株表现出主根较长,叶子萎焉少,POD和SOD活性较高,MDA含量和电解质渗漏率较少。主要功能验证表明,GmWRKY35基因在烟草中表达增强了干旱胁迫耐受性。这项研究表明大豆RING-H2型锌指蛋白在植物逆境中有重要作用,同时也为大豆抗性育种提供一个优良的候选基因。
[Abstract]:Soybean is vulnerable to drought, salt, low temperature and other abiotic stresses, which seriously lead to the decline of yield. It is an effective way to improve crop resistance by using molecular biology technology. Zinc finger protein is an important transcription factor in plants, which can regulate the expression of various stress-induced genes and effectively enhance the comprehensive resistance. In this study, the Glycine max L.)GmWRKY35 gene was cloned by RT-PCR method. Its cDNA is 1 500bp, which encodes a 499-amino acid protein. The predicted molecular weight is 54.89kD and the isoelectric point I) is 6.74. Subcellular localization analysis showed that GmWRKY35 was located in the nucleus. Real-time fluorescence quantitative PCR analysis showed that GmWRKY35 transcription could be induced by drought. Transformation of GmWRKY35 Gene into Nicotiana tabacum L. by Agrobacterium tumefaciens In. Under drought stress, transgenic tobacco plants showed longer main roots, higher POD and SOD activities and less electrolyte leakage. The main function test showed that the expression of GmWRKY35 gene in tobacco enhanced tolerance to drought stress. This study indicates that RING-H2 zinc finger protein plays an important role in plant stress and provides a good candidate gene for soybean resistance breeding.
【作者单位】：黄淮学院生物与食品工程学院;
【基金】：河南省科技发展计划(112300410042)
【分类号】：S565.1

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