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大豆E3连接酶基因GmPLR-2 的克隆及抗旱功能鉴定
引用本文:张靓,刘添祎,冀采凤,郭轩麟,邵瑞超,周靖萱,王丕武.大豆E3连接酶基因GmPLR-2 的克隆及抗旱功能鉴定[J].中国油料作物学报,2020,42(5):835.
作者姓名:张靓  刘添祎  冀采凤  郭轩麟  邵瑞超  周靖萱  王丕武
作者单位:吉林农业大学农学院,吉林长春,130118
基金项目:国家自然科学基金面上项目(31571689);吉林省教育厅科学技术研究项目(JJKH20190935KJ)
摘    要:PEG模拟干旱胁迫条件下E3连接酶GmPLR-2 基因在大豆根茎叶中呈上调表达,推测其可能参与大豆抗 旱调节。本研究克隆GmPLR-2 基因,构建植物过表达载体pCAMBIA3301-GmPLR-2、RNA干扰表达载体pCAM? BIA3301-GmPLR-2-RNAi并转化到吉农38中,对T2转基因植株进行分子检测,连续7 d不浇水后测定转基因叶片 中相关生理生化指标。结果表明:共获得T2过表达阳性植株12株,RNA干扰阳性植株11株。干旱胁迫7 d后过表 达植株中相对含水率、POD活性、SOD活性、Pro含量均高于未转化植株,而RNA干扰植株中则低于未转化植株;另 一方面过表达植株中相对电导率、MDA含量均低于未转化植株而RNA干扰植株中含量则高于未转化植株,且差异 极显著,说明GmPLR-2 基因的表达与大豆中相关抗旱指标的变化有关。

关 键 词:基因克隆  载体构建  遗传转化  抗旱性验证  

Cloning of soybean E3 ligase gene GmPLR-2 and identification of its function in drought resistance
ZHANG Liang,LIU Tian-yi,JI Cai-feng,GUO Xuan-lin,SHAO Rui-chao,ZHOU Jing-xuan,WANG Pi-wu.Cloning of soybean E3 ligase gene GmPLR-2 and identification of its function in drought resistance[J].Chinese Journal of Oil Crop Sciences,2020,42(5):835.
Authors:ZHANG Liang  LIU Tian-yi  JI Cai-feng  GUO Xuan-lin  SHAO Rui-chao  ZHOU Jing-xuan  WANG Pi-wu
Institution:College of Agriculture, Jilin Agricultural University, Changchun 130118, China
Abstract:Under drought stress, GmPLR-2 gene was up-regulated expression in soybean roots, stems, and leaves. The verification of this gene on drought resistance effection of soybean was necessary to develop the new drought-resistant soybean variety.In this study, the E3 ligase GmPLR-2 gene was cloned, and the plant overexpres? sion vector pCAMBIA3301-GmPLR-2 and the RNA interference expression vector pCAMBIA3301-GmPLR-2- RNAi were transformed into the recipient Jinong 38, and molecular detection was erformed on T2 generation trans?genic plants. The relevant physiological and biochemical indexes in the transgenic leaves were measured after seven days without watering. The results showed that the plant overexpression vector pCAMBIA3301-GmPLR-2 and the RNA interference expression vector pCAMBIA3301-GmPLR-2-RNAi were successfully constructed, and 12 T2 generation overexpression positive plants and 11 RNA interference positive plants were obtained. The relative water content, POD, SOD, and Pro content of over-expressed plants were higher than those of untransformed plants and the content of RNA interference plants was lower than that of untransformed plants after seven days of drought stress. Both are lower than untransformed plants and the content of RNA interference plants is higher than untrans? formed plants, and the difference is extremely significant, which indicates that the expression of GmPLR-2 gene is related to changes in related drought resistance indexes in soybean, and affects soybean drought resistance.
Keywords:gene cloning  vector construction  genetic transformation  drought resistance verification    
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