大豆E3连接酶基因GmPLR-2的克隆及抗旱功能鉴定

Cloning of soybean E3 ligase gene GmPLR-2 and identification of its function in drought resistance

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pCAMBIA3301-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。

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 overexpression 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 transgenic 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 untransformed 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.