GmVP1基因克隆与转GmVP1/GmNHX1双价基因的大豆发状根耐盐性分析

Cloning and transformation of GmVP1 gene and salt tolerance analysis of transgenic soybean hairy roots with GmVP1 and GmNHX1 gene

将Na^+外排和区隔化作用的Na^+/H^+逆向转运蛋白(NHX)的基因和能够为其提供跨液泡膜H^+梯度驱动力的液泡膜H^+-焦磷酸酶(VP)的基因共转化能够提高植物耐盐性。本研究从抗盐大豆品种‘冀豆7号’中克隆得到液泡膜H^+-焦磷酸酶基因GmVP1,构建了GmVP1单价以及GmVP1与GmNHX1双价转化载体,利用发根农杆菌介导的大豆子叶转化体系验证其功能。结果表明,GmVP1在转录水平响应盐胁迫,GmVP1基因在一定条件下可以提高转基因大豆发状根对NaCl的耐受性,GmVP1/GmNHX1双价基因过表达比GmVP1单基因过表达更能提高大豆发状根相对生长量及其耐盐性。

GmNHX1 has been proved to improve salt tolerance of Arabidopsis thaliana and restore salt tolerance of yeast mutants. In addition, studies have shown that the expression of vacuolar membrane H^+-pyrophosphatase gene(VP) can increase the H^+-transmembrane driving force, thus facilitating the transport of harmful ions by ion transporters. Therefore, the vacuolar membrane H^+-pyrophosphatase gene GmVP1 was cloned from salt-tolerant soybean variety Jidou7, and its transcription level was detected under salt stress. The monovalent and bivalent transformation vectors of GmVP1 and GmVP1/GmNHX1 were constructed. The cotyledon transformation system mediated by Agrobacterium rhizogenes was used to express GmVP1 or GmVP1/GmNHX1 separately. The results showed that GmVP1 responded to salt stress at the transcriptional level.GmVP1 gene could improve the tolerance of transgenic soybean hairy roots to NaCl under certain conditions. Overexpression of GmVP1/GmNHX1 bivalent gene could improve the relative growth and salt tolerance of soybean hairy roots more than overexpression of GmVP1 single gene.