广谱抗病基因chi和hrpZpsta双价表达载体的构建及其在大豆中的遗传转化

Construction of Bivalent Plant Expression Vector with Two Broad-Spectrum Disease Resistance Genes chi and hrp Zpsta and Transformation into Soybean

大豆的真菌性病害已经成为了限制大豆产量增长及品质提高的主要原因,利用基因工程技术培育出具有广谱性抗病能力的大豆新品种,已成为目前提高大豆抗性的有效途径之一。本研究构建了含2种广谱抗病基因chi和hrp Zpsta的双价植物表达载体,利用农杆菌介导技术导入大豆中,获得了能够在转录水平上表达2种外源抗病性基因的转基因大豆。对经抗性筛选得到的阳性苗及后代植株进行PCR检测、Southern杂交、荧光定量PCR检测,共得到T0代阳性植株7株,T1代阳性植株27株。转化植株的基因组在整合外源基因时出现不同情况,chi-hrp Zpsta双价载体分别以单价和双价两种形式随机整合到受体大豆基因组中并且能稳定遗传,同时在转录水平上亦有不同。

Soybean fungous disease has become the main cause that limited soybean yield increasing and quality enhancement,one of the effective ways to enhance the resistance of soybean is using genetic engineering technology to cultivate soybean varieties with broad-spectrum disease resistance. In this research,two broad-spectrum disease resistance genes were built into bivalent plant expression vector,imported into soybeans by Agrobacterium-mediated method,and obtained transgenic soybeans with gene expression at the transcriptional level. After PCR,southern blot analysis and Q-PCR detection on resistant positive plants and their offspring,7 T0 positive plants and 27 T1 positive plants were obtained. Different situation occurred when exogenous gene integrated with transformation of plant genome,chi-hrp Zpsta bivalent expression vector in the form of monovalent and bivalent random integrated into the recipient soybean genome and could be genetic stabilized,meanwhile at the transcriptional level were different as well.