论文部分内容阅读
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。
Fungal diseases of soybean have become the main reason to limit the increase of soybean yield and quality. Using genetic engineering to breed new varieties of soybean with broad-spectrum disease resistance has become an effective way to increase soybean resistance. In this study, a bivalent plant expression vector containing two broad-spectrum resistance genes, chi and hrp Zpsta, was constructed and introduced into soybean using Agrobacterium tumefaciens-mediated technique. Two transgenic plants expressing the exogenous genes at the transcriptional level GM soy. The positive seedlings and progeny plants obtained by resistance screening were detected by PCR, Southern blot and fluorescence quantitative PCR. Seven T0 positive plants and 27 T1 positive plants were obtained. Different genotypes of transformed plants appeared when integrating foreign genes. The chi-hrp Zpsta binary vector was integrated into the receptor soybean genome randomly and in both monovalent and bivalent formats, and was stable at the transcription level different.