根癌农杆菌介导AtNHX1基因转化小麦

Agrobacterium Tumefaciens Mediated AtNHX1 Gene Transformation to Wheat

利用根癌农杆菌对 3种基因型小麦进行了转化研究 .发现基因型对转化有明显影响 .将小麦品种烟优 36 1、烟 10 3、核生 3号的胚性愈伤组织用含报告基因NPTⅡ和抗盐基因Na+ H+反向转运AtNHX1的农杆菌菌株pROK2AT GV310 1感染和共培养后 ,用巴龙霉素 5 0 - 15 0mg L筛选抗性愈伤组织及转化植株 .对抗性植株的总DNA用AtNHX1基因的特异引物进行PCR检测 ,目的基因的PCR阳性频率为 1.3(烟 10 3)和 2 .9% (核生 3号 ) .

Three wheat ( Tritivum aestivum L. ) varieties were transferred by Agrobacterium tumefaciens . It was discovered that genotypes of wheat has impacted on the transformation. The strain of A .tumefaciens pROK2AT/GV3101 carrying NPTⅡ gene and AtNHX1 gene was used to transfer wheat cv. Yanyou 361, Yan103 and Hesen no.3. Calli derived from young embryos of wheat were infected and co cultured with A .tumefaciens. The transformed calli were selected by Paromycin 50-150?mg/L and 26 resistant plants from 92 calli used for transformation were identified by PCR amplifycation with the primer of AtNHX1 genes. The positive frequency of PCR was about 1.3(Yan103) and 2.9%(Hensen no.3.) respectively, and the integration of the foreign genes in the wheat genome was confirmed by Southern hybridization.