根癌农杆菌介导转化水稻成熟胚及转Metr基因植株的获得

Agrobacterium-mediated Transformation of Rice Mature Embryos and Regeneration of Transgenic Plants with Metr Gene

以粳稻品种广陵香糯为材料,用携带有双元载体pBB的根癌农杆菌EHA105为载体,研究了根癌农杆菌介导转化粳稻成熟胚愈伤组织的几个影响因素,建立了合适的粳稻成熟胚愈伤组织转化系统.一种基于MS为基本培养基的商品培养基(HRM)较适合于作为粳稻成熟胚愈伤组织诱导培养基,合适的愈伤组织诱导培养天数为7～8 d,合适的筛选培养基为CC培养基.在此基础上,将Metr基因导入粳稻品种广陵香糯中,获得了一批转基因水稻植株,PCR分析表明外源基因已经整合进了水稻的基因组中.部分转基因植株后代遗传分析表明外源基因的分离符合3:1的理论比例.

The mature embryos of a japonica rice, Guanglingxiangnuo, were used for the study on optimization of Agrobacterium-mediated transformation. Several factors affecting the transformation were investigated and a suitable transformation system was set up. For this transformation system, the HRM medium, based on the MS medium, was suitable for inducing callus from rice mature embryos. The suitable time span of initial culture in this medium was 7-8 days before co-culturing with Agrobacterium and suitable medium for selection was CO medium. Using this transformation system, the Metr gene was introduced into Guanglingxiangnuo, and many transgenic plants were obtained. Most of these transgenic rice plants were confirmed by PCR technique and basta resistance, indicating the T-DNA had been integrated into the genome of transgenic rice plants.