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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。
Taking Guanglingxiangnuo as a material, several factors influencing the transformation of callus from Japonica rice mediated by Agrobacterium tumefaciens were studied by using Agrobacterium tumefaciens EHA105 harboring binary vector pBB as the vector, and the suitable Japonica Mature Embryo Callus Transformation System. A commercial medium based on MS (HRM) is more suitable as a medium for callus induction of mature embryo callus of Japonica rice. Suitable callus induction culture days are 7-8 days. A suitable screening medium is CC Medium. Based on this, we obtained a batch of transgenic rice plants by introducing the Metr gene into the rice variety Guangling Xiangnuo, and PCR analysis indicated that the foreign gene has been integrated into the genome of rice. The genetic analysis of progenies of some transgenic plants showed that the isolation of exogenous genes conformed to the theoretical ratio of 3: 1.