农杆菌介导籼稻中籼3037转化体系的优化及其应用  

Application and Optimization Agrobacterium Mediated Transformation System on Indica Variety Zhongxian 3037

作　　者：王歆[1] 于恒秀[1] 曾燕楠[1] 刘巧泉[1] 龚志云[1] 程祝宽[2] 

机构地区：[1]扬州大学教育部植物功能基因组学重点实验室,江苏省作物遗传生理重点实验室,扬州225009 [2]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《分子植物育种》2008年第3期457-464,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(30770131);江苏省自然科学基金项目(BK2007510);省教育厅项目(06KJA21018,k05015)

摘　　要：以籼稻品种中籼3037为材料,研究了影响农杆菌介导转化籼稻的几个重要因素,结果表明:MSⅡ培养基作为愈伤组织的诱导培养基好于N6D2;以幼胚为外植体,愈伤组织的诱导率和转化率明显比成熟胚和幼穗高;根癌农杆菌菌株EHA105介导的基因转化率高于AGLO;当以幼胚为外植体,根癌农杆菌菌株E-HA105介导的中籼3037的转化率为18.0%,而AGLO为5.5%;EHA105菌液OD值为0.6,浸染时间14min为中籼3037合适的转化参数;在分化培养基MSR中添加20g/L的山梨醇有利于抗性愈伤组织的分化。在此基础上,采用优化的农杆菌介导法转化中籼3037及其来源的突变体A和B,获得了转化植株,PCR、Southern杂交和T1代遗传分析表明外源基因已经整合到水稻基因组中。An indica rice variety, Zhongxian 3037, was used for optimization of Agrobacterium-mediated transformation, and several factors which affected the transformation were studied and a suitable transformation system was created. For this transformation system, immature embryo was suitable as explants. The MS II medium based on the MS medium, was suitable for inducing callus. Agrobacterium tumefaciens strain EHA105 was more efficient than that of AGLO for indica rice cultivar Zhongxian 3037 transformation, and its transformation frequency was 18.0 percent while AGLO＇s transformation frequency was 5.5 percent when using immature embryo as explants; While the suitable concentration of Agrobacterium was OD600=0.6 and the incursion time was 14 min. The frequen- cy of callus regeneration was much higher when 20 g/L sorbitol was added to the regeneration medium MSR than that of the control. Using this transformation system, the HPT gene was introduced into Zhongxian 3037 and its mutant A and B, and many transgenic plants were obtained. Some of these transgenic rice plants were confirmed by PCR and Southern blotting techniques which indicating the T-DNA had been integrated into the genome of transgenic rice plants, Genetic analysis showed that the transgenes were segregated in a mendelian fashion in the T1 generation.

关 键 词：根癌农杆菌 转化 籼稻 

分 类 号：S511.21[农业科学—作物学]