根癌农杆菌介导籼稻遗传转化影响因素研究  

Study on Factors Related to Agrobacterium-mediated Transformation of Indica Rice

作　　者：刘元风[1] 刘彦卓[1] 王金花[1] 罗文永[1] 毛兴学[1] 李玲[2] 李晓方[1] 

机构地区：[1]广东省农业科学院水稻研究所,广州510640 [2]华南师范大学生命科学学院,广州510631

出　　处：《分子植物育种》2005年第5期737-743,748,共8页Molecular Plant Breeding

摘　　要：研究了抗生素种类、继代和菌液浓度对农杆菌介导籼稻转化的影响,并将反义蜡质基因导入4个高产、直链淀粉含量高的籼稻品种中。结果显示,羧苄青霉素的抑菌效果优于头胞霉素,其适宜浓度为250￣350mg/L;4个品种继代愈伤比初代愈伤能获得更高的抗性愈伤得率;各个品种转化有其最佳的农杆菌菌液浓度(OD600值),茉莉新占和绿黄占为1.0,粤香占和矮秀占为0.6。应用此优化的农杆菌转化体系,我们得到了转化植株;PCR扩增和Southern杂交结果证明外源基因已经整合到转基因水稻的基因组中。The mature-embryo-derived calli from 4 high-yielding indica rice cultivars with high amylose content were transformed with Agrobacterium tumefacien strain, EHA105 harboring anti-waxy gene, and some factors including antibiotics, bacterial cell density and subculturing of calli were studied in relation to transformation efficiency. We found that carbenicillin was superior to cefotaxime in controlling the growth of bacteria, with optimal concentration of 250-350mg/L. Higher frequency of hygromycine-resistant calli was obtained when subcultured calli of 4 cultivars were transformed in comparison to primary calli. The optimal bacterial cell density for transformation was 0.6 of OD600 for cultivars of Yue-Xiang-Zhan and Ai-Xiu-Zhan, and it was 1.0 of OD600 for cultivars of Mo-Li-Xin-Zhan and Lv-Huang-Zhan. Transgenic plants were acquired in 3 cultivars by using a transformation procedure based on the optimization of the 3 factors. The integration of foreign DNA into the genome of transgenic plants was confirmed by PCR amplification and Southern Blotting.

关 键 词：籼稻 根癌农杆菌 基因转化 反义WX基因 根癌农杆菌介导 籼稻品种 遗传转化 羧苄青霉素 直链淀粉含量 菌液浓度 

分 类 号：S511.21[农业科学—作物学]