农杆菌介导的玉米转化后分化和生根条件优化  

Maize Genetic Transformation Mediated by Agrobacterium tumefaciens and Optimization of the Differentiation and Rhizogenesis Conditions

作　　者：刘丽霞[1] 胡晓丽[1] 宗晓娟[1] 刘玉鲲[1] 李德全[1] 

机构地区：[1]山东农业大学生命科学学院作物生物学国家重点实验室,山东泰安271018

出　　处：《西北植物学报》2007年第11期2188-2194,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金项目(30471052)

摘　　要：通过诱导5种自交系玉米授粉10～12d的未成熟幼胚,产生胚性愈伤组织,以农杆菌介导法,分别转化‘18-599(H)’、‘齐319’、‘综31’、‘Z674’和‘郑58’等5种玉米自交系。以GUS基因作为报告基因进行组织化学染色和潮霉素磷酸转移酶基因片段PCR特异扩增等方法检测都证明了外源基因的成功转化。对5种自交系玉米的出愈率、抗性愈伤率、转化率等指标进行了详细统计,并对分化培养基中附加不同细胞分裂素的分化效果以及对不同生根培养基的生根效果进行比较、优化,结果发现,通过优化抗性愈伤的最佳分化培养基(N6+1.0mg·L-1KT)和最佳生根培养基(1/2MS),使‘18-599(H)’的转化效率最高达6.1%,‘齐319’次之为3.5%。The maize immature embryos of five inbred lines,including ‘18-599（H）＇, ‘Qi 319＇, ‘Zong 31 ＇,‘Z674＇ and ‘Zheng 58＇ ,were used to initiate embryogenic calli. Then, these calli were used for the maize transformation mediated by Agrobacterium tumefaciens. The stable expression of GUS gene in transgenic plants was confirmed by histochemical assay and the Hpt gene was identified by PCR. The results indicated that the five maize inbred lines were different in the embryogenic calli initiation, the rate of resistant calli and the transformation frequency. Our results suggest that transformation efficiencies of ‘18-599（H）＇ and ‘Qi 319＇ were significantly improved up to 6.1% and 3.5% with the optimizing differentiation and rhizo- Renesis conditions, respectively.

关 键 词：玉米 农杆菌 遗传转化 生根条件优化 转基因玉米 

分 类 号：Q789[生物学—分子生物学]