摘要
以水稻品种中花11的幼胚、成熟胚和幼穗的愈伤组织为材料，经携带Ti质粒pDs－Bar1300的根癌农杆菌EHA105感染和共培养后，通过50mg／L和25mg／L潮霉素的二次筛选，结果表明34％幼胚愈伤组织、17％成熟胚愈伤组织和16．5％幼穗愈伤组织表现潮霉素抗性。这些抗性愈伤组织转入分化和再生培养基培养，获得了14棵转基因植株。经Southern杂交分析，证明Ds和Bar基因都已整合进转基因植株的染色体组。对T1代植株的PPT抗性测定表明，外源Bar基因的分离符合孟德尔法则。
We have developed an Agrobacterium-mediated gene transfer system in rice (Oryza sativa ssp. japonica ). Using this system, immature embryos, mature embryos and young panicles of rice variety Zhonghua No. 11 were used as explants to induce calli,respectively. The calli were infected and transformed by A. tumefacien strain EHA105 harboring Ti plasmid pDsBar1300 carrying Ds transposon and Bar gene. The results showed that infected immature embryos,mature embryos and young panicle calli gave 34 ％, 17％ and 16. 5 ％ resistant calli, respectively, after cocultivation and two successive hygromycin resistant selections. The resistant calli were successively transferred to differentiation and regeneration media, 14 transgenic plants were obtained. Southern blotting analysis of the total DNA extracted from transgenic plants indicated that the T-DNA had integrated into the rice genome,the result of PPT resistant genetic analysis showed that some of transgenic plants were segregated in a Mendelian fashion in the T1, progenies.
出处
《上海农业学报》
CSCD
1998年第4期7-12,共6页
Acta Agriculturae Shanghai
基金
国家科委863项目
关键词
水稻
根癌农杆菌
遗传转化
Ds转座因子
Oryza sativa ssp. japonica
Agrobacterium-mediated gene transformation
Ds transposon
Transgenic plant
