抗黄萎病基因StVe转化番茄的研究  

Transformation of Tomato with the Verticillium dahliae Resistance Gene StVe from Solanum torvum

作　　者：陈玉辉[1] 赵凌侠[1] 柴友荣[2] 崔丽洁[1] 董仲琦[3] 钱虹妹[1] 唐克轩[1] 

机构地区：[1]上海交通大学复旦-交大-诺丁汉植物生物技术研发中心,上海200240 [2]西南大学农学与生物科技学院,重庆400716 [3]上海交通大学生命科学技术学院,上海200240

出　　处：《园艺学报》2008年第5期693-700,共8页Acta Horticulturae Sinica

基　　金：上海市科技攻关重大项目(03DZ19310);国家‘863’项目(2007AA100503);上海市国内科技合作项目(073158202)

摘　　要：为了验证克隆自水茄(Solanum torvum Swartz)的StVe基因功能,将StVe连入中间载体p35S-2300::gus::noster的BamHⅠ和SacⅠ位点,取代gus基因构建植物双元表达载体p35S-2300::StVe::noster;用农杆菌介导法转化樱桃番茄22号,获得了72株卡那霉素抗性植株,PCR和Southern blot检测确认有5株为阳性植株;RT-PCR结果显示,StVe在转基因番茄植株间的转录水平表达存在差异;PDA平板抑菌试验显示,转StVe基因番茄叶片总可溶蛋白具有抑制番茄黄萎病菌生理小种1(Verticillium dahliae race1)生长的作用。In order to verify the function of StVe gene （3.4 kb） which was isolated from Solanum torvum, we substituted the StVe gene for gus in plasmid p35S-2300 ：： gus ：： noster, and constructed plant bina- ry expression vector p35S-2300 ：： StVe ：： noster. Subsequently, the resulting vector was introduced into tomato plants via Agrobacterium tumefaciens-mediated method. In total, we obtained 72 tomato plants resistant to kanamycin and 5 independent transgenic tomato plants were confirmed by PCR and Southern blot analysis. The five transgenic tomato plants were further analyzed the expression of StVe gene at transcription level through semi-quantitative one-step RT-PCR. StVe gene was found to express with variable levels between different transgenic lines. The crude proteins were extracted from the transgenic tomato leaves and proved to have an effect of restraining the mycelia growth of Verticillium dahliae race 1 in vitro.

关 键 词：番茄 抗病 StVe基因 大丽轮枝菌 转化 

分 类 号：S641.2[农业科学—蔬菜学]