表达dsRNA的转基因烟草能阻止烟草花叶病毒的侵染  

Transgenic Tobacco Plants Expressed dsRNA Can Prevent Tobacco mosaic virus Infection

作　　者：张凯[1] 牛颜冰[1] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所,杭州310029

出　　处：《农业生物技术学报》2005年第2期226-229,共4页Journal of Agricultural Biotechnology

基　　金：国家重点基础研究规划(973)项目(No.G200016204)资助。

摘　　要：将烟草花叶病毒(Tobaccomosaicvirus,TMV)的部分运动蛋白基因(ΔMP)构建成反向重复结构,插入植物表达载体pBin438上,通过三亲交配法导入根癌农杆菌(Agrobacteriumtumefaciens)菌株EH105后通过农杆菌介导法转化烟草(Nico-tianatabacam)品种云烟87,获得了50株转基因烟草。PCR和Southernblot分析表明目标基因已整合到烟草植株的基因组内。用TMV对转基因烟草进行挑战接种,症状观察和病毒浓度的TAS-ELISA检测表明,转基因烟草对TMV的抗性包括3种类型:免疫型占10%;抗病型占4%;感病型占86%。Northernblot分析表明,目标基因mRNA在不同抗病类型植株中的积累量存在明显差异,目标基因mRNA的积累量与抗病性成负相关。Inverted repeats of the partial movement protein gene (ΔMP) of Tobacco mosaic virus (TMV) were introduced into plant expression vector pBin438, the recombinant plasmids were transformed into Agrobacterium tumefaciens EHA105 by tri-parental mating method. Fifty transgenic Nicotiana tabacum cv. Yunyan 87 plants were obtained with Agrobacterium-mediated transformation. PCR and Southern blotting analyses showed that target gene had integrated into tobacco genome. When transgenic tobacco plants were challenged with TMV, symptom observation and TAS-ELISA showed that transgenic plants had three kinds of phenotypes to the virus infection: immune (10%), resistant (4%) and susceptible (86%). Northern blotting analysis showed that the levels of target mRNA accumulation varied among transgenic tobacco lines, and inverse correlation between target mRNA accumulation and virus resistance was found.

关 键 词：转基因烟草 烟草花叶病毒 DSRNA Southern Northern ELISA检测 侵染 mosaic 植物表达载体 农杆菌介导法 目标基因 blot mRNA virus 根癌农杆菌 重复结构 蛋白基因 云烟87 烟草植株 挑战接种 分析表 TMV 积累量 分运动 PCR 

分 类 号：S572[农业科学—烟草工业] S435.72[农业科学—作物学]