摘要
以大白菜栽培品种“福山大包头”的子叶柄为供试材料 ,对影响大白菜植株再生和基因转化频率的因素进行了研究。在此基础上 ,建立了大白菜高效再生体系和有效的基因转化体系 ,并将芜菁花叶病毒的 CP基因 (Tu MV- CP)导入大白菜中 ,获得转化植株。 PCR检测和 Southern杂交分析证明Tu MV- CP基因已整合于大白菜的基因组中 ;Northern杂交分析及 EL ISA检测表明 Tu MV- CP基因在转录和翻译水平上进行了有效表达。转基因植株 T1代的遗传分析表明 ,外源基因在转基因植株后代中遵循 3∶ 1的分离规律。抗病性测定结果显示 ,转基因植株具有明显的抗病毒侵染能力。
Using cotyledonary petioles of Chinese cabbage [ Brassica campestris L. ssp. pekinensis (Lour) Olsson] 'Fushan Dabaotou' as material, we investigated the factors affec ting plant regeneration and genetic transformation mediated by Agrobacterium tumefaciens . As a result, an efficient system for plant regeneration and genetic transformation was established, and the TuMV CP gene was introduced into Chinese cabbage. Preliminary molecular analysis indicated that five of the 32 putative transgenic plants were PCR positive. The five plants were then subsequently analyzed by Southern blot. The results showed that the TuMV CP gene was integrated into the chromosomes of the plants. Northern blot analysis and ELISA test indicated that the TuMV CP gene was expressed at both transcriptional and translational levels. Further studies indicated that the TuMV CP gene was segregated at a 3∶1 ratio in T 1 generation. Resistance assay revealed that the plants of T 1 generation showed high level of resistance to TuMV infection.
出处
《植物病理学报》
CAS
CSCD
北大核心
2001年第3期257-264,共8页
Acta Phytopathologica Sinica
基金
国家"九五"科技攻关项目 (0 1- 0 2 - 0 3- 0 1)
山东省"三0工程"资助项目
关键词
大白菜
TuMV-CP基因
根癌
农杆菌
基因转化
Chinese cabbage
TuMV CP gene
Agrobacterium tumefaciens
genetic transformation
