【摘要】 通过根癌农杆菌(Agrobacterium tume faciens)介导的叶盘法,将马铃薯X病毒(PVX)外壳蛋白基因(CP)导入烟草细胞并获得大量再生的转基因烟草植株。经胭脂碱检测、酶联免疫分析和Western blot分析,证明已获得了具有马铃薯X病毒外壳蛋白基因表达的转基因烟草。用2μg/ml的PVX磨擦接种烟草,8天后进行观察,发现对照植株开始发病,而转基因烟草植株在20天后才开始发病,有的转基因烟草植株在整个生长期中都不得病。对PVX CP基因表达量较高的4株植株的自交后代进行了胭脂碱的检测。发现其分离比都近似于3:1。说明PVX CP基因插入到染色体上的拷贝数均为1。用15μg/ml的病毒液接种烟草,两周后发现含有胭脂碱的转基因烟草后代大部分没有发病,而对照植株大都发病。更多还原

【Abstract】 A major commercial cultivar of tobacco was transformed via Agrobacterium mediated procedure. Tobacco leaves started to form shoots on shoot inducing medium containing kanamycin after infected by Agrobacterium containing the plasmid with PVX CP gene. Regenerated plants were obtained in two weeks on hormone-free MS medium containing kanamycin. The transgenic tobacco plants were identified with nopaline detection, enzyme -linked immunosorbent assay and western blot analysis, symptom appearance was sig- nificantly delayed and virus accumulation was either absent or reduced in PVX CP gene transformed plants. Progenies of transgenic tobacco plants also gained resistance to PVX infection to a certain degree. These experiments demonstrate that CP protection is effective a-gainst PVX.更多还原