摘要
通过根癌农杆菌介导的叶盘法，将马铃薯Ｘ病毒外壳蛋白基因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。
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.
关键词
马铃薯X病毒
外壳蛋白基因
烟草
Potato virus X
Coat protein gene
Plant virus resistance
Tobacco
