用激光微束穿刺法将PAP cDNA导入油菜获得抗病毒转基因植株

Introduction of PAP cDNA into Brassica napus L. and Acquisition of Tansgenic Plants via Laser Microbeam Puncture

采用激光微束穿刺法将损伤诱导型启动子控制之下的商陆抗病毒蛋白（PAP）CDNA导入了甘蓝型油菜（BrassicanapusL．）双低品种H165，经庆大霉素筛选，获得了抗性再生植株。PCR扩增检测及Southern杂交分析表明，PAPcDNA连同损伤诱导型启动子已稳定整合至受体基因组，转化效率为1．7％。攻毒实验表明，转基因油菜植株对供试病毒芜菁花叶叶（TuMV）具有较高抵抗能力。

The PAP cDNA controlled by a wound-inducible promoter was introduced into Brassica napus via laser microbeam puncture, and plantlets resistant to Gentamycin were obtained. The transgenicity of these plantlets was verified by PCR amplification and the Southern blotting analysis, which showed that PAP cDNA together with the woundinducible promoter and a terminator have been integrated into the rape genome, with the transformation frequency being 1. 7%. The test of virus challenge showed that all these transgenic rape plants were resistant to virus TuMV mechanically inoculated in higher degree.