多抗转基因马铃薯植株的获得及农杆菌介导试管薯遗传转化体系优化

Obtaining of Multiple Resistance Transgenic Potato Plant and Optimization of Agrobacterium-mediated Genetic Transformation System of Potato in Vitro

以马铃薯品种陇薯11号试管薯为受体材料,通过农杆菌介导法,将PVX、PVS、PVY和PLRV 4种病毒CP融合基因导入马铃薯,并对影响遗传转化的因素进行了优化。结果表明,薯片分化和生根阶段的选择压分别为Kan 50、75 mg/L,薯片分化和生根阶段有效抑菌浓度分别为Cb 500、200 mg/L,农杆菌活化时间为4.5 h、侵染时间为7 min、共培养时间为2 d时利于遗传转化。通过该体系将4种病毒CP融合基因导入马铃薯,获得了具卡那霉素抗性的转化植株,经PCR检测,外源基因已导入马铃薯基因组中。

The CP fusion genes of PVX,PVS,PVY and PLRV were transferred into Longshu 11 test tube potato through the agrobacterium-mediated method,and condtions of which was also optimized.The results showed the optimal Kan selective concentration is 50 mg/L and 75 mg/L for potato chip differentiation and root germination respectively,and Cb is 500 mg/L and 200 mg/L.The optimal transformation conditions:agrobacterium should be actived for 4.5 hours,transfection for 7 minutes,and co-cultivation time for 2 days,it was beneficial to genetic transformation.Via this system,four viral CP fusion gene were introduced into potatoes,and later transformed plants with kanamycin resistance were obtained.PCR detection confirmed the fusion gene had been successfully introduced into those potato genome.