利用农杆菌介导法获得转ipt基因半夏植株

以贵州珍珠半夏Pinellia ternate(Thunb.)Breit为材料,采用根癌农杆菌介导法,探索了影响半夏遗传转化效率的主要因子.结果表明,卡那霉素的最佳筛选浓度是100 mg/L,侵染时间、共培养时间、乙酰丁香酮浓度对转化频率都有一定的影响.其中侵染时间为20 min,共培养时间为3 d,乙酰丁香酮的浓度控制在60～100 mg/L时可以有效提高遗传转化效率.在此基础上辅以超声波处理5 min,可以有效提高叶柄转化的瞬时表达率.对抗性转化植株进行Gus染色和PCR检测,结果表明外源基因ipt已经整合到半夏基因组中.

Transgenic Pinellia ternate(Thunb.)Breit Plants Containing ipt Obtained by Agrobacterium-Mediated Transformation

Several factors affecting genetic transformation of Pinellia ternate(Thunb.)Breit mediated by Agrobacterium tume faciens were investigated.The results showed that the infection time,acetosyringone(As)concentration and co-cultivation time had different effects on transformation efficiency.The highest frequency was observed when the explants without pre-culture were infected for 20 min in medium added with 60～100 mg/L As,followed by co-cultivateon for 3 d.Treatment with 5-min sonication may significantly enhance the transient expression of petiole transformation.The Gus staining and PCR verification of the regenerated plants proved that the exogenous gene has been successfully integrated into the genome of Pinellia ternate(Thunb.)Breit.