四倍体菘蓝转抗虫基因研究Ⅰ.根癌农杆菌介导的转基因菘蓝

Agrobacterium tumefaciens mediated transgenic tetraploid Isatis indigotica carrying anti-insect gene from Pinella Ternata Agglutinin

目的 将外源半夏凝集素基因 ( PTA )转入四倍体菘蓝以获得鳞翅目昆虫抗性。方法 构建了以 Ca MV 35 S为启动子 ,新霉素磷酸转移酶 ( Npt- )为选择标记 ,带有半夏凝集素基因 PTA的 p CAMB A2 2 0 0植物双元表达载体 ,应用根癌农杆菌 EHA 10 5遗传转化四倍体菘蓝。结果 菘蓝外植体植株再生率达到 95 %以上 ,转化率有10 %。结论 建立了以 6 - BA和

Object Pinella Ternata Agglutinin (PTA) gene was conducted into tetraploid Isatis indigotica Fort. to get anti-pest species.Methods Agrobacterium tumefaciens mediated transformation in which the “high toxic” strain EHA105 and the plant binary expression vector pCAMB I A2200 were adopted under CaMV 35S promotor and neomycin phosphorus transferred enzyme (NptⅡ) served as the selection marker.Results The plant regeneration rate of transgenic explants was up to 95% and transgenic frequency was 10%.Conclusion An in vitro highly effcient plantlet regeneration system and transgenic system mediated by A.tumefaciens in the main combination of 6-BA and NAA was established via cotyledon and hypocotyl segments in tetraploid I.indigotica.