根癌农杆菌介导大岩桐遗传转化体系探究

Research on Gloxinia Genetic Transformation System Mediated by Agrobacterium tumefaciens

本研究以大岩桐叶片为外植体,以LEA基因为目的基因,为实施根癌农杆菌介导转化大岩桐,研究了卡那霉素和头孢霉素、预培养时间、共培养时间和延迟筛选时间等因素对大岩桐外植体及转化的影响,结果表明,外植体分化卡那霉素浓度为50mg/L,头孢霉素浓度为250mg/L,生根筛选压浓度为10mg/L,外植体预培养时间为2d,共培养时间为4d,延迟筛选时间为6d;以上述条件下,将农杆菌与大岩桐叶盘共培养,共获得16株抗性植株,GUS染色有3株转LEA基因的阳性植株,初步建立了外源基因LEA导入大岩桐植株遗传转化体系,以遗传改良大岩桐品种的抗旱性。

In this study,the effects of kanamycin and cephalosporin,pre-culture time,co-culture time and delayed screening time on the explants and transformation of gloxinia were studied by using leaves of gloxinia as explants and LEA gene as target gene and Agrobacterium tumefaciens mediated transformation.The results show that kanamycin concentration was 50 mg/L,cephalosporin concentration was 250 mg/L,rooting screening pressure concentration was 10 mg/L,explant pre-culture time was 2 d,co-culture time was 4 d,and delayed screening time was 6 d.Under the above conditions,Agrobacterium tumefaciens and leaf disc of gloxinia were co-culture,and 16 resistant plants were obtained.Three LEA transgenic plants were stained by GUS.The foreign gene LEA was introduced into the genetic transformation system of gloxinia to improve the drought resistance of gloxinia varieties.