甘薯胚性悬浮细胞遗传转化的研究

Studies on the Genetic Transformation of Embryogenic Suspension Cultures in Sweetpotato

以甘薯品种栗子香的胚性悬浮细胞为材料 ,以A2 0 8SE(pROA93)为供体菌株 ,对甘薯胚性悬浮细胞遗传转化的基本条件进行了研究。结果表明 ,对胚性悬浮细胞预培养 1～ 3d有利于转化 ,适宜的共培养时间为 4～ 5d ,适宜的Kan浓度为 5 0～ 75mg·L-1(悬浮培养阶段 )和 10 0mg·L-1(增殖培养阶段和植株再生阶段 ) ,适宜的Carb浓度为 10 0mg·L-1。获得了经GUS检测及PCR检测的转基因植株。

Genetic transformation of embryogenic suspension cultures of sweetpotato cv. Lizixiang was conducted by using Agribacterium tumefaciens strain A208SE harboring the binary vectors pROA93 with nptⅡ gene and gus gene. The results indicated that embryogenic suspension cultures precultured for 1-3 days were suitable for the transformation. The optimal cocultivation time was 4-5 days. The optimal concentration of kanamycin was 50-75 mg·L -1 for suspension culture and 100 mg·L -1 for embryogenic callus formation and plant regeneration. The optimal concentration of carbencillin was 100 mg·L -1 . Transgenic plants identified with GUS assays and PCR analysis were obtained.