农杆菌介导玉米幼胚愈伤组织遗传转化体系的优化

以玉米骨干自交系‘郑58’的幼胚为材料,用农杆菌介导法对诱导培养出的胚性愈伤组织进行三价植物表达载体的遗传转化,同时对影响转化效率的主要因素进行研究.结果表明,当浸染液和共培养培养基中乙酰丁香酮(AS)的浓度为150mg·L-1时,抗性愈伤组织获得率显著提高17.39%;在菌液浓度D600为0.3、浸染时间为20min、共培养时间为60h、共培养温度为22℃、共培养培养基pH为5.4以及头孢霉素浓度为400mg·L-1的试验体系下,有利于提高玉米抗性愈伤组织的获得率,其获得率最高可达17.26%.

Optimization of genetic transformation system for maize immature embryo calli mediated by Agrobacterium tume faciens

The immature embryo of excellent maize inbred "Zheng-58" was used in the experiment,taking the embryogenic callus as the receptor for the transformation of Trivalent plant expression vector which was mediated by Agrobacterium tumefaciens.Important factors which influenced the efficiency of the transformation were identified and an efficient transformation system was developed for maize immature embryo calli.The results showed that the resistant calli rate was remarkably improved (17.39 % higher than other concentration) when the concentration of acetosyringone (AS) was 150 mg·L-1 both in the infection solution and co-culture medium.A higher efficiency system for transformation was established with the combination of 0.3 D×20 minutes for A.tumefaciens concentration and incubation-time,20 ℃×60 h for the co-culture temperature and time,400 mg·L-1 for the cefotaxime (Cef) concentration,and the pH was 5.4.By this optimized system,the rate of the resistant calli increased by 17.26 %.