农杆菌介导的玉米愈伤组织遗传转化研究  

Study on Genetic Transformation of Maize Callus Mediated by Agrobacterium tumefaciens

作　　者：李海霞[1] 谢久凤 孙金花[1] LI Hai-xia;XIE Jiu-feng;SUN Jin-hua(Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南农业大学,河南郑州450002

出　　处：《河北农业科学》2021年第2期99-103,共5页Journal of Hebei Agricultural Sciences

基　　金：2019年河南农业大学教学质量工程项目。

摘　　要：为优化玉米农杆菌介导法的转化体系,以玉米杂交种昌7-2×HiⅡA和郑22×87-1的胚性愈伤组织为受体材料,对转化过程中抗菌素抑制效果、侵染液类型、侵染时间、共培养处理方法等条件进行了优化。结果表明:100 mg/L头孢霉素可完全抑制农杆菌生长,不同侵染液类型对转化率影响不大;侵染转化过程中加入乙酰丁香酮(As)可以明显提高转化效率,加入As的LB和YEP培养基染色率分别较不加As分别提高40.0%和33.3%;侵染时间为10 min时抗性愈伤率最高;在培养基中加1层灭菌滤纸,既能实现共同培养,又能减少农杆菌对愈伤组织的伤害,得到质量好的抗性愈伤。To improve the agrobacterium-mediated transformation system of maize,the embryogenic callus of Chang 7-2×HiⅡA and Zheng 22×87-1 were used as the acceptor materials,and the antibacterial inhibition effect,types of impregnation solution,infection time and co-culture method were optimized.The results showed that 100 mg/L cephalosporin could completely inhibit the growth of Agrobacterium tumefaciens,and the types of immersion solution had little effect on the conversion rate.The addition of AS could significantly improve the transfection efficiency during the transfection process,and the dyeing rate of LB and YEP medium with AS increased by 40.0%and 33.3%,respectively.The resistant callus rate was the highest when the infection time was 10 min.Adding a layer of sterilized filter paper in the medium can not only realize co-culture,but also reduce the damage of Agrobacterium to callus,and obtain high quality resistant callus.

关 键 词：玉米 胚性愈伤组织 农杆菌 遗传转化 

分 类 号：S513[农业科学—作物学]