农杆菌菌株及其侵染浓度和时间对基于菜豆黄矮病毒表达载体瞬时表达外源基因的影响  

Effects of Agrobacterium tumefaciens strain and its infection time and concentration on transient expression of foreign genes based on expression vector of bean yellow dwarf virus

作　　者：李颖 张悦婧 王馨 庞海龙[1] 贾凌云[1] 冯汉青[1] Li Ying;Zhang Yue-Jing;Wang Xin;Pang Hai-Long;Jia Ling-Yun;Feng Han-Qing(College of Life Sciences, Northwest Normal University, Lanzhou 730070, China)

机构地区：[1]西北师范大学生命科学学院,兰州730070

出　　处：《植物科学学报》2021年第3期297-305,共9页Plant Science Journal

基　　金：国家自然科学基金(31870246,31560070);甘肃省高等学校科研项目(2015A-007);甘肃省重点研发计划项目(18YF1NA051);甘肃省高校基本科研业务费;西北师范大学青年创新团队项目。

摘　　要：以3种常见的农杆菌菌株(GV3101、EHA105、LBA4404)和基于菜豆黄矮病毒的复制型植物表达载体为材料,利用农杆菌介导的瞬时转化技术,将外源绿色荧光蛋白(GFP)基因导入本氏烟(Nicotiana benthamiana L.)叶片中实现瞬时表达,并对不同农杆菌菌株、侵染浓度及侵染时间对于瞬时表达水平的影响进行比较。结果显示,3种不同的农杆菌菌株在介导转化本氏烟叶片瞬时表达GFP积累水平之间存在显著差异,其中EHA105菌株转化效率最高,LBA4404次之,GV3101最低。此外,GV3101、EHA105和LBA4404最适侵染浓度的OD600值分别为0.5、0.3和0.3;最佳侵染时间均为第4 d。研究结果表明农杆菌菌株染色体结构和Ti质粒的差异是影响瞬时转化过程中农杆菌侵染浓度及其外源基因瞬时表达效率的重要因素。To explore the effects of different Agrobacterium tumefaciens strains and their infection time and concentration on the transient expression of foreign genes based on geminivirus expression vectors,three common A.tumefaciens strains and replicative plant expression vectors based on the bean yellow dwarf virus(BeYDV)were used as experimental materials.The exogenous green fluorescent protein(GFP)gene was introduced into Nicotiana benthamiana L.leaves to achieve transient expression by transient transformation mediated by A.tumefaciens.The effects of different A.tumefaciens strains,infection concentration,and infection time on transient expression were compared.Results showed significant differences among the three different strains of A.tumefaciens in the transient expression of GFP in N.benthamiana leaves.Among them,the transformation efficiency of strain EHA105 was the highest,followed by LBA4404,and then GV3101.In addition,the optimum infection concentrations of GV3101,EHA105,and LBA4404 were 0.5,0.3,and 0.3,respectively,and the best infection time was 4 d.These results suggest that the differences in chromosome structure and Ti plasmid of the A.tumefaciens strains are important factors affecting the infection concentration of A.tumefaciens and the instantaneous expression efficiency of foreign genes during transient transformation.

关 键 词：根癌农杆菌菌株 农杆菌介导转化 双生病毒衍生载体 瞬时表达 

分 类 号：Q943.2[生物学—植物学]