摘要
以本氏烟草(Nicotiana benthamiana)为植物材料,分析了不同农杆菌菌株(LBA4404菌株、EHA105菌株、GV3101菌株)、菌液浓度以及侵染时间在瞬时转化过程中对报告基因GFP荧光表达量的影响。结果显示,不同的农杆菌菌株瞬时表达外源基因的最适浓度和时间均有所不同:LBA4404菌株在菌悬液OD_(600)值为0.8时所介导的瞬时表达效率最高;而EHA105和GV3101菌株在菌悬液OD_(600)值为0.6时可达到最高瞬时表达效率。LBA4404菌株所介导的瞬时表达在农杆菌注射后第2天时表达量最高,而EHA105和GV3101菌株所介导的瞬时表达在农杆菌注射后第4天时表达量最高。不同菌株间比较分析表明,LBA4404菌株所介导的瞬时表达效率最高。上述结果表明,农杆菌菌株以及浓度和侵染时间等转化条件均是影响瞬时表达效率的重要因素。
By using Nicotiana benthamiana as plant material,the influence of the different Agrobacterium strains(LBA4404,EHA105,GV3101),bacterial concentration and infection time on GFP(as the reporter gene)the fluorescence transient expression after transformation were analyzed respectively.The results showed that the expression of GFP with different Agrobacterium strains were various in the optimal concentration and time for transient:the highest transient expression efficiency with LBA4404 strain was achieved when the OD_(600) of bacterial suspension was 0.8;while the highest transient expression efficiency with EHA105 and GV3101 strains was achieved when the OD_(600) of the bacterial suspension reached 0.6.The level of transient expression by strain LBA4404 was the highest at 2 days later Agrobacterium infiltration while the level of transient expression by strains EHA105 and GV3101 was the highest at 4 days later Agrobacterium infiltration.The comparative analysis among these different strains showed that the transient expression efficiency by using LBA4404 strain was the highest.The above results indicated that Agrobacterium strains and transformation conditions such as bacterial concentration and infection time were important factors that affected the transient expression efficiency.
作者
张悦婧
李颖
王娟娟
庞海龙
贾凌云
冯汉青
Zhang Yuejing;Li Ying;Wang Juanjuan;Pang Hailong;Jia Lingyun;Feng Hanqing(College of Life Science,Northwest Normal University,Lanzhou 730070)
出处
《植物研究》
CAS
CSCD
北大核心
2022年第1期121-129,共9页
Bulletin of Botanical Research
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。
关键词
本氏烟草
注射渗透法
瞬时表达
GFP
Nicotiana benthamiana
syringe infiltration
transient expression
GFP
