不同转化条件对3种农杆菌GFP基因在本氏烟草中瞬时表达的影响

doi:10.7525/j.issn.1673-5102.2022.01.013

植物研究 ›› 2022, Vol. 42 ›› Issue (1): 121-129.doi: 10.7525/j.issn.1673-5102.2022.01.013

不同转化条件对3种农杆菌GFP基因在本氏烟草中瞬时表达的影响

张悦婧, 李颖, 王娟娟, 庞海龙, 贾凌云, 冯汉青()

西北师范大学生命科学学院，兰州 730070

收稿日期:2021-03-11 出版日期:2022-01-20 发布日期:2021-12-30
通讯作者: 冯汉青 E-mail:fenghanq@nwnu.edu.cn
作者简介:张悦婧（1995—），女，硕士研究生，主要从事植物生理生态方面研究工作。
基金资助:
国家自然科学基金(31870246);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

Effects of Three Kinds of Agrobacterium and Different Transformation Conditions on the Transient Expression of GFP in Nicotiana benthamiana

Yuejing Zhang, Ying Li, Juanjuan Wang, Hailong Pang, Lingyun Jia, Hanqing Feng()

College of Life Science，Northwest Normal University，Lanzhou 730070

Received:2021-03-11 Online:2022-01-20 Published:2021-12-30
Contact: Hanqing Feng E-mail:fenghanq@nwnu.edu.cn
About author:Zhang Yuejing（1995—），female，senior engineer，mainly engaged in plantphysiology research work.
Supported by:
National Natural Science Foundation of China(31870246);University Scientific Research Project of Gansu Province(2015A-007);Key Research and Development Project of Gansu Province(18YF1NA051);Fundamental Research Funds for the Gansu Universities of Gansu Provincial Department of Finance;Youth Innovation Team of Northwest Normal University

摘要/Abstract

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OD₆₀₀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OD₆₀₀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。

关键词: 本氏烟草, 注射渗透法, 瞬时表达, GFP

Abstract:

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 OD₆₀₀ of bacterial suspension was 0.8； while the highest transient expression efficiency with EHA105 and GV3101 strains was achieved when the OD₆₀₀ 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.

Key words: Nicotiana benthamiana, syringe infiltration, transient expression, GFP

中图分类号:

S572

张悦婧, 李颖, 王娟娟, 庞海龙, 贾凌云, 冯汉青. 不同转化条件对3种农杆菌GFP基因在本氏烟草中瞬时表达的影响[J]. 植物研究, 2022, 42(1): 121-129.

Yuejing Zhang, Ying Li, Juanjuan Wang, Hailong Pang, Lingyun Jia, Hanqing Feng. Effects of Three Kinds of Agrobacterium and Different Transformation Conditions on the Transient Expression of GFP in Nicotiana benthamiana[J]. Bulletin of Botanical Research, 2022, 42(1): 121-129.

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不同转化条件对3种农杆菌GFP基因在本氏烟草中瞬时表达的影响

Effects of Three Kinds of Agrobacterium and Different Transformation Conditions on the Transient Expression of GFP in Nicotiana benthamiana

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