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

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。

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

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.