利用农杆菌注射法鉴定番茄黄化曲叶病毒的沉默抑制子

Determination of Silencing Suppressor of Tomato Yellow Leaf Curl Virus by Agroinfiltration

利用农杆菌注射法鉴定了番茄黄化曲叶病毒的沉默抑制子V2。将分别携带目标基因与GFP表达载体的农杆菌共注射到本氏烟草中,2d后GFP瞬时高量表达。西瓜褪绿矮化病毒BV1基因表达载体和空载体pBIN分别与GFP共侵染7d后,GFP表达量由于寄主沉默机制而急剧减弱。V2基因表达载体与GFP表达载体共侵染叶片7d后,GFP仍保持高水平表达,说明V2作为番茄黄化曲叶病毒沉默抑制子可阻止寄主的沉默机制。农杆菌注射法可快速方便筛选植物病毒沉默抑制子。

We described the utilization of agroinfiltration to screen and verify the silencing suppressor,V2,of Tomato yellow leaf curl virus(TYLCV).The target gene together with the GFP gene was transiently co-expressed in leaves of Nicotiana benthamiana by agroinfiltrations.In all tests of agroinfiltrations,the GFP showed high expression at 2 days post inoculation(dpi),showing the infiltration and expression was successful.On the leaves co-infected with the BV1 gene of Watermelon chlorotic stunt virus or the empty vector pBIN,the GFP fluorescence almost disappeared at 7 dpi due to silencing mechanism of the host plant.The leaves co-infected with V2,showed continuously high expression of GFP,indicating that V2 is the silencing suppressor of TYLCV.Agroinfiltration can be utilized as potent tool for screening the silencing suppressor of plant virus.