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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。
Agrobacterium tumefaciens injection was used to identify Tomato yellow leaf curl virus silencing inhibitor V2. Agrobacterium tumefaciens harboring the target gene and GFP expression vector were co-injected into N. benthamiana. After 2 days, GFP was transiently overexpressed. After co-infection of watermelon chlorotic dwarf virus BV1 gene vector and empty vector pBIN with GFP for 7 days respectively, the GFP expression level abruptly decreased due to the host silencing mechanism. V2 gene expression vector and GFP expression vector co-infection leaf 7d, GFP remained high level expression, indicating that V2 as Tomato yellow leaf curl inhibitor of silencing can prevent the host silence mechanism. Agrobacterium injection can be quickly and conveniently screened for plant virus silencing suppressor.