Wang Lan-Lan, Wei Xue-Mei, Ye Xiao-Dong, Xu Hong-Xing, Zhou Xue-Ping, Liu Shu-Sheng, Wang Xiao-Wei
Ministry of Agriculture Key Laboratory of Agricultural Entomology, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
Pest Manag Sci. 2014 Oct;70(10):1624-31. doi: 10.1002/ps.3750. Epub 2014 Mar 17.
Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus within the family Geminiviridae, is an important pathogen of tomato in many tropical, subtropical and temperate regions. TYLCV is exclusively transmitted by the whitefly Bemisia tabaci in a circulative manner. The viral coat protein (CP) has been assumed to play important roles in the entry of TYLCV into the insect midgut cells.
Testing the hypothesis that CP plays an important role in TYLCV acquisition by B. tabaci, a soluble form of the CP was expressed and purified. The purified recombinant CP made it possible to examine the function of TYLCV CP without other viral proteins. In an in vivo binding assay, specific binding of TYLCV CP to B. tabaci midguts was detected when purified CP was fed to B. tabaci. In addition, real-time polymerase chain reaction analysis of virus titre revealed that B. tabaci fed with purified CP had reduced the level of virus in their midgut compared with those fed with bovine serum albumin or maltose-binding protein. These results suggest that binding of TYLCV CP to the B. tabaci midgut specifically inhibits virus acquisition.
The findings that TYLCV CP binds to B. tabaci midguts and decreases virus acquisition provide direct evidence that CP mediates the attachment of TYLCV to receptors on the epithelial cells of the B. tabaci midgut.
番茄黄化曲叶病毒(TYLCV)是双生病毒科菜豆金色花叶病毒属的成员,是许多热带、亚热带和温带地区番茄的重要病原体。TYLCV仅由烟粉虱以循环方式传播。病毒外壳蛋白(CP)被认为在TYLCV进入昆虫中肠细胞过程中起重要作用。
为验证CP在烟粉虱获取TYLCV过程中起重要作用这一假设,表达并纯化了一种可溶性形式的CP。纯化的重组CP使得在无其他病毒蛋白的情况下研究TYLCV CP的功能成为可能。在体内结合试验中,当将纯化的CP喂给烟粉虱时,检测到TYLCV CP与烟粉虱中肠有特异性结合。此外,病毒滴度的实时聚合酶链反应分析表明,与喂食牛血清白蛋白或麦芽糖结合蛋白的烟粉虱相比,喂食纯化CP的烟粉虱中肠内病毒水平降低。这些结果表明,TYLCV CP与烟粉虱中肠的结合特异性抑制病毒获取。
TYLCV CP与烟粉虱中肠结合并减少病毒获取的研究结果提供了直接证据,证明CP介导TYLCV与烟粉虱中肠上皮细胞受体的附着。