Department of Zoology, University of Delhi, Delhi, 110007, India.
Division of Entomology, Indian Agricultural Research Institute, New Delhi, 110012, India.
Cell Microbiol. 2016 May;18(5):663-78. doi: 10.1111/cmi.12538. Epub 2015 Nov 17.
Begomoviruses are a major group of plant viruses, transmitted exclusively by Bemisia tabaci (Gennadius) in a persistent circulative non-propagative manner. The information regarding molecular and cellular basis underlying Begomovirus - whitefly interaction is very scarce. Evidences have suggested that the insect gut possesses some crucial protein receptors that allow specific entry of virus into the insect haemolymph. We have performed yeast two hybrid gut cDNA expression library screening against coat protein of Tomato leaf curl New Delhi virus (ToLCV) and Cotton leaf curl Rajasthan virus (CLCuV) as bait. Midgut protein (MGP) was the common protein found interacting with both ToLCV and CLCuV. MGP was localized in whole mount B. tabaci as well as in dissected guts through confocal microscopy. Pull down and dot blot assays confirmed in vitro interaction between ToLCV/CLCuV coat protein and MGP. Immunolocalization analysis also showed colocalization of ToLCV/CLCuV particles and MGP within insect's gut. Finally, anti-MGP antibody fed B. tabaci, exhibited 70% reduction in ToLCV transmission, suggesting a supportive role for MGP in virus transmission.
双生病毒是一类重要的植物病毒，仅通过烟粉虱（Bemisia tabaci）以持久循回非增殖的方式传播。有关双生病毒-粉虱相互作用的分子和细胞基础的信息非常有限。有证据表明，昆虫肠道具有一些关键的蛋白受体，允许病毒特异性进入昆虫血淋巴。我们已经针对番茄黄曲叶病毒（ToLCV）和棉花曲叶病毒（CLCuV）的外壳蛋白进行了酵母双杂交肠道 cDNA 表达文库筛选。中肠蛋白（MGP）是与 ToLCV 和 CLCuV 都相互作用的共同蛋白。通过共聚焦显微镜，MGP 被定位在整个 B. tabaci 以及解剖肠道中。下拉和斑点印迹分析证实了 ToLCV/CLCuV 外壳蛋白与 MGP 之间的体外相互作用。免疫定位分析还显示，ToLCV/CLCuV 颗粒与 MGP 在昆虫肠道内共定位。最后，用抗-MGP 抗体喂养的烟粉虱，ToLCV 的传播减少了 70%，表明 MGP 在病毒传播中起支持作用。
