State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
Division of Crop Protection and Plant Health, Crop Research Institute, Praha 6, Czech Republic.
Cell Microbiol. 2019 Sep;21(9):e13047. doi: 10.1111/cmi.13047. Epub 2019 Jun 2.
Many plant viruses are vectored by insects in a persistent circulative manner. The insect gut and salivary gland are important barriers limiting virus spread, but the mechanisms by which viruses are able to cross the gut escape barriers of the insect remain largely unknown. Wheat dwarf virus (WDV), transmitted by Psammotettix alienus in a persistent, circulative, and nonpropagative manner, causes the most economically important virus disease in wheat. In this study, ADP ribosylation factor 1 (ARF1) was found to interact with the coat protein of WDV in a yeast two-hybrid, pull-down assay and to colocalise with virions in the gut and salivary glands of P. alienus. When transcription of ARF1 was suppressed by RNA interference, the WDV titre decreased in the haemolymph and salivary glands, and transmission efficiency decreased, but titre in the gut did not differ from that of the control. These data suggest that ARF1 of P. alienus binds to the WDV virion and helps virus spread from gut to haemolymph. Our study provides direct experimental evidence that WDV can use the existing membrane trafficking mechanism to aid its spread within the insect vector. This first analysis of the molecular interaction between WDV and its vector P. alienus contributes to understanding the mechanisms involved in circulative transmission of the virus by the leafhopper vector.
许多植物病毒通过昆虫以持久循环的方式传播。昆虫的肠道和唾液腺是限制病毒传播的重要屏障,但病毒能够穿越肠道逃避昆虫屏障的机制在很大程度上仍不清楚。小麦丛矮病毒(WDV)由叶蝉 Psammotettix alienus 以持久、循环和非增殖的方式传播,是小麦上最重要的经济病毒病。在这项研究中,发现 ADP 核糖基化因子 1(ARF1)在酵母双杂交、下拉测定中与 WDV 的外壳蛋白相互作用,并在 P. alienus 的肠道和唾液腺中与病毒粒子共定位。当 ARF1 的转录被 RNA 干扰抑制时,WDV 在血淋巴和唾液腺中的滴度降低,传播效率降低,但肠道中的滴度与对照没有差异。这些数据表明,P. alienus 的 ARF1 与 WDV 病毒粒子结合,有助于病毒从肠道传播到血淋巴。我们的研究提供了直接的实验证据,证明 WDV 可以利用现有的膜运输机制来帮助其在昆虫载体中传播。这是首次分析 WDV 与其载体 P. alienus 之间的分子相互作用,有助于理解叶蝉载体循环传播病毒的机制。