Pathogenic viruses of bees and aphids: Are plants a common reservoir?

第1-2年利用威斯利和其他RHS站点以及剑桥大学植物园的生物多样性，发现和检查植物，蚜虫，蜜蜂（蜜蜂和许多熊蜂和独居蜜蜂）和蚜虫相关昆虫（蚜虫养殖蚂蚁，寄生蜂，瓢虫）中双顺反子病毒的多样性。方法将包括逆转录PCR与双顺反子病毒特异性引物，以检测和测序双顺反子病毒存在。建立蚜虫致病性双顺反子病毒的培养物（第1-2年）我们将从WP 1A中调查的蚜虫中建立携带双顺反子病毒的蚜虫和植物种群。双链病毒在蚜虫之间水平传播，通过植物传播，或垂直传播给后代（Ban等人，2007，2008），促进受感染种群的建立。第3年-分析双顺反子病毒对蚜虫的表现，行为，生存和与宿主植物的相互作用以及植物病毒的传播的影响我们将确定控制蚜虫致病性双顺反子病毒在昆虫中引起疾病的能力的病毒序列，并利用植物作为水库。如果拟南芥是一个储库（WP 2A），我们将使用诱变群体的无偏筛选或关注已知支持病毒运动的特定基因来鉴定调节双顺反子病毒驻留的植物基因（Chang et al. 2016）。

Years 1-2 Exploit the biodiversity of Wisley and other RHS sites and Cambridge University Botanic Garden to discover examine the diversity of dicistroviruses in plants, aphids, bees (honeybees and many bumblebees and solitary bees), and aphid-associated insects (aphid-farming ants, parasitoid wasps, ladybirds). Methods will include reverse transcription PCR with dicistrovirus-specific primers to detect and sequence dicistroviruses present. Establish cultures of aphid-pathogenic dicistroviruses (Years 1-2) We will establish dicistrovirus-carrying aphid and plant populations from aphids surveyed in WP1A. Dicistroviruses pass horizontally between aphids, through plants, or vertically to offspring (Ban et al. 2007, 2008), facilitating the establishment of infected stocks. Year 3 - Analyse effects of dicistroviruses on aphid performance, behaviour, survival and interactions with host plants and transmission of plant viruses We will identify viral sequences controlling the ability of aphid-pathogenic dicistroviruses to cause disease in insects and exploit plants as reservoirs. If Arabidopsis is a reservoir (WP2A) we will identify plant genes conditioning dicistrovirus residency using either unbiased screens of mutagenized populations, or focusing on specific genes known to support virus movement (Chang et al. 2016).