Study aimed to determine how the transmission routes of viruses (Deformed wing virus (DWV), closely related Varroa destructor virus-1 (VDV-1) and their recombinants) impact on virus diversity. DWV/VDV-1 is transmitted both vertically and orally in the mite free colonies. The mite Varroa destructor

A honeybee (Apis mellifera) colony with established Varroa-free infestation (August 2013, Warwickshire, UK) was used. Samples included worker Varroa-free pupae, day 15 of development, injected with DWV/VDV-1 preparation, control Varroa-free pupae, Varroa-exposed newly emerged adult bees with symptoms of deformed wing disease, and Varroa -free asymptomatic newly emerged adults. Poly(A) RNA fraction from whole individual honeybees, which included honeybee mRNA and viral (DWV/VDV-1) genomic RNA, was sequenced by RNA-seq (Illumina HiSeq-2000). We also sequenced RNA isolated form the virus preparation from the symptomatic DWV/VDV-1-infected bees, which was used in the experimental pupae injection, and the control mixture of in vitro transcripts from DWV/VDV-1 clones were also sequenced.

本研究采用2013年8月于英国沃里克郡建立的西方蜜蜂（Apis mellifera）无瓦螨（Varroa）寄生蜂群。样本涵盖以下类群：经DWV/VDV-1制剂注射处理的发育至第15天的无瓦螨寄生工蜂蛹、空白对照无瓦螨寄生工蜂蛹、表现出变形翅病症的瓦螨侵染新羽化成虫蜂，以及无瓦螨寄生且无明显症状的新羽化成虫蜂。从单只整蜂中提取的聚腺苷酸RNA（Poly(A) RNA）组分（包含蜜蜂mRNA与病毒（DWV/VDV-1）基因组RNA）通过RNA测序（RNA-seq，Illumina HiSeq-2000平台）完成测序。本研究同时对两类样本进行测序：一类是用于实验性蛹注射的、源自症状性DWV/VDV-1感染蜂的病毒制剂中提取的RNA，另一类是DWV/VDV-1克隆的体外转录本对照混合物。