Schoonvaere Karel, De Smet Lina, Smagghe Guy, Vierstraete Andy, Braeckman Bart P, de Graaf Dirk C
Laboratory of Molecular Entomology and Bee Pathology, Department of Biochemistry and Microbiology, Faculty of Sciences, Ghent University, Ghent, Belgium.
Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
PLoS One. 2016 Dec 22;11(12):e0168456. doi: 10.1371/journal.pone.0168456. eCollection 2016.
The diversity of eukaryote organisms and viruses associated with wild bees remains poorly characterized in contrast to the well-documented pathosphere of the western honey bee, Apis mellifera. Using a deliberate RNA shotgun metagenomic sequencing strategy in combination with a dedicated bioinformatics workflow, we identified the (micro-)organisms and viruses associated with two bumble bee hosts, Bombus terrestris and Bombus pascuorum, and two solitary bee hosts, Osmia cornuta and Andrena vaga. Ion Torrent semiconductor sequencing generated approximately 3.8 million high quality reads. The most significant eukaryote associations were two protozoan, Apicystis bombi and Crithidia bombi, and one nematode parasite Sphaerularia bombi in bumble bees. The trypanosome protozoan C. bombi was also found in the solitary bee O. cornuta. Next to the identification of three honey bee viruses Black queen cell virus, Sacbrood virus and Varroa destructor virus-1 and four plant viruses, we describe two novel RNA viruses Scaldis River bee virus (SRBV) and Ganda bee virus (GABV) based on their partial genomic sequences. The novel viruses belong to the class of negative-sense RNA viruses, SRBV is related to the order Mononegavirales whereas GABV is related to the family Bunyaviridae. The potential biological role of both viruses in bees is discussed in the context of recent advances in the field of arthropod viruses. Further, fragmentary sequence evidence for other undescribed viruses is presented, among which a nudivirus in O. cornuta and an unclassified virus related to Chronic bee paralysis virus in B. terrestris. Our findings extend the current knowledge of wild bee parasites in general and addsto the growing evidence of unexplored arthropod viruses in valuable insects.
与西方蜜蜂（Apis mellifera）记录详尽的病原体群相比，与野生蜜蜂相关的真核生物和病毒的多样性仍鲜为人知。我们采用了一种精心设计的RNA鸟枪法宏基因组测序策略，并结合专门的生物信息学工作流程，鉴定了与两种熊蜂宿主（熊蜂（Bombus terrestris）和帕斯夸熊蜂（Bombus pascuorum））以及两种独居蜂宿主（角额壁蜂（Osmia cornuta）和黄地蜂（Andrena vaga））相关的（微）生物和病毒。Ion Torrent半导体测序产生了约380万个高质量读数。最显著的真核生物关联是熊蜂体内的两种原生动物，即蜜蜂微孢子虫（Apicystis bombi）和克氏锥虫（Crithidia bombi），以及一种线虫寄生虫球型蜂线虫（Sphaerularia bombi）。在独居蜂角额壁蜂中也发现了锥虫原生动物克氏锥虫。除了鉴定出三种蜜蜂病毒——黑蜂王台病毒、囊雏病病毒和瓦螨毁灭病毒1型以及四种植物病毒外，我们还根据其部分基因组序列描述了两种新型RNA病毒——斯卡尔迪斯河蜜蜂病毒（SRBV）和甘达蜜蜂病毒（GABV）。这两种新型病毒属于负链RNA病毒类别，SRBV与单股负链RNA病毒目相关，而GABV与布尼亚病毒科相关。本文结合节肢动物病毒领域的最新进展，讨论了这两种病毒在蜜蜂中的潜在生物学作用。此外，还提供了其他未描述病毒的片段序列证据，其中包括角额壁蜂体内的一种裸病毒和熊蜂体内一种与慢性蜜蜂麻痹病毒相关的未分类病毒。我们的研究结果总体上扩展了目前对野生蜜蜂寄生虫的认识，并进一步证明了在珍贵昆虫中存在尚未探索的节肢动物病毒。
