Abstract
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.
Highlights
Bees are valuable insects because of their dedicated role in the pollination of flowering plants [1, 2]
Using a deliberate RNA shotgun metagenomic sequencing strategy in combination with a dedicated bioinformatics workflow, we identified theorganisms and viruses associated with two bumble bee hosts, Bombus terrestris and Bombus pascuorum, and two solitary bee hosts, Osmia cornuta and Andrena vaga
The profiles of polyA+ mRNA samples showed a strong reduction of rRNA peak sizes and a general decrease in the rRNA/mRNA ratio compared to the total RNA samples (S6 File)
Summary
Bees are valuable insects because of their dedicated role in the pollination of flowering plants [1, 2]. The domesticated western honey bee, Apis mellifera, has a central role in commercial pollination and is traditionally employed by farmers to enhance crop yields [3]. Bumble bees and even solitary bees—which are collectively referred to as wild bees—find their way into commercial environments as alternative or supplemental crop pollinators. Solitary bees are a speciesrich group of valuable wild insects in terms of pollination ecosystem services [1] and the conservation of plant diversity [5]. A Tubulinosema parasite was discovered infecting bumble bees in Argentina [16]
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