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Development of a Honey Bee RNA Virus Vector Based on the Genome of a Deformed Wing Virus
Open AccessArticle

Green Bees: Reverse Genetic Analysis of Deformed Wing Virus Transmission, Replication, and Tropism

1
Biomedical Sciences Research Complex, University of St. Andrews, St. Andrews KY16 9ST, UK
2
Centre for Inflammation Research, Queen‘s Medical Research Institute, University of Edinburgh, Edinburgh EH16 4TJ, UK
3
Institute of Biological and Environmental Sciences, School of Biological Sciences, University of Aberdeen, Aberdeen AB24 3FX, UK
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(5), 532; https://doi.org/10.3390/v12050532
Received: 31 March 2020 / Revised: 6 May 2020 / Accepted: 8 May 2020 / Published: 12 May 2020
(This article belongs to the Special Issue Advances in Honey Bee Virus Research)
Environmental and agricultural pollination services by honey bees, Apis mellifera, and honey production are compromised by high levels of annual colony losses globally. The majority are associated with disease caused by deformed wing virus (DWV), a positive-strand RNA virus, exacerbated by the ectoparasitic mite Varroa destructor. To improve honey bee health, a better understanding of virus transmission and pathogenesis is needed which requires the development of tools to study virus replication, transmission, and localisation. We report the use of reverse genetic (RG) systems for the predominant genetically distinct variants of DWV to address these questions. All RG-recovered viruses replicate within 24 h post-inoculation of pupae and could recapitulate the characteristic symptoms of DWV disease upon eclosion. Larvae were significantly less susceptible but could be infected orally and subsequently developed disease. Using genetically tagged RG DWV and an in vitro Varroa feeding system, we demonstrate virus replication in the mite by accumulation of tagged negative-strand viral replication intermediates. We additionally apply a modified DWV genome expressing a fluorescent reporter protein for direct in vivo observation of virus distribution in injected pupae or fed larvae. Using this, we demonstrate extensive sites of virus replication in a range of pupal tissues and organs and in the nascent wing buds in larvae fed high levels of virus, indicative of a direct association between virus replication and pathogenesis. These studies provide insights into virus replication kinetics, tropism, transmission, and pathogenesis, and produce new tools to help develop the understanding needed to control DWV-mediated colony losses. View Full-Text
Keywords: insect viruses; honey bee; pollination; virus vector; Varroa; RNA viruses; DWV; reverse genetics insect viruses; honey bee; pollination; virus vector; Varroa; RNA viruses; DWV; reverse genetics
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Gusachenko, O.N.; Woodford, L.; Balbirnie-Cumming, K.; Campbell, E.M.; Christie, C.R.; Bowman, A.S.; Evans, D.J. Green Bees: Reverse Genetic Analysis of Deformed Wing Virus Transmission, Replication, and Tropism. Viruses 2020, 12, 532.

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