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dc.contributor.authorGusachenko, Olesya N.
dc.contributor.authorWoodford, Luke
dc.contributor.authorBalbirnie-Cumming, Katharin
dc.contributor.authorCampbell, Ewan M.
dc.contributor.authorChristie, Craig R.
dc.contributor.authorBowman, Alan S.
dc.contributor.authorEvans, David J.
dc.date.accessioned2020-05-14T14:30:05Z
dc.date.available2020-05-14T14:30:05Z
dc.date.issued2020-05-12
dc.identifier.citationGusachenko , O N , Woodford , L , Balbirnie-Cumming , K , Campbell , E M , Christie , C R , Bowman , A S & Evans , D J 2020 , ' Green bees : reverse genetic analysis of deformed wing virus transmission, replication, and tropism ' , Viruses , vol. 12 , no. 5 , 532 . https://doi.org/10.3390/v12050532en
dc.identifier.issn1999-4915
dc.identifier.otherPURE: 267943346
dc.identifier.otherPURE UUID: 5b788904-0ae2-47f8-9432-45add356d022
dc.identifier.otherBibtex: v12050532
dc.identifier.otherORCID: /0000-0003-2530-2120/work/74118085
dc.identifier.otherScopus: 85084786541
dc.identifier.urihttp://hdl.handle.net/10023/19943
dc.descriptionThis work was supported by grant funding from Biotechnology and Biological Sciences Research Council (BBSRC): BBSRC BB/M00337X/2 and BB/I000828/1. C.R.C. was supported by a KTN BBSRC CASE studentship BB/M503526/1 (http://www.bbsrc.ac.uk). C.R.C was part-funded by the Scottish Beekeeping Association (https://www.scottishbeekeepers.org.uk/) and the Animal Health and Welfare program by the Scottish Government. E.M.C. was supported by the Veterinary Medicines Directorate, Department for Environment Food & Rural Affairs (Project # VM0517) (https://www.gov.uk/government/organisations/veterinary-medicines-directorate).en
dc.description.abstractEnvironmental 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.
dc.format.extent17
dc.language.isoeng
dc.relation.ispartofVirusesen
dc.rightsCopyright © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).en
dc.subjectInsect virusesen
dc.subjectHoney beeen
dc.subjectPollinationen
dc.subjectVirus vectoren
dc.subjectVarroaen
dc.subjectRNA virusesen
dc.subjectDWVen
dc.subjectReverse geneticsen
dc.subjectQR355 Virologyen
dc.subjectQH301 Biologyen
dc.subject3rd-DASen
dc.subject.lccQR355en
dc.subject.lccQH301en
dc.titleGreen bees : reverse genetic analysis of deformed wing virus transmission, replication, and tropismen
dc.typeJournal articleen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews.School of Biologyen
dc.contributor.institutionUniversity of St Andrews.Sir James Mackenzie Institute for Early Diagnosisen
dc.contributor.institutionUniversity of St Andrews.Biomedical Sciences Research Complexen
dc.identifier.doihttps://doi.org/10.3390/v12050532
dc.description.statusPeer revieweden


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