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

Olesya N. Gusachenko*, Luke Woodford, Katharin Balbirnie-Cumming, Ewan M. Campbell, Craig R. Christie, Alan S. Bowman, David J. Evans

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)
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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.
Original languageEnglish
Article number532
Number of pages17
Issue number5
Early online date12 May 2020
Publication statusPublished - May 2020

Bibliographical note

Funding: This 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 ( C.R.C was part-funded by the Scottish Beekeeping Association ( 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) (
Acknowledgments: We would like to express our gratitude to Robert J Paxton (Martin Luther University Halle-Wittenberg) for providing a reference sequence for the construction of DWV-B RG system, Marcus Bischoff and Gill McVee (University of St Andrews) for helping with microscopic imaging set up, Javier Tello (University of St Andrews) for help and advice on sample cryosectioning, Ivan Gusachenko (University of St Andrews) for obtaining a photo of EGFP-expressing pupa, Ashley Pearson (University of St Andrews) for assistance in molecular biology assays, and Kirsten Bentley (University of St Andrews) for the high practical and moral support throughout the project.


  • insect viruses
  • honey bee
  • pollination
  • virus vector
  • Varroa
  • RNA viruses
  • DWV
  • reverse genetics
  • Reverse genetics
  • Pollination
  • Insect viruses
  • Virus vector
  • Honey bee


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