Construction and Rescue of a Molecular Clone of Deformed Wing Virus (DWV)

PLoS One. 2016 Nov 9;11(11):e0164639. doi: 10.1371/journal.pone.0164639. eCollection 2016.

Abstract

European honey bees are highly important in crop pollination, increasing the value of global agricultural production by billions of dollars. Current knowledge about virulence and pathogenicity of Deformed wing virus (DWV), a major factor in honey bee colony mortality, is limited. With this study, we close the gap between field research and laboratory investigations by establishing a complete in vitro model for DWV pathogenesis. Infectious DWV was rescued from a molecular clone of a DWV-A genome that induces DWV symptoms such as crippled wings and discoloration. The expression of DWV proteins, production of infectious virus progeny, and DWV host cell tropism could be confirmed using newly generated anti-DWV monoclonal antibodies. The recombinant RNA fulfills Koch's postulates circumventing the need of virus isolation and propagation of pure virus cultures. In conclusion, we describe the development and application of a reverse genetics system for the study of DWV pathogenesis.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Base Sequence
  • Bees / virology
  • Blotting, Western
  • Capsid Proteins / immunology
  • Genome, Viral / genetics
  • Host-Pathogen Interactions
  • Immunohistochemistry
  • Insect Viruses / genetics*
  • Insect Viruses / metabolism
  • Insect Viruses / physiology
  • Mice, Inbred BALB C
  • Microscopy, Electron, Transmission
  • Phylogeny
  • Picornaviridae / classification
  • Picornaviridae / genetics*
  • Picornaviridae / metabolism
  • Polyproteins / genetics
  • Polyproteins / metabolism
  • Pupa / virology
  • RNA Viruses / genetics*
  • RNA Viruses / metabolism
  • RNA Viruses / ultrastructure
  • RNA, Viral / genetics*
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Viral Proteins / metabolism
  • Wings, Animal / virology

Substances

  • Antibodies, Monoclonal
  • Capsid Proteins
  • Polyproteins
  • RNA, Viral
  • Viral Proteins

Grants and funding

This study was funded by “BIOS Science Austria” (http://bios-science.at/index.php/en/; grant: 2012_vetmediuni_2) and the “University of veterinary medicine” (http://www.vetmeduni.ac.at/en/; grant: PP23014275). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.