Replication of Agrotis segetum granulovirus in continuous insect cell lines

Research output: Chapter in Book/Report/Conference proceedingAbstract

Abstract

The Agrotis ipsilon cell lines, IPLB-AiE1611T and AiEd6T, were inoculated with occlusion-body derived virions (ODV) of the A. segetum granulovirus (AgseGV). The capsules were dissolved by treatment with 0.1M NaH2CO3/0.1M NaCl for 30 min. to release the occluded virions followed by neutralization with an acidic salt solution and inoculation onto cells in 24-well tissue culture plates. Within a few days, numerous cells in the culture developed cytopathic effects consisting of darkened nuclei and enlarged cells when examined with phase contrast or Nomarski microscopy optics. After one week dense particles were seen within the nuclei of infected cells and the infection extended in the culture. Both cell lines replicated AgseGV equally well. Transfer of filtered supernatant from the ODV-inoculated culture to fresh cells led to similar cytopathic effects in the new culture suggesting the production of infectious (budded) virus from the infected cells. Electron microscopy of cells inoculated with the 2nd passage virus showed typical granulovirus capsules in the nuclei of many cells and in great numbers. The capsules are being tested for in vivo infectivity in A. segetum larvae. This is the first successful replication of this virus in cell cultures and one of the few examples of in vitro replication and passaging of granuloviruses. It opens up the possibility to study the molecular genetics and functional biology of AgseGV in much detail and to engineer this virus for better insecticidal properties. This work was made possible through a visitors grant from the Wageningen Graduate School ‘Production Ecology & Resource Conservation’
Original languageEnglish
Title of host publicationAbstract Book of the 42nd Annual Meeting of the Society for Invertebrate Pathology, Park City, Utah, USA, 16 - 20 August, 2009
Place of PublicationPark City, Utah
Pages55-55
Publication statusPublished - 2009
Event42nd Annual Meeting of the Society for Invertebrate Pathology, Park City, Utah, USA -
Duration: 16 Aug 200920 Aug 2009

Conference

Conference42nd Annual Meeting of the Society for Invertebrate Pathology, Park City, Utah, USA
Period16/08/0920/08/09

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    Lynn, D. R., van Lent, J. W. M., van Oers, M. M., & Vlak, J. M. (2009). Replication of Agrotis segetum granulovirus in continuous insect cell lines. In Abstract Book of the 42nd Annual Meeting of the Society for Invertebrate Pathology, Park City, Utah, USA, 16 - 20 August, 2009 (pp. 55-55).