In vitro culture of Lambdina fiscellaria lugubrosa nucleopolyhedrovirus in heterologous cell lines

  • Beatrixe H. Whittome-Waygood
  • John C. Fraser
  • Christopher J. Lucarotti
  • Imre S. Otvos
  • Nicholas Conder
  • David B. Levin
Article
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Abstract

Infections of two heterologous insect cell lines derived from Malacosoma disstria (Md108) and Choristoneura fumiferana (Cf70) by the Lambdina fiscellaria lugubrosa nucleopolyhedrovirus (LafiNPV-W) were characterized. Cytopathic effects characteristic of LafiNPV-W infection, including rounding of cells, nuclear hypertrophy, and occlusion body (OB) production, were observed in both cell lines. Budded virus titers were slightly higher in Md108 cells than Cf70 cells (5.8 × 107 versus 3.1 × 107 TCID50 units mL−1). Viral replication kinetics and cytopathic effects induced by LafiNPV-W infection were very similar in both cell lines. Actin rearrangements and redistribution of heterochromatin and euchromatin were observed within 24 h post-inoculation (hpi), and large quantities of nucleocapsids and virions were observed by electron microscopy at 48 hpi in both cell lines. Cf70 cultures produced OBs with numerous embedded virions, while OBs in Md108 cultures contained few virions or were empty with nucleocapsids packed in the nucleoplasm between OBs. In bioassays against second instar L. fiscellaria lugubrosa, OBs derived from LafiNPV-W-infected Md108 cells induced significantly lower levels of mortality than OBs derived from LafiNPV-W-infected Cf70 cells or from infected L. fiscellaria fiscellaria larvae.

Keywords

Baculovirus Hemlock looper Insect cell culture Virion occlusion 
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Notes

Acknowledgments

We are grateful to Dr. Sadar Sohi and the late Dr. John C. Cunningham for the cell lines and LafiNPV-W. The time course assays and host specificity assays were performed with the aid of Mandy Miller. We also thank Anne van de Raadt, Holly Armstrong, Joanna McGarvie, Meaghan Complin, and Karolina Monge-Monge for their assistance with the bioassays. Statistical advice was provided by the University of Victoria, Statistical Help Center. This research was supported by funds from the Biocontrol Network (Natural Sciences and Engineering Research Council of Canada), the Canadian Forest Service-Canadian Biotechnology Strategy (Natural Resources Canada), and Forest Protection Limited (Lincoln, New Brunswick).

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Copyright information

© The Society for In Vitro Biology 2009

Authors and Affiliations

  • Beatrixe H. Whittome-Waygood
    • 1
  • John C. Fraser
    • 1
  • Christopher J. Lucarotti
    • 2
  • Imre S. Otvos
    • 3
  • Nicholas Conder
    • 3
  • David B. Levin
    • 1
    • 4
  1. 1.Department of BiologyUniversity of VictoriaVictoriaCanada
  2. 2.Natural Resources Canada, Canadian Forest ServiceAtlantic Forestry CentreFrederictonCanada
  3. 3.Natural Resources Canada, Canadian Forest ServicePacific Forestry CentreVictoriaCanada
  4. 4.Department of Biosystems EngineeringUniversity of ManitobaWinnipegCanada

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