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Successful co-infection of two different baculovirus species in the same cell line reveals a potential strategy for large in vitro production

  • Biotechnology and Industrial Microbiology - Research Paper
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Abstract

Baculoviruses have been applied for biocontrol of agricultural pests, such as velvetbean caterpillar (Anticarsia gemmatalis) and fall armyworm (Spodoptera frugiperda). Cell culture is an interesting approach for large-scale production of these viruses. Co-infection of a host cell with two distinct viruses can contribute to reduce costs due to saving cell culture media, bioreactor space and the resulting co-occluded polyhedra may help to reduce final biopesticide costs. The baculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) and Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) were chosen to test a model for in vitro co-infection in SF21 cells. Different proportions of SfMNPV/AgMNPV were evaluated along three in vitro passages by optical microscopy analysis of cells and real-time PCR (qPCR) of DNA obtained from budded viruses (BVs) and occlusion bodies (OBs). The kinetics of viral protein synthesis was carried out for analysis of the co-infection in first passage and bioassays with the resulting OBs were performed against A. gemmatalis and S. frugiperda larvae. The results demonstrated successful co-infection in these cells. The quantity of SfMNPV and AgMNPV in supernatants and sediments tends to be maintained stable during the three passages, although the amount of AgMNPV was higher than SfMPNV in most of the experiments. Analysis of the kinetics of radiolabed proteins showed that the cell protein synthesis was shut off and two distinct bands of about 30 kDa, regarded to be the polyhedrin of each virus, were strongly detected at 48 and 72 hp.i. Although the pathogenicity of the produced viruses was not completely satisfactory, the bioassays confirmed occurrence of co-infected larvae with disproportional amount of each virus.

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Acknowledgements

We are grateful to Rick Alan Harder for the language review. ALR Santos (UniCeub, Undergraduate student) for the help with lab analysis. FGV Schmidt (Embrapa) provided the A. gemmatalis and S. frugiperda specimens.

Funding

The study received financial support from Fundação de apoio à pesquisa do DF (FAP-DF) (PPP/FAPDF/CNPq), grant number 193.000.661/2015 and Embrapa (grant number 11.15.02.001.03.05).

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Authors and Affiliations

  1. Embrapa Recursos Genéticos e Biotecnologia, Brasilia, DF, 70770-917, Brazil

    M. M. Sanches, W. Sihler & M. L. Souza

  2. Centro Universitário de Brasília, Brasilia, DF, Brazil

    G. C. Guimarães

Authors
  1. M. M. Sanches
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  2. G. C. Guimarães
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  3. W. Sihler
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  4. M. L. Souza
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Contributions

Conceptualization: MMS, MLS; Methodology: MMS, WS, MLS; Formal analysis and investigation: MMS, GCG, WS; Writing — original draft preparation: MMS, MLS; Writing — review and editing: all authors; Funding acquisition: MMS; Resources: MMS, MLS, WS; Supervision: MMS.

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Correspondence to M. M. Sanches.

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Responsible Editor: Fernando R. Spilki

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Sanches, M.M., Guimarães, G.C., Sihler, W. et al. Successful co-infection of two different baculovirus species in the same cell line reveals a potential strategy for large in vitro production. Braz J Microbiol 52, 1835–1843 (2021). https://doi.org/10.1007/s42770-021-00622-z

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  • DOI: https://doi.org/10.1007/s42770-021-00622-z

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