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Protocols for Microapplicator-assisted Infection of Lepidopteran Larvae with Baculovirus
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Spatially Segregated Transmission of Co-Occluded Baculoviruses Limits Virus-Virus Interactions Mediated by Cellular

Verónica Pazmiño-Ibarra1, Salvador Herrero2, Rafael Sanjuan1

  • 1Institute for Integrative Systems Biology (I2SysBio), Consejo Superior de Investigaciones Científicas-Universitat de València, C/Catedrático Agustín Escardino 9, 46980 Paterna, Spain.

Viruses
|August 26, 2022
PubMed

Insights

Baculoviruses transmit multiple viral particles together, but coinfection of insect cells with different variants is limited. However, viral recombination within cells later in infection allows for interactions between baculovirus variants.

Area of Science:

  • Virology
  • Insect Pathology
  • Molecular Biology

Background:

  • Alphabaculoviruses form occlusion bodies (OBs) for efficient insect host transmission.
  • Nucleopolyhedroviruses exhibit nucleocapsid aggregation, suggesting coinfection potential.
  • Coinfection may promote stable interactions between different baculovirus variants.

Purpose of the Study:

  • To quantify the joint transmission of different baculovirus variants.
  • To investigate the role of occlusion-derived virions (ODVs) in promoting coinfection.
  • To understand the mechanisms of baculovirus variant interaction within insect hosts.

Main Methods:

  • Coinfection of insect cells with two fluorescent reporter-tagged baculovirus constructs.
  • Generation of occlusion bodies (OBs) from coinfected cells.
  • Inoculation of Spodoptera exigua larvae with OBs and subsequent midgut analysis via microscopy.
  • Analysis of a polyhedrin-deficient mutant's transmission and recombination.

Main Results:

  • Microscopy revealed segregation of reporter genes into different infection foci, indicating limited ODV co-transmission to the same host cell.
  • A polyhedrin-deficient mutant successfully transmitted by exploiting the OBs of a functional virus.
  • Recombination allowed the polyhedrin-deficient mutant to re-acquire a lost gene, demonstrating cellular coinfection.
  • Spatial segregation during transmission and primary infection limits baculovirus variant interactions.

Conclusions:

  • Baculovirus transmission and initial infection lead to spatial segregation of variants, limiting direct interactions.
  • Cellular coinfection and recombination within infected insect cells facilitate interactions between baculovirus variants later in infection.
  • This study provides insights into the evolutionary dynamics of baculovirus populations within hosts.
Keywords:
baculoviruscollective infectious unitscooperationocclusion bodiessocial evolution

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