Abstract

Polydnaviruses are dsDNA viruses associated with endoparasitoid wasps. Delivery of the virus during parasitization of a caterpillar and subsequent virus gene expression is required for production of an amenable environment for parasitoid offspring development. Consequently, understanding of Polydnavirus gene function provides insight into mechanisms of host susceptibility and parasitoid wasp host range. Polydnavirus genes predominantly are arranged in multimember gene families, one of which is the vinnexins, which are virus homologues of insect gap junction genes, the innexins. Previous studies of Campoletis sonorensis Ichnovirus Vinnexins using various heterologous systems have suggested the four encoded members may provide different functionality in the infected caterpillar host. Here, we expressed two of the members, vnxG and vnxQ2, using recombinant baculoviruses in susceptible host, the caterpillar Heliothis virescens. Following intrahemocoelic injections, we observed that >90% of hemocytes (blood cells) were infected, producing recombinant protein. Larvae infected with a vinnexin-recombinant baculovirus exhibited significantly reduced molting rates relative to larvae infected with a control recombinant baculovirus and mock-infected larvae. Similarly, larvae infected with vinnexin-recombinant baculoviruses were less likely to survive relative to controls and showed reduced ability to encapsulate chromatography beads in an immune assay. In most assays, the VnxG protein was associated with more severe pathology than VnxQ2. Our findings support a role for Vinnexins in CsIV and more broadly Ichnovirus pathology in infected lepidopteran hosts, particularly in disrupting multicellular developmental and immune physiology.

Highlights

  • Our findings support a role for Vinnexins in Campoletis sonorensis Ichnovirus (CsIV) and more broadly Ichnovirus pathology in infected lepidopteran hosts, in disrupting multicellular developmental and immune physiology

  • We utilized recombinant baculoviruses that were previously characterized [21]. These include two experimental recombinant viruses that encode Campoletis sonorensis Ichnovirus (CsIV) vinnexinG and CsIV vinnexinQ2, each with C-terminus 6x-His epitope, and a third recombinant virus encoding a fish melanocortin-4-receptor (Mc4r) with an N-terminus FLAG epitope to serve as control for effects of recombinant virus and overexpression of an exogenous membrane protein

  • Anti-His (VnxG-His, VnxQ2-His; Figure 1B) and anti-FLAG (FLAG-Mc4r; Figure 1C) immunomicroscopy results corresponded to patterns observed Sf9 cells infected with the recombinant viruses [21], and Sf9 and High Five cells transfected with vinnexin expression plasmids [30]

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Summary

Introduction

Polydnaviruses (PDVs) are a remarkable virus family These large dsDNA viruses are obligately and mutualistically associated with specific families of parasitoid wasps, a relationship that generates unusual selection pressures on the viruses. PDVs are a paraphyletic lineage: Bracoviruses (BVs), associated with PDVs are associated with the microgastroid complex of the wasp family Braconidae, while Ichnoviruses (IVs) are associated with Campopleginae and Banchinae wasp subfamilies of the wasp family Ichneumonidae. Despite their evolutionary independence, the different lineages of PDVs exhibit similar genome organization and are integrated as a provirus into the genome of the host wasp. There is no evidence of PDV replication in the infected secondary host during this period [5]

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