Systemic and in vitro infection process of Bombyx mori nucleopolyhedrovirus

Abstract

To analyse the systemic progression of infection by Bombyx mori nucleopolyhedrovirus (BmNPV) through oral ingestion by the silkworm larvae, a recombinant virus (vBmp10GFP) expressing the green fluorescent protein (GFP) under the control of the very late, viral p10 promoter (which still forms the polyhedral occlusion bodies) was constructed. Infection of B. mori derived BmN cells with the recombinant virus resulted in the expression of GFP from 12 h post infection (hpi), with maximal accumulation of the expressed protein by 60 hpi. B. mori larvae that ingested the polyhedra containing vBmp10GFP showed localized expression of GFP in the midgut epithelial cells within 24 hpi, indicating virus replication. The primary spread of the virus infection occurred through the tracheae. Viral multiplication was subsequently detected in nearly all the larval tissues including the neurons and regions of silk-glands that were in contact with the tracheae. Infection in fat bodies was widespread by 48 hpi, by which time the haemocytes also showed infection. In vitro infection of isolated organs/tissues from B. mori with the budded virions (BV) of vBmp10GFP also showed viral multiplication in the cells that were associated with the tracheae, confirming the role of tracheae in spreading the infection.