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Background   Bombyx mori  nucleopolyhedrovirus (BmNPV)  orf64  ( Bm64 , a homologue of  ac78 ) is a core baculovirus gene. Recently, Li et al. reported that Ac78 was not essential for budded viruses (BVs) production and occlusion-derived viruses (ODVs) formation (Virus Res 191:70–82, 2014). Conversely, Tao et al. demonstrated that Ac78 was localized to the BV and ODV envelopes and was required for BV production and ODV formation (J Virol 87:8441–50, 2013). In this study, the function of  Bm64  was characterized to determine the role of  Bm64  in the BmNPV infection cycle.    Method  The temporal expression of  Bm64  was examined using total RNA extracted from BmNPV-infected BmN cells at different time points by reverse-transcription PCR (RT-PCR) and 5’ RACE analysis. To determine the functions of  Bm64  in viral replication and the viral phenotype throughout the viral life cycle, a deletion virus (vBm 64KO ) was generated via homologous recombination in  Escherichia coli . Viral replication and BV production were determined by real-time PCR. Electron microscopy was used to detect virion morphogenesis. The subcellular localization of Bm64 was determined by microscopy, and  per os  infectivity was used to determine its role in the baculovirus oral infection cycle.    Results  Viral plaque and titer assay results showed that a few infectious BVs were produced by vBm 64KO , suggesting that deletion of  Bm64  affected BV production. Viral DNA replication was detected and polyhedra were observed in vBm 64KO -transfected cells. Microscopy analysis revealed that Bm64 was predominantly localized to the ring zone of the nuclei during the infection cycle. Electron microscopy showed that  Bm64  was not essential for the formation of ODVs or the subsequent occlusion of ODV into polyhedra. The  per os  infectivity results showed that the polyhedra of vBm 64KO  were unable to infect silkworm larvae.    Conclusion  In conclusion, our results suggest that  Bm64  plays an important role in BV production and  per os  infection, but is not required for viral DNA replication or ODV maturation.    Electronic supplementary material  The online version of this article (doi:10.1186/s12985-015-0399-9) contains supplementary material, which is available to authorized users.

Bombyx mori nucleopolyhedrovirus (BmNPV) Bm64 is required for BV production and per os infection