title: Disruption of Bombyx mori nucleopolyhedrovirus ORF71 (Bm71) results in inefficient budded virus production and decreased virulence in host larvae.
abstract: The Bombyx mori nucleopolyhedrovirus (BmNPV) is a baculovirus that selectively infects domestic silkworm. BmNPV ORF71 (Bm71) is not a core set gene in baculovirus and shares 92 % amino acid sequence identity with Autographa californica multinucleocapsid NPV ORF88 (Ac88/cg30). Previously, it has been reported that virus lacking Ac88 had no striking phenotypes in cell lines or host larvae. However, the exact role of Bm71 during BmNPV life cycle remains unknown. In the present study, we constructed a Bm71-disrupted (Bm71-D) virus and assessed the effect of the Bm71 disruption on viral replication and viral phenotype throughout the viral life cycle. Results showed that the Bm71-D bacmid could successfully transfect Bm5 cell lines and produce infectious budded virus (BV). But the BV titer was 10- to 100-fold lower than that of the wild-type (WT) virus during infection, and the decreased BV titer was rescued by Bm71 gene repair virus (Bm71-R). A larval bioassay showed that Bm71-D virus took 7.5 h longer than the WT to kill Bombyx mori larvae. Transmission electron microscopy analysis indicated that the Bm71-D virus-infected cells had typical virogenic stroma, bundles of nucleocapsids and polyhedra. Taken together, these results suggest that Bm71 has important implications for determining BV yield and virulence in viral life cycle even though it is not an essential gene for replication of BmNPV.
authors.affiliation: Ministry of Agriculture Key Laboratory of Agricultural Entomology, Institute of Insect Science, Zhejiang University, Yuhangtang Road 866, Hangzhou, 310058 Zhejiang, China.
authors.forename: Min-Juan
authors.initials: MJ
authors.lastname: Zhang
authors.forename: Ruo-Lin
authors.initials: RL
authors.lastname: Cheng
authors.forename: Yi-Han
authors.initials: YH
authors.lastname: Lou
authors.forename: Wan-Lu
authors.initials: WL
authors.lastname: Ye
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Xiao-Ying
authors.initials: XY
authors.lastname: Fan
authors.forename: Hai-Wei
authors.initials: HW
authors.lastname: Fan
authors.forename: Chuan-Xi
authors.initials: CX
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001681:Biological Assay; D012831:Bombyx / Q000254:growth & development / Q000821:virology*; D002460:Cell Line; D059765:Homologous Recombination; D007814:Larva / Q000821:virology; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D016366:Open Reading Frames / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014774:Virulence; D057074:Virus Release