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1 Department of Biochemistry and Biophysics
and2 Department of Entomology, Texas A&M University and the Texas Agricultural Experiment Station,
and The3 Center for Advanced Invertebrate Molecular Sciences, Institute of Biosciences and Technology, College Station, TX 77843-2475, USA
Serial passage of nuclear polyhedrosis viruses (NPVs) through cultured cell lines results in the appearance of mutants with a complex phenotype referred to as the ‘few polyhedra’ (FP) phenotype. The altered plaque morphology and reduced occlusion production associated with the FP phenotype have been observed in Autographa californica multinucleocapsid nuclear polyhedrosis virus (AcMNPV) bearing mutations in the gene encoding the 25 kDa protein (25K gene). In this study, we sequenced the 25K genes of four spontaneously occurring AcMNPV FP mutants. These mutants, together with an artificially generated FP mutant (AcFP
gal, in which the gene for -galactosidase is fused in frame with the 25K ORF), were examined at the ultrastructural level to see if they exhibited the reduced virion occlusion and intranuclear envelopment which is associated with the FP phenotype. Observations on Spodoptera frugiperda Sf9 cells infected with the FP mutants revealed that all five mutants were impaired in virion occlusion and intranuclear nucleocapsid envelopment. The 25K mutants were also found to release two- to fivefold more infectious virus (p.f.u.) into the media of infected Sf9 cells. Marker rescue of AcFPgal restored wild-type virion occlusion, intranuclear envelopment and levels of infectious virus production.
* Author for correspondence. Fax +1 409 845 8934. e-mail mds7162@zeus.tamu.edu
Received 22 September 1994;
accepted 20 January 1995.
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