Physical Maps and Comparative DNA Hybridization of Mamestra Brassicae and Panolis Flammea Nuclear Polyhedrosis Virus Genomes

Abstract

SUMMARYThe genome DNAs from the Panolis flammea (Pf) multiple nucleocapsid nuclear polyhedrosis virus (MNPV) and the Mamestra brassicae (Mb) MNPV were analysed with the restriction endonucleases BamHI, BgllI, KpnI, HindlII, Sinai and XhoI. The profiles produced by each enzyme for the two virus genomes were quite dissimilar with very few comigrating fragments. The size of PfMNPV DNA was calculated to be 145 kilobase pairs (kbp) and that of MbMNPV 150 kbp. Physical maps of the two genomes were constructed utilizing the above enzymes. The two maps were oriented in relation to their putative polyhedrin genes. Alignment of the two restriction maps for PfMNPV and MbMNPV was achieved by performing cross blot hybridization between XhoI digests of the two virus genomes. This showed that despite differences in the physical maps the two genomes shared overall similarity in gene organization. The two viruses were also compared using dot blot hybridization analysis to quantify homology with Autographa californica (AC) MNPV. These data showed that both PfMNPV and MbMNPV were distantly related to AcMNPV but exhibited a high degree of homology to each other (nearly 100% in 20% formamide, 70% in 50% formamide).

Characteristically distinct isolates of the nuclear polyhedrosis virus from Spodoptera litura

“…DNA restriction enzyme analysis and hybridization experiments have shown genetic relatedness between baculoviruses (Knudson & Tinsley, 1978;Smith & Summers, 1978Jewell & Miller, 1980;Vlak & Gr6ner, 1980;Harvey & Tanada, 1985;Rohrmann.et al, 1982;Wiegers & Vlak, 1984;Kislev, 1985;Possee & Kelly, 1988) and restriction endonuclease analysis of the viral genome has revealed that wild stocks of NPVs are often composed of several genetically different isolates (Lee & Miller, 1978;Miller & Dawes, 1978;Smith & Summers, 1978, 1979Miller et al, 1980;Knell & Summers, 1981 ;Gettig & McCarthy, 1982;Kislev & Edelman, 1982; t Present address: Odawara Research Institute, Nippon Soda Co. Ltd, Odawara 250-02, Japan.…”

Evidence for the presence of a low-level, persistent baculovirus infection of Mamestra brassicae insects.

“…This virus was activated by feeding the M. brassicae larvae with either the closely related Panolis flammea multiple nucleocapsid nucleopolyhedrovirus (PaflMNPV), or the more distantly related Autographa californica (Ac) MNPV. DNA profiles from the activated virus showed that it was very closely related, if not identical, to M. brassicae (Mb) MNPV (Possee & Kelly, 1988). PCR amplification of polyhedrin gene sequences demonstrated that the MbMNPV-like virus was present in the egg, larval, pupal and adult stages of the insect life-cycle.…”

Characterization of the ecdysteroid UDP-glucosyltransferase gene from Mamestra brassicae nucleopolyhedrovirus

“…It has been released in the UK on a large scale for the control of the pine beauty moth, Panolis flammea (Entwistle & Evans, 1987). However, the molecular biology of this virus is poorly understood and polyhedrin is the only MbMNPV gene sequenced to date (Possee & Kelly, 1988;Cameron & Possee, 1989). MbMNPV is therefore an attractive subject for both genetic analysis and recombinant studies with a view to future field studies.…”