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Applied and Environmental Microbiology, November 2001, p. 5204-5209, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5204-5209.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Persistence of an Occlusion-Negative Recombinant Nucleopolyhedrovirus in Trichoplusia ni Indicates High Multiplicity of Cellular Infection

James C. Bull,1 H. C. J. Godfray,2 and David R. O'Reilly1,*

Department of Biology, Imperial College of Science, Technology, and Medicine, London SW7 2AZ,1 and NERC Centre for Population Biology, Imperial College at Silwood Park, Ascot, Berkshire SL5 7PY,2 United Kingdom

Received 7 May 2001/Accepted 30 July 2001

We use data from the serial passage of co-occluded recombinant Autographa californica nuclear polyhedrosis virus (AcMNPV) to estimate the viral multiplicity of infection of cells within infected insects. Co-occlusion, the incorporation of wild-type and mutant virus genomes in the same occlusion body, has been proposed as a strategy to deliver genetically modified viruses as insecticides in a way that contains their spread in the environment. It may also serve as a means whereby naturally occurring mutant forms of NPVs can be maintained in a stable polymorphism. Here, a recombinant strain of AcMNPV was constructed with a deletion of its polyhedrin gene, rendering it incapable of producing occlusion bodies (i.e., occlusion negative). This was co-occluded with wild-type AcMNPV and used to infect fifth-instar Trichoplusia ni larvae. The fate of both genotypes was monitored over several rounds of insect infection. Levels of the occlusion-negative virus genome declined slowly over successive rounds of infection. We applied these data to a model of NPV population genetics to derive an estimate of 4.3 ± 0.3 viral genomes per occlusion body-producing cell.

* Corresponding author. Mailing address: Department of Biology, Imperial College of Science, Technology, and Medicine, Imperial College Road, London SW7 2AZ, United Kingdom. Phone: 44(0)20-75945376. Fax: 44(0)20-75842056. E-mail: dor{at}ic.ac.uk.

Applied and Environmental Microbiology, November 2001, p. 5204-5209, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5204-5209.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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