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Molecular comparisons of alphabaculovirus-based products: Gypchek with Disparvirus (Lymantria dispar) and TM BioControl-1 with Virtuss (Orgyia pseudotsugata)

Published online by Cambridge University Press:  02 April 2012

Jianhua Zhang ,
Renée Lapointe ,
David Thumbi ,
Benoit Morin  and
Christopher J. Lucarotti
Jianhua Zhang
Affiliation:
Sylvar Technologies Inc., P.O. Box 636, Station A Fredericton, New Brunswick, Canada E3B 5A6
Renée Lapointe
Affiliation:
Sylvar Technologies Inc., P.O. Box 636, Station A Fredericton, New Brunswick, Canada E3B 5A6
David Thumbi
Affiliation:
Sylvar Technologies Inc., P.O. Box 636, Station A Fredericton, New Brunswick, Canada E3B 5A6
Benoit Morin
Affiliation:
Natural Resources Canada, Canadian Forest Service-Atlantic Forestry Centre, P.O. Box 4000, Fredericton, New Brunswick, Canada E3B 5P7
Christopher J. Lucarotti*
Affiliation:
Natural Resources Canada, Canadian Forest Service-Atlantic Forestry Centre, P.O. Box 4000, Fredericton, New Brunswick, Canada E3B 5P7
*
1 Corresponding author (e-mail: [email protected]).
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Abstract

Gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae), multicapsid nucleopolyhedrovirus (LdMNPV) has been registered as a microbial pest-control product in the United States (Gypchek®) and Canada (Disparvirus®). Similarly, Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough) (Lepidoptera: Lymantriidae), multicapsid nucleopolyhedrovirus (OpMNPV) is registered in the United States and Canada as TM BioControl-1® and a product derived from TM BioControl-1 (Virtuss®) is also registered in Canada. To determine changes that may have occurred in these products over time, we compared DNA from Gypchek with Disparvirus and DNA from TM BioControl-1 with Virtuss using restriction fragment length polymorphism (RFLP) analysis. Gypchek and Disparvirus showed the same RFLP banding patterns when viral genomic DNA was digested with BamH I, EcoR V, and Hind III and only a single band difference at approximately 1.6 kilobase (kb) when digested with Bgl II. TM BioControl-1 and Virtuss showed no differences in genomic DNA when digested with Bgl II, Sam I or Hind III. Twelve viral open reading frames (ORFs) were amplified from Gypchek and Disparvirus and nine from TM BioControl-1 and Virtuss by polymerase chain reactions (PCR). The amplified ORFs ranged from highly conserved (polyhedrin) to least conserved (vp91 capsid associated protein). The products were sequenced and the deduced protein products compared. Amino acid sequences deduced from the sequenced PCR products indicated that 8 of the 12 proteins were identical in the two LdMNPV products. The four proteins showing minor sequence variations were DNA polymerase, LEF-8, P74 envelope protein, and VP 91 capsid associated protein. No differences were detected in the protein products deduced from the nine sequenced ORFs from TM BioControl-1 and Virtuss. Comparative RFLP and protein phylogenetic analyses of Gypchek with Disparvirus and TM BioControl-1 with Virtuss revealed little difference between the respective LdMNPV and OpMNPV populations that make up these product pairs.

Résumé

Le virus à capsides multiples de la nucléopolyhédrose de la spongieuse, Lymantria dispar (L.) (Lepidoptera : Lymantriidae), (LdMNPV) est enregistré comme produit microbien antiparasitaire aux États-Unis (GypcheckMD) et au Canada (DisparvirusMD). De même, le virus à capsides multiples de la nucléopolyhédrose de la chenille à houppes du sapin Douglas, Orgyia pseudotsugata (McDunnough) (Lepidoptera : Lymantriidae), (OpMNPV) est enregistré aux États-Unis et au Canada sous le nom de TM BioControl-1MD; il existe aussi un produit dérivé de TM BioControl-1 (VirtussMD) au Canada. Afin de déterminer les changements qui ont pu survenir dans ces produits dans le temps, nous comparons l'ADN de Gypcheck et de Disparvirus ainsi que l'ADN de TM BioControl-1 et de Virtuss par une analyse du polymorphisme de la longueur des fragments de restriction (RFLP). Gypcheck et Disparvirus possèdent les mêmes patrons de bandes lorsque l'ADN viral génomique est digéré avec BamH I, EcoR V et Hind III et une seule différence de bande à approximativement 1,6 kb avec une digestion avec Bgl II. TM BioControl-1 et Virtuss ne montrent aucune différence d'ADN génomique lors de digestions avec Bgl II, Sam I ou Hind III. Douze cadres ouverts de lecture (ORF) viraux ont été amplifiés chez Gypcheck et Disparvirus et neuf cadres chez TM BioControl-1 et Virtuss par amplification en chaîne par polymérase (PCR). Les ORF amplifiés varient de fortement conservés (polyhédrine) à faiblement conservés (protéine vp91 associée à la capside). Les produits ont été séquencés et les produits protéiniques obtenus comparés. Les séquences d'acides aminés obtenues des produits séquencés de la PCR montrent que huit des 12 protéines sont identiques dans les deux produits de LdMNPV. Les quatre protéines qui montrent des variations mineures de séquence sont l'ADN polymérase, LEF-8, la protéine P74 de l'enveloppe et la protéine VP 91 associée à la capside. Aucune différence n’est détectée dans les produits protéiniques des neuf ORF séquencés de TM BioControl-1 et Virtuss. Des analyses comparatives de RFLP et des analyses phylogénétiques des protéines de Gypcheck et de Disparvirus, de même que de TM BioControl-1 et Virtuss, montrent peu de différences entre les populations de LdMNPV et de OpMNPV qui constituent ces paires de produits.

[Traduit par la Rédaction]

Type
Articles
Information
The Canadian Entomologist , Volume 142 , Issue 6 , December 2010 , pp. 546 - 556
Copyright
Copyright © Entomological Society of Canada 2010

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