Physical and Partial Genetic Map of Spodoptera frugiperda Nucleopolyhedrovirus (SfMNPV) Genome

A Nicaraguan isolate of Spodoptera frugiperda multicapsid nucleopolyhedrovirus (SfMNPV) is undergoing field trials for control of this pest in the Americas. This isolate is composed of multiple genotypes, some of which are deletion mutants. Identification of the genetic changes in deleted genotypes cannot be accomplished without the construction of a detailed physical map. In the present study, combinations of restriction endonuclease analysis and Southern blot analysis was performed. This map was refined by sequencing the termini of cloned restriction fragments. The SfMNPV genome was estimated to be 129.3 kb, 8 kb larger than the previously characterized Sf-2 variant from the United States, due to a deletion between 14.8 and 21.0 m.u. in the physical map described in this study. A total of 27.92 kb were sequenced, which represented 21.5% of the whole genome and included 38 ORFs. Comparison with other sequenced baculoviruses revealed that SfMNPV displayed the highest sequence identity (66%) and gene arrangement (78%) with Spodoptera exigua MNPV, sharing 36 putative ORFs. In addition, the genome organization was similar to that of SeMNPV, with minor differences. Phylogenetic analysis confirmed the close relatedness between SeMNPV and SfMNPV, suggesting they evolved from a common ancestor.

[1]  D. Rock,et al.  Genome Sequence of a Baculovirus Pathogenic forCulex nigripalpus , 2001, Journal of Virology.

[2]  M. Summers,et al.  Investigation of genetic heterogeneity in wild isolates of Spodoptera frugiperda nuclear polyhedrosis virus by restriction endonuclease analysis of plaque-purified variants. , 1981, Virology.

[3]  T. Luque,et al.  The complete sequence of the Cydia pomonella granulovirus genome. , 2001, Journal of General Virology.

[4]  K. Okano,et al.  Sequence analysis of the Xestia c-nigrum granulovirus genome. , 1999, Virology.

[5]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[6]  T. Williams,et al.  Virus entry or the primary infection cycle are not the principal determinants of host specificity of Spodoptera spp. nucleopolyhedroviruses. , 2004, The Journal of general virology.

[7]  E. Herniou,et al.  Use of Whole Genome Sequence Data To Infer Baculovirus Phylogeny , 2001, Journal of Virology.

[8]  J. Fuxa.  Insect control with baculoviruses. , 1991, Biotechnology advances.

[9]  S. Maeda,et al.  Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. , 1999, The Journal of general virology.

[10]  E. Herniou,et al.  Whole genome analysis of the Epiphyas postvittana nucleopolyhedrovirus. , 2002, The Journal of general virology.

[11]  F. Moscardi,et al.  Assessment of the application of baculoviruses for control of Lepidoptera. , 1999, Annual review of entomology.

[12]  I. Smith,et al.  In vivo isolation of baculovirus genotypes. , 1988, Virology.

[13]  Xinwen Chen,et al.  Comparative analysis of the complete genome sequences of Helicoverpa zea and Helicoverpa armigera single-nucleocapsid nucleopolyhedroviruses. , 2002, The Journal of general virology.

[14]  J. Vlak,et al.  A highly conserved genomic region in baculoviruses: sequence analysis of an 11.3 kbp DNA fragment (46.5-55.1 m.u.) of the Spodoptera exigua multicapsid nucleopolyhedrovirus. , 1998, Virus research.

[15]  R. Matthews.  The classification and nomenclature of viruses. Summary of results of meetings of the International Committee on Taxonomy of Viruses in The Hague, September 1978. , 1979, Intervirology.

[16]  D. Goulson,et al.  Evaluation of a Baculovirus Bioinsecticide for Small-Scale Maize Growers in Latin America , 1999 .

[17]  The Mosaic Structure of the Polyhedrin Gene of the Autographa californica Nucleopolyhedrovirus (AcMNPV) , 2004, Virus Genes.

[18]  Songnian Hu,et al.  Sequence analysis of the Spodoptera litura multicapsid nucleopolyhedrovirus genome. , 2001, Virology.

[19]  T. Matsumoto,et al.  Sequence analysis of the Plutella xylostella granulovirus genome. , 2000, Virology.

[20]  C. Ahrens,et al.  The sequence of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. , 1997, Virology.

[21]  V. Georgiev.  Virology , 1955, Nature.

[22]  J. Maruniak,et al.  Nucleotide sequence and transcriptional analysis of the gp41 gene of Spodoptera frugiperda nuclear polyhedrosis virus. , 1995, The Journal of general virology.

[23]  R. Possee,et al.  The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. , 1994, Virology.

[24]  W. Pearson.  Rapid and sensitive sequence comparison with FASTP and FASTA. , 1990, Methods in enzymology.

[25]  G. Rohrmann,et al.  Patterns of genome organization and content in lepidopteran baculoviruses. , 2000, Virology.

[26]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[27]  D. Goulson,et al.  Selection of a nucleopolyhedrovirus for control of Spodoptera frugiperda (Lepidoptera: Noctuidae): structural, genetic, and biological comparison of four isolates from the Americas. , 1999, Journal of economic entomology.

[28]  A. Sparks.  Fall Armyworm Symposium: A Review of the Biology of the Fall Armyworm , 1979 .

[29]  D. Kelly.  The DNA contained by nuclear polyhedrosis viruses isolated from four Spodoptera sp. (Lepidoptera, Noctuidae): genome size and homology assessed by DNA reassociation kinetics. , 1977, Virology.

[30]  M. Summers,et al.  Insertion of the SfMNPV polyhedrin gene into an AcMNPV polyhedrin deletion mutant during viral infection. , 1989, Virology.

[31]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[32]  J. Fuxa,et al.  DNA restriction polymorphism in wild isolates of Spodoptera frugiperda nuclear polyhedrosis virus. , 1991, Journal of invertebrate pathology.

[33]  H. A. Wood,et al.  A synchronous peroral technique for the bioassay of insect viruses , 1981 .

[34]  S. E. Brown,et al.  Physical maps of SfMNPV baculovirus DNA and its genomic variants. , 1984, Virology.

[35]  R. Page.  Extracting species trees from complex gene trees: reconciled trees and vertebrate phylogeny. , 2000, Molecular phylogenetics and evolution.

[36]  E. Herniou,et al.  The genome sequence and evolution of baculoviruses. , 2003, Annual review of entomology.

[37]  D. Winstanley,et al.  The complete sequence of the Adoxophyes orana granulovirus genome. , 2003, Virology.

[38]  Qianjun Li,et al.  Identification and genomic analysis of a second species of nucleopolyhedrovirus isolated from Mamestra configurata. , 2002, Virology.

[39]  D. Levin,et al.  Genotypic Variation Among Wild Isolates of Douglas-Fir Tussock Moth (Lepidoptera: Lyntantriidae) Nuclear Polyhedrosis Virus , 1996 .

[40]  T. Williams,et al.  Defective or effective? Mutualistic interactions between virus genotypes , 2003, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[41]  G. Croizier,et al.  Recombination as a possible major cause of genetic heterogeneity in Anticarsia gemmatalis nuclear polyhedrosis virus wild populations , 1992 .

[42]  Qianjun Li,et al.  Sequence and organization of the Mamestra configurata nucleopolyhedrovirus genome. , 2002, Virology.

[43]  J. Hoeffler,et al.  Gene Expression Systems: Using Nature For The Art Of Expression , 2006 .

[44]  T. Luque,et al.  Sequence Analysis of a 5.1 kbp Region of the Spodoptera frugiperda Multicapsid Nucleopolyhedrovirus Genome that Comprises a Functional Ecdysteroid UDP-Glucosyltransferase (egt) Gene , 2003, Virus Genes.

[45]  C. Funk,et al.  Sequence and analysis of the genome of a baculovirus pathogenic for Lymantria dispar. , 1999, Virology.

[46]  J. Vlak,et al.  Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. , 1999, The Journal of general virology.

[47]  J. Vlak,et al.  Biochemical and biological characterization of four isolates of Spodoptera exigua nuclear polyhedrosis virus , 1992 .

[48]  W. Dickerson,et al.  Velvetbean Caterpillar: A Rearing Procedure and Artificial Medium , 1976 .

[49]  Trevor Williams,et al.  Genetic Structure of a Spodoptera frugiperda Nucleopolyhedrovirus Population: High Prevalence of Deletion Genotypes , 2004, Applied and Environmental Microbiology.

[50]  M. Kool,et al.  Functional mapping of regions of the Autographa californica nuclear polyhedrosis viral genome required for DNA replication. , 1994, Virology.

[51]  J. Vlak,et al.  Orientation of the Genome of Autographa californica Nuclear Polyhedrosis Virus: a Proposal , 1982, Journal of virology.

[52]  E. Huang,et al.  Spodoptera frugiperda Nuclear Polyhedrosis Virus Genome: Physical Maps for Restriction Endonucleases BamHI and HindIII , 1981, Journal of virology.

[53]  J. Fuxa,et al.  Pathogens for the Suppression of the Fall Armyworm , 1980 .

[54]  Xinwen Chen,et al.  The sequence of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus genome. , 2001, The Journal of general virology.