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High levels of genetic variation within Helicoverpa armigera nucleopolyhedrovirus populations in individual host insects

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Abstract

It has been well documented that baculovirus populations exhibit high levels of genetic variation. Due to the lack of sensitivity of the techniques currently used to study baculovirus genetic variation, relatively little is known about baculovirus genetic diversity at the individual insect level. Since denaturing gradient gel electrophoresis (DGGE) has key advantages over other methods used to study genetic variation in baculoviruses, DGGE assays were used to obtain a better understanding of the genetic variation within baculovirus populations in individual host insects. Helicoverpa armigera nucleopolyhedrovirus (HearNPV) was used as a model baculovirus system, and neonate H. armigera larvae were infected with one of two geographically distinct HearNPV isolates. DGGE assays for two lepidopteran-specific baculovirus genes, me53 and dbp1, detected many HearNPV genetic variants within individual host larvae, with up to 20 genetic variants detected in a 434-bp fragment of the dbp1 gene in a single neonate larva. High levels of HearNPV genetic diversity were detected in individual host larvae irrespective of the HearNPV isolate used to infect the larvae. This study sets a benchmark for HearNPV genetic variation in individual H. armigera larvae. The levels of HearNPV genetic diversity detected are higher than reported previously for a baculovirus population at the individual insect level.

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References

  1. Baillie VL, Bouwer G (2011) Development of highly sensitive assays for detection of genetic variation in key Helicoverpa armigera nucleopolyhedrovirus genes. J Virol Methods 178:179–185

    Article  PubMed  CAS  Google Scholar 

  2. Baillie VL, Bouwer G (2011) High levels of genetic variation within core Helicoverpa armigera nucleopolyhedrovirus genes. Virus Genes 44:149–162

    Article  PubMed  Google Scholar 

  3. Bot J (1966) Rearing Helothis armigera Hübner and Prodemia litura F. on an artificial diet. S Afr J Agric Sci 9:535–538

    Google Scholar 

  4. Boutte C, Grubisic S, Balthasart P, Wilmotte A (2006) Testing of primers for the study of cyanobacterial molecular diversity by DGGE. J Microbiol Methods 65:542–550

    Article  PubMed  CAS  Google Scholar 

  5. Bouwer G, Avyidi D (2006) Application of the droplet feeding assay method to two economically important African lepidopteran pests. Afr Entomol 14:195–198

    Google Scholar 

  6. Chen X, IJkel WFT, Tarchini R, Sun X, Sandbrink H, Wang H, Peters S, Zuidema D, Lankhorst RK, Vlak JM, Hu Z (2001) The sequence of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus genome. J Gen Virol 82:241–257

    PubMed  CAS  Google Scholar 

  7. Christian PD, Gibb N, Kasprzak AB, Richards A (2001) A rapid method for the identification and differentiation of Helicoverpa nucleopolyhedroviruses (NPV Baculoviridae) isolated from the environment. J Virol Methods 96:51–65

    Article  PubMed  CAS  Google Scholar 

  8. Cory JS, Myers JH (2003) The ecology and evolution of insect baculoviruses. Annu Rev Ecol Evol Sys 34:239–272

    Article  Google Scholar 

  9. Cory JS, Green BM, Paul RK, Hunter-Fujita F (2005) Genotypic and phenotypic diversity of a baculovirus population within an individual insect host. J Invertebr Pathol 89:101–111

    Article  PubMed  Google Scholar 

  10. Crook NE, Payne CC (1980) Comparison of three methods of ELISA for baculoviruses. J Gen Virol 46:29–37

    Article  Google Scholar 

  11. Crook NE, Spencer RA, Payne CC, Leisy DJ (1985) Variation in Cydia pomonella granulosis virus isolates and physical maps of the DNA from three variants. J Gen Virol 66:2423–2430

    Article  CAS  Google Scholar 

  12. Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS (1991) ‘Touchdown’ PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19:4008

    Article  PubMed  CAS  Google Scholar 

  13. Duffy S, Shackelton LA, Holmes EC (2008) Rates of evolutionary change in viruses: patterns and determinants. Nature Rev Genet 9:267–276

    Article  PubMed  CAS  Google Scholar 

  14. Evans H, Shapiro M (1997) Viruses. In: Lacey LA (ed) Manual of techniques in insect pathology. Academic Press, San Diego, pp 17–53

    Chapter  Google Scholar 

  15. Federici BA (1997) Baculovirus pathogenesis. In: Miller LK (ed) The baculoviruses. Plenum Press, New York, pp 33–60

    Google Scholar 

  16. Figueiredo E, Muñoz D, Murillo R, Mexia A, Caballero P (2009) Diversity of Iberian nucleopolyhedrovirus wild-type isolates infecting Helicoverpa armigera (Lepidoptera: Noctuidae). Biol Control 50:43–49

    Article  Google Scholar 

  17. Finney DJ (1971) Probit analysis, 3rd edn. Cambridge University Press, London

    Google Scholar 

  18. Fischer SG, Lerman LS (1980) Separation of random fragments of DNA according to properties of their sequences. Proc Natl Acad Sci USA 77:4420–4424

    Article  PubMed  CAS  Google Scholar 

  19. Fischer SG, Lerman LS (1983) DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory. Proc Natl Acad Sci USA 80:1579–1583

    Article  PubMed  CAS  Google Scholar 

  20. Funk CJ, Braunagel SC, Rohrmann GF (1997) Baculovirus structure. In: Miller LK (ed) The Baculoviruses. Plenum Press, New York, pp 7–32

    Google Scholar 

  21. Fuxa JR (2004) Ecology of insect nucleopolyhedroviruses. Agric Ecosyst Environ 103:27–43

    Article  Google Scholar 

  22. Gettig RR, McCarthy WJ (1982) Genotypic variation among wild isolates of Heliothis spp nuclear polyhedrosis viruses from different geographical regions. Virology 117:245–252

    Article  PubMed  CAS  Google Scholar 

  23. Graham RI, Tyne WI, Possee RD, Sait SM, Hails RS (2004) Genetically variable nucleopolyhedroviruses isolated from spatially separate populations of the winter moth Operophtera brumata (Lepidoptera: Geometridae) in Orkney. J Invertebr Pathol 87:29–38

    Article  PubMed  Google Scholar 

  24. Green SJ, Leigh MB, Neufeld JD (2009) Denaturing gradient gel electrophoresis (DGGE) for microbial community analysis. In: Timmis KN (ed) Microbiology of hydrocarbons, oils, lipids, and derived compounds. Springer, Heidelberg, pp 4137–4158

    Google Scholar 

  25. Harris KA, Teo CG (2001) Diversity of hepatitis C virus quasispecies evaluated by denaturing gradient gel electrophoresis. Clin Diagn Lab Immunol 8:62–73

    PubMed  CAS  Google Scholar 

  26. Harrison RL (2009) Structural divergence among genomes of closely related baculoviruses and its implications for baculovirus evolution. J Invertebr Pathol 101:181–186

    Article  PubMed  CAS  Google Scholar 

  27. Herniou EA, Jehle JA (2007) Baculovirus phylogeny and evolution. Curr Drug Targets 8:1043–1050

    Article  PubMed  CAS  Google Scholar 

  28. Hitchman RB, Hodgson DJ, King LA, Hails RS, Cory JS, Possee RD (2007) Host mediated selection of pathogen genotypes as a mechanism for the maintenance of baculovirus diversity in the field. J Invertebr Pathol 94:153–162

    Article  PubMed  CAS  Google Scholar 

  29. Hodgson DJ, Vanbergen AJ, Hartley SE, Hails RS, Cory JS (2002) Differential selection of baculovirus genotypes mediated by different species of host food plant. Ecol Lett 5:512–518

    Article  Google Scholar 

  30. Hughes PR, Wood HA (1981) A synchronous peroral technique for the bioassay of insect viruses. J Invertebr Pathol 37:154–159

    Article  Google Scholar 

  31. Hughes PR, Gettig RR, McCarthy WJ (1983) Comparison of the time-mortality response of Heliothis zea to 14 isolates of Heliothis nuclear polyhedrosis virus. J Invertebr Pathol 41:256–261

    Article  Google Scholar 

  32. Jehle JA, Blissard GW, Bonning BC, Cory JS, Herniou EA, Rohrmann GF, Theilmann DA, Thiem SM, Vlak JM (2006) On the classification and nomenclature of baculoviruses: a proposal for revision. Arch Virol 151:1257–1266

    Article  PubMed  CAS  Google Scholar 

  33. Kamiya K, Okimoto N, Ikeda M, Kunimi Y, Kobayashi M, Kawamura S (2003) Genotypic variation of a wild isolate of Hyphantria cunea nucleopolyhedrovirus. J Insect Biotechnol Sericol 72:57–64

    CAS  Google Scholar 

  34. Khan S, Sneddon K, Fielding B, Ward V, Davison S (2003) Functional characterization of the ecdysteroid UDP-glucosyl transferase gene of Helicoverpa armigera single-enveloped nucleopolyhedrovirus isolated in South Africa. Virus Genes 27:17–27

    Article  PubMed  CAS  Google Scholar 

  35. Knebel-Mörsdorf D, Kremer A, Jahnel F (1993) Baculovirus gene ME53, which contains a putative zinc finger motif, is one of the major early-transcribed genes. J Virol 67:753–758

    PubMed  Google Scholar 

  36. Lee HH, Miller LK (1978) Isolation of genotypic variants of Autographa californica nuclear polyhedrosis virus. J Virol 27:754–767

    PubMed  CAS  Google Scholar 

  37. Motta FC, Rosado AS, Couceiro JN (2002) Standardization of denaturing gradient gel electrophoresis for mutant screening of influenza A (H3N2) virus samples. J Virol Methods 101:105–115

    Article  PubMed  CAS  Google Scholar 

  38. Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59:695–700

    PubMed  CAS  Google Scholar 

  39. Muyzer G (1999) DGGE/TGGE a method for identifying genes from natural ecosystems. Curr Opin Microbiol 2:317–322

    Article  PubMed  CAS  Google Scholar 

  40. Myers RM, Fischer SG, Lerman LS, Maniatis T (1985) Nearly all single base substitutions in DNA fragments joined to a GC-clamp can be detected by denaturing gradient gel electrophoresis. Nucleic Acids Res 13:3131–3145

    Article  PubMed  CAS  Google Scholar 

  41. Parnell M, Grzywacz D, Jones KA, Brown M (2002) The strain variation and virulence of granulovirus of diamondback moth (Plutella xylostella Linnaeus, Lep., Yponomeutidae) isolated in Kenya. J Invertebr Pathol 79:192–196

    Article  PubMed  Google Scholar 

  42. Pijlman GP, van den Born E, Martens DE, Vlak JM (2001) Autographa californica baculoviruses with large genomic deletions are rapidly generated in infected insect cells. Virology 283:132–138

    Article  PubMed  CAS  Google Scholar 

  43. Potter KN, Faulkner P, MacKinnon EA (1976) Strain selection during serial passage of Trichoplusia ni nuclear polyhedrosis virus. J Virol 18:1040–1050

    PubMed  CAS  Google Scholar 

  44. Richards AR, Christian PD (1999) A rapid bioassay screen for quantifying nucleopolyhedroviruses (Baculoviridae) in the environment. J Virol Methods 82:63–75

    Article  PubMed  CAS  Google Scholar 

  45. Rowley DL, Popham HJR, Harrison RL (2011) Genetic variation and virulence of nucleopolyhedroviruses isolated worldwide from the heliothine pests Helicoverpa armigera, Helicoverpa zea, and Heliothis virescens. J Invertebr Pathol 107:112–126

    Article  PubMed  Google Scholar 

  46. Sheffield VC, Cox DR, Lerman LS, Myers RM (1989) Attachment of a 40-base-pair G+C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. Proc Natl Acad Sci USA 86:232–236

    Article  PubMed  CAS  Google Scholar 

  47. Short SM, Suttle CA (1999) Use of the polymerase chain reaction and denaturing gradient gel electrophoresis to study diversity in natural virus communities. Hydrobiologia 401:19–32

    Article  CAS  Google Scholar 

  48. Simón O, Williams T, López-Ferber M, Caballero P (2004) Genetic structure of a Spodoptera frugiperda nucleopolyhedrovirus population: high prevalence of deletion genotypes. Appl Environ Microbiol 70:5579–5588

    Article  PubMed  Google Scholar 

  49. Simón O, Williams T, López-Ferber M, Taulemesse J, Caballero P (2008) Population genetic structure determines speed of kill and occlusion body production in Spodoptera frugiperda multiple nucleopolyhedrovirus. Biol Control 44:321–330

    Article  Google Scholar 

  50. Smith IR, Crook NE (1988) In vivo isolation of baculovirus genotypes. Virology 166:240–244

    Article  PubMed  CAS  Google Scholar 

  51. Sun X, Chen X, Zhang Z, Wang H, Bianchi FJJA, Peng H, Vlak JM, Hu Z (2002) Bollworm responses to release of genetically modified Helicoverpa armigera nucleopolyhedroviruses in cotton. J Invertebr Pathol 81:63–69

    Article  PubMed  Google Scholar 

  52. Sun X, Wang H, Sun X, Chen X, Peng C, Pan D, Jehle JA, van der Werf W, Vlak JM, Hu Z (2004) Biological activity and field efficacy of a genetically modified Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus expressing an insect-selective toxin from a chimeric promoter. Biol Control 29:124–137

    Article  Google Scholar 

  53. Theilmann DA, Blissard GW (2008) Baculoviruses: molecular biology of nucleopolyhedroviruses. In: Mahy BWJ, van Regenmortel MHV (eds) Encylopedia of virology. Elsevier Academic Press, Oxford, pp 254–265

    Chapter  Google Scholar 

  54. van Oers MM, Vlak JM (2007) Baculovirus genomics. Curr Drug Targets 8:1051–1068

    Article  PubMed  Google Scholar 

  55. Vasuki V, Hoti SL, Sadanandane C, Jambulingam P (2003) A simple and rapid DNA extraction method for the detection of Wuchereria bancrofti infection in the vector mosquito, Culex quinquefasciatus by Ssp I PCR assay. Acta Trop 86:109–114

    Article  PubMed  CAS  Google Scholar 

  56. Zhang CX, Ma XC, Guo ZJ (2005) Comparison of the complete genome sequence between C1 and G4 isolates of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus. Virology 333:190–199

    Article  PubMed  CAS  Google Scholar 

  57. Zhang Y, Ma F, Wang Y, Yang B, Chen S (2008) Expression of v-cath gene from HearNPV in tobacco confers an antifeedant effect against Helicoverpa armigera. J Biotechnol 138:52–55

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was partially funded by an Innovation Fund (Department of Science and Technology, South Africa) research grant awarded to Gustav Bouwer. We thank anonymous reviewers for comments that improved this manuscript.

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  1. School of Molecular and Cell Biology, University of the Witwatersrand, Private Bag 3, Wits, Johannesburg, 2050, South Africa

    Vicky Lynne Baillie & Gustav Bouwer

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  1. Vicky Lynne Baillie
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  2. Gustav Bouwer
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Correspondence to Gustav Bouwer.

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Baillie, V.L., Bouwer, G. High levels of genetic variation within Helicoverpa armigera nucleopolyhedrovirus populations in individual host insects. Arch Virol 157, 2281–2289 (2012). https://doi.org/10.1007/s00705-012-1416-6

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  • DOI: https://doi.org/10.1007/s00705-012-1416-6

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