1887

Abstract

Ultrastructural analysis of the polydnavirus of the braconid wasp revealed that virions consist of one cylindrical nucleocapsid enveloped by a single unit membrane. Nucleocapsids have a constant diameter of 33·7 ± 1·4 nm and a variable length of between 8 and 46 nm. Spreading of viral DNA showed that the genome consists of circular dsDNA molecules of variable sizes and measurement of the contour lengths indicated sizes of between 7 and 31 kbp. When virions were exposed to osmotic shock conditions to release the DNA, only one circular molecule was released per particle suggesting that the various DNA molecules are singly encapsidated in this bracovirus. The viral genome was seen to consist of at least 10 different segments and the aggregate genome size is in the order of 200 kbp. By partial digestion of viral DNA with dIII or RI in the presence of ethidium bromide and subsequent ligation with dIII-cut pSP65 or RI-cut pSP64 and transfection into , libraries of 103 and 23 RI clones were obtained. Southern blots revealed that complete and unrearranged segments were cloned with this approach, and restriction maps for five segments were obtained. Part of a 16·8 kbp segment was sequenced, found to be AT-rich (73%) and to contain six copies of a 17 bp repeated sequence. The development of the female reproductive tract in the course of pupal-adult development of the wasp was investigated and seen to be strictly correlated with the pigmentation pattern. By the use of a semiquantitative PCR, replication of viral DNA was observed to initiate at a specific stage of pupal-adult development.

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1994-12-01
2024-03-28
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References

  1. Blissard G. W., Fleming J. G. W., Vinson S. B., Summers M. D. 1986; Campoletis sonorensis virus: expression in Heliothis virescens and identification of expressed sequences. Journal of Insect Physiology 32:351–359
    [Google Scholar]
  2. Bühler A., Hanzlik T. N., Hammock B. D. 1985; Effects of parasitization of Trichoplusia ni by Chelonus sp . Physiological Entomology 10:383–394
    [Google Scholar]
  3. Coudron T. A. 1991; Host-regulating factors associated with parasitic hymenoptera. In American Chemical Society Symposium Series : Naturally Occurring Pest Bioregulators 449 pp 41–65 Hedin P. A. Edited by Washington D.C.: American Chemical Society;
    [Google Scholar]
  4. Davis R. W., Simon M., Davidson M. 1971; Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. Methods in Enzymology 21:413–428
    [Google Scholar]
  5. De Buron I., Beckage N. E. 1992; Characterization of a polydnavirus (PDV) and virus-like filamentous particle (VLFP) in the braconid wasp Contesia congregata (Hymenoptera: Braconidae). Journal of Invertebrate Pathology 59:315–327
    [Google Scholar]
  6. Feinberg A. P., Vogelstein B. 1985; A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132:6–13
    [Google Scholar]
  7. Fleming J. A. G. 1992; Polydnaviruses - mutualists and pathogens. Annual Review of Entomology 37:401–425
    [Google Scholar]
  8. Fleming J. A. G., Krell P. J. 1993; Polydnavirus genome organization. In Parasites and Pathogens of Insects 1 pp 189–225 Beckage N. E., Thompson S. N., Federici B. A. Edited by San Diego: Academic Press;
    [Google Scholar]
  9. Fleming J. A. G., Summers M. D. 1986; Campoletis sonorensis endoparasitic wasps contain forms of Campoletis sonorensis virus-DNA suggestive of integrated and extrachromosomal polydnavirus DNAs. Journal of Virology 57:552–562
    [Google Scholar]
  10. Fleming J. A. G., Summers M. D. 1991; Polydnavirus DNA is integrated in the DNA of its parasitoid wasp host. Proceedings of the National Academy of Sciences U.S.A: 889770–9774
    [Google Scholar]
  11. Francki R. I. B., Fauquet C. M., Knudson D. L., Brown F. 1991 Classification and Nomenclature of Viruses, Fifth Report of the International Committee on Taxonomy of Viruses pp 129–131 Wien & New York; Springer-Verlag:
    [Google Scholar]
  12. Grossniklaus-Bürgin C., Lanzrein B. 1990; Endocrine interrelationship between the parasitoid Chelonus sp. and its host Trichoplusia ni . Archives of Insect Biochemistry and Physiology 14:201–216
    [Google Scholar]
  13. Grossniklaus-Bürgin C., Wyler T., Pfister-Wilhelm R., Lanzrein B. 1994; Biology and morphology of the parasitoid Chelonus inanitus (Braconidae, Hymenoptera) and effects on the development of its host Spodoptera littoralis (Noctuidae, Lepidop-tera). Invertebrate Reproduction and Development 25:143–158
    [Google Scholar]
  14. Innis M. A., Gelfand D. H. 1990; Optimization of PCRs. In PCR Protocols pp 1–12 Innis M. A., Gelfand D. H., Sninsky J. J., White T. J. Edited by San Diego: Academic Press, Harcourt Brace Jovanovich Publishers;
    [Google Scholar]
  15. Inoue H., Nojima H., Okayama H. 1990; High efficiency transformation of Escherichia coli with plasmids. Gene 96:23–28
    [Google Scholar]
  16. Jones D., Jones G., Hammock B. D. 1981; Developmental and behavioral responses of larval Trichoplusia ni to parasitization by an imported braconid parasite Chelonus sp . Physiological Entomology 6:387–394
    [Google Scholar]
  17. Jones D., Sreekrishna S., Iwaya M., Yang J. 1986; Comparison of viral ultrastructure and DNA banding patterns from the reproductive tracts of eastern and western hemisphere Chelonus spp. (Braconidae: Hymenoptera). Journal of Invertebrate Pathology 47:105–115
    [Google Scholar]
  18. Kleinschmidt A. K., Lang D., Jacherts D., Zahn R. K. 1962; Darstellung und Längenmessungen des gesamten Desoxyribonuclein-saure-lnhaltes von T2 -Bakteriophagen. Biochimica et Biophysica Acta 61:857–864
    [Google Scholar]
  19. Krell P. J., Stoltz D. B. 1979; Unusual baculovirus of the parasitic wasp Apanteles melanoscelus: isolation and preliminary characterization. Journal of Virology 29:1118–1130
    [Google Scholar]
  20. Krell P. J., Summers M. D., Vinson S. B. 1982; Virus with a multipartite superhelical DNA genome from the ichneumonid parasitoid Campoletis sonorensis . Journal of Virology 43:859–870
    [Google Scholar]
  21. Lawrence P. O., Lanzrein B. 1993; Hormonal interactions between insect endoparasites and their host insects. In Parasites and Pathogens of Insects 1 pp 59–86 Beckage N. E., Thompson S. N., Federici B. A. Edited by San Diego: Academic Press;
    [Google Scholar]
  22. Mulligan R. C., Berg P. 1980; Expression of a bacterial gene in mammalian cells. Science 209:1422–1427
    [Google Scholar]
  23. Norton W. N., Vinson S. B. 1983; Correlating the initiation of virus replication with a specific pupal developmental phase of an ichneumonid parasitoid. Cell and Tissue Research 231:387–398
    [Google Scholar]
  24. Rechav Y., Orion T. 1975; The development of the immature stages of Chelonus inanitus . Annals of the Entomological Society of America 68:457–462
    [Google Scholar]
  25. Robberson D., Aloni Y., Attardi G. 1971; Expression of the mitochondrial genome in HeLa cells VI. Size determination of mitochondrial ribosomal RNA by electron microscopy. Journal of Molecular Biology 60:473–484
    [Google Scholar]
  26. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Ehrlich H. A. 1988; Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  27. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn.. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences U.S.A: 745463–5467
    [Google Scholar]
  29. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  30. Spurr A. R. 1969; A low-viscosity epoxy resin embedding medium for electron microscopy. Journal of Ultrastructure Research 26:31–33
    [Google Scholar]
  31. Stoltz D. B. 1990; Evidence for chromosomal transmission of polydnavirus DNA. Journal of General Virology 71:1051–1056
    [Google Scholar]
  32. Stoltz D. B., Vinson S. B. 1977; Baculovirus-like particles in the reproductive tracts of female parasitoid wasps II: the genus Apanteles. Canadian Journal of Microbiology 23:28–37
    [Google Scholar]
  33. Stoltz D. B., Vinson S. B. 1979; Viruses and parasitism in insects. Advances in Virus Research 24:125–171
    [Google Scholar]
  34. Stoltz D. B., Xu D. 1990; Polymorphism in polydnavirus genomes. Canadian Journal of Microbiology 36:538–543
    [Google Scholar]
  35. Stoltz D. B., Vinson S. B., Mackinnon E. 1976; Baculovirus-like particles in the reproductive tracts of female parasitoid wasps. Canadian Journal of Microbiology 22:1013–1023
    [Google Scholar]
  36. Stoltz D. B., Krell P. J., Vinson S. B. 1981; Polydisperse viral DNAs in ichneumonid ovaries: a survey. Canadian Journal of Microbiology 27:123–130
    [Google Scholar]
  37. Stoltz D. B., Guzo D., Cook D. 1986; Studies on polydnavirus transmission. Virology 155:120–131
    [Google Scholar]
  38. Strand M. R., Mckenzie D. I., Grassl V., Dover B. A., Aiken J. M. 1992; Persistence and expression of Microplitis demolitor polydnavirus in Pseudoplusia includens . Journal of General Virology 73:1627–1635
    [Google Scholar]
  39. Theilmann D. A., Summers M. D. 1987; Physical analysis of the Campoletis sonorensis virus multipartite genome and identification of a family of tandemly repeated elements. Journal of Virology 61:2589–2598
    [Google Scholar]
  40. Vinson S. B., Iwantsch G. F. 1980; Host regulation by insect parasitoids. Quarterly Review of Biology 55:143–165
    [Google Scholar]
  41. Webb B. A., Summers M. D. 1992; Stimulation of polydnavirus replication by 20-hydroxyecdysone. Experientia 48:1018–1022
    [Google Scholar]
  42. Wellauer P. K., Dawid I. B. 1974; Secondary structure maps of ribosomal RNA and DNA. I. Processing of Xenopus laevis ribosomal RNA and structure of single-stranded ribosomal DNA. Journal of Molecular Biology 89:379–395
    [Google Scholar]
  43. Xu D. M., Stoltz D. 1991; Evidence for a chromosomal location of polydnavirus DNA in the ichneumonid parasitoid Hyposoter fugitivus . Journal of Virology 65:6693–6704
    [Google Scholar]
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