1887

Abstract

A combination of the polymerase chain reaction (PCR), asymmetric PCR (A-PCR) and DNA sequencing was used to determine the nucleotide sequence of a hypervariable region of the bipartite genome of bean golden mosaic geminivirus (BGMV). This region, which was part of the intergenic region of the DNA-B component, was amplified using primers designed from the nucleotide sequence of a DNA-B component clone (pDRB1) of an isolate of BGMV from the Dominican Republic (BGMV-DR). pDRB1 is infectious on beans when coinoculated with the DNA-A component of BGMV-DR (pDRA1), and typical bean golden mosaic symptoms are observed on infected plants. Bean leaf tissue infected with BGMV was collected at five separate field locations in the Dominican Republic and the hypervariable region was amplified by PCR, ssDNA was produced using A-PCR, and partial nucleotide sequences were determined. The sequences of the hypervariable region from the field-collected samples ranged from 95% (one sample) to 98% (four samples) identical to the sequence of pDRB1. This contrasts with sequence identities of 86, 75 and 46% between the pDRB1 hypervariable region and the hypervariable regions of BGMV isolates from Guatemala, Puerto Rico and Brazil respectively, and 42% with bean dwarf mosaic geminivirus. These results indicate that Dominican Republic isolates of BGMV are very similar and should be considered isolates of the same virus (BGMV-DR), and that the infectious clones of BGMV-DR are representative of BGMV isolates in the Dominican Republic. The procedures described for DNA extraction from leaf tissue and for production of high quality ssDNA using PCR and A-PCR are rapid and efficient and could be applied to studies of variability and epidemiology of other viruses.

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1991-11-01
2022-01-22
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References

  1. Brown J. K., Chapman M. A., Nelson M. R. 1990; Bean calico mosaic, a new disease of common bean caused by a whitefly-transmitted geminivirus. Plant Disease 74:81
    [Google Scholar]
  2. Cohen S., Duffus J. E., Larsen R. C., Liu H. Y., Flock R. A. 1983; Purification, serology, and vector relationships of squash leaf curl virus, a whitefly-transmitted geminivirus. Phytopathology 73:1669–1673
    [Google Scholar]
  3. Dellaporta S. L., Wood J., Hicks J. B. 1983; A plant DNA minipreparation: version II. Plant Molecular Biology Reporter 1:19–21
    [Google Scholar]
  4. Demmler G. J., Buffone G. J., Schimbor C. M., May R. A. 1988; Detection of cytomegalovirus in urine from newborns using polymerase chain reaction DNA amplification. Journal of Infectious Diseases 158:1177–1184
    [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12:387–395
    [Google Scholar]
  6. Ehrlich G. D., Greenberg S., Abbott M. A. 1990; Detection of human T-cell lymphoma/leukemia viruses. In PCR Protocols: A Guide to Methods and Applications pp 325–336 Edited by Innis M. A., Gelfand D. H., Sninsky J. J., White T. J. New York: Academic Press;
    [Google Scholar]
  7. Faria J. C., Gilbertson R. L., Morales F. J., Russell D. R., Ahlquist P. G., Hanson S. F., Maxwell D. P. 1990; Sequence of infectious clones of two mechanically transmissible isolates of bean golden mosaic geminivirus. Phytopathology 80:983
    [Google Scholar]
  8. Galvez G. E., Morales F. J. 1989; Whitefly-transmitted viruses. In Bean Production Problems in the Tropics, 2nd. edn, pp 379–406 Edited by Schwartz H. F., Pastor-Corrales M. A. Cali, Colombia: Centro Intemacional de Agricultura Tropical;
    [Google Scholar]
  9. Gilbertson R. L., Faria J. C., Morales F. J., Leong S. A., Maxwell D. P., Ahlquist P. G. 1988; Molecular characterization of geminiviruses causing bean golden mosaic. Phytopathology 78:1568
    [Google Scholar]
  10. Gilbertson R. L., Faria J. C., Hanson S. F., Hendrastuti E., Ahlquist P. G., Morales F. J., Maxwell D. P. 1990; Relationships among bean-infecting geminiviruses. Annual Report of the Bean Improvement Cooperative 33:72–73
    [Google Scholar]
  11. Gilbertson R. L., Hidayat S. H., Martinez R. T., Leong S. A., Faria J. C., Morales F. J., Maxwell D. P. 1991a; Differentiation of bean-infecting geminiviruses by nucleic acid hybridization probes and aspects of bean golden mosaic in Brazil. Plant Disease 75:336–342
    [Google Scholar]
  12. Gilbertson R. L., Faria J. C., Hanson S. F., Morales F. J., Ahlquist P. G., Maxwell D. P., Russell D. R. 1991b; Cloning of the complete DNA genomes of four bean-infecting geminiviruses and determining their infectivity by electric discharge particle acceleration. Phytopathology (in press)
    [Google Scholar]
  13. Howarth A. J., Caton J., Bossert M., Goodman R. M. 1985; Nucleotide sequence of bean golden mosaic virus and a model for gene regulation in geminiviruses. Proceedings of the National Academy of Sciences, U.S.A. 82:3572–3576
    [Google Scholar]
  14. Kaneko S., Miller R. H., Feinstone S. N., Unoura M., Kobayashi K., Hattori N., Purcell R. H. 1989; Detection of serum hepatitis B virus DNA in patients with chronic hepatitis using the polymerase chain reaction. Proceedings of the National Academy of Sciences, U.S.A. 86:312–316
    [Google Scholar]
  15. Kellogg D. E., Kwok S. 1990; Detection of human immunodeficiency virus. In PCR Protocols: A Guide to Methods and Applications pp 337–347 Edited by Innis M. A., Gelfand D. H., Sninsky J. J., White T. J. New York: Academic Press;
    [Google Scholar]
  16. Larzul D., Guigue F., Sninsky J. J., Mack D. H., Brechot C., Guesdon J. L. 1988; Detection of hepatitis B virus sequences in serum by using in vitro enzymatic amplification. Journal ofVirological Methods 20:227–237
    [Google Scholar]
  17. McCabe P. C. 1990; Production of single-stranded DNA by asymmetric PCR. In PCR Protocols: A Guide to Methods and Applications pp 76–84 Edited by Innis M. A., Gelfand D. H., Sninsky J. J., White T. J. New York: Academic Press;
    [Google Scholar]
  18. Manos M. M., Ting Y., Wright D. K., Lewis A. J., Broker T. R., Wolinsky S. M. 1989; The use of the polymerase chain reaction for the detection of genital human papillomaviruses. Cancer Cells 7:209–214
    [Google Scholar]
  19. Morales F. J., Niessen A. I. 1988; Comparative responses of selected Phaseolus vulgaris germ plasm inoculated artificially and naturally with bean golden mosaic virus. Plant Disease 72:1020–1023
    [Google Scholar]
  20. Morales F. J., Niessen A., Ramirez B., Castano M. 1990; Isolation and partial characterization of a geminivirus causing bean dwarf mosaic. Phytopathology 90:96–101
    [Google Scholar]
  21. Puchta H., Sanger H. L. 1989; Sequence analysis of minute amounts of viroid RNA using the polymerase chain reaction (PCR). Archives of Virology 106:335–340
    [Google Scholar]
  22. Robertson N. L., French R., Gray S. M. 1991; Use of group-specific primers and the polymerase chain reaction for the detection and identification of luteoviruses. Journal of General Virology 72:1473–1477
    [Google Scholar]
  23. Rybicki E. P., Hughes F. L. 1990; Detection and typing of maize streak virus and other distantly related geminiviruses of grasses by polymerase chain reaction amplification of a conserved viral sequence. Journal of General Virology 71:2519–2526
    [Google Scholar]
  24. 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]
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