1887

Abstract

SUMMARY

Because it contains the gene, potato cv. ‘Maris Piper’ reacts hypersensitively to inoculation with group three strains of potato virus X (PVX). However, protoplasts prepared from aseptically grown shoot cultures of potato cv. ‘Maris Piper’ were infected reproducibly with either PVX or PVX RNA using inocula containing polyethylene glycol. The extent of infection of protoplasts was assessed by fluorescent antibody staining, and by assaying extracts for antigen by ELISA, for infectivity and for PVX RNA content by cDNA hybridization. Infection occurred in 43 to 65% of the protoplasts infected with PVX and in 57 to 70% of the protoplasts infected with PVX RNA. In comparison, when high quality protoplasts were obtained from leaves of pot-grown plants 55 to 65% became infected when inoculated with PVX RNA. However, protoplasts obtained from leaves of whole plants varied greatly in both quantity and quality, whereas shoot cultures reproducibly gave high quality protoplast preparations in large numbers. In protoplasts from either source, yields of progeny virus were between 10 and 30 pg per infected protoplast. The multiplication of a group 3 strain of PVX in protoplasts containing the gene did not induce necrosis and confirms that hypersensitivity is not always expressed in isolated protoplasts.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-66-6-1341
1985-06-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/66/6/JV0660061341.html?itemId=/content/journal/jgv/10.1099/0022-1317-66-6-1341&mimeType=html&fmt=ahah

References

  1. Bailey J. M., Davidson D. 1976; Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Analytical Biochemistry 70:75–85
    [Google Scholar]
  2. Barker H., Harrison B. D. 1982; Infection of potato mesophyll protoplasts with five plant viruses. Plant Cell Reports 1:247–249
    [Google Scholar]
  3. Bercks R. 1970; Potato virus X. Commonwealth Mycological Institute/Association of Applied Biologists Descriptions of Plant Viruses no. 4
    [Google Scholar]
  4. Clark M. F., Adams A. N. 1977; Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 24:475–483
    [Google Scholar]
  5. Cockerham G. 1955; Strains of potato virus. X. Proceedings of the Second Conference on Potato Virus Diseases Lisse-Wageningen 1954 pp 89–92
    [Google Scholar]
  6. Cockerham G. 1970; Genetical studies on resistance to potato viruses X and Y. Heredity 25:309–348
    [Google Scholar]
  7. Gonda T. J., Symons R. H. 1979; Cucumber mosaic virus replication in cowpea protoplasts: time course of virus, coat protein and RNA synthesis. Journal of General Virology 45:723–726
    [Google Scholar]
  8. Gould A. R., Symons R. H. 1978; Determination of the sequence homology between four RNA species of cucumber mosaic virus by hybridization analysis with complementary DNA. Nucleic Acids Research 4:3787–3802
    [Google Scholar]
  9. Harrison B. D., Mayo M. A. 1983; The use of protoplasts in plant virus research. In Use of Tissue Culture and Protoplasts in Plant Pathology pp 69–137 Edited by Helgeson J. P., Deverall B. J. London: Academic Press;
    [Google Scholar]
  10. Jones R. A. C. 1982; Breakdown of potato virus X resistance gene Nx: selection of a group four strain from strain group three. Plant Pathology 31:325–331
    [Google Scholar]
  11. Koike M., Hibi T., Yaor K. 1977; Infection of cowpea mesophyll protoplasts with cucumber mosaic virus. Virology 83:413–416
    [Google Scholar]
  12. Kubo S., Harrison B. D., Robinson D. J., Mayo M. A. 1975; Tobacco rattle virus in tobacco mesophyll protoplasts: infection and virus multiplication. Journal of General Virology 27:293–304
    [Google Scholar]
  13. Maule A. J. 1983; Infection of protoplasts from severalBrassica species with cauliflower mosaic virus following inoculation using polyethylene glycol. Journal of General Virology 64:2655–2660
    [Google Scholar]
  14. Maule A. J., Boulton M. I., Edmunds C., Wood K. R. 1980; Polyethylene glycol-mediated infection of cucumber protoplasts by cucumber mosaic virus and virus RNA. Journal of General Virology 47:199–203
    [Google Scholar]
  15. Moreira A., Jones R. A. C., Fribourg C. E. 1980; Properties of a resistance-breaking strain of potato virus X. Annals of Applied Biology 95:93–103
    [Google Scholar]
  16. Motoyoshi F., Oshima N. 1977; Expression of genetically controlled resistance to tobacco mosaic virus infection in isolated tomato leaf protoplasts. Journal of General Virology 34:499–506
    [Google Scholar]
  17. Murashige T., Skoog F. 1962; A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia plantarum 15:473–497
    [Google Scholar]
  18. Nelson R. S., Creissen G. P., Bright S. W. J. 1983; Plant regeneration from protoplasts of Solanum brevidens. Plant Science Letters 30:355–362
    [Google Scholar]
  19. O’hara J. F., Henshaw G. G. 1982; The preparation of protoplasts from potato and related Solanumspecies Plant Tissue Culture 1982. Proceedings of the 5th International Congress of Plant Tissue and Cell Culture pp 591–592
    [Google Scholar]
  20. Oldfield S. J., Coutts R. H. A. 1980; Isolation and infection of cowpea primary leaf protoplasts with tobacco necrosis virus. In Tissue Culture Methods for Plant Pathologists pp 87–93 Edited by Ingram D. S., Helgeson J. P. Oxford: Blackwells;
    [Google Scholar]
  21. Otsuki Y., Takebe I. 1969; Fluorescent antibody staining of tobacco mosaic virus antigen in tobacco mesophyll protoplasts. Virology 38:497–499
    [Google Scholar]
  22. Otsuki Y., Shimomura T., Takebe I. 1972; Tobacco mosaic virus multiplication and expression of the N gene in necrotic responding tobacco varieties. Virology 50:45–50
    [Google Scholar]
  23. Otsuki Y., Takebe I., Honda Y., Kajita S., Matsui C. 1974; Infection of tobacco mesophyll protoplasts by potato virus X. Journal of General Virology 22:375–385
    [Google Scholar]
  24. Reichmann M. E. 1959; Potato virus X. Part III. Light scattering studies. Canadian Journal of Chemistry 37:384–388
    [Google Scholar]
  25. Shalla T. E., Peterson L. J. 1973; Infection of isolated plant protoplasts with potato virus X. Phytopathology 63:1125–1130
    [Google Scholar]
  26. Shepard J. F., Totten R. E. 1977; Mesophyll cell protoplasts of potato. Plant physiology 60:313–316
    [Google Scholar]
  27. Takebe I. 1977; Protoplasts in the study of plant virus replication. In Comprehensive Virology vol 11: pp 237–282 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  28. Takebe I. 1983; Protoplasts in plant virus research. International Review of Cytology suppl 16:89–111
    [Google Scholar]
  29. Taylor I. M., Illmensee R., Summers J. 1976; Efficient transcription of RNA into DNA by avian sarcoma virus polymerase. Biochimica et biophysica acta 442:324–330
    [Google Scholar]
  30. Thomas E. 1981; Plant regeneration from shoot culture derived protoplasts of tetraploid potato(Solanum tuberosum cv. ‘Maris Bard’). Plant Science Letters 23:81–88
    [Google Scholar]
  31. Thomas P. S. 1980; Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proceedings of the National Academy of Sciences, U,. S,. A 77:5201–5205
    [Google Scholar]
  32. Widholm J.M. 1972; Use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technology 47:189–194
    [Google Scholar]
  33. Zassenhaus H. P., Butow R. A., Hannon Y. P. 1982; Rapid electroelution of nucleic acids from agarose and acrylamide gels. Analytical Biochemistry 125:125–130
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-66-6-1341
Loading
/content/journal/jgv/10.1099/0022-1317-66-6-1341
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error