1887

Abstract

SUMMARY

Protoplasts were isolated from leaves of three tomato cultivars and inoculated with citrus exocortis viroid (CEV), potato spindle tuber viroid (PSTV) and cucumber pale fruit viroid (CPFV), respectively, using glycine-KOH-buffered mannitol at pH 9. CPFV multiplied detectably in protoplasts from the tomato cultivar Hilda 72 but replication of the other two viroids was rarely detectable. The minimum viroid concentration for infection was 10 µg RNA/ml using 10 protoplasts/ml. Newly synthesized viroid RNA was detected 36 h after inoculation by bioassay, and 48 h after inoculation by H-uridine incorporation into the viroid RNA band obtained by polyacrylamide gel electrophoresis. Incorporation of radioactive uridine into CPFV in inoculated protoplasts reached about 0.6% of that into tRNA during 72 h after inoculation. In protoplasts isolated from systemically infected leaves, however, incorporation of H-uridine into CPFV was only 0.1% of that into tRNA. In inoculated protoplasts, incorporation of H-uridine into CPFV was much more rapid at 35 °C than at 25 °C but incorporation into tRNA was similar at the two temperatures.

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/content/journal/jgv/10.1099/0022-1317-35-2-377
1977-05-01
2024-04-26
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References

  1. Aoki S., Takebe I. 1969; Infection of tobacco mesophyll protoplasts by tobacco mosaic virus ribonucleic acid. Virology 39:439–448
    [Google Scholar]
  2. Aoki S., Takebe I. 1975; Replication of tobacco mosaic virus RNA in tobacco mesophyll protoplasts inoculated in vitro. Virology 65:343–354
    [Google Scholar]
  3. Diener T. O. 1971; Potato spindle tuber ‘virus’. IV. A replicating, low molecular weight RNA. Virology 45:411–428
    [Google Scholar]
  4. Diener T. O., Lawson R. H. 1973; Chrysanthemum stunt: a viroid disease. Virology 51:94–101
    [Google Scholar]
  5. Diener T. O., Smith D. R. 1975; Potato spindle tuber viroid. XIII. Inhibition of replication by actinomycin D. Virology 63:421–427
    [Google Scholar]
  6. Henco K., Riesner D., Sänger H. L. 1977; Conformations of viroids. Nucleic Acids Research 4:177–194
    [Google Scholar]
  7. Hollings M., Stone O. M. 1973; Some properties of chrysanthemum stunt, a virus with the characteristics of an uncoated ribonucleic acid. Annals of Applied Biology 74:333–348
    [Google Scholar]
  8. Motoyoshi F., Bancroft J. B., Watts J. W., Burgess J. 1973; The infection of tobacco protoplasts with cowpea chlorotic mottle virus and its RNA. Journal of General Virology 20:177–193
    [Google Scholar]
  9. Motoyoshi F., Oshima N. 1975; Infection with tobacco mosaic virus of leaf mesophyll protoplasts from susceptible and resistant lines of tomato. Journal of General Virology 29:81–91
    [Google Scholar]
  10. Mühlbach H.-P., Camacho-Henriquez A., Sänger H. L. 1977; Isolation and properties of protoplasts from leaves of healthy and viroid infected tomato plants. Plant Science Letters 8: (in the press)
    [Google Scholar]
  11. Otsuki Y., Takebe I. 1969; Fluorescent antibody staining of tobacco mosaic virus antigen in tobacco mesophyll protoplasts. Virology 38:497–499
    [Google Scholar]
  12. Paterson R., Knight C. A. 1975; Protein synthesis in tobacco protoplasts infected with tobacco mosaic virus. Virology 64:10–22
    [Google Scholar]
  13. Randles J. W. 1975; Association of two ribonucleic acid species with cadang-cadang disease of coconut palm. Phytopathology 65:163–167
    [Google Scholar]
  14. Randles J. W., Rillo E. P., Diener T. O. 1976; The viroid-like structure and cellular location of anomalous RNA associated with the cadang-cadang disease. Virology 74:128–139
    [Google Scholar]
  15. Romaine C. P., Horst R. K. 1975; Suggested viroid etiology for chrysanthemum chlorotic mottle disease. Virology 64:86–95
    [Google Scholar]
  16. Sakai F., Takebe I. 1974; Protein synthesis in tobacco mesophyll protoplasts induced by tobacco mosaic virus infection. Virology 63:426–433
    [Google Scholar]
  17. Sänger H. L. 1972; An infectious and replicating RNA of low molecular weight: the agent of the exocortis disease of citrus. Advances in the Biosciences 8:103–116
    [Google Scholar]
  18. Sänger H. L., Klotz G., Riesner D., Gross H. J., Kleinschmidt A. K. 1976; Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures. Proceedings of the National Academy of Sciences of the United States of America 78:3852–3856
    [Google Scholar]
  19. Sänger H. L., Mühlbach H.-P. 1976; Replication of viroid RNA in tomato protoplasts. In Proceedings of the Symposium on ‘Nucleic acid and protein synthesis in plants’ Strasbourg C. N. R. S.: (in the press)
    [Google Scholar]
  20. Sänger H. L., Ramm K. 1975; Radioactive labelling of viroid-RNA. In Modification of the information content of plant cells pp 229–252 Edited by Markham R., Davies D. R., Hopwood D. A., Home R. W. Amsterdam: North-Holland Publishing Company;
    [Google Scholar]
  21. Sänger H. L., Riesner D. 1977; Purification and some different properties of five different viroids. Journal of General Virology (in the press)
    [Google Scholar]
  22. Sarkar S., Upadhya M. D., Melchers G. 1974; A highly efficient method of inoculation of tobacco mesophyll protoplasts with ribonucleic acid of tobacco mosaic virus. Molecular and General Genetics 135:1–9
    [Google Scholar]
  23. Semancik J. S., Weathers L. G. 1972; Exocortis disease: evidence for a new species of‘infectious’ low molecular weight RNA in plants. Nature New Biology 237:242–244
    [Google Scholar]
  24. Sutton D. W., Kemp J. D. 1976; Calculation of absolute rates of RNA synthesis, accumulation, and degradation in tobacco callus in vivo. Biochemistry 15:3153–3157
    [Google Scholar]
  25. Suzuki M., Takebe L. 1976; Uptake of single-stranded bacteriophage DNA by isolated tobacco protoplasts. Zeitschrift für Pflanzenphysiologie 78:421–433
    [Google Scholar]
  26. Takahashi T., Diener T. O. 1975; Potato spindle tuber viroid. XIV. Replication in nuclei isolated from infected leaves. Virology 64:106–114
    [Google Scholar]
  27. Van Dorst H. J. M., Peters D. 1974; Some biological observations on pale fruit, a viroid-incited disease of cucumber. Netherlands Journal of Plant Pathology 80:85–96
    [Google Scholar]
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