A Study of Rice Dwarf Virus in Vector Cell Monolayers by Fluorescent Antibody Focus Counting Free

Abstract

Summary

Infectivity assays of rice dwarf virus (RDV) were done by the fluorescent antibody focus counting technique on vector cell monolayers of the green rice leafhopper, . The focus count method was shown to be an accurate and quantitative method for determining RDV infectivities. The optimal pH value for inoculation was about 6.0 in a solution containing 0.1 -histidine HCl and 0.01 -MgCl. Below pH 5.5 and above 6.5, the infectivity of RDV dropped rapidly. The optimal adsorption period at 28 °C was dependent upon the RDV concentration. Optimal periods for adsorption with relative RDV concentrations of 10, 10 and 10 were about 60, 90 and 120 min, respectively. The period from virion adsorption to penetration into the cell was about 90 to 120 min. Infective progeny virions were first detected 12 h after the initial inoculation. From 12 to 20 h, the growth rate of the virus in the cells was exponential with a doubling time of about 96 min, and then from 20 to 28 h there was little or no further increase in infective virus. When the infectivities of the same inocula were compared by using the focus count and vector insect injection methods, the dilution endpoints were approx. 10 and 10, respectively. The focus count assay method was thus about 100 times more sensitive than vector injection.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-67-10-2119
1986-10-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/67/10/JV0670102119.html?itemId=/content/journal/jgv/10.1099/0022-1317-67-10-2119&mimeType=html&fmt=ahah

References

  1. Black L. M. 1941; Further evidence for multiplication of the aster-yellows virus in the aster leafhopper. Phytopathology 31:120–135
    [Google Scholar]
  2. Chiu R. J., Black L. M. 1967; Monolayer cultures of insect cell lines and their inoculation with a plant virus. Nature, London 215:1076–1078
    [Google Scholar]
  3. Chiu R. J., Black L. M. 1969; Assay of wound tumor virus by the fluorescent cell counting technique. Virology 37:667–677
    [Google Scholar]
  4. Chiu R. J., Liu H. Y., Macleod R., Black L. M. 1970; Potato yellow dwarf virus in leafhopper cell culture. Virology 40:387–396
    [Google Scholar]
  5. Fukushi T. 1940; Further studies on the dwarf disease of rice plant. Journal of the Faculty of Agriculture, Hokkaido Imperial University 45:83–154
    [Google Scholar]
  6. Hsu H. T., Black L. M. 1973; Inoculation of vector cell monolayers with potato yellow dwarf virus. Virology 52:187–198
    [Google Scholar]
  7. Kimura I. 1984; Establishment of new cell lines from leafhopper vector and inoculation of its cell monolayers with rice dwarf virus. Proceedings of the Japan Academy 60(B):198–201
    [Google Scholar]
  8. Kimura I. 1985; Quick detection of rice dwarf virus infection employing its vector cell monolayers. Japan Agricultural Research Quarterly 19:109–114
    [Google Scholar]
  9. Kimura I., Black L. M. 1971; Some factors affecting infectivity assay of wound-tumor virus on cell monolayers from an insect vector. Virology 46:266–276
    [Google Scholar]
  10. Kimura I., BLACK L. M. 1972; Growth of wound-tumor virus in vector cell monolayers. Virology 48:852–854
    [Google Scholar]
  11. Kimura I., Fukushi T. 1960; Studies on the rice dwarf virus. Annals of the Phytopathological Society ofJapan 25:131–135 (In Japanese with English summary.)
    [Google Scholar]
  12. Liu H. Y., Black L. M. 1976; Improvements in establishing and growing leafhopper cell cultures. Proceedings of the American Phytopathological Society 3:234–235 (Abstract.)
    [Google Scholar]
  13. Liu H. Y., Kimura I., Black L. M. 1973; Specific infectivity of different wound tumor virus isolates. Virology 51:320–326
    [Google Scholar]
  14. Maramorosch K. 1955; Mechanical transmission of curly top virus to its insect vector by needle inoculation. Virology 1:286–300
    [Google Scholar]
  15. Reddy D. V. R. 1977; Techniques of invertebrate tissue culture for the study of plant viruses. Methods in Virology 6:393–434
    [Google Scholar]
  16. Reddy D. V. R., Black L. M. 1972; Increase of wound tumor virus in leafhoppers as assayed on vector cell monolayers. Virology 50:412–421
    [Google Scholar]
  17. Spendlove R. S. 1967; Microscopic techniques. Methods in Virology 3:475–520
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-67-10-2119
Loading
/content/journal/jgv/10.1099/0022-1317-67-10-2119
Loading

Data & Media loading...

Most cited Most Cited RSS feed