1887

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Volume 72, Issue 12

Change in Phenotype and Encapsidated RNA Segments of an Isolate of Alfalfa Mosaic Virus: An Influence of Host Passage Free

Abstract

A local lesion isolate of alfalfa mosaic virus (AMV-N20) from lucerne was found to encapsidate two extra RNAs in addition to the four major RNAs (RNA1, -2, -3 and -4). These were resolved by gel electrophoresis both under native conditions and after glyoxal denaturation. The RNA with an electrophoretic mobility between that of RNAs 2 and 3 was designated RNA3l, that between RNAs 3 and 4 was designated RNA3s. Sucrose density gradient centrifugation analysis of AMV-N20 showed six instead of the normal four nucleoprotein components, the additional two presumably representing encapsidated RNAs 3l and 3s. RNAs 3l and 3s were both shown by Northern blot hybridization to be unrelated to host plant RNA and to contain the AMV coat protein gene sequence, which resides in RNA3. Primer extension of the RNAs 3l and 3s using a primer complementary to the 3′ common terminus of all genomic AMV RNAs provided further evidence that they contained AMV sequences. RNA31 represents an addition of about 255 nucleotides, compared with RNA3, and RNA3s represents a loss of about 308 nucleotides. The coat proteins of variants encapsidating either RNA3l, -3 or -3s had the same indicating that the addition or deletion of the nucleotides was outside the coat protein gene. Serial mechanical passage of AMV-N20 over 5 years in four host species led both to changes in the composition of the RNA3 mixture, and to changes in symptom severity. For example, following passage in , RNA3 was lost whereas passage in either or resulted in the loss of RNA3l. No association was found between the changes in RNA3 and phenotypic changes that resulted from continuous passage for 5 years. Phenotypic changes with passage are thus presumably determined by mutations elsewhere in the virus genome.

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© Journal of General Virology 1991
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1991-12-01
2024-04-25
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References

  1. Almond J. W., Westrop G. D., Wareham K. A., Skinner M. A., Evans D. M. A., Magrath D., Schild G. C., Minor P. D. 1987; Studies on the genetic basis of attenuation of the Sabin type 3 poliovirus vaccine. Biochemical Society Symposium 53:85–90
     [Google Scholar]
  2. Bol J. F., Lak-Kaashoek M. 1974; Composition of alfalfa mosaic virus nucleoproteins. Virology 60:476–484
     [Google Scholar]
  3. Brederode F. Th., KoperZ-warthoff E. C., Bol J. F. 1980; Complete nucleotide sequence of alfalfa mosaic virus RNA4. Nucleic Acids Research 8:2213–2223
     [Google Scholar]
  4. Burgyan J., Rubino L., Russo M. 1991; De novo generation of cymbidium ringspot virus defective interfering RNA. Journal of General Virology 72:505–509
     [Google Scholar]
  5. Carrington J. C., Morris T. J. 1984; Complementary DNA cloning and analysis of carnation mottle virus RNA. Virology 139:22–31
     [Google Scholar]
  6. Crill E., Hagedorn N. J., Hanson E. W. 1971; An artificial system for differentiating strains of alfalfa mosaic virus. Plant Disease Reporter 55:127–130
     [Google Scholar]
  7. Dietzgen R. G., Francki R. I. B. 1988; Analysis of lettuce necrotic yellows virus structural proteins with monoclonal antibodies and concanavalin A. Virology 166:486–494
     [Google Scholar]
  8. Dore J.-M., Pinck L. 1988; Plasmid DNA containing a copy of RNA3 can substitute for RNA3 in alfalfa mosaic virus RNA inocula. Journal of General Virology 69:1331–1338
     [Google Scholar]
  9. Dore J.-M., Pinck M., Pinck L. 1989; Competitive multiplication of RNA 3 species of different strains of alfalfa mosaic virus. Journal of General Virology 70:777–782
     [Google Scholar]
  10. Ey P. L., Ashman L. K. 1986; The use of alkaline phosphatase conjugated anti-immunoglobulin with immunoblots for determining the specificity of monoclonal antibodies to protein mixtures. Methods in Enzymology 121:497–509
     [Google Scholar]
  11. Francki R. I. B., Hu J., Palukaitis P. 1986; Taxonomy of cucurbit-infecting tobamoviruses as determined by serological and molecular hybridization analyses. Intervirology 26:156–163
     [Google Scholar]
  12. García-Arenal F., Palukaitis P., Zaitlin M. 1984; Strains and mutants of tobacco mosaic virus are both found in virus derived from single-lesion passaged inoculum. Virology 132:131–137
     [Google Scholar]
  13. Gonsalves D., Garnsey S. M. 1989; Cross-protection techniques for control of plant virus diseases in the tropics. Plant Disease 73:592–597
     [Google Scholar]
  14. Gottlieb M., Chavko M. 1987; Silver staining of native and denatured eucaryotic DNA in agarose gels. Analytical Biochemistry 165:33–37
     [Google Scholar]
  15. Gunn M. R., Symons R. H. 1980; Sequence homology at the 3′-termini of the four RNAs of alfalfa mosaic virus. FEBS Letters 109:145–150
     [Google Scholar]
  16. Hajimorad M. R., Francki R. I. B. 1988; Alfalfa mosaic virus isolates from lucerne in South Australia: biological variability and antigenic similarity. Annals of Applied Biology 113:45–54
     [Google Scholar]
  17. Hajimorad M. R., Francki R. I. B. 1989; Preparation of soluble, biologically active alfalfa mosaic virus coat protein and its CaCl2-induced degradation. Journal of Virological Methods 25:49–61
     [Google Scholar]
  18. Hajimorad M. R., Francki R. I. B. 1991a; Effect of glutaraldehyde-fixation on the immunogenicity, particle stability and antigenic reactivity of alfalfa mosaic virus, and the specificity of elicited antibodies. Journal of Virological Methods 33:13–25
     [Google Scholar]
  19. Hajimorad M. R., Francki R. I. B. 1991b; Serological differentiation of some strains of alfalfa mosaic virus with polyclonal antibodies. Phytopathology 81:603–610
     [Google Scholar]
  20. Hajimorad M. R., Dietzgen R. G., Francki R. I. B. 1990; Differentiation and antigenic characterization of closely related alfalfa mosaic virus strains with monoclonal antibodies. Journal of General Virology 71:2809–2816
     [Google Scholar]
  21. Hatta T., Francki R. I. B. 1984; Differences in the morphology of isometric particles of some plant viruses stained with uranyl acetate as an aid to their identification. Journal of Virological Methods 9:237–247
     [Google Scholar]
  22. Heijtink R. A., Jaspars E. M. J. 1974; RNA contents of abnormally long particles of certain strains of alfalfa mosaic virus. Virology 59:371–382
     [Google Scholar]
  23. Hiruki C., Miczynski K. A. 1987; Severe isolate of alfalfa mosaic virus and its impact on alfalfa cultivars grown in Alberta. Plant Disease 71:1014–1018
     [Google Scholar]
  24. Huisman M. J., Cornelissen B. J. C., Groenendijk C. F. M., Bol J. F., Van Vloten-Doting L. 1989; Alfalfa mosaic virus temperature-sensitive mutants. V. The nucleotide sequence of Tbts 7 RNA 3 shows limited nucleotide changes and evidence for heterologous recombination. Virology 171:409–416
     [Google Scholar]
  25. Hull R. 1968; Virus diseases of garden lupin in Great Britain. Annals of Applied Biology 61:373–380
     [Google Scholar]
  26. Hull R. 1970; Studies on alfalfa mosaic virus. IV. An unusual strain. Virology 42:283–292
     [Google Scholar]
  27. Hull R., Hill G. J., Markham R. 1969; Studies on alfalfa mosaic virus. II. The structure of the virus components. Virology 37:416–428
     [Google Scholar]
  28. Knorr D. A., Mullin R. H., Hearne P. Q., Morris T. J. 1991; De novo generation of defective interfering RNAs of tomato bushy stunt virus by high multiplicity passage. Virology 181:193–202
     [Google Scholar]
  29. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
     [Google Scholar]
  30. Langereis K., Mugnier M. -A., Cornelissen B. J. C., Pinck L., Bol J. F. 1986; Variable repeats and poly(A)-stretches in the leader sequence of alfalfa mosaic virus RNA 3. Virology 154:409–414
     [Google Scholar]
  31. McMaster G. K., Carmichael G. 1977; Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proceedings of the National Academy of Sciences, U.S.A. 74:4835–4838
     [Google Scholar]
  32. Neeleman L., Van Der Kuyl A. C., Bol J. F. 1991; Role of alfalfa mosaic virus coat protein gene in symptom formation. Virology 181:687–693
     [Google Scholar]
  33. Paliwal Y. C. 1982; Virus diseases of alfalfa and biology of alfalfa mosaic virus in Ontario and western Quebec. Canadian Journal of Plant Pathology 4:175–179
     [Google Scholar]
  34. Palukaitis P. 1984; Detection and characterization of subgenomic RNA in plant viruses. Methods in Virology 7:259–317
     [Google Scholar]
  35. Palukaitis P., Symons R. H. 1980; Nucleotide sequence homology of thirteen tobamovirus RNAs as determined by hybridization analysis with complementary DNA. Virology 107:354–361
     [Google Scholar]
  36. Palukaitis P., Cotts S., Zaitlin M. 1985; Detection and identification of viroids and viral nucleic acids by ′dot blot′ hybridization. Acta horticulturae 164:109–118
     [Google Scholar]
  37. Peden K. W. C., Symons R. H. 1973; Cucumber mosaic virus contains a functionally divided genome. Virology 53:487–492
     [Google Scholar]
  38. Quadt R., Jaspars E. M. J. 1989; RNA polymerases of plus-strand RNA viruses of plants. Molecular Plant-Microbe Interactions 2:219–223
     [Google Scholar]
  39. Ravelonandro M., Godefroy-Colburn T., Pinck L. 1983; Structure of the 5′-terminal untranslated region of the genomic RNAs from two strains of alfalfa mosaic virus. Nucleic Acids Research 11:2815–2826
     [Google Scholar]
  40. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  41. Van Beynum G. M. A., De Graaf J. M., Castel A., Kraal B., Bosch L. 1977; Structural studies on the coat protein of alfalfa mosaic virus. The complete primary structure. European Journal of Biochemistry 72:63–78
     [Google Scholar]
  42. Van Dun C. M. P., Bol J. F., Van Vloten-Doting L. 1987; Expression of alfalfa mosaic virus and tobacco rattle virus coat protein genes in transgenic tobacco plants. Virology 159:299–305
     [Google Scholar]
  43. Walter B., Kuszala J., Ravelonandro M., Pinck L. 1985; Alfalfa mosaic virus isolated from Buddleia davidii compared with other strains. Plant Disease 69:266–267
     [Google Scholar]
  44. Watanabe Y., Morita N., Nishiguchi M., Okada Y. 1987; Attenuated strains of tobacco mosaic virus. Reduced synthesis of a viral protein with a cell-to-cell movement function. Journal of Molecular Biology 194:699–704
     [Google Scholar]
  45. Wray W., Boulikas T., Wray V. P., Hancock R. 1981; Silver staining of proteins in polyacrylamide gels. Analytical Biochemistry 118:197–203
     [Google Scholar]
  46. Yarwood C. E. 1979; Host passage effects with plant viruses. Advances in Virus Research 25:169–190
     [Google Scholar]
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Change in Phenotype and Encapsidated RNA Segments of an Isolate of Alfalfa Mosaic Virus: An Influence of Host Passage
J. Gen. Virol. 72, 2885 (1991); https://doi.org/10.1099/0022-1317-72-12-2885
/content/journal/jgv/10.1099/0022-1317-72-12-2885
/content/journal/jgv/10.1099/0022-1317-72-12-2885
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