1887

Abstract

Potato virus Y (PVY) full-length cDNA has been found to be refractory to cloning in cells. A full-length 9.7 kb PVY cDNA was obtained by reverse transcription polymerase chain reaction (RT-PCR) from the RNA of PVY (tuber necrotic strain, PVY). Double-stranded DNA fragments were used as primers (ds megaprimers), to include signals for transcription (a cauliflower mosaic virus 35S RNA promoter and a nopaline synthase terminator) in the final PCR product. Biolistic bombardment with a helium particle gun was used to inoculate the amplified product to detached tobacco leaves. Inoculation of tobacco plants with ground inoculated leaves followed by northern blot, ELISA and immuno-electron microscopy demonstrated that the DNA was highly infectious with up to 90% of bombarded leaves containing the virus. This methodology will allow the use of reverse genetics in the study of PVY-plant interactions and will also be useful for obtaining infectious cDNA from other viruses with large RNA genomes.

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1996-03-01
2021-10-27
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References

  1. Barnes W. M. 1994; PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proceedings of the National Academy of Sciences, USA 91:2216–2220
    [Google Scholar]
  2. Boyer J. C., Haenni A. L. 1994; Infectious transcripts and cDNA clones of plant viruses. Virology 198:415–426
    [Google Scholar]
  3. Cheng S., Fockler C., Barnes W. M., Hiouchi R. 1994; Effective amplification of long targets from cloned inserts and human genomic DNA. Proceedings of the National Academy of Sciences, USA 91:5695–5699
    [Google Scholar]
  4. De Bokx J. A., Huttinga H. 1981; Potato virus Y. CMI/AAB Descriptions of Plant Viruses no. 242
    [Google Scholar]
  5. Dolja V. V., Mcbride H. J., Carrington J. C. 1992; Tagging of plant potyvirus replication and movement by insertion of β-glucuronidase into the viral polyprotein. Proceedings of the National Academy of Sciences, USA 89:10208–10212
    [Google Scholar]
  6. Domier L. L., Franklin K. M., Hunt A. G., Rhoads R. E., Shaw J. G. 1989; Infectious in vitro transcripts from cloned cDNA of a potyvirus, tobacco vein mottling virus. Proceedings of the National Academy of Sciences, USA 86:3509–3513
    [Google Scholar]
  7. Fromm M. E., Taylor L. P., Walbot V. 1986; Stable transformation of maize after gene transfer by electroporation. Nature 319:791–793
    [Google Scholar]
  8. Gal-On A., Antignus Y., Rosner A., Raccah B. 1991; Infectious in vitro RNA transcripts derived from the cloned cDNA of the cucurbit potyvirus, zucchini yellow mosaic. Journal of General Virology 72:2639–2643
    [Google Scholar]
  9. Garzon-Tiznado J. A., Torres-Pacheco I., Ascencio-Ibanez J. T., Herrera-Estrella L., Rivera-Bustamante R. F. 1993; Inoculation of peppers with infectious clones of a new geminivirus by a biolistic procedure. Phytopathology 83:514–521
    [Google Scholar]
  10. Gébré Sélassié K., Marchoux G., Delecolle B., Pochard E. 1985; Variabilite naturelle des souches du virus Y de la pomme de terre dans les cultures de piment du sud-est de la France. Caractérisation et classification en pathotypes. Agronomic 5:621–630
    [Google Scholar]
  11. Godon C., Caboche M., Daniel-Vedele F. 1993; Transient plant gene expression: a simple and reproducible method based on flowing particle gun. Biochimie 75:591–595
    [Google Scholar]
  12. Gooding G. V. J. R., Tolin S. A. 1973; Strains of potato virus Y affecting flue-cured tobacco in the southeastern United States. Plant Disease Report 57:200–204
    [Google Scholar]
  13. Hagen L. S., Lot H., Godon C., Tepfer M., Jacquemond M. 1994; Infection of Theobroma cacao using cloned DNA of cacao swollen shoot virus and particle bombardment. Phytopathology 10:1239–1243
    [Google Scholar]
  14. Higuchi R. 1990; Recombinant PCR. In PCR Protocols: A Guide to Methods and Applications pp 177–183 Edited by Innis M. A., Gelfand D. H., Sninsky J. J., White T. J. San Diego: Academic Press;
    [Google Scholar]
  15. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. 1989; Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59
    [Google Scholar]
  16. Hollings M., Brunt A. A. 1981; Potyviruses. In Handbook of Plant Virus Infections and Comparative Diagnosis pp 731–807 Edited by Kurstak E. New York: Elsevier/North-Holland.;
    [Google Scholar]
  17. Kay R., Chan A., Daly M., McPherson J. 1987; Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302
    [Google Scholar]
  18. Le Romancer M., Kerlan C., Nedellec M. 1994; Biological characterization of various geographical isolates of potato virus Y inducing superficial necrosis on potato tubers. Plant Pathology 43:138–144
    [Google Scholar]
  19. Maiss E., Timpe U., Brisske-Rode A., Lesemann D.-E., Casper R. 1992; Infectious in vivo transcripts of plum pox potyvirus fulllength cDNA clone containing the cauliflower mosaic virus 35S RNA promoter. Journal of General Virology 73:709–713
    [Google Scholar]
  20. Murphy J. F., Rhoads R. E., Hunt A. G., Shaw J. G. 1990; The VPg of tobacco etch virus RNA is the 49-kDa proteinase or the N-terminal 24-kDa part of the proteinase. Virology 178:285–288
    [Google Scholar]
  21. Odell J. T., Nagy F., Chua N. H. 1985; Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313:810–812
    [Google Scholar]
  22. Riechmann J. L., Lain S., Garcia J. A. 1990; Infectious in vitro transcripts from a plum pox potyvirus cDNA clone. Virology 177:710–716
    [Google Scholar]
  23. Robaglia C., Durand-Tardif M., Tronchet M., Boudazin G., Astier-Manifacier S., Casse-Delbart F. 1989; Nucleotide sequence of potato virus Y (N strain) genomic RNA. Journal of General Virology 70:935–947
    [Google Scholar]
  24. Robaglia C., Bruening G., Haseloff J., Gerlach G. L. 1993; Evolution and replication of tobacco ringspot virus satellite RNA mutants. EMBO Journal 12:2969–2976
    [Google Scholar]
  25. Sarnow P. 1989; Role of 3′ end sequences in infectivity of poliovirus transcripts made in vitro . Journal of Virology 63:467–470
    [Google Scholar]
  26. Thole V., Dalmay T., Burgyan J., Balazs E. 1993; Cloning and sequencing of potato virus Y (Hungarian isolate) genomic RNA. Gene 123:149–156
    [Google Scholar]
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