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Protection Against Detrimental Effects of Potyvirus Infection in Transgenic Tobacco Plants Expressing the Papaya Ringspot Virus Coat Protein Gene

Abstract

We obtained transgenic tobacco plants expressing the papaya ringspot virus (PRV) coat protein (CP) gene by transformation via Agrobacterium tumefaciens. Expression was effectively monitored by enzyme-linked immunosorbent assays (ELISA) of crude tissue extracts. Subcloned plants derived from eight original Ro transformants were inoculated with potyviruses: tobacco etch (TEV), potato virus Y (PVY), and pepper mottle (PeMV). Plants that accumulated detectable levels of the PRV CP showed significant delay in symptom development and the symptoms were attenuated. Similar results were obtained with inoculated R1 plants. We conclude that the expression of the PRV CP-gene imparts protection against infection by a broad spectrum of potyviruses.

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References

  1. Powell-Abel, P., Nelson, R.S., De, B., Hoffmann, N., Rogers, S.G., Fraley, R.T. and Beachy, R.N. 1986. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232: 738–743.

    Article  Google Scholar 

  2. Tumer, N.E., O'Connell, K.M., Nelson, R.S., Sanders, P.R., Beachy, R.N., Fraley, R.T. and Shah, D.M. 1987. Expression of alfalfa mosaic virus coat protein gene confers cross-protection in transgenic tobacco and tomato plants. EMBO J. 6: 1181–1188.

    Article  CAS  Google Scholar 

  3. Loesch-Fries, L.S., Merlo, D., Zinnen, T., Burhop, L., Hill, K., Krahn, K., Jarvis, N., Nelson, S. and Halk, E. 1987. Expression of alfalfa mosaic virus RNA 4 in transgenic plants confers virus resistance. EMBO J. 6: 1845–1851.

    Article  CAS  Google Scholar 

  4. Van Dun, C.M.P., Bol, J.F. and 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.

    Article  CAS  Google Scholar 

  5. Cuozzo, M., O'Connell, K.M., Kaniewski, W., Fang, R.-X., Chua, N.-H. and Tumer, N.E. 1988. Viral protection in transgenic tobacco plants expressing the cucumber mosaic virus coat protein or its antisense RNA. Bio-Technology 6: 549–557.

    CAS  Google Scholar 

  6. Quemada, H.D., Gonsalves, D. and Slightom, J.L. 1991. CMV-C coat protein gene expression in tobacco: Protection against infection by CMV strains transmitted mechanically or by aphids. Phytopathology 81: in press.

    Google Scholar 

  7. Hemenway, C., Fang, R.-X., Kaniewski, W.K., Chua, N.-H. and Tumer, N.E. 1988. Analysis of the mechanism of protection in transgenic plants expressing the potato virus X coat protein or its antisense RNA. EMBO J. 7: 1273–1280.

    Article  CAS  Google Scholar 

  8. Hoekema, A., Huisman, M.J., Molendijk, L. Van den Elzen, P.J.M. and Cornelissen, B.J.C. 1989. The genetic engineering of two commercial potato cultivars for resistance to potato virus X. Bio/Technology 7: 273–278.

    Google Scholar 

  9. Lawson, C., Kaniewski, W., Haley, L., Rozman, R., Newell, C., Sanders, P. and Tumer, N.E. 1990. Engineering resistance to mixed virus infection in a commercial potato cultivar: Resistance to potato virus X and potato virus Y in transgenic russet burbank. Bio/Technology 8: 127–134.

    CAS  Google Scholar 

  10. Van Dun, C.M.P., Overduin, B., Van Vloten-Doting, L. and Bol, J.F. 1988. Transgenic tobacco expressing tobacco streak virus or mutated alfalfa mosaic virus coat protein does not cross-protect against alfalfa mosaic virus infection. Virology 164: 383–389.

    Article  CAS  Google Scholar 

  11. Van Dun, C.M.P. and Bol, J.F. 1988. Transgenic tobacco plants accumulating tobacco rattle virus coat protein resist infection with tobacco rattle virus and pea early browning virus. Virology 167: 649–652.

    Article  CAS  Google Scholar 

  12. Stark, D.M. and Beachy, R.N. 1989. Protection against potyvirus infection in transgenic plants: Evidence for broad spectrum resistance. Bio/Technology 7: 1257–1262.

    Google Scholar 

  13. Quemada, H., L'Hostis, B., Gonsalves, D., Reardon, I., Heinrickson, R., Hiebert, E.L., Sieu, L.C. and Slightom, J.L. 1990. The nucleotide sequences of the 3′-terminal regions of papaya ringspot virus strains W and P. J. Gen. Virol. 71: 203–210.

    Article  CAS  Google Scholar 

  14. Fitch, M.M.M., Manshardt, R.M., Gonsalves, D., Slightom, J.L. and Sanford, J.C. 1990. Stable transformation of papaya via microprojectile bombardment. Plant Cell Reports 9: 189–194.

    CAS  PubMed  Google Scholar 

  15. Sligthom, J.L. 1991. Custom polymerase-chain-reaction engineering of a plant expression vector. Gene 100: In press.

    Google Scholar 

  16. Yeh, S.-D. and Gonsalves, D. 1985. Translation of papaya ringspot virus RNA in vitro: detection of a possible polyprotein that is processed for capsid protein, cylindrical-inclusion protein, and amorphous-inclusion protein. Virology 143: 260–271.

    Article  CAS  Google Scholar 

  17. Quemada, H., Kearney, C., Gonsalves, D. and Slightom, J.L. 1989. Nucleotide sequences of the coat protein genes and flanking regions of cucumber mosaic virus strains C and WL RNA 3. J. Gen. Virol. 70: 1065–1073.

    Article  CAS  Google Scholar 

  18. Carrington, J.C. and Dougherty, W.G. 1987. Processing of the tobacco etch virus 49 K protease requires autoproteolysis. Virology. 160: 355–362.

    Article  CAS  Google Scholar 

  19. An, G. 1987. Binary Ti vectors for plant transformation and promoter analysis. Methods Enzymol. 153: 292–305.

    Article  CAS  Google Scholar 

  20. Horsch, R.B., Fry, J.E., Hoffman, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.T. 1985. A simple and general method for transferring genes into plants. Science 227: 1229–1231.

    Article  CAS  Google Scholar 

  21. Anderson, E.J., Stark, D.M., Nelson, R.S., Powell, P.A., Tumer, N.E. and Beachy, R.N. 1989. Transgenic plants that express the coat protein genes of tobacco mosaic virus or alfalfa mosaic virus interfere with disease development of some nonrelated viruses. Phytopathology 79: 1284–1290.

    Article  CAS  Google Scholar 

  22. Quemada, H., Sieu, L.C., Siemieniak, D.R., Gonsalves, D. and Slightom, J.L. 1990. Watermelon mosaic virus II and zucchini yellow mosaic virus: cloning of 3′-terminal regions, nucleotide sequences, and phylogenetic comparisons. J. Gen. Virol. 71: 1451–1460.

    Article  CAS  Google Scholar 

  23. Shukla, D.D. and Ward, C.W. 1989. Identification and classification of potyviruses on the basis of coat protein sequence data and serology. Archives of Virology 106: 171–200.

    Article  CAS  Google Scholar 

  24. Beachy, R.N., Loesch-Fries, S. and Tumer, N.E. 1990. Coat protein-mediated resistance against virus infection. Ann. Rev. Phytopathol. 28: 451–474.

    Article  CAS  Google Scholar 

  25. Gonsalves, D. and Ishii, M. 1980. Purification and serology of papaya ringspot virus. Phytopathology 70: 1028–1032.

    Article  CAS  Google Scholar 

  26. Moghal, S.M. and Francki, R.I.B. 1976. Towards a system for the identification and classification of potyviruses I. Serology and amino acid composition of six distinct viruses. Virology 73: 350–362.

    Article  CAS  Google Scholar 

  27. Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S., Higuchi, R., Horn, G.T., Mullis, K.B. and Erlich, H.A. 1988. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487–491.

    Article  CAS  Google Scholar 

  28. Maniatis, T., Fritsch, E. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  29. Morelle, G. 1989. A plasmid extraction procedure on a miniprep scale. Focus 11: 7–8.

    Google Scholar 

  30. Sanger, F., Nicklen, S. and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Nat. Acad. Sci. USA 74: 5463–5467.

    Article  CAS  Google Scholar 

  31. Zagurski, R.J., Baumeister, K., Lomax, N. and Berman, M.L. 1985. Rapid and easy sequencing of large double-stranded DNA and supercoiled plasmid DNA. Gene Analy. Tech. 2: 89–94.

    Article  Google Scholar 

  32. Jefferson, R.A. 1987. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Reporter 5: 387–405.

    Article  CAS  Google Scholar 

  33. Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.

    Article  CAS  Google Scholar 

  34. Berger, P.H., Hunt, A.G., Domier, L.L., Hellmann, G.M., Stram, Y., Thornbury, D.W. and Pirone, T.P. 1989. Expression in transgenic plants of a viral gene product that mediates insect transmission of potyviruses. Proc. Nat. Acad. Sci. USA 86: 8402–8406.

    Article  CAS  Google Scholar 

  35. Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Nat. Acad. Sci. USA 76: 4350–4354.

    Article  CAS  Google Scholar 

  36. Hu, J.S., Gonsalves, D., Boscia, D. and Namba, S. 1990. Use of monoclonal antibodies to characterize grapevine leafroll associated closteroviruses. Phytopathology 80: 920–925.

    Article  CAS  Google Scholar 

  37. Van Dun, C.M.P., Van Vloten-Doting, L. and Bol, J.F. 1988. Expression of alfalfa mosaic virus cDNA 1 and 2 in transgenic tobacco plants. Virology 163: 572–578.

    Article  CAS  Google Scholar 

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Ling, K., Namba, S., Gonsalves, C. et al. Protection Against Detrimental Effects of Potyvirus Infection in Transgenic Tobacco Plants Expressing the Papaya Ringspot Virus Coat Protein Gene. Nat Biotechnol 9, 752–758 (1991). https://doi.org/10.1038/nbt0891-752

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