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Engineered resistance to tomato spotted wilt virus in transgenic peanut expressing the viral nucleocapsid gene

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Abstract

The nucleocapsid gene of tomato spotted wilt virus Hawaiian L isolate in a sense orientation, and the GUS and NPTII marker genes, were introduced into peanut (Arachis hypogaea cv. New Mexico Valencia A) using Agrobacterium-mediated transformation. Modifications to a previously defined transformation protocol reduced the time required for production of transformed peanut plants. Transgenes were stably integrated into the peanut genome and transmitted to progeny. RNA expression and production of nucleocapsid protein in transgenic peanut were observed. Progeny of transgenic peanut plants expressing the nucleocapsid gene showed a 10- to 15-day delay in symptom development after mechanical inoculations with the donor isolate of tomato spotted wilt virus. All transgenic plants were protected from systemic tomato spotted wilt virus infection. Inoculated non-transformed control plants and plants transformed with a gene cassette not containing the nucleocapsid gene became systemically infected and displayed typical tomato spotted wilt virus symptoms. These results demonstrate that protection against tomato spotted wilt virus can be achieved in transgenic peanut plants by expression of the sense RNA of the tomato spotted wilt virus nucleocapsid gene

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References

  • Beachy, R.N., Loesch-Fries, S. and Tumer, N.E. (1990) Coat protein-mediated resistance against virus infection. Annu. Rev. Phytopath. 28, 451-74.

    Google Scholar 

  • Burow, M.D., Chlan, C.A., Sen, P. and Murai, N. (1990) High frequency generation of transgenic tobacco plants after modified leaf disk cocultivation with Agrobacterium tumefaciens. Plant Mol. Biol. Rep. 8, 124-39.

    Google Scholar 

  • Camann, M.A., Culbreath, A.K., Pickering, J., Todd, J.W. and Demski, J.W. (1995) Spatial and temporal patterns of spotted wilt epidemics in peanut. Phytopath. 85, 879-85.

    Google Scholar 

  • Cheng, M., Jarret, R.L., Li, Z., Xing, A. and Demski, J.W. (1996) Production of fertile transgenic peanut (Arachis hypogaeaL.) plants using Agrobacterium tumefaciens. Plant Cell Rep. 15, 653-7.

    Google Scholar 

  • Culbreath, A.K., Todd, J.W., Demski, J.W. and Chamberlin, J.R. (1992) Disease progress of spotted wilt in peanut cultivars Florunner and Southern Runner. Phytopath. 82, 766-71.

    Google Scholar 

  • Culbreath, A.K., Todd, J.W., Branch, W.D., Brown, S.L., Demski, J.W. and Beasley, J.P. (1994) Effects of new peanut cultivar Georgia Brown on epidemics of spotted wilt. Plant Dis. 78, 1185-9.

    Google Scholar 

  • De Haan, P., Wagemakers, L., Peters, D. and Goldbach, R. (1990) The S RNA segment of tomato spotted wilt virus has an ambisense character. J. Gen. Virol. 71, 1001-7.

    Google Scholar 

  • De Haan, P., Gielen, J.J.L., Prins, M., Wijkamp, I.G., van Schepen, A., Peters, D., van Grinsven, M.Q.J.M. and Goldbach, R. (1992) Characterization of RNA-mediated resistance to tomato spotted wilt virus in transgenic tobacco plants. Bio/ Technology 10, 1133-7.

    Google Scholar 

  • Flavell, R.B. (1994) Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc. Natl. Acad. Sci. USA 91, 3490-96.

    Google Scholar 

  • Gamborg, O.L. (1966) Aromatic metabolism in plants. II. Enzymes of the shikimate pathway in suspension cultures of plant cells. Can. J. Biochem. 44, 791-9.

    Google Scholar 

  • Gielen, J.J.L., de Haan, P., Kool, A.J., Peters, D., Grinsven, M.Q.J.M. van and Goldbach, R.W. (1991) Engineered resistance to tomato spotted wilt virus, a negative-strand RNA virus. Bio/Technology 9, 363-7.

    Google Scholar 

  • Hood, E.E., Chilton, W.S., Chilton, M.D. and Fraley, R.T. (1986) T-DNA and opine synthetic loci in tumors incited by Agrobacterium tumefaciensA281 on soybean and alfalfa plants. J. Bacter. 168, 1283-90.

    Google Scholar 

  • Jefferson, R.A. (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387-405.

    Google Scholar 

  • Kim, J.W., Sun, S.S.M. and German, T.L. (1994) Disease resistance in tobacco and tomato plants transformed with the tomato spotted wilt virus nucleocapsid gene. Plant Disease 78, 615-21.

    Google Scholar 

  • MacKenzie, D.J. and Ellis, P.J. (1992) Resistance to tomato spotted wilt virus infection in transgenic tobacco expressing the viral nucelocapsid gene. Mol. Plant Microbe Interact. 5, 34-40.

    Google Scholar 

  • Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473-97.

    Google Scholar 

  • Pang, S.Z., Nagpala, P., Wang, M., Slightom, J.L. and Gonsalves D. (1992) Resistance to heterologous isolates of tomato spotted wilt virus in transgenic tobacco expressing its nucleocapsid protein gene. Phytopath. 82, 1223-9.

    Google Scholar 

  • Pang, S.Z., Slightom, J.L. and Gonsalves, D. (1993) Different mechanisms protect transgenic tobacco against tomato spotted wilt and impatiens necrotic spot Tospoviruses. Bio/Technology 11, 819-24.

    Google Scholar 

  • Pang, S.Z., Bock, J.H., Gonsalves, C., Slightom, J.L. and Gonsalves, D. (1994) Resistance of transgenic Nicotinana benthamianaplants to tomato spotted wilt and impatiens necrotic spot tospoviruses: evidence of involvement of the N protein and N gene RNA in resistance. Phytopath. 84, 243-9.

    Google Scholar 

  • Peters, D. and Goldbach, R. (1995) The biology of Tospoviruses. In: Singh, R.P., Singh, U.S. and Kohmoto, K. (eds.) Pathogenesis and Host Specificity in Plant Diseases. Vol III. Viruses and Viroids, Oxford, UK: Pergamon Press, pp. 199-210.

    Google Scholar 

  • Rogers, S.O. and Bendich, A.J. (1985) Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol. Biol. 5, 69-76.

    Google Scholar 

  • Wadsworth, G.J., Redinbaugh, M.C. and Scandalios, J.G. (1980) A procedure for the small-scale isolation of plant RNA suitable for RNA blot analysis. Anal. Biochem. 172, 279-83.

    Google Scholar 

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Li, Z., Jarret, R.L. & Demski, J.W. Engineered resistance to tomato spotted wilt virus in transgenic peanut expressing the viral nucleocapsid gene. Transgenic Res 6, 297–305 (1997). https://doi.org/10.1023/A:1018462729127

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