Abstract
Transgenic inbred squash lines containing various combinations of the cucumber mosaic virus (CMV), watermelon mosaic virus 2 (WMV 2) or zucchini yellow mosaic virus (ZYMV) coat protein (CP) genes were produced using Agrobacterium-mediated transformation. Progeny from lines transformed with single or multiple CP gene constructs were tested for virus resistance under field conditions, and exhibited varying levels of resistance to infection by CMV, WMV 2 or ZYMV. Most transgenic lines remained nonsymptomatic throughout the growing seasons and produced marketable fruits, while other lines showed a delay in the onset of symptoms and/or a reduction in symptom severity. A few lines failed to display any level of resistance. Depending on the CP gene used, 40 to 95% of the transgenic lines containing single CP constructs of either CMV, WMV 2 or ZYMV were resistant to the virus from which the CP gene was derived. Transgenic lines transformed with a double CP construct containing the CP genes from CMV and WMV 2, designated CW, displayed high level of resistance to CMV and WMV 2. A transgenic line, designated ZW-20, which contained the CP genes from ZYMV and WMV 2 displayed excellent resistance to ZYMV and WMV 2 in that most of the plants showed complete resistance. A few plants developed localized chlorotic dots or blotches, yet fruits remained asymptomatic. Southern blot analysis revealed that the CP inserts of some resistant plants of line ZW-20 were no longer linked to the neomycin phosphotransferase II (NPT II) gene. This loss of linkage allowed the marker gene to be separated from the virus resistance trait by Mendelian segregation. Further analysis of these plants showed that they contained multiple WMV 2 inserts which were designated B and H, the latter consisting of two hybridization signals. Analysis of inoculated plants showed that plants with the H inserts were symptomless or developed only chlorotic dots, while those without the H insert developed more prominent chlorotic blotches. In addition to lines with resistance to two viruses, a line with resistance to three viruses was also identified. Transgenic line CZW-3, transformed with the triple CP construct containing the CMV, WMV 2 and ZYMV CP genes, exhibited resistance to all three viruses. These two transgenic inbred lines, ZW-20 and CZW-3, have allowed for the development of commercial squash hybrids with multiple virus resistance.
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References
Lisa, V. and Lecoq, H. 1984. Zucchini yellow mosaic virus. CMI/AAB. Description of plant viruses, No. 282.
Purcifull, D., Hiebert, E. and Edwardson, J. 1984. Watermelon mosaic virus 2. CMI/AAB. Description of plant viruses, No. 293.
Purcifull, D., Edwardson, J., Hiebert, E. and Gonsalves, D. 1984. Papaya ringspot virus. CMI/AAB. Description of plant viruses, No. 292.
Palukaitis, P., Roossinck, M., Dietzgen, R.G. and Francki, R.I.B. 1992. Cucumber mosaic virus. Advances in Virus Research 41: 281–348.
Gibbs, A. and Harrison, B. 1976. Transmission by vectors and in other natural ways. Plant Virology, The Principles. Halsted Press, N.Y. p 186–187.
Zitter, T.A. and Simons, J.N. 1980. Management of viruses by alteration of vector efficiency and by cultural practices. Ann. Rev. Phytopathol. 18: 289–310.
Brown, J.E., Dangler, J.M., Woods, F.M., Tilt, K.M., Henshaw, M.D., Griffey, W.A. and West, M.S. 1993. Delay in mosaic virus onset and aphid vector reduction in summer squash grown on reflective mulches. HortScience 28: 895–896.
Gibson, R.W. and Rice, A.D. 1986. The combined use of mineral oils and pyrethroids to control plant viruses transmitted non- and semi- persistently by Myzus persicae. Ann Appl. Biol. 109: 465–472.
Loebenstein, G. and Raccah, B. 1980. Control of non-persistently transmitted aphid-bome viruses. Phytoparasitica 8: 221–235.
Marco, S. 1986. Incidence of aphid-transmitted virus infections reduced by whitewash sprays on plants. Phytopathology 76: 1344–1348.
Marco, S. 1993. Incidence of nonpersistently transmitted viruses, p. 331–407. In: The plant viruses, vol. 4, The filamentous viruses, R.G. Milne (Ed.). Plenum Press, Inc., New York.
Simons, J.N. and Zitter, T.A. 1980. Use of oils to control aphid-borne viruses. Plant Disease 64: 542–546.
Webb, S.E. and Linda, S.B. 1993. Effect of oil and insecticide on epidemics of potyviruses in watermelon in Florida. Plant Disease 77: 869–874.
Provvidenti, R. 1993. Resistance to viral diseases of Cucurbits, p. 8–43. In: Resistance to viral diseases of vegetables: genetics and breeding. Timber Press, Portland, Oregon.
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.
Beachy, R.N. 1993. Virus resistance through expression of coat protein genes, p. 89–104. In: Biotechnology in Plant Disease Control, Wiley-Liss, Inc.
Scholthof, K.B.G., Scholthof, H.D. and Jackson, A.O. 1993. Control of plant virus diseases by pathogen-derived resistance in transgenic plants. Plant Physiol. 102: 7–12.
Wilson, T.M.A. 1993. Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms. Proc. Natl. Acad. Sci. USA 90: 3134–3141.
Gonsalves, D., Chee, P., Provvidenti, R., Seem, R. and Slightom, J. 1992. Comparison of coat protein-mediated and genetically derived resistance in cucumber to infection by cucumber mosaic virus under field conditions with natural challenge inoculations by vectors. Biotechnology 10: 1562–1570.
Fang, G. and Grumet, R. 1993. Genetic engineering of potyvirus resistance using constructs derived from the zucchini yellow mosaic virus coat protein gene. Molec. Plant Microbe Interact. 6: 358–367.
Slightom, J.L. 1991. Custom polymerase-chain-reaction engineering of a plant expression vector. Gene 100: 251–255.
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: 1065–1074.
Quemada, H.D., Gonsalves, D. and Slightom, J.L. 1991. Expression of coat protein gene from cucumber mosaic virus strain C in tobacco: protection against infections by CMV strains transmitted mechanically or by aphids. Phytopathology 81: 794–802.
Namba, S., Ling, K., Gonsalves, C., Slightom, J.L. and Gonsalves, D. 1992. Protection of transgenic plants expressing the coat protein gene of watermelon mosaic virus II or zucchini yellow mosaic virus against six potyviruses. Phytopathology 82: 940–946.
Kanieswki, W., Lawson, C., Sammons, B., Haley, L., Hart, J., Delannay, X. and Turner, N.E. 1990. Field resistance of transgenic Russet Burbank potato to effects of infection by potato virus X and potato virus Y. Bio/Technology 8: 750–754.
Fuchs, M. and Gonsalves, D. 1995. Resistance of transgenic hybrid squash ZW-20 expressing the coat protein genes of zucchini yellow mosaic virus and watermelon mosaic virus 2 to mixed infections by both potyviruses. Bio/Technology 13: 1466–1473.
An, G. 1986. Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol. 81: 86–91.
Jefferson, R.A., Kavanaugh, T.A. and Bevan, M.W. 1987. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO Journal 6: 3901–3907.
Allmansberger, R., Brau, B. and Piepersberg, W. 1985. Genes for gentamicin-(3)-N-acetyl-transferases III and IV. II. Nucleotide sequences of three AAC(3) genes and evolutionary aspects. Molecular General Genetics 198: 514–520.
Kay, R. and MCPherson, J. 1987. Hybrid pUC vectors for addition of new restriction enzyme sites to the ends of DNA fragments. Nucleic Acids Res. 15: 2778.
Quemada, H.D., 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.
Horsch, R.B., Fry, J.E., Hoffmann, 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.
Maniatis, X., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning. Cold Spring Harbor, New York, Cold Spring Harbor Laboratory.
Clark, M.L. and Adams, A.N. 1977. Characteristics of the microplate method for enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virol. 34: 475–483.
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Tricoll, D., Carney, K., Russell, P. et al. Field Evaluation of Transgenic Squash Containing Single or Multiple Virus Coat Protein Gene Constructs for Resistance to Cucumber Mosaic Virus, Watermelon Mosaic Virus 2, and Zucchini Yellow Mosaic Virus. Nat Biotechnol 13, 1458–1465 (1995). https://doi.org/10.1038/nbt1295-1458
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DOI: https://doi.org/10.1038/nbt1295-1458
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