1887

Abstract

We showed previously that 218 and 110 bp N gene segments of tomato spotted wilt virus (TSWV) that were fused to the non-target green fluorescent protein (GFP) gene were able to confer resistance to TSWV via post-transcriptional gene silencing (PTGS). N gene segments expressed alone did not confer resistance. Apparently, the GFP DNA induced PTGS that targetted N gene segments and the incoming homologous TSWV for degradation, resulting in a resistant phenotype. These observations suggested that multiple resistance could be obtained by replacing the GFP DNA with a viral DNA that induces PTGS. The full-length coat protein (CP) gene of turnip mosaic virus (TuMV) was linked to 218 or 110 bp N gene segments and transformed into . A high proportion (4 of 18) of transgenic lines with the 218 bp N gene segment linked to the TuMV CP gene were resistant to both viruses, and resistance was transferred to R plants. Nuclear run-on and Northern experiments confirmed that resistance was via PTGS. In contrast, only one of 14 transgenic lines with the TuMV CP linked to a 110 bp N gene segment yielded progeny with multiple resistance. Only a few R plants were resistant and resistance was not observed in R plants. These results clearly show the applicability of multiple virus resistance through the fusion of viral segments to DNAs that induce PTGS.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-8-2103
2000-08-01
2019-12-06
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/8/0812103a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-8-2103&mimeType=html&fmt=ahah

References

  1. Baulcombe, D. C. ( 1996; ). Mechanisms of pathogen-derived resistance to viruses in transgenic plants. Plant Cell 8, 1833-1844.[CrossRef]
    [Google Scholar]
  2. Baulcombe, D. C. & English, J. J. ( 1996; ). Ectopic pairing of homologous DNA and post-transcriptional gene silencing in transgenic plants. Current Opinion in Biotechnology 7, 173-180.[CrossRef]
    [Google Scholar]
  3. Beachy, R. N. ( 1997; ). Mechanisms and applications of pathogen-derived resistance in transgenic plants. Current Opinion in Biotechnology 8, 215-220.[CrossRef]
    [Google Scholar]
  4. Crété, P. & Vaucheret, H. ( 1999; ). Expression and sequence requirements for nitrite reductase co-suppression. Plant Molecular Biology 41, 105-114.[CrossRef]
    [Google Scholar]
  5. Dawson, W. D. ( 1996; ). Gene silencing and virus resistance: a common mechanism. Trends in Plant Science 1, 107-108.[CrossRef]
    [Google Scholar]
  6. de Haan, P., Wagemakers, L., Peters, D. & Goldbach, R. ( 1990; ). The S RNA segment of tomato spotted wilt virus has an ambisense character. Journal of General Virology 71, 1001-1007.[CrossRef]
    [Google Scholar]
  7. de Haan, P., Kormelink, R., Resende, R. de O., van Poelwijk, F., Peters, D. & Goldbach, R. ( 1991; ). Tomato spotted wilt virus L RNA encodes a putative RNA polymerase. Journal of General Virology 72, 2207-2216.[CrossRef]
    [Google Scholar]
  8. Dehio, C. & Schell, J. ( 1994; ). Identification of plant genetic loci involved in a posttranscriptional mechanism for meiotically reversible transgene silencing. Proceedings of the National Academy of Sciences, USA 91, 5538-5542.[CrossRef]
    [Google Scholar]
  9. Dougherty, W. G. & Carrington, J. C. ( 1988; ). Expression and function of potyviral gene products. Annual Review of Phytopathology 26, 123-143.[CrossRef]
    [Google Scholar]
  10. Dougherty, W. G. & Parks, T. D. ( 1995; ). Transgenes and gene suppression: telling us something new? Current Opinion in Cell Biology 7, 399-405.[CrossRef]
    [Google Scholar]
  11. Feinberg, A. P. & Vogelstein, B. ( 1983; ). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132, 6-13.[CrossRef]
    [Google Scholar]
  12. Fuchs, M. & 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.[CrossRef]
    [Google Scholar]
  13. Fuchs, M., McFerson, J. R., Tricoli, D. M., McMaster, J. R., Deng, R. Z., Boeshore, M. L., Reynolds, J. F., Russell, P. F., Quemada, H. D. & Gonsalves, D. ( 1997; ). Cantaloupe line CZW-30 containing coat protein genes of cucumber mosaic virus, zucchini yellow mosaic virus, and watermelon mosaic virus-2 is resistant to these three viruses in the field. Molecular Breeding 3, 279-290.[CrossRef]
    [Google Scholar]
  14. Fuchs, M., Tricoli, D. M., Carney, K. J., Schesser, M., McFerson, J. R. & Gonsalves, D. ( 1998; ). Comparative virus resistance and fruit yield of transgenic squash with single and multiple coat protein genes. Plant Disease 82, 1350-1356.[CrossRef]
    [Google Scholar]
  15. German, T. L., Ullman, D. E. & Moyer, J. W. ( 1992; ). Tospoviruses (i): diagnosis, molecular biology, phylogeny, and vector relationships. Annual Review of Phytopathology 30, 315-348.[CrossRef]
    [Google Scholar]
  16. Green, S. K. & Deng, T. C. ( 1985; ). Turnip mosaic virus strains in cruciferous hosts in Taiwan. Plant Disease 69, 28-31.[CrossRef]
    [Google Scholar]
  17. Horsch, R. B., Fry, J. E., Hoffmann, N. L., Eichholtz, D., Rogers, S. G. & Fraley, R. T. ( 1985; ). A simple and general method for transferring genes into plants. Science 227, 1229-1231.[CrossRef]
    [Google Scholar]
  18. Jacobs, J. J. M. R., Sanders, M., Bots, M., Andriessen, M., van Eldik, G. J., Litière, K., Van Montagu, M. & Cornelissen, M. ( 1999; ). Sequences throughout the basic β-1,3-glucanase mRNA coding region are targets for homology dependent post-transcriptional gene silencing. Plant Journal 20, 143-152.[CrossRef]
    [Google Scholar]
  19. Jan, F.-J., Pang, S.-Z., Fagoaga, F. & Gonsalves, D. ( 1999; ). Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post-transcriptional gene silencing. Transgenic Research 8, 203-213.[CrossRef]
    [Google Scholar]
  20. Jan, F.-J., Fagoaga, C., Pang, S.-Z. & Gonsalves, D. ( 2000; ). A minimum length of N gene sequence in transgenic plants is required for RNA-mediated tospovirus resistance. Journal of General Virology 81, 235-242.
    [Google Scholar]
  21. Kaniewski, W., Lawson, C., Sammons, B., Haley, L., Hart, J., Delannay, X. & Tumer, 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.[CrossRef]
    [Google Scholar]
  22. Kaniewski, W., Ilardi, V., Tomassoli, L., Mitsky, T., Layton, J. & Barba, M. ( 1999; ). Extreme resistance to cucumber mosaic virus (CMV) in transgenic tomato expressing one or two viral coat proteins. Molecular Breeding 5, 111-119.[CrossRef]
    [Google Scholar]
  23. Kormelink, R., de Haan, P., Peters, D. & Goldbach, R. ( 1992a; ). Viral RNA synthesis in tomato spotted wilt virus-infected Nicotiana rustica plants. Journal of General Virology 73, 687-693.[CrossRef]
    [Google Scholar]
  24. Kormelink, R., de Haan, P., Meurs, C., Peters, D. & Goldbach, R. ( 1992b; ). The nucleotide sequence of the M RNA segment of tomato spotted wilt virus, a bunyavirus with two ambisense RNA segments. Journal of General Virology 73, 2795-2804.[CrossRef]
    [Google Scholar]
  25. Kormelink, R., Storms, M., van Lent, J., Peters, D. & Goldbach, R. ( 1994; ). Expression and subcellular location of the NSM protein of tomato spotted wilt virus (TSWV), a putative viral movement protein. Virology 200, 56-65.[CrossRef]
    [Google Scholar]
  26. Lawson, C., Kaniewski, W., Haley, L., Rozman, R., Newell, C., Sanders, P. & 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.[CrossRef]
    [Google Scholar]
  27. Ling, K. S., Provvidenti, R. & Gonsalves, D. ( 1995; ). Detection of turnip mosaic virus isolates using an antiserum to coat protein breakdown products. Plant Disease 79, 809-812.[CrossRef]
    [Google Scholar]
  28. Maiss, E., Ivanova, L., Breyel, E. & Adam, G. ( 1991; ). Cloning and sequencing of the S RNA from a Bulgarian isolate of tomato spotted wilt virus. Journal of General Virology 72, 461-464.[CrossRef]
    [Google Scholar]
  29. Marano, M. R. & Baulcombe, D. ( 1998; ). Pathogen-derived resistance targeted against the negative-strand RNA of tobacco mosaic virus: RNA strand-specific gene silencing? Plant Journal 13, 537-546.[CrossRef]
    [Google Scholar]
  30. Napoli, C., Lemieux, C. & Jorgensen, R. ( 1990; ). Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2, 279-290.[CrossRef]
    [Google Scholar]
  31. Pang, S.-Z., Nagpala, P., Wang, M., Slightom, J. L. & Gonsalves, D. ( 1992; ). Resistance to heterologous isolates of tomato spotted wilt virus in transgenic tobacco expressing its nucleocapsid protein gene. Phytopathology 82, 1223-1229.[CrossRef]
    [Google Scholar]
  32. Pang, S.-Z., Jan, F.-J., Carney, K., Stout, J., Tricoli, D. M., Quemada, H. D. & Gonsalves, D. ( 1996; ). Post-transcriptional transgene silencing and consequent tospovirus resistance in transgenic lettuce are affected by transgene dosage and plant development. Plant Journal 9, 899-909.[CrossRef]
    [Google Scholar]
  33. Pang, S.-Z., Jan, F.-J. & Gonsalves, D. ( 1997; ). Nontarget DNA sequences reduce the transgene length necessary for RNA-mediated tospovirus resistance in transgenic plants. Proceedings of the National Academy of Sciences, USA 94, 8261-8266.[CrossRef]
    [Google Scholar]
  34. Prins, M. & Goldbach, R. ( 1996; ). RNA-mediated virus resistance in transgenic plants. Archives of Virology 141, 2259-2276.[CrossRef]
    [Google Scholar]
  35. Prins, M., de Haan, P., Luyten, R., van Veller, M., van Grinsven, M. Q. J. M. & Goldbach, R. ( 1995; ). Broad resistance to tospoviruses in transgenic tobacco plants expressing three tospoviral nucleoprotein gene sequences. Molecular Plant–Microbe Interactions 8, 85-91.[CrossRef]
    [Google Scholar]
  36. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  37. Seymour, G. B., Fray, R. G., Hill, P. & Tucker, G. A. ( 1993; ). Down-regulation of two non-homologous tomato genes with a single chimaeric sense gene construct. Plant Molecular Biology 23, 1-9.[CrossRef]
    [Google Scholar]
  38. Shukla, D. D., Ward, C. W. & Brunt, A. A. (1994). The Potyviridae. Wallingford, UK: CAB International.
  39. Stam, M., de Bruin, R., Kenter, S., van der Hoorn, R. A. L., van Blokland, R., Mol, J. N. M. & Kooter, J. M. ( 1997; ). Post-transcriptional silencing of chalcone synthase in Petunia by inverted transgene repeats. Plant Journal 12, 63-82.[CrossRef]
    [Google Scholar]
  40. Tomlinson, J. A. ( 1987; ). Epidemiology and control of virus diseases of vegetables. Annals of Applied Biology 110, 661-681.[CrossRef]
    [Google Scholar]
  41. Tricoli, D. M., Carney, K. J., Russell, P. F., McMaster, J. R., Groff, D. W., Hadden, K. C., Himmel, P. T., Hubbard, J. P., Boeshore, M. L. & Quemada, H. D. ( 1995; ). 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. Bio/Technology 13, 1458-1465.[CrossRef]
    [Google Scholar]
  42. Van Blokland, R., Van der Geest, N., Mol, J. N. M. & Kooter, J. M. ( 1994; ). Transgene-mediated suppression of chalcone synthase expression in Petunia hybrida results from an increase in RNA turnover. Plant Journal 6, 861-877.[CrossRef]
    [Google Scholar]
  43. van den Boogaart, T., Lomonossoff, G. P. & Davies, J. W. ( 1998; ). Can we explain RNA-mediated virus resistance by homology-dependent gene silencing? Molecular Plant–Microbe Interactions 11, 717-723.[CrossRef]
    [Google Scholar]
  44. Wang, M. & Gonsalves, D. ( 1990; ). ELISA detection of various tomato spotted wilt virus isolates using specific antisera to structural proteins of the virus. Plant Disease 74, 154-158.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-8-2103
Loading
/content/journal/jgv/10.1099/0022-1317-81-8-2103
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error