1887

Abstract

Transgenic papaya cultivars SunUp and Rainbow express the coat protein (CP) gene of the mild mutant of papaya ringspot virus (PRSV) HA. Both cultivars are resistant to PRSV HA and other Hawaii isolates through homology-dependent resistance via post-transcriptional gene silencing. However, Rainbow, which is hemizygous for the CP gene, is susceptible to PRSV isolates from outside Hawaii, while the CP-homozygous SunUp is resistant to most isolates but susceptible to the YK isolate from Taiwan. To investigate the role of CP sequence similarity in overcoming the resistance of Rainbow, PRSV HA recombinants with various CP segments of the YK isolate were constructed and evaluated on Rainbow, SunUp and non-transgenic papaya. Non-transgenic papaya were severely infected by all recombinants, but Rainbow plants developed a variety of symptoms. On Rainbow, a recombinant with the entire CP gene of YK caused severe symptoms, while recombinants with only partial YK CP sequences produced a range of milder symptoms. Interestingly, a recombinant with a YK segment from the 5′ region of the CP gene caused very mild, transient symptoms, whereas recombinants with YK segments from the middle and 3′ parts of the CP gene caused prominent and lasting symptoms. SunUp was resistant to all but two recombinants, which contained the entire CP gene or the central and 3′-end regions of the CP gene and the 3′ non-coding region of YK, and the resulting symptoms were mild. It is concluded that the position of the heterologous sequences in the recombinants influences their pathogenicity on Rainbow.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-82-11-2827
2001-11-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/82/11/0822827a.html?itemId=/content/journal/jgv/10.1099/0022-1317-82-11-2827&mimeType=html&fmt=ahah

References

  1. Anandalakshmi R., Pruss G. J., Ge X., Marathe R., Mallory A. C., Smith T. H., Vance V. B. 1998; A viral suppressor of gene silencing in plants. Proceedings of the National Academy of Sciences, USA 95:13079–13084
    [Google Scholar]
  2. Bass B. L. 2000; Double-stranded RNA as a template for gene silencing. Cell 101:235–238
    [Google Scholar]
  3. Baulcombe D. C. 1996; Mechanisms of pathogen-derived resistance to viruses in transgenic plants. Plant Cell 8:1833–1844
    [Google Scholar]
  4. Baulcombe D. 1999a; Viruses and gene silencing in plants. Archives of Virology Supplementum 15:189–201
    [Google Scholar]
  5. Baulcombe D. C. 1999b; Fast forward genetics based on virus-induced gene silencing. Current Opinion in Plant Biology 2:109–113
    [Google Scholar]
  6. 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
    [Google Scholar]
  7. Beachy R. N. 1993; Introduction: transgenic resistance to plant viruses. Seminars in Virology 4:327–328
    [Google Scholar]
  8. Brigneti G., Voinnet O., Li W. X., Ji L. H., Ding S. W., Baulcombe D. C. 1998; Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO Journal 17:6739–6746
    [Google Scholar]
  9. Chiang C.-H., Yeh S.-D. 1997; Infectivity assays of in vitro and in vivo transcripts of papaya ringspot potyvirus. Botanical Bulletin of Academia Sinica 38:153–163
    [Google Scholar]
  10. Clark M. F., Adams A. N. 1977; Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 34:475–483
    [Google Scholar]
  11. Dalmay T., Hamilton A., Mueller E., Baulcombe D. C. 2000a; Potato virus X amplicons in Arabidopsis mediate genetic and epigenetic gene silencing. Plant Cell 12:369–379
    [Google Scholar]
  12. Dalmay T., Hamilton A., Rudd S., Angell S., Baulcombe D. C. 2000b; An RNA-dependent RNA polymerase gene in Arabidopsis is required for posttranscriptional gene silencing mediated by a transgene but not by a virus. Cell 101:543–553
    [Google Scholar]
  13. Dougherty W. G., Parks T. D. 1995; Transgenes and gene suppression: telling us something new?. Current Opinion in Cell Biology 7:399–405
    [Google Scholar]
  14. English J. J., Mueller E., Baulcombe D. C. 1996; Suppression of virus accumulation in transgenic plants exhibiting silencing of nuclear genes. Plant Cell 8:179–188
    [Google Scholar]
  15. English J. J., Davenport G. F., Elmayan T., Vaucheret H., Baulcombe D. C. 1997; Requirement of sense transcription for homology-dependent virus resistance and trans-inactivation. Plant Journal 12:597–603
    [Google Scholar]
  16. Fitch M. M. M., Manshardt R. M., Gonsalves D., Slightom J. L., Sanford J. C. 1992; Virus resistant papaya derived from tissues bombarded with the coat protein gene of papaya ringspot virus. Bio/Technology 10:1466–1472
    [Google Scholar]
  17. Gonsalves D. 1998; Control of papaya ringspot virus in papaya: a case study. Annual Review of Phytopathology 36:415–437
    [Google Scholar]
  18. Gonsalves D., Ishii M. 1980; Purification and serology of papaya ringspot virus. Phytopathology 70:1028–1032
    [Google Scholar]
  19. Goodwin J., Chapman K., Swaney S., Parks T. D., Wernsman E. A., Dougherty W. G. 1996; Genetic and biochemical dissection of transgenic RNA-mediated virus resistance. Plant Cell 8:95–105
    [Google Scholar]
  20. Hamilton A. J., Baulcombe D. C. 1999; A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286:950–952
    [Google Scholar]
  21. Jacobs J. J. M. R., Sanders M., Bots M., Andriessen M., van Eldik G. J., Litiere 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
    [Google Scholar]
  22. Jan F.-J., Pang S.-Z., Tricoli D. M., Gonsalves D. 2000; Evidence that resistance in squash mosaic comovirus coat protein-transgenic plants is affected by plant developmental stage and enhanced by combination of transgenes from different lines. Journal of General Virology 81:2299–2306
    [Google Scholar]
  23. Kasschau K. D., Carrington J. C. 1998; A counterdefensive strategy of plant viruses: suppression of posttranscriptional gene silencing. Cell 95:461–470
    [Google Scholar]
  24. Levy L., Lee I. M., Hadidi A. 1994; Simple and rapid preparation of infected plant tissue extracts for PCR amplification of virus, viroid, and MLO nucleic acids. Journal of Virological Methods 49:295–304
    [Google Scholar]
  25. Lindbo J. A., Silva-Rosales L., Proebsting W. M., Dougherty W. G. 1993; Induction of a highly specific antiviral state in transgenic plants: implications for regulation of gene expression and virus resistance. Plant Cell 5:1749–1759
    [Google Scholar]
  26. Ling K., Namba S., Gonsalves C., Slightom J. L., Gonsalves D. 1991; Protection against detrimental effects of potyvirus infection in transgenic tobacco plants expressing the papaya ringspot virus coat protein gene. Bio/Technology 9:752–758
    [Google Scholar]
  27. Lomonossoff G. P. 1995; Pathogen-derived resistance to plant viruses. Annual Review of Phytopathology 33:323–343
    [Google Scholar]
  28. Manshardt R. M. 1999 ‘UH Rainbow’ papaya . University of Hawaii College of Tropical Agriculture and Human Resources, New Plants for Hawaii-1 Honolulu: University of Hawaii College of Tropical Agriculture and Human Resources;
    [Google Scholar]
  29. Meins F. Jr 2000; RNA degradation and models for post-transcriptional gene silencing. Plant Molecular Biology 43:261–273
    [Google Scholar]
  30. Metzlaff M., O’Dell M., Cluster P. D., Flavell R. B. 1997; RNA-mediated RNA degradation and chalcone synthase A silencing in petunia. Cell 88:845–854
    [Google Scholar]
  31. Montgomery M. K., Fire A. 1998; Double-stranded RNA as a mediator in sequence-specific genetic silencing and co-suppression. Trends in Genetics 14:255–258
    [Google Scholar]
  32. Mourrain P., Beclin C., Elmayan T., Feuerbach F., Godon C., Morel J. B., Jouette D., Lacombe A. M., Nikic S., Picault N., Remoue K., Sanial M., Vo T. A., Vaucheret H. 2000; Arabidopsis SGS2 and SGS3 genes are required for posttranscriptional gene silencing and natural virus resistance. Cell 101:533–542
    [Google Scholar]
  33. 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
    [Google Scholar]
  34. Pang S. Z., Jan F. J., Tricoli D. M., Russell P. F., Carney K. J., Hu J. S., Fuchs M., Quemada H. D., Gonsalves D. 2000; Resistance to squash mosaic comovirus in transgenic squash plants expressing its coat protein genes. Molecular Breeding 6:87–93
    [Google Scholar]
  35. Prins M., Goldbach R. 1996; RNA-mediated virus resistance in transgenic plants. Archives of Virology 141:2259–2276
    [Google Scholar]
  36. Quemada H., L’Hostis B., Gonsalves D., Reardon I. M., Heinrikson R., Hiebert E. L., Sieu L. C., Slightom J. L. 1990; The nucleotide sequences of the 3′-terminal regions of papaya ringspot virus strains W and P. Journal of General Virology 71:203–210
    [Google Scholar]
  37. Sanford J. C., Johnston S. A. 1985; The concept of parasite-derived resistance – deriving resistance genes from the parasite’s own genome. Journal of Theoretical Biology 113:395–405
    [Google Scholar]
  38. Shukla D. D., Strike P. M., Tracy S. L., Gough K. H., Ward C. W. 1988; The N and C termini of the coat proteins of potyviruses are surface-located and the N terminus contains the major virus-specific epitopes. Journal of General Virology 69:1497–1508
    [Google Scholar]
  39. Sijen T., Wellink J., Hiriart J.-B., van Kammen A. 1996; RNA-mediated virus resistance: role of repeated transgenes and delineation to targeted regions. Plant Cell 8:2277–2294
    [Google Scholar]
  40. Smith H. A., Swaney S. L., Parks T. D., Wernsman E. A., Dougherty W. G. 1994; Transgenic plant virus resistance mediated by untranslatable sense RNAs: expression, regulation, and fate of nonessential RNAs. Plant Cell 6:1441–1453
    [Google Scholar]
  41. Sonoda S., Mori M., Nishiguchi M. 1999; Homology-dependent virus resistance in transgenic plants with the coat protein gene of sweet potato feathery mottle potyvirus: target specificity and transgene methylation. Phytopathology 89:385–391
    [Google Scholar]
  42. Tennant P. F., Gonsalves C., Ling K. S., Fitch M., Manshardt R., Slightom J. L., Gonsalves D. 1994; Differential protection against papaya ringspot virus isolates in coat protein gene transgenic papaya and classically cross-protected papaya. Phytopathology 84:1359–1366
    [Google Scholar]
  43. Tennant P., Fermin G., Fitch M. M., Manshardt R. M., Slightom J. L., Gonsalves D. 2001; Papaya ringspot virus resistance of transgenic Rainbow and SunUp is affected by gene dosage, plant development, and coat protein homology. European Journal of Plant Pathology (in press
    [Google Scholar]
  44. Voinnet O., Pinto Y. M., Baulcombe D. C. 1999; Suppression of gene silencing: a general strategy used by diverse DNA and RNA viruses of plants. Proceedings of the National Academy of Sciences, USA 96:14147–14152
    [Google Scholar]
  45. Wang C.-H., Yeh S.-D. 1992; Nucleotide sequence comparison of the 3′-terminal regions of severe, mild, and non-papaya infecting strains of papaya ringspot virus. Archives of Virology 127:345–354
    [Google Scholar]
  46. Wang C.-H., Yeh S.-D. 1997; Divergence and conservation of the genomic RNAs of Taiwan and Hawaii strains of papaya ringspot potyvirus. Archives of Virology 142:271–285
    [Google Scholar]
  47. Wassenegger M., Pelissier T. 1998; A model for RNA-mediated gene silencing in higher plants. Plant Molecular Biology 37:349–362
    [Google Scholar]
  48. Waterhouse P. M., Graham M. W., Wang M. B. 1998; Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proceedings of the National Academy of Sciences, USA 95:13959–13964
    [Google Scholar]
  49. Yeh S. D., Gonsalves D. 1984; Evaluation of induced mutants of papaya ringspot virus for control by cross protection. Phytopathology 74:1086–1091
    [Google Scholar]
  50. Yeh S.-D., Jan F.-J., Chiang C.-H., Doong T.-J., Chen M.-C., Chung P.-H., Bau H.-J. 1992; Complete nucleotide sequence and genetic organization of papaya ringspot virus RNA. Journal of General Virology 73:2531–2541
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-82-11-2827
Loading
/content/journal/jgv/10.1099/0022-1317-82-11-2827
Loading

Data & Media loading...

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