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Post-transcriptional gene silencing in plum pox virus resistant transgenic European plum containing the plum pox potyvirus coat protein gene

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Abstract

Transgenic plums containing the plum pox potyvirus coat protein (PPV-CP) gene were inoculated with PPV. Infection was monitored by evaluating symptoms, ELISA, and IC-RT-PCR. Transgenic clone C5 was highly resistant to PPV during four years of testing and displayed characteristics typical of post-transcriptional gene silencing (PTGS), including a high level of transgene transcription in the nucleus, low levels of transgene mRNA in the cytoplasm, a complex multicopy transgene insertion with aberrant copies, and methylation of the silenced PPV-CP transgene. The PPV-CP transgene was also methylated in seedlings of C5 and these seedlings were resistant to PPV. Our results show, for the first time, that PTGS functions as a mechanism for virus resistance in a woody perennial species.

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References

  • Adams AN and Patterson VSA (1980) Plum pox virus. East Malling Research Station Report for 1979: 104.

  • Adams AN, Guise CM and Crossley SJ (1998) Detection of plum pox virus in root and bark samples from dormant plum trees. Acta Hort 472: 469–474.

    Google Scholar 

  • Balandin T and Castresana C (1997) Silencing of a ß -1,3-glucanase transgene is overcome during seed formation. Plant Mol Biol 34: 125–137.

    Google Scholar 

  • Baulcombe DC (1996) Mechanisms of pathogen-derived resistance to viruses in transgenic plants. Plant Cell 8 1833–1844.

    Google Scholar 

  • Beachy RN, Loesch-Fries S and Tumer NE (1990) Coat proteinmediated resistance against virus infection. Ann Rev Phytopathol 28: 451–474.

    Google Scholar 

  • Carrington JC and Whitham SA (1998) Viral invasion and host defense: strategies and counter strategies. Current Opinions Plant Biol 1: 336–341.

    Google Scholar 

  • Damsteegt VD, Waterworth HE, Mink GI, Howell WE and Levy L (1997) Prunus tomentosa as a diagnostic host for detection of plum pox virus and other Prunus viruses. Plant Dis 81: 329–332.

    Google Scholar 

  • Davies GJ, Sheikh MA, Ratcliff OJ, Coupland G and Furner IJ (1997) Genetics of homology-dependent gene silencing in Arabidopsis; a role for methylation. Plant J 12: 791–804.

    Google Scholar 

  • Dehio C and Schell J (1994) Identification of plant genetic loci involved in posttranscriptional mechanism for meiotically reversible transgene silencing. Proc Natl Acad Sci USA 91: 5538–5542.

    Google Scholar 

  • Depicker A and Van Montagu M (1997) Post-transcriptional gene silencing in plants. Curr Opin Cell Biol 9: 373–382.

    Google Scholar 

  • Dougherty WG and Parks TD (1995) Transgenes and gene suppression: Telling us something new? Curr Opin Cell Biol 7: 399–405.

    Google Scholar 

  • English JJ, Mueller E and Baulcomb DC (1996) Suppression of virus accumulation in transgenic plants exhibiting silencing of nuclear genes. Plant Cell 8: 179–188.

    Google Scholar 

  • Fitch MMM, Manshardt RM, Gonsalves D, Slightom JL and Sanford JC (1990) Stable transformation of papaya via microprojectile bombardment. Plant Cell Repts 9: 189–194.

    Google Scholar 

  • Guo HS, Lopez-Moya JJ and Garcia JA (1999) Mitotic stability of infection-induced resistance to plum pox potyvirus associated with transgene silencing and DNA methylation. MPMI 12: 103–111.

    Google Scholar 

  • Ingelbrecht IL, Van Houdt H, Van Montagu M and Depicker A (1994) Post-transcriptional silencing of reporter transgenes in tobacco correlates with DNA methylation. Proc Natl Acad Sci USA 91: 10502–10506.

    Google Scholar 

  • Jones L, Hamilton AJ, Voinnet O, Thomas CL, Maule AJ and Baulcombe DC (1999) RNA-DNA interactions and DNA methylation in post-transcriptional gene silencing. Plant Cell 11: 2291–2301.

    Google Scholar 

  • Kobayashi N, Horikoshi T, Katsuyama H, Handa T and Takayanagi K (1998) A simple and efficient DNA extraction method for plants, especially woody plants. Plant Tissue Culture and Biotech 4(2) 76–80.

    Google Scholar 

  • Kohli A, Gahakawa D, Vain P, Laurie DA and Christou P (1999) Transgene expression in rice engineered through particle bombardment: molecular factors controlling stable expression and transgene silencing. Planta 208: 88–97.

    Google Scholar 

  • Kunz C, Hanspeter S, Stam M, Kooter JM and Meins F Jr. (1996) Developmentally regulated silencing and reactivation of tobacco chitinase transgene expression. Plant J 10: 437–450.

    Google Scholar 

  • Levy L and Hadidi A (1994) A simple and rapid method for processing tissue infected with plum pox potyvirus for use with specific 3′ non-coding RT-PCR assays. OEPP/EPPO Bull 24: 595–604.

    Google Scholar 

  • Ling KS, Namba S, Gonsalves C, Slightom JL and 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 

  • Malinowski T, Zawadzka B, Ravelonandro M and Scorza R (1998) Preliminary report on the apparent breaking of resistance of a transgenic plum by chip bud inoculation of plum pox virus PPVS. Acta Virol 42: 241–243.

    Google Scholar 

  • Maniatis T, Fritsch EF and Sambrook J (1982) Molecular cloning: A Laboratory Manual. Cold Springs Harbor Laboratory, NY.

    Google Scholar 

  • Nemeth M (1994) History and importance of plum pox in stone-fruit production. EPPO Bul 24: 525–536.

    Google Scholar 

  • Palauqui JC, Elmayan T, Pollien JM and Vaucheret J (1997) Systemic acquired silencing: transgene-specific post-transcriptional silencing is transmitted by grafting from silenced stocks to non-silenced scions. EMBO J 16: 4738–4745.

    Google Scholar 

  • Palauqui JC and Vaucheret V (1998) Transgenes are dispensable for the RNA degradation step of cosuppression. Proc Natl Acad Sci USA 95: 9675–9680.

    Google Scholar 

  • Pang SZ, Jan FJ, Carney K, Stiout J, Tricoli DM, Quemada HD et al. (1996) Post-transcriptional transgene silencing and consequent tospovirus resistance in transgenic lettuce are affected by transgene dosage and plant development. Plant J 9: 899–909.

    Google Scholar 

  • Ravelonandro M, Monsion M, Teycheney PY, Delbos R and Dunez J (1992) Construction of a chimeric viral gene expressing plum pox coat protein. Gene 120: 167–173.

    Google Scholar 

  • Ravelonandro M, Scorza R, Bachelier JC, Labonne G, Levy L, Damsteegt V et al. (1997) Resistance of transgenic Prunus domestica to plum pox virus infection. Plant Dis 81: 1231–1235.

    Google Scholar 

  • Saxena PK, Fowke LC and King J (1985) An efficient procedure for isolation of nuclei from plant protoplasts. Protoplasma 128: 184–189.

    Google Scholar 

  • Scorza R, Ravelonandro M, Callahan AM, Cordts JM, Fuchs M, Dunez J et al. (1994) Transgenic plums (Prunus domestica L.) express the plum pox virus coat protein gene. Plant Cell Repts 14: 18–22.

    Google Scholar 

  • Scorza R, Levy L, Damsteegt V, Yepes LM, Cordts J, Hadidi A et al. (1995) Transformation of plum with the papaya ringspot virus coat protein gene and reaction of transgenic plants to plum pox virus. J Am Soc Hort Sci 120: 943–952.

    Google Scholar 

  • Scorza R, Callahan AM, Levy L, Damsteegt V and Ravelonandro M (1998) Transferring potyvirus coat protein genes through hybridization of transgenic plants to produce plum pox virus resistant plums (Prunus domestica L.). Acta Hort 472: 421–427.

    Google Scholar 

  • Sijen T, Wellink J, Hiriart J-B and van Kammen AE (1996) RNA-mediated virus resistance: role of repeated transgenes and delineation of targeted regions. Plant Cell 8: 2277–2294.

    Google Scholar 

  • Sonoda S, Mori M and 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. Phytopath 89: 385–391.

    Google Scholar 

  • Stam M, de Bruin R, Kenter S, van der Hoorn RAL, van Blokland R, Mol JNM et al. (1997) Post-transcriptional silencing of chalcone synthase in Petunia by inverted transgene repeats. Plant J 12: 63–82.

    Google Scholar 

  • Tanzer MM, Thompson WF, Law MD, Wernsman EA and Uknes S (1997) Characterization of post-transcriptionally suppressed transgene expression that confers resistance to tobacco etch virus infection in tobacco. Plant Cell 9: 1411–1423.

    Google Scholar 

  • Wassenegger M and Pélissier T (1998) A model for RNA-mediated gene silencing in higher plants. Plant Mol Biol 37: 349–362.

    Google Scholar 

  • Waterhouse PM, Graham MW and Wang M-B (1998) Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA 95: 13959–13964.

    Google Scholar 

  • Wilson TMA (1993) Strategies to protect crop plants against viruses: Pathogen-derived resistance blossoms. Proc Natl Acad Sci USA 90: 3134–3141.

    Google Scholar 

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Authors and Affiliations

  1. United States Department of Agriculture, Agricultural Research Service, Appalachian Fruit Research Station, 45 Wiltshire Road, Kearneysville, WV, 25430, USA

    Ralph Scorza, Ann Callahan & Kevin Webb

  2. United States Department of Agriculture, Animal Plant Health Inspection Service, Plant Protection and Quarantine, Beltsville Agricultural Research Center, East Building 580, Beltsville, MD, 20705, USA

    Laurene Levy

  3. United States Department of Agriculture, Agricultural Research Service, Foreign Disease-Weed Science Research Unit, 1301 Ditto Ave., Ft. Detrick, MD, 21702, USA

    Vern Damsteegt

  4. Station de Pathologie Vegetale-INRA, Bordeaux, BP 81, 33883, Villenave d'Ornon, France

    Michel Ravelonandro

Authors
  1. Ralph Scorza
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  2. Ann Callahan
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  3. Laurene Levy
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  4. Vern Damsteegt
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  5. Kevin Webb
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  6. Michel Ravelonandro
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Scorza, R., Callahan, A., Levy, L. et al. Post-transcriptional gene silencing in plum pox virus resistant transgenic European plum containing the plum pox potyvirus coat protein gene. Transgenic Res 10, 201–209 (2001). https://doi.org/10.1023/A:1016644823203

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