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Transgenic expression of tomato bushy stunt virus silencing suppressor P19 via the pOp/LhG4 transactivation system induces viral-like symptoms in tomato

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Abstract

During natural infection, the Tomato bushy stunt virus (TBSV) silencing suppressor protein P19 is expressed at high levels, which are required for optimum viral pathogenicity and silencing suppression. To date, expression of P19 in transgenic host plants has failed to achieve comparable expression levels and thus has provided only limited information on its in planta effects. To obtain high P19 expression and study its effects on host plant development in the absence of virus infection, we generated HA-tagged P19 (P19HA)-transgenic tomato reporter plants using the pOp/LhG4 transactivation system, which separates transformation from transgene expression. Upon reporter plant activation with a strong constitutive promoter, the transactivated F1 plants expressed high levels of a functional P19HA protein and displayed multiple abnormal phenotypes, some of which were highly reminiscent of the symptoms described previously for TBSV-infected tomato. Moreover, phenotype severity correlated with P19HA expression level, amount of bound miRNA/miRNA* duplexes, and accumulation of miRNA target transcripts. Together our results demonstrate that the tomato miRNA pathway is markedly compromised by P19, in particular when this protein is relatively abundant, as occurs during natural infection. We suggest that such interference with endogenous silencing may be responsible for at least some of the symptoms characteristic of TBSV-infected tomato.

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References

  1. P. Brodersen, O. Voinnet, Trends Genet. 22, 268 (2006)

    Article  CAS  Google Scholar 

  2. O. Voinnet, Nat. Rev. Genet. 6, 206 (2005)

    Article  CAS  Google Scholar 

  3. P. Dunoyer, O. Voinnet, Curr. Opin. Plant Biol. 8, 415 (2005)

    Article  CAS  Google Scholar 

  4. E.J. Chapman, A.I. Prokhnevsky, K. Gopinath, V.V. Dolja, J.C. Carrington, Genes Dev. 18, 1186 (2004)

    Article  Google Scholar 

  5. D. Garcia, Semin. Cell Dev. Biol. 19, 586 (2008)

    Article  CAS  Google Scholar 

  6. K.D. Kasschau, Z. Xie, E. Allen, C. Llave, E.J. Chapman, K.A. Krizan, J.C. Carrington, Dev. Cell 4, 205 (2003)

    Article  CAS  Google Scholar 

  7. J.A. Diaz-Pendon, S.W. Ding, Annu. Rev. Phytopathol. 46, 303 (2008)

    Article  CAS  Google Scholar 

  8. R. Anandalakshmi, G.J. Pruss, X. Ge, R. Marathe, A.C. Mallory, T.H. Smith, V.B. Vance, Proc. Natl. Acad. Sci. USA 95, 13079 (1998)

    Article  CAS  Google Scholar 

  9. G. Brigneti, O. Voinnet, W.X. Li, L.H. Ji, S.W. Ding, D.C. Baulcombe, EMBO J. 17, 6739 (1998)

    Article  CAS  Google Scholar 

  10. K.D. Kasschau, J.C. Carrington, Cell 95, 461 (1998)

    Article  CAS  Google Scholar 

  11. Y.M. Shiboleth, E. Haronsky, D. Leibman, T. Arazi, M. Wassenegger, S.A. Whitham, V. Gaba, A. Gal-On, J. Virol. 81, 13135 (2007)

    Article  CAS  Google Scholar 

  12. W. Qiu, J.W. Park, H.B. Scholthof, Mol. Plant Microbe Interact. 15, 269 (2002)

    Article  CAS  Google Scholar 

  13. F. Qu, T.J. Morris, Mol. Plant Microbe Interact. 15, 193 (2002)

    Article  CAS  Google Scholar 

  14. O. Voinnet, Y.M. Pinto, D.C. Baulcombe, Proc. Natl. Acad. Sci. USA 96, 14147 (1999)

    Article  CAS  Google Scholar 

  15. H.B. Scholthof, Nat. Rev. Genet. 4, 405 (2006)

    Article  CAS  Google Scholar 

  16. T. Dalmay, L. Rubino, J. Burgyan, A. Kollar, M. Russo, Virology 194, 697 (1993)

    Article  CAS  Google Scholar 

  17. H.B. Scholthof, K.B. Scholthof, M. Kikkert, A.O. Jackson, Virology 213, 425 (1995)

    Article  CAS  Google Scholar 

  18. H.B. Scholthof, K.B. Scholthof, A.O. Jackson, Plant Cell 7, 1157 (1995)

    Article  CAS  Google Scholar 

  19. K.B. Scholthof, H.B. Scholthof, A.O. Jackson, Virology 211, 324 (1995)

    Article  CAS  Google Scholar 

  20. H.B. Scholthof, B. Desvoyes, J. Kuecker, E. Whitehead, Mol. Plant Microbe Interact. 12, 670 (1999)

    Article  CAS  Google Scholar 

  21. R. Omarov, K. Sparks, L. Smith, J. Zindovic, H.B. Scholthof, J. Virol. 80, 3000 (2006)

    Article  CAS  Google Scholar 

  22. D. Silhavy, A. Molnar, A. Lucioli, G. Szittya, C. Hornyik, M. Tavazza, J. Burgyan, EMBO J. 21, 3070 (2002)

    Article  CAS  Google Scholar 

  23. J.M. Vargason, G. Szittya, J. Burgyan, T.M. Hall, Cell 115, 799 (2003)

    Article  CAS  Google Scholar 

  24. K. Ye, L. Malinina, D.J. Patel, Nature 426, 874 (2003)

    Article  CAS  Google Scholar 

  25. L. Lakatos, T. Csorba, V. Pantaleo, E.J. Chapman, J.C. Carrington, Y.P. Liu, V.V. Dolja, L.F. Calvino, J.J. Lopez-Moya, J. Burgyan, EMBO J. 25, 2768 (2006)

    Article  CAS  Google Scholar 

  26. I. Papp, M.F. Mette, W. Aufsatz, L. Daxinger, S.E. Schauer, A. Ray, J. van der Winden, M. Matzke, A.J. Matzke, Plant Physiol. 132, 1382 (2003)

    Article  CAS  Google Scholar 

  27. P. Dunoyer, C.H. Lecellier, E.A. Parizotto, C. Himber, O. Voinnet, Plant Cell 16, 1235 (2004)

    Article  CAS  Google Scholar 

  28. S.A. Siddiqui, C. Sarmiento, E. Truve, H. Lehto, K. Lehto, Mol. Plant Microbe Interact. 21, 178 (2008)

    Article  CAS  Google Scholar 

  29. I. Moore, L. Galweiler, D. Grosskopf, J. Schell, K. Palme, Proc. Natl. Acad. Sci. USA 95, 376 (1998)

    Article  CAS  Google Scholar 

  30. J.P. Alvarez, I. Pekker, A. Goldshmidt, E. Blum, Z. Amsellem, Y. Eshed, Plant Cell 18, 1134 (2006)

    Article  CAS  Google Scholar 

  31. A.P. Gleave, Plant Mol. Biol. 20, 1203 (1992)

    Article  CAS  Google Scholar 

  32. S. McCormick, Transformation of Tomato with Agrobacterium tumefaciens (Kluwer Academic Publishers, Dordrecht, The Netherlands, 1991)

    Book  Google Scholar 

  33. G.S. Pall, C. Codony-Servat, J. Byrne, L. Ritchie, A. Hamilton, Nucleic Acids Res. 35, e60 (2007)

    Article  Google Scholar 

  34. K.R. Markham, D.J. Ofman, Phytochemistry 34, 679 (1993)

    Article  CAS  Google Scholar 

  35. S. Moxon, R. Jing, G. Szittya, F. Schwach, R.L. Rusholme Pilcher, V. Moulton, T. Dalmay, Genome Res. 18, 1602 (2008)

    Article  CAS  Google Scholar 

  36. Y.C. Hsieh, R.T. Omarov, H.B. Scholthof, J. Virol. 83, 2188 (2009)

    Article  CAS  Google Scholar 

  37. M.L. Alvarez, H.L. Pinyerd, E. Topal, G.A. Cardineau, Plant Mol. Biol. 68, 61 (2008)

    Article  CAS  Google Scholar 

  38. D.S. Sutic, R.E. Ford, M.T. Tosic, Handbook of Plant Virus Diseases (CRC Press, Boca Raton, 1999)

    Google Scholar 

  39. L. Lakatos, G. Szittya, D. Silhavy, J. Burgyan, EMBO J. 23, 876 (2004)

    Article  CAS  Google Scholar 

  40. B. Yu, E.J. Chapman, Z. Yang, J.C. Carrington, X. Chen, FEBS Lett. 580, 3117 (2006)

    Article  CAS  Google Scholar 

  41. Y. Berger, S. Harpaz-Saad, A. Brand, H. Melnik, N. Sirding, J.P. Alvarez, M. Zinder, A. Samach, Y. Eshed, N. Ori, Development 136, 823 (2009)

    Article  CAS  Google Scholar 

  42. T. Blein, A. Pulido, A. Vialette-Guiraud, K. Nikovics, H. Morin, A. Hay, I.E. Johansen, M. Tsiantis, P. Laufs, Science 322, 1835 (2008)

    Article  CAS  Google Scholar 

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Acknowledgments

We thank the labs of Ori N. and Eshed Y. for their help with the pOp/LhG4 plasmids and for the tomato transformation guidance. We thank Ovadia R. for the anthocyanin measurements. We thank A. Gal-On for critical reading of the manuscript. This is contribution no. 130/2009 series, from the Agricultural Research Organization, the Volcani Center, Israel.

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Authors and Affiliations

  1. Institute of Plant Sciences, Agricultural Research Organization, The Volcani Center, P.O. Box 6, 50250, Bet Dagan, Israel

    Ran Stav, Anat Hendelman, Kobi Buxdorf & Tzahi Arazi

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  1. Ran Stav
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  2. Anat Hendelman
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  3. Kobi Buxdorf
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  4. Tzahi Arazi
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Correspondence to Tzahi Arazi.

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Control and P19HA-expressing fruit phenotypes. Representative whole and halved red tomato fruits of greenhouse-grown 35S:LhG4 and 35SÂ>>P19HA F1 plants (as indicated). Bar = 2.5 cm (TIF 15100 kb)

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Stav, R., Hendelman, A., Buxdorf, K. et al. Transgenic expression of tomato bushy stunt virus silencing suppressor P19 via the pOp/LhG4 transactivation system induces viral-like symptoms in tomato. Virus Genes 40, 119–129 (2010). https://doi.org/10.1007/s11262-009-0415-5

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