Skip to main content

Mungbean yellow mosaic virus (MYMV) AC4 suppresses post-transcriptional gene silencing and an AC4 hairpin RNA gene reduces MYMV DNA accumulation in transgenic tobacco

Abstract

Mungbean yellow mosaic virus (MYMV) is a legume-infecting geminivirus that causes yellow mosaic disease in blackgram, mungbean, soybean, Frenchbean and mothbean. AC4/C4, which is nested completely within the Rep gene, is less conserved among geminiviruses. Much less is known about its role in viral pathogenesis other than its known role in the suppression of host-mediated gene silencing. Transient expression of MYMV AC4 by agroinfiltration suppressed post-transcriptional gene silencing in Nicotiana benthamiana 16c expressing green fluorescence protein, at a level comparable to MYMV TrAP expression. AC4 full-length gene and an inverted repeat of AC4 (comprising the full-length AC4 sequence in sense and antisense orientations with an intervening intron) which makes a hairpin RNA (hpRNA) upon transcription were introduced into tobacco by Agrobacterium-mediated leaf disc transformation. Leaf discs of the transgenic plants were agroinoculated with partial dimers of MYMV and used to study the effect of the AC4-sense and AC4 hpRNA genes on MYMV DNA accumulation. Leaf discs of two transgenic plants that express the AC4-sense gene displayed an increase in MYMV DNA accumulation. Leaf discs of six transgenic plants containing the AC4 hpRNA gene accumulated small-interfering RNAs (siRNAs) specific to AC4, and upon agroinoculation with MYMV they exhibited a severe reduction in the accumulation of MYMV DNA. Thus, the MYMV AC4 hpRNA gene has emerged as a good candidate to engineer resistance against MYMV in susceptible plants.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. 1.

    L. Hanley-Bowdoin, S.B. Settlage, B.M. Orozco, S. Nagar, D. Robertson, Crit. Rev. Plant Sci. 18, 71–106 (1999)

    Article  CAS  Google Scholar 

  2. 2.

    M.R. Rojas, C. Hagen, W.J. Lucas, R.L. Gilbertson, Annu. Rev. Phytopathol. 43, 361–394 (2005)

    PubMed  Article  CAS  Google Scholar 

  3. 3.

    C.M. Fauquet, R.W. Briddon, J.K. Brown, E. Moriones, J. Stanley, M. Zerbini, X. Zhou, Arch. Virol. 153, 783–821 (2008)

    PubMed  Article  CAS  Google Scholar 

  4. 4.

    A.S. Karthikeyan, R. Vanitharani, V. Balaji, S. Anuradha, P. Thillaichidambaram, P.V. Shivaprasad, C. Parameswari, V. Balamani, M. Saminathan, K. Veluthambi, Arch. Virol. 149, 1643–1652 (2004)

    PubMed  Article  CAS  Google Scholar 

  5. 5.

    A. Varma, V.G. Malathi, Ann. Appl. Biol. 142, 145–164 (2003)

    Article  CAS  Google Scholar 

  6. 6.

    J.S. Elmer, L. Brand, G. Sunter, W.E. Gardiner, D.M. Bisaro, S.G. Rogers, Nucleic Acids Res. 16, 7043–7060 (1988)

    PubMed  Article  CAS  Google Scholar 

  7. 7.

    W. Pooma, I.T.D. Petty, J. Gen. Virol. 77, 1947–1951 (1996)

    PubMed  Article  CAS  Google Scholar 

  8. 8.

    Y.K. Sung, R.H.A. Coutts, J. Gen. Virol. 76, 2809–2815 (1995)

    PubMed  Article  CAS  Google Scholar 

  9. 9.

    R.A. Hoogstraten, S.F. Hanson, D.P. Maxwell, Mol. Plant Microbe Interact. 9, 594–599 (1996)

    PubMed  Article  CAS  Google Scholar 

  10. 10.

    R. VanWezel, X. Dong, P. Blake, J. Stanley, Y. Hong, Mol. Plant Pathol. 3, 461–471 (2002)

    Article  CAS  Google Scholar 

  11. 11.

    J. Stanley, J.R. Latham, Virology 190, 506–509 (1992)

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    L.R. Krake, M.A. Rezaian, I.B. Dry, Mol. Plant Microbe Interact. 11, 413–417 (1998)

    Article  CAS  Google Scholar 

  13. 13.

    J.E. Rigden, L.R. Krake, M.A. Rezaian, I.B. Dry, Virology 204, 847–850 (1994)

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    L.A. Selth, J.W. Randles, M.A. Rezaian, Mol. Plant Microbe Interact. 17, 27–33 (2004)

    PubMed  Article  CAS  Google Scholar 

  15. 15.

    M. Saeed, S. Mansoor, M.A. Rezaian, R.W. Briddon, J.W. Randles, Arch. Virol. 153, 1367–1372 (2008)

    PubMed  Article  CAS  Google Scholar 

  16. 16.

    I. Jupin, F. De Kouchkovsky, F. Jouanneau, B. Gronenborn, Virology 204, 82–90 (1994)

    PubMed  Article  CAS  Google Scholar 

  17. 17.

    K. Teng, H. Chen, J. Lai, Z. Zhang, Y. Fang, R. Xia, X. Zhou, H. Guo, Q. Xie, PLoS ONE 5, e11280 (2010)

    PubMed  Article  Google Scholar 

  18. 18.

    K. Mills-Lujan, C.M. Deom, Protoplasma 239, 95–110 (2010)

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    N. Piroux, K. Saunders, A. Page, J. Stanley, Virology 362, 428–440 (2007)

    PubMed  Article  CAS  Google Scholar 

  20. 20.

    J. Lai, H. Chen, K. Teng, Q. Zhao, Z. Zhang, Y. Li, L. Liang, R. Xia, Y. Wu, H. Guo, Q. Xie, Plant J. 57, 905–917 (2009)

    PubMed  Article  CAS  Google Scholar 

  21. 21.

    J. Park, H.S. Hwang, K.J. Buckley, J.B. Park, C.K. Auh, D.G. Kim, S. Lee, K.R. Davis, Plant Cell Rep. 29, 1377–1389 (2010)

    PubMed  Article  CAS  Google Scholar 

  22. 22.

    R. Vanitharani, P. Chellappan, J.S. Pita, C.M. Fauquet, J. Virol. 78, 9487–9498 (2004)

    PubMed  Article  CAS  Google Scholar 

  23. 23.

    P. Chellappan, R. Vanitharani, C.M. Fauquet, Proc. Natl. Acad. Sci. USA. 102, 10381–10386 (2005)

    PubMed  Article  CAS  Google Scholar 

  24. 24.

    V.F. Fondong, R.V.C. Reddy, C. Lu, B. Hankoua, C. Felton, K. Czymmek, F. Achenjang, Mol. Plant Microbe Interact. 20, 380–391 (2007)

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    P. Gopal, P.P. Kumar, B. Sinilal, J. Jose, K.A. Yadunandam, R. Usha, Virus Res. 123, 9–18 (2007)

    PubMed  Article  CAS  Google Scholar 

  26. 26.

    S.C. Dogra, O. Eini, M.A. Rezaian, J.W. Randles, Plant Mol. Biol. 71, 25–38 (2009)

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    M.T. Ruiz, O. Voinnet, D.C. Baulcombe, Plant Cell 10, 937–946 (1998)

    PubMed  CAS  Google Scholar 

  28. 28.

    R.P. Hellens, E.A. Edwards, N.R. Leyland, S. Bean, P.M. Mullineaux, Plant Mol. Biol. 42, 819–832 (2000)

    PubMed  Article  CAS  Google Scholar 

  29. 29.

    T. Komari, Y. Hiei, Y. Saito, N. Murai, T. Kumashiro, Plant J. 10, 165–174 (1996)

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    R. Rajeswaran, S. Sunitha, P.V. Shivaprasad, M.M. Pooggin, T. Hohn, K. Veluthambi, Mol. Plant Microbe Interact. 20, 1545–1554 (2007)

    PubMed  Article  CAS  Google Scholar 

  31. 31.

    S. Sunitha, P.V. Shivaprasad, K. Sujata, K. Veluthambi, Plant Mol. Biol. Rep. 30, 158–167 (2012)

    Article  CAS  Google Scholar 

  32. 32.

    C. Llave, K.D. Kasschau, J.C. Carrington, Proc. Natl. Acad. Sci. USA 97, 13401–13406 (2000)

    PubMed  Article  CAS  Google Scholar 

  33. 33.

    G. Sunilkumar, K. Vijayachandra, K. Veluthambi, Plant Sci. 141, 51–58 (1999)

    Article  CAS  Google Scholar 

  34. 34.

    T. Murashige, F. Skoog, Physiol. Plant. 15, 473–497 (1962)

    Article  CAS  Google Scholar 

  35. 35.

    S.O. Rogers, A.J. Bendich, in Plant Molecular Biology Manual, vol.D1, ed. by S.B. Gelvin, R.A. Schilperoort (Kluwer Academic Publishers, Dordrecht, 1994), pp. 1–8

    Google Scholar 

  36. 36.

    Y. Hong, J. Stanley, Mol. Plant Microbe Interact. 9, 219–225 (1996)

    Article  CAS  Google Scholar 

  37. 37.

    K. Pawlowski, R. Kunze, S. DeVries, T. Bisseling, in Plant Molecular Biology Manual, vol. D5, ed. by S.B. Gelvin, R.A. Schilperoort (Kluwer Academic Publishers, Dordrecht, 1994), pp. 1–13

    Google Scholar 

  38. 38.

    S.S. Jacob, R. Vanitharani, A.S. Karthikeyan, Y. Chinchore, P. Thillaichidambaram, K. Veluthambi, Plant Dis. 87, 247–251 (2003)

    Article  CAS  Google Scholar 

  39. 39.

    P.V. Shivaprasad, P. Thillaichidambaram, V. Balaji, K. Veluthambi, Virus Genes 33, 365–374 (2006)

    PubMed  Article  CAS  Google Scholar 

  40. 40.

    D. Trinks, R. Rajeswaran, P.V. Shivaprasad, R. Akbergenov, E.J. Oakeley, K. Veluthambi, T. Hohn, M.M. Pooggin, J. Virol. 79, 2517–2527 (2005)

    PubMed  Article  CAS  Google Scholar 

  41. 41.

    Y. Hong, K. Saunders, J. Stanley, Virology 228, 383–387 (1997)

    PubMed  Article  CAS  Google Scholar 

  42. 42.

    G. Sunter, J.L. Sunter, D.M. Bisaro, Virology 285, 59–70 (2001)

    PubMed  Article  CAS  Google Scholar 

  43. 43.

    R. van Wezel, X. Don, H. Liu, P. Tien, J. Stanley, Y. Hong, Mol. Plant Microbe Interact. 15, 203–208 (2002)

    Article  Google Scholar 

  44. 44.

    L. Wang, S.J. Brown, Nucleic Acids Res. 34, W243–W248 (2006)

    PubMed  Article  CAS  Google Scholar 

  45. 45.

    A.S. Karthikeyan, K.S. Sarma, K. Veluthambi, Plant Cell Rep. 15, 328–331 (1996)

    Article  CAS  Google Scholar 

  46. 46.

    R. Saini, S. Jaiwal, P.K. Jaiwal, Plant Cell Rep. 21, 851–859 (2003)

    PubMed  CAS  Google Scholar 

  47. 47.

    S. Sunitha, D. Marian, B. Hohn, K. Veluthambi, Virus Genes 43, 445–453 (2011)

    PubMed  Article  CAS  Google Scholar 

  48. 48.

    M. Prins, M. Laimer, E. Noris, J. Schubert, M. Wassenegger, M. Tepfer, Mol. Plant Pathol. 9, 73–83 (2008)

    PubMed  CAS  Google Scholar 

  49. 49.

    D.N. Shepherd, D.P. Martin, J.A. Thomson, Plant Sci. 176, 1–11 (2009)

    Article  CAS  Google Scholar 

  50. 50.

    H. Vanderschuren, M. Stupak, J. Futterer, W. Gruissem, P. Zhang, Plant Biotechnol. J. 5, 207–220 (2007)

    PubMed  Article  CAS  Google Scholar 

  51. 51.

    Z. Luo, Z. Chen, Plant Cell 19, 943–958 (2007)

    PubMed  Article  CAS  Google Scholar 

  52. 52.

    A.B. Rose, R.L. Last, Plant J. 11, 455–464 (1997)

    PubMed  Article  CAS  Google Scholar 

  53. 53.

    P. Pandey, N.C. Choudhury, S.K. Mukherjee, Virology J. 6, 152 (2009)

    Article  Google Scholar 

  54. 54.

    R. Anandalakshmi, R. Marathe, X. Ge, J.M. Jr Herr, C. Mau, A. Mallory, G. Pruss, L. Bowman, V.B. Vance, Science 290, 142–144 (2000)

    PubMed  Article  CAS  Google Scholar 

  55. 55.

    S.A. Siddiqui, C. Sarmiento, E. Truve, H. Lehto, K. Lehto, Mol. Plant Microbe Interact. 21, 178–187 (2008)

    PubMed  Article  CAS  Google Scholar 

  56. 56.

    K. Bonfim, J.C. Faria, E.O.P. Nogueira, E.A. Mendes, F.J. Araga˜o, Mol. Plant Microbe Interact. 20, 717–726 (2007)

    PubMed  Article  CAS  Google Scholar 

  57. 57.

    A. Fuentes, P.L. Ramos, E. Fiallo, D. Callard, Y. Sanchez, R. Peral, R. Rodriguez, M. Pujol, Transgenic Res. 15, 291–304 (2006)

    PubMed  Article  CAS  Google Scholar 

  58. 58.

    S.V. Ramesh, A.K. Mishra, S. Praveen, Oligonucleotide 17, 251–257 (2007)

    Article  CAS  Google Scholar 

  59. 59.

    H. Vanderschuren, A. Alder, P. Zhang, W. Gruissem, Plant Mol. Biol. 70, 265–272 (2009)

    PubMed  Article  CAS  Google Scholar 

  60. 60.

    M. Pooggin, P.V. Shivaprasad, K. Veluthambi, T. Hohn, Nat. Biotechnol. 21, 131–132 (2003)

    PubMed  Article  CAS  Google Scholar 

  61. 61.

    H. Vanderschuren, R. Akbergenov, M.M. Pooggin, T. Hohn, W. Gruissem, P. Zhang, Plant Mol. Biol. 64, 549–557 (2007)

    PubMed  Article  CAS  Google Scholar 

  62. 62.

    A. Zrachya, P.P. Kumar, U. Ramakrishnan, Y. Levy, A. Loyter, T. Arazi, M. Lapidot, Y. Gafni, Transgenic Res. 16, 385–398 (2007)

    PubMed  Article  CAS  Google Scholar 

  63. 63.

    S. Praveen, S.V. Ramesh, A.K. Mishra, V. Koundal, P. Palukaitis, Transgenic Res. 19, 45–55 (2010)

    PubMed  Article  CAS  Google Scholar 

  64. 64.

    S.V. Wesley, C.A. Helliwell, N.A. Smith, M.B. Wang, D.T. Rouse, Q. Liu, P.S. Gooding, S.P. Singh, D. Abbott, P.A. Stoutjesdijk, S.P. Robinson, A.P. Gleave, A.G. Green, P.M. Waterhouse, Plant J. 27, 581–590 (2001)

    PubMed  Article  CAS  Google Scholar 

  65. 65.

    A.P. Gleave, Plant Mol. Biol. 20, 1203–1207 (1992)

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Prof. Thomas Hohn, University of Basel for providing Nicotiana benthamiana 16c seeds. We thank Dr. Donna Bond, University of Cambridge for her comments on the manuscript. We are thankful to Dr. K. Dharmalingam, Madurai Kamaraj University for permitting us to use the Liquid Scintillation Counter. Ms. Jasvinder Kaur and Ms. K. Bhagyalakshmi are acknowledged for the construction of pCAM-AC4-gus. This study was financially supported by the Department of Biotechnology (Project Number-BT/PR9823/AGR/36/10/2007 dt 7/12/2007), Government of India and University Grants Commission, Government of India. S.S. acknowledges the Council of Scientific and Industrial Research, Government of India for the research fellowship.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Karuppannan Veluthambi.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (TIFF 430 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sunitha, S., Shanmugapriya, G., Balamani, V. et al. Mungbean yellow mosaic virus (MYMV) AC4 suppresses post-transcriptional gene silencing and an AC4 hairpin RNA gene reduces MYMV DNA accumulation in transgenic tobacco. Virus Genes 46, 496–504 (2013). https://doi.org/10.1007/s11262-013-0889-z

Download citation

Keywords

  • Geminivirus resistance
  • Hairpin RNA
  • Legume-infecting begomovirus
  • RNA silencing
  • Silencing suppression
  • siRNA