The efficiency of the partial genome of Turnip mosaic virus (TuMV) on resistance induction in canola

Document Type : Research Paper

Authors

1 Former Ph. D. Student, Plant Pathology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

2 Associate Professor, Plant Breeding Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

3 Professor, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

Abstract

Turnip mosaic virus (TuMV) a species of the genus Potyvirus in the family Potyviridae, is one of the most prevalent viruses of canola fields in Iran. So far, commercial resistant lines or varieties against TuMV have not been known in Iran. In this research, the possibility of inducing resistant to TuMV in transgenic canola lines carrying a short fragment of the coat protein gene of the virus was evaluated. Therefore, a 130 nucleotide fragment of TuMV coat protein gene, as sense or antisense orientations, were cloned in the pFGC5941 vector independently and then were transformed to canola cotyledonary (Hayola R-Line 401 variety) explants using Agrobacterium tumefaciens strain LBA4404. Selection and confirmation of transgenic T0 and T1 lines were carried out via PCR and glufosinate ammonium herbicide (selective marker) resistance assay under greenhouse conditions. After virus inoculation to T1 lines, resistance assay was done by the scoring system. The obtained results confirmed that transgenic lines with sense construction did not show any resistant. However, several transgenic lines with antisense construction showed a different level of resistant to TuMV including delay in symptoms appearance and a decrease in symptoms severity (0-30% of T1 progeny) or symptoms recovery (10-66% of T1 progeny). This is the first report of increased resistant against TuMV via transformation of a short viral fragment sequence with the antisense orientation in canola.

Keywords


  1. Baulcombe, D. C. (1996). Mechanisms of pathogen-derived resistance to viruses in transgenic plants. Plant Cell, 8, 1833-1844.
  2. Brodersen, P. & Voinnet, O. (2006). The diversity of RNA silencing pathways in plants. Trends in Genetics,22, 268-280.
  3. Dellaporta, S. L., Wood, J. & Hicks, J. B. (1983). A plant DNA minipreparation: Version II. Plant Molecular Biology Reporter, 1, 19-21.
  4. Ding, S-W. & Voinnet, O. (2007). Antiviral immunity directed by small RNAs. Cell, 130, 413–426.
  5. Farzadfar, S., Ohshima, K., Pourrahim, R., Golnaraghi, A. R., Jalali, S. & Ahoonmanesh, A.  (2005). Occurrence of Turnip mosaic virus on ornamental crops in Iran. Plant Pathology, 54, 261.
  6. Ghorbani, S., Shahraeen, N., Dehghanyar, H., Sahandi, A. & Pourrahim, R. (2007). Serological identification and Purification of Turnip mosaic virus. (TuMV) in the oil seed rape. Iranian Journal of Biology, 20, 61-71.
  7. Goldbach, R., Bucher, E. & Prins, M. (2003). Resistance mechanisms to plant viruses: an overview. Virus Research, 92, 207-212.
  8. Hu, Q., Niu, Y., Zhang, K., Liu, Y. & Zhou, X. (2011). Virus-derived transgenes expressing hairpin RNA give immunity to Tobacco mosaic virus and Cucumber mosaic virus. Virology Journal, 8, 41.
  9. Jafari, M., Shams-Bakhsh, M. & Moieni, A. (2016). Reaction of commercial canola varieties and lines against Turnip mosaic virus (TuMV) isolate. Iranian Journal of Plant Pathology, 52, 147-159.
  10. Jan, F.J. Fagoaga, C., Pang, S.-Z. & Gonsalves, D. (2000). A single chimeric transgene drived from two distinct viruses confers multi-virus resistance in transgenic plants through homology-dependent gene silencing. Journal of General Virology, 81, 2103-2109.
  11. Jan, F. J., Pang, S. Z., Fagoaga, C. & Gonsalves, D. (1999).  Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post-transcriptional gene silencing. Transgenic Research, 8, 203-213.
  12. Lindbo, J. A. & Dougherty, W. G. (1992a). Pathogen-derived resistance to a potyvirus: immune and resistant phenotypes in transgenic tobacco expressing altered forms of a potyvirus coat nucleotide sequence. Molecular Plant–Microbe Interactions, 5, 144-153.
  13. Lindbo, J. A. & Dougherty, W. G. (1992b) Untranslatable transcripts of the tobacco etch virus coat protein gene sequence can interfere with tobacco etch virus replication in transgenic plants and protoplasts. Virology, 189, 725-733.
  14. Moloney, M. M., Walker, J. M. & Sharma, K. K. (1989). High efficiency transformation of Brassica napus using Agrobacterium vectors. Plant Cell Reports, 8, 238-242.
  15. Nomura, K., Ohshima, K., Anai, T., Uekusa, H. & Kita, N. (2004). RNA silencing of the introduced coat protein gene of Turnip mosaic virus confers broad-spectrum resistance in transgenic Arabidopsis. Phytopathology, 94, 730-736.
  16. Prins, M., Laimer, M., Noris, E., Schubert, J., Wassenegger, M. & Tepfer, M. (2008). Strategies for antiviral resistance in transgenic plants. Molecular Plant Pathology, 9, 73-83.
  17. Roohi, L., Zamani, M. R. & Motallebi, M. (2013). Transgenic canola plants harboring beta 1, 3 glucanase (bgnI) gene from Trichoderma virens inhibit mycelial growth of Sclerotinia sclerotiorum. Iranian Journal of Biology, 26, 28-40.
  18. Sabokkhiz, M. A., Jafarpour, B., Shahriari, A. & Tarighi, S. (2012).  Identification of Turnip mosaic virus isolated from canola in northeast area of Iran. African Journal of Biotechnology, 11, 14553-14560.
  19. Sambrook, J. & Russell, D. W. (2001). Molecular cloning: a laboratory manual. 3rd ed., Cold Spring Harbor Laboratory Press, New York. 2100 pp.
  20. Savenkov, E. I. & Valkonen, J. P. T. (2001). Coat protein gene-mediated resistance to Potato virus A in transgenic plants is suppressed following infection with another potyvirus. Journal of General virology, 82, 2275-2278.
  21. Shahraeen, N. (2012). An overview of oilseed rape (canola) virus diseases in Iran. International Research Journal of Microbiology, 3, 24-28.
  22. Shahraeen, N., Farzadfar, S. & Lesemann, D.E. (2003). Incidence of viruses infecting winter oilseed rape (Brassica napus ssp. oleifera) in Iran. Journal of Phytopathology, 151, 614-616
  23. Simon-Mateo, C. and Garcia, J. A. (2011). Antiviral sterategies in plants based on RNA silencing. Biochimica et Biophysica Acta, 1809, 722-731.
  24. Stam, M., Mol, J. N. M. & Kooter, J. M. (1997). Review Article: The Silence of Genes in Transgenic Plants, Annals of Botany, 79, 3-12.
  25. Vollmann, J. & Rajkan, I. (2009). Oil crops. Speringer. 548 pp.
  26. Waterhouse, P. M. & Upadhyaya, M. N. (1998). Genetic engineering of virus resistance. In: Molecular biology of rice. Tokyo: Springer-Verlag, 257-281 pp.
  27. 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 of the United States of Americ, 95, 13959-13964.
  28. Watson, J. M. & Wang, M. (2012). Antiviral resistance in plants. Methods and prptocols. Humana Press. 380 pp.
  29. Zahedi Tabarestani, A., Shamsbakhsh, M. & Safaei, N. (2010). Distribution of three important aphid borne canola viruses in Golestan province, Iranian journal of Plant Protection Science, 41, 251-259.
  30. Zhandong, Y., Shuangyi, Z. & Qiwei, H. (2007). High level resistance to Turnip mosaic virus in Chinese cabbage (Brassica campestris ssp. pekinensis (Lour) Olsson) transformed with the antisense NIb gene using marker-free Agrobacterium tumefaciens inīŦltration. Plant Science, 172, 920-929.
  31. Zhao, S. & Hao, X. (2010). High level resistance to TuMV (Turnip mosaic virus) in transgenic Mustard with the Antisense NIb Gene of the Virus. (2010). Forth International Conference on Computational and Information Sciences, 1080-1082.