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Virus-induced Gene Silencing as Tool for Functional Genomics in a Glycine max

  • Jeong, Rae-Dong (Department of Agricultural Biotechnology and Center for Plant Molecular Genetics and Breeding Research, Seoul National University) ;
  • Hwang, Sung-Hyun (Department of Agricultural Biotechnology and Center for Plant Molecular Genetics and Breeding Research, Seoul National University) ;
  • Kang, Sung-Hwan (Department of Agricultural Biotechnology and Center for Plant Molecular Genetics and Breeding Research, Seoul National University) ;
  • Choi, Hong-Soo (Plant Pathology Div., National Institute of Agricultural Science and Technology, RDA) ;
  • Park, Jin-Woo (Plant Pathology Div., National Institute of Agricultural Science and Technology, RDA) ;
  • Kim, Kook-Hyung (Department of Agricultural Biotechnology and Center for Plant Molecular Genetics and Breeding Research, Seoul National University)
  • Published : 2005.01.01

Abstract

Virus-induced gene silencing (VIGS) is a recently developed gene transcript suppression technique for characterizing the function of plant genes. However, efficient VIGS has only been studied in a few plant species. In order to extend the application of VIGS, we examined whether a VIGS vector based on TRV would produce recognizable phenotypes in soybean. Here, we report that VIGS using the Tobacco rattle virus (TRV) viral vector can be used in several soybean cultivars employing various agro-inoculation methods including leaf infiltration, spray inoculation, and agrodrench. cDNA fragments of the soybean phytoene desaturase(PDS) was inserted into TRV RNA-2 vector. By agrodrench, we successfully silenced the expression of PDS encoding gene in soybean. The silenced phenotype of PDS was invariably obvious 3 weeks after inoculation with the TRV-based vector. Real-time RT-PCR analyses showed that the endogenous level of GmPDS transcripts was dramatically reduced in the silenced leaf tissues. These observations confirm that the silenced phenotype is closely correlated with the pattern of tissue expression. The TRV-based VIGS using agrodrench can be applied to functional genomics in a soybean plants to study genes involved in a wide range of biological processes. To our knowledge, this is the first high frequency VIGS method in soybean plants.

References

  1. Andersen, S. U., Cvitanich, C., Hougaard, B. K., Roussis, A., Gronlund, M., Jensen, D. B., Frokjaer, L. A. and Jensen, E. O. 2003. The glucocorticoid-inducible GVG system causes severe growth defects in both root and shoot of the model legume Lotus japonicus. Mol. Plant-Microbe Interact. 16:1069-1076 https://doi.org/10.1094/MPMI.2003.16.12.1069
  2. Baulcombe, D. C. 1999. Fast forward genetics based on virusinduced gene silencing. Curr. Opin. Plant Biol. 2: 109-113 https://doi.org/10.1016/S1369-5266(99)80022-3
  3. Brigneti, G., Martin-Hernandez, A.M., Jin, H., Chen, J., Baulcombe, D. C., Baker, B. and Jones, J. D. 2004. Virus-induced gene silencing in Solanum species. Plant J. 39: 264-272 https://doi.org/10.1111/j.1365-313X.2004.02122.x
  4. Chung, E., Seong, E., Kim, Y., C., Chung, E., Oh, S. K., Lee, S., Park, J.-M. and Choi, D. 2004. A method of high frequency virus-induced gene silencing in chill pepper (Capsicum annuum L. cv. Bukang). Mol. Cells 17:377-385
  5. Dalmay, T., Hamilton, A, Mueller, E. and Baulcombe, D. C. 2000. Potato virus X amplicons in arabidopsis mediate genetic and epigenetic gene silencing. Plant Cell 12:369-379 https://doi.org/10.1105/tpc.12.3.369
  6. Demmig-Adams, B., Gilmore, A M. and Adams, W W, III. 1996. Carotenoids 3: in vivo function of carotenoids in higher plants. FASEB J. 10:41-45
  7. Dinesh-Kumar, S. P., Anandalakshmi, R., Marathe, R., Schiff, M. and Liu, Y. 2003. Virus-induced gene silencing. Methods Mol. Biol. 236:287-294
  8. Ekengren, S. K., Liu, Y., Schiff, M., Dinesh-Kumar, S. P. and Martin, G B. 2003. Two MAPK cascades, NPR1, and TGA transcription factors playa role in Pto-mediated disease resistance in tomato. Plant J. 36:905-917 https://doi.org/10.1046/j.1365-313X.2003.01944.x
  9. English, J. D. and Sweatt, J. D. 1997. A requirement for the mitogen-activated protein kinase cascade in hippocampal long term potentiation. J. Biol. Chem. 272: 19103-19706 https://doi.org/10.1074/jbc.272.31.19103
  10. Gabriela, D., Britta N., Stuart, A., Mogens, N., Ida, E. J., Elisabeth J. and Ole, S. 2004. Virus-induced gene silencing as a tool for functional genomics in a legume species. Plant J. 40:622-631 https://doi.org/10.1111/j.1365-313X.2004.02233.x
  11. Holzberg, S., Brosio, P., Gross, C. and Pogue, G. P. 2002. Barley stripe mosaic virus-induced gene silencing in a monocot plant. Plant J. 30:315-327 https://doi.org/10.1046/j.1365-313X.2002.01291.x
  12. Jeong, R.-D., Lim, W-S., Kwon, S.-W. and Kim, K.-H. 2005. Identification of Glycine max genes expressed in response to Soybean mosaic virus infection. Plant Pathol. J. 21 :47-54 https://doi.org/10.5423/PPJ.2005.21.1.047
  13. Jin, H., Axtell, M. J., Dahlbeck, D., Ekwenna, O., Zhang, S., Staskawicz, B. and Baker, B. 2002. NPK1, an MEKK1-like mitogen-activated protein kinase kinase kinase, regulates innate immunity and development in plants. Dev. Cell 3:291-297 https://doi.org/10.1016/S1534-5807(02)00205-8
  14. Kumagai, H. and Kouchi, H. 2003. Gene silencing by expression of hairpin RNA in Lotus japonicus roots and root nodules. Mol. Plant Microbe Interact. 16:663-668 https://doi.org/10.1094/MPMI.2003.16.8.663
  15. Lindbo, J. A., Fitzmaurice, W. P. and Della-Cioppa, G. 2001. Virus-mediated reprogramming of gene expression in plants. Curr. Opin. Plant Biol. 4: 181-185 https://doi.org/10.1016/S1369-5266(00)00158-8
  16. Liu, Y., Schiff, M. and Dinesh-Kumar, S. P. 2002a. Virus-induced gene silencing in tomato. Plant J. 31 : 777-786 https://doi.org/10.1046/j.1365-313X.2002.01394.x
  17. Liu, Y., Schiff, M., Marathe, R. and Dinesh-Kumar, S. P. 2002b. Tobacco Rarl, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to Tobacco mosaic virus. Plant J. 415-249
  18. Lu, R., Martin-Hernandez, A. M., Peart, J. R., Malcuit, I. and Baulcombe, D. C. 2003. Virus-induced gene silencing in plants. Methods 30:296-303 https://doi.org/10.1016/S1046-2023(03)00037-9
  19. Matthews, R. E. F. 1991. Plant Virology, 3rd ed. San Diego: Academic Press
  20. MacFarlane, S. A. and Popovich, A. H. 2000. Efficient expression of foreign proteins in roots from tobravirus vectors. Virology 267:29-35 https://doi.org/10.1006/viro.1999.0098
  21. Peart, J. R., Cook, G., Feys, B. J., Parker, J. E. and Baulcombe, D. C. 2002. An EDS1 orthologue is required for N-mediated resistance against tobacco mosaic virus. Plant J. 29:569-579 https://doi.org/10.1046/j.1365-313X.2002.029005569.x
  22. Ratcliff, F., Martin-Hernandez, A. M. and Baulcombe, D. C. 2001. Technical Advance. Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J. 25:237-245 https://doi.org/10.1046/j.0960-7412.2000.00942.x
  23. Ryu, C. M., Anand, A., Kang, L. and Mysore, K. S. 2004. Agrodrench: a novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse Solanaceous species. Plant J. 40:322-331 https://doi.org/10.1111/j.1365-313X.2004.02211.x
  24. Sharma, P. C., Ito, A., Shimizu, T., Terauchi, R., Kamoun, S. and Saitoh, H. 2003. Virus- I induced silencing of WIPK and SIPK genes reduces resistance to a bacterial pathogen, but has no effect on the INF1-induced hypersensitive response (HR) in Nicotiana benthamiana. Mol. Genet. Genomics 269:583-591 https://doi.org/10.1007/s00438-003-0872-9
  25. Slaymaker, D. H., Navarre, D. A., Clark, D., del Pozo, O., Martin, G. B. and Klessig, D. F. 2002. The tobacco salicylic acid-binding protein 3 (SABP3) is the chloroplast carbonic anhydrase, which exhibits antioxidant activity and plays a role in the hypersensitive defense response. Proc. Natl. Acad. Sci. USA 99:11640-11645 https://doi.org/10.1073/pnas.182427699
  26. Stiller, J., Martirani, L., Tuppale, S., Chian, R., Chiurazzi, M. and Gresshoff, P. 1997. High frequency transformation and regeneration of transgenic plants in the model legume Lotus japonicus. J. Exp. Bot. 48:1357-1365 https://doi.org/10.1093/jxb/48.7.1357
  27. Trieu, A. T., Burleigh, S. H. , Kardailsky, I. V, Maldonado-Mendoza, I. E., Versaw, W. K., Blaylock, L. A., Shin, H., Chiou, T. J., Katagi, H., Dewbre, G.R., Weigel, D. and Harrison, M. J. 2000. Transformation of Medicago truncatula via infiltration of seedlings or flowering plants with Agrobacterium. Plant J. 22:531-541 https://doi.org/10.1046/j.1365-313x.2000.00757.x
  28. Turnage, M. A., Muangsan, N., Peele, C. G. and Robertson, D. 2002. Geminivirus-based vectors for gene silencing in Arabidopsis. Plant J. 30: 107 -114 https://doi.org/10.1046/j.1365-313X.2002.01261.x
  29. Visser, P. B. and Bol, J. F. 1999. Nonstructural proteins of Tobacco rattle virus which have a role in nematode-transmission: expression pattern and interaction with viral coat protein. J. Gen. Virol. 80:3273-3280 https://doi.org/10.1099/0022-1317-80-12-3273

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