Abstract
Primary transformants of SR1 Nicotiana tabacum plants with RNA interference-based silencing of the gene for extracellular ribonuclease Nk1 were obtained. It was demonstrated that the profiles of ribonuclease activities of leaf protein extracts from these plants lacked ribonuclease with electrophoretic mobility corresponding to that of the Nk1 protein. Primary transformants did not differ phenotypically from control plants. They represent a new model for investigation of the biological role of extracellular ribonucleases, including the molecular mechanisms of resistance to pathogens.
References
Trifonova, E.A, Kochetov, A.V., and Shumnyi, V.K., The Role of Nucleases in Physiological Processes of Higher Plants, Usp. Sovrem. Biol., 2000, vol. 120, pp. 395–405.
Ye, Z. and Droste, D., Isolation and Characterization of cDNAs Encoding Xylogenesis-Associated and Wounding-Induced Ribonucleases in Zinnia elegans, Plant Mol. Biol., 1996, vol. 30, pp. 697–709.
Kariu, T., Sano, K., Shimokawa, H., et al., Isolation and Characterization of a Wound-Inducible Ribonuclease from Nicotiana glutinosa Leaves, Boisci. Biotechnol. Biochem., 1998, vol. 62, pp. 1144–1151.
Ohno, H. and Ehara, Y., Expression of Ribonuclease Gene in Mechanically Injured or Virus-Inoculated Nicotiana tabacum Leaves, Tohoku J. Agric. Res., 2005, vol. 55, pp. 99–109.
Galiana, E., Bonnet, P., Conrod, S., et al., RNase Activity Prevents the Growth of a Fungal Pathogen in Tobacco Leaves and Increases upon Induction of Systemic Acquired Resistance with Elicitin, Plant Physiol., 1997, vol. 115, pp. 1557–1567.
Begam, M., Narwal, S., Roy, S., et al., An Antiviral Protein Having Deoxyribonuclease and Ribonuclease Activity from Leaves of the Post-Flowering Stage of Celosia cristata, Biochemistry (Moscow), 2006, vol. 71, pp. 44–48.
Trifonova, E.A., Sapotsky, M.V., Komarova, M.L., et al., Protection of Transgenic Tobacco Plants Expressing Bovine Pancreatic Ribonuclease against Tobacco Mosaic Virus, Plant Cell Rep., 2007, vol. 26, pp. 1121–1126.
Sangaev, S.S., Trifonova, E.A., Titov, S.E., et al., Effective Expression of the Gene Encoding an Extracellular Ribonuclease of Zinnia elegans in the SR1 Nicotiana tabacum Plants, Russ. J. Genet., 2007, vol. 43, no. 2, pp. 1002–1005.
LeBrasseur, N.D., MacIntosh, G.C., Perez-Amador, M.A., et al., Local and Systemic Wound-Induction of RNase and Nuclease Activities in Arabidopsis: RNS1 as a Marker for a JA-Independent Systemic Signaling Pathway, Plant J., 2002, vol. 29, pp. 393–403.
Alting-Mees, M.A. and Short, J.M., pBluescript II: Gene Mapping Vectors, Nucleic Acid Res., 1987, vol. 17, p. 9494.
Alliotte, T., Zhu, L.H., Van Montagu, M., et al., Plant Expression Vectors with the Origin of Replication of the W-Type Plasmid Sa, Plasmid, 1988, vol. 19, pp. 251–254.
An, G., Ebert, P.R., Mitra, A., and Ha, S.B., Binary Vectors, Plant Molecular Biology: Manual, Gelvin, S.B., and Schilperoort, R.A., Eds., Kluwer, 1988, pp. 1–19.
Deblaere, R., Reynaerts, A., Hofte, H., et al., Vectors for Cloning in Plant Cells, Methods Enzymol., 1987, vol. 153, pp. 277–292.
Yen, Y. and Green, P.J., Identification and Properties of the Major Ribonucleases of Arabidopsis thaliana, Plant Physiol., 1991, vol. 97, pp. 1487–1493.
Bradford, M.M., A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Anal. Biochem., 1976, vol. 72, pp. 248–254.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © S.S. Sangaev, E.A. Trifonova, S.E. Titov, A.V. Romanova, Ya.S. Kolodyazhnaya, M.V. Sapotsky, V.I. Malinovsky, A.V. Kochetov, 2010, published in Genetika, 2010, Vol. 46, No. 1, pp. 131–134.
Rights and permissions
About this article
Cite this article
Sangaev, S.S., Trifonova, E.A., Titov, S.E. et al. Silencing of the Nk1 gene in the SR1 Nicotiana tabacum plants by RNA interference. Russ J Genet 46, 117–119 (2010). https://doi.org/10.1134/S1022795410010187
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1022795410010187