The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

Development of a Screening System for Plant Defense-Inducing Agent using Transgenic Tobacco Plant with PR-1a Promoter and GUS Gene

  • Oh, Sang-Keun (Plant Genomics Lab., Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Seon-Woo (Division of Applied Biology, Dong-A University) ;
  • Kwon, Suk-Yoon (Lab of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology) ;
  • Choi, Do-Il (Plant Genomics Lab., Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2005.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

Pathogenesis-related protein-1a (PR-1a) is strongly induced in tobacco plants by pathogen attack, exogenous salicylic acid (SA) application and by other developmental processes. In order to develop a rapid screening system for the selection of plant defense-inducing compounds originated from various sources, we have transformed tobacco Samsun NN plants with a chimeric construct consisting of GUS $(\beta-glucuronidase)$. In the $T_1$ generation, three transgenic lines having stable GUS expression were selected for further promoter analysis. Using GUS histochemical assay, we observed strong GUS induction driven by PR-1a promoter in PR1a-GUS transgenic tobacco leaves in response to the exogenous application of SA or benzol (1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (BTH), a SA­derivative compound. In addition, GUS expression was maintained locally or systemically in PR1a-GUS transgenic line $\#5\;T_2$ generation) until after 3 days when they were treated with same chemicals. Our results suggested that the PR1a-GUS reporter gene system in tobacco plants may be applicable for the large-scale screening of defense-inducing substances.

Keywords

References

  1. Church, G. M. and Gilbert, W. 1984. Genomic sequencing. Proc. Natl. Acad. Sci. USA. 81:1991-1995
  2. Dangl, I. L., Dietrich, R. A. and Richberg, M. H. 1996. Death don't have no mercy: Cell death programs in plant-microbe interactions. Plant Cell 8:1793-1807 https://doi.org/10.1105/tpc.8.10.1793
  3. Friedrich, L., Vernooij, B., Gaffuey, T., Morse, A. and Ryals, I. 1995. Characterization of tobacco plants expressing a bacterial salicylate hydroxylase gene. Plant Mol. Biol. 29:959-969 https://doi.org/10.1007/BF00014969
  4. Glazebrook, J., Chen, W., Esters, B., Chang, H.-S., Nawrath, C., Metraux, J.-P., Zhu, T. and Katagiri, F. 2003. Topology of the network integrating salicylate and jasmonate signal transduction derived from global expression phenotyping. Plant J. 34:217-228 https://doi.org/10.1046/j.1365-313X.2003.01717.x
  5. Gerlach, J., Volrath, S., Knauf-Beiter, G., Hengy, G., Beckhove, U., Kogel, K-H., Oostendorp, M., Staub, T., Ward, E., Kessmann, H. and Ryals, J. 1996. Benzothiadiazole, a novel class of inducers of systemic acquired resistance, activates gene expression and disease resistance in wheat. Plant Cell 8:629-643 https://doi.org/10.1105/tpc.8.4.629
  6. Gruner, R. and Pfitzner, U. M. 1994. The upstream region of the gene for the pathogenesis-related protein 1a from tobacco responds to environmental as well as to developmental signals in transgenic plants. Eur. J. Biochem. 220:247-255 https://doi.org/10.1111/j.1432-1033.1994.tb18620.x
  7. Gruner, R., Strompent, G., Pfitzner, A. J. P. and Pfitzner, U. M. 2003. Salicylic acid and the hypersensitive response initiate distinct signal transduction pathways in tobacco that converge on the as-1-like element of the PR-1a promoter. Eur. J. Biochem. 270:4876-4886 https://doi.org/10.1046/j.1432-1033.2003.03888.x
  8. Jefferson, R. A., Kavanagh, T. A. and Bevan, M. W. 1987. GUS fusions: $\beta$-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6:3901-3907
  9. Metraux, J.-P., Ahl Goy, P., Staub, T., Speich, J., Steinemann, A., Ryals, J. and Ward, E. 1991. Induced resistance in cucumber in response to 2,6-dichloroisonicotinic acid and pathogens. In: Advances in Molecular Genetics of Plant-Microbe Interactions, Vol. 1, H. Hennecke and D. P. D. Verma, eds (Dordrecht, The Netherlands: Kluwer), pp. 432-439
  10. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant. 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  11. Oh, S-K., Cheong, I.-J., Hwang, I. and Choi, D. 1999. Similarities of tobacco mosaic virus-induced hypersensitive cell death and copper-induced abiotic cell death in tobacco. Plant Pathol. J. 15:8-13
  12. Oh, S-K., Park, J. M., Joung, Y. H., Lee, S., Chung, E., Kim, S-Y., Yu, S. H. and Choi, D. 2005. A plant EPF-type zinc-finger protein, CaPIFI, involves in defense against pathogens. Mol. Plant. Pathol. 6:269-285 https://doi.org/10.1111/j.1364-3703.2005.00284.x
  13. Rushton, P. J., Torres, T. J., Pamiske, M., Wernert, P., Hahlbrock, K. and Somssich, I. E. 1996. Interaction of elicitor-induced DNA-binding proteins with elicitor response elements in the promoters of parsley PR1 genes. EMBO J. 15:5690-5700
  14. Ryals, J. A., Neuenschwander, U. H., Willits, M. G., Molina, A., Steiner, H-Y. and Hunt, M. D. 1996. Systemic acquired resistance. Plant Cell 8:1809-1819 https://doi.org/10.1105/tpc.8.10.1809
  15. Schurter, R., Kunz, W. and Nyfeler, R. 1987. Process and a composition for immunizing plants against diseases. U.S. Patent No. 4,931,581. Issued June 5, 1990
  16. Strompen, G., Gruner, R. and Pfitzner, U. M. 1998. An as-1-like motif controls the level of expression of the gene for the pathogenesis-related protein la from tobacco. Plant Mol. Biol. 37:871-883 https://doi.org/10.1023/A:1006003916284
  17. Suh, M. C., Choi, D. and Liu, J. R. 1998. Cadmium resistance in transgenic tobacco plants expressing the Nicotiana glutinosa L. metallothionein-like gene. Mol. Cells 8:678-684
  18. Van de Rhee, M. D., Van Kan, J. A. L., Gonzalez-Jaen M. T. and Bol, J. F. 1990. Analysis of regulatory elements involved in the induction oftwo tobacco genes by salicylate treatment and virus infection. Plant Cell 2:357-366 https://doi.org/10.1105/tpc.2.4.357
  19. Van Loon, L. C. and Van Kammen, A. 1970. Polyacrylamide disk electrophoresis of the soluble proteins from Nicotiana tabacum var. 'Samsun' and 'Samsun NN'. II. Changes in protein constitution after infection with tobacco mosaic virus. Virology 40: 199-211 https://doi.org/10.1016/0042-6822(70)90395-8
  20. Van Loon, L. C. and Van Strien, E. A. 1999. The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol. Mol. Plant Pathol. 55:85-97 https://doi.org/10.1006/pmpp.1999.0213
  21. Ward, E. R., Ukenes, S. J., Williams, S. C., Dincher, S. S., Wiederhold, D. L., Alexander, D. C., Ahl-Goy, P., Metraux, J. P. and Ryals, J. A. 1991. Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell 3:1085-1094 https://doi.org/10.1105/tpc.3.10.1085

Cited by

  1. Functional analysis of the promoter of the pepper pathogen-induced gene, CAPIP2, during bacterial infection and abiotic stresses vol.172, pp.2, 2007, https://doi.org/10.1016/j.plantsci.2006.08.015
  2. Expression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection in peanut (Arachis hypogaea L.) plants vol.67, pp.2, 2011, https://doi.org/10.1002/ps.2041
  3. Functional roles of the pepper pathogen-induced bZIP transcription factor, CAbZIP1, in enhanced resistance to pathogen infection and environmental stresses vol.224, pp.5, 2006, https://doi.org/10.1007/s00425-006-0302-4
  4. Identification and functional expression of the pepper pathogen-induced gene, CAPIP2, involved in disease resistance and drought and salt stress tolerance vol.62, pp.1-2, 2006, https://doi.org/10.1007/s11103-006-9010-5