Acta Biologica Hungarica

, Volume 67, Issue 2, pp 148–158 | Cite as

Salicylic Acid Induced Cysteine Protease Activity During Programmed Cell Death in Tomato Plants

  • Judit Kovács
  • Péter Poór
  • Ágnes Szepesi
  • Irma TariEmail author


The hypersensitive response (HR), a type of programmed cell death (PCD) during biotic stress is mediated by salicylic acid (SA). The aim of this work was to reveal the role of proteolysis and cysteine proteases in the execution of PCD in response of SA. Tomato plants were treated with sublethal (0.1 mM) and lethal (1 mM) SA concentrations through the root system. Treatment with 1 mM SA increased the electrolyte leakage and proteolytic activity and reduced the total protein content of roots after 6 h, while the proteolytic activity did not change in the leaves and in plants exposed to 0.1 mM SA. The expression of the papain-type cysteine protease SlCYP1, the vacuolar processing enzyme SlVPE1 and the tomato metacaspase SlMCA1 was induced within the first three hours in the leaves and after 0.5 h in the roots in the presence of 1 mM SA but the transcript levels did not increase significantly at sublethal SA. The Bax inhibitor-1 (SlBI-1), an antiapoptotic gene was over-expressed in the roots after SA treatments and it proved to be transient in the presence of sublethal SA. Protease inhibitors, SlPI2 and SlLTC were upregulated in the roots by sublethal SA but their expression remained low at 1 mM SA concentration. It is concluded that in contrast to leaves the SA-induced PCD is associated with increased proteolytic activity in the root tissues resulting from a fast up-regulation of specific cysteine proteases and down-regulation of protease inhibitors.


Bax Inhibitor 1 cysteine protease programmed cell death salicylic acid tomato 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Benchabane, M., Schlüter, U., Vorster, J., Goulet, M.-C., Michoud, D. (2010) Plant cystatins. Biochemie 92, 1657–1666.CrossRefGoogle Scholar
  2. 2.
    Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.CrossRefGoogle Scholar
  3. 3.
    Chomczynski, P., Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate- phenol-chloroform extraction. Anal. Biochem. 162, 156–159.CrossRefPubMedGoogle Scholar
  4. 4.
    Hatsugai, N., Kuroyanagi, M., Yamada, K., Meshi, T., Tsuda, S., Kondo, M., Hara-Nishimura, I. (2004) A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death. Science 305, 855–858.CrossRefGoogle Scholar
  5. 5.
    Hayat, Q., Hayat, S., Irfan, M., Ahmad, A. (2010) Effect of exogenous salicylic acid under changing environment: a review. Environ. Exp. Bot. 68, 14–25.CrossRefGoogle Scholar
  6. 6.
    Hoeberichts, F. A., Ten Have, A., Woltering, E. J. (2003) A tomato metacaspase gene is upregulated during programmed cell death in Botrytis cinerea-infected leaves. Planta 217, 517–522.CrossRefGoogle Scholar
  7. 7.
    Horváth, E., Csiszár, J., Gallé, Á., Poór, P., Szepesi, Á., Tari, I. (2015) Hardening with salicylic acid induces concentration-dependent changes in abscisic acid biosynthesis of tomato under salt stress. J. Plant Physiol. 183, 54–63.CrossRefGoogle Scholar
  8. 8.
    Ishikawa, T., Watabane, N., Nagano, M., Kawai-Yamada, M., Lam, E. (2011) Bax inhibitor-1: a highly conserved endoplasmic reticulum–resident cell death suppressor. Cell Death Diff. 18, 1271–1278.CrossRefGoogle Scholar
  9. 9.
    Jones, J. D., Dangl, J. L. (2006) The plant immune system. Nature 444, 323–329.CrossRefPubMedGoogle Scholar
  10. 10.
    Kawai-Yamada, M., Ohori, Y., Uchimiya, H. (2004) Dissection of Arabidopsis Bax inhibitor-1 suppressing Bax-, hydrogen peroxide-, and salicylic acid-induced cell death. Plant Cell 16, 21–32.CrossRefPubMedGoogle Scholar
  11. 11.
    Kim, I., Xu, W., Reed, J. C. (2008) Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities. Nature reviews drug discovery 7, 1013–1030.CrossRefGoogle Scholar
  12. 12.
    Livak, K. J., Schmittgen, T. D. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25, 402–408.CrossRefGoogle Scholar
  13. 13.
    Lu, S., Faris, J. D., Sherwood, R., Edwards, M. C. (2013) Dimerization and protease resistance: new insight into the function of PR-1. J. Plant Physiol. 170, 105–110.CrossRefGoogle Scholar
  14. 14.
    Morris, K., Mackerness, S. A. H., Page, T., John, C. F., Murphy, A. M., Carr, J. P., Buchanan-Wollaston, V. (2000) Salicylic acid has a role in regulating gene expression during leaf senescence. Plant J. 23, 677–685.CrossRefGoogle Scholar
  15. 15.
    Pereira, D. A., Ramos, M. V., Souza, D. P., Portela, T. C., Guimarães, J. A., Madeira, S. V., Freitas, C. D. (2010) Digestibility of defense proteins in latex of milkweeds by digestive proteases of Monarch butterflies, Danaus plexippus L.: a potential determinant of plant–herbivore interactions. Plant Sci. 179, 348–355.CrossRefGoogle Scholar
  16. 16.
    Poór, P. (2013) Investigation of salt stress- and salicylic acid-induced physiological changes in tomato plants: acclimation or programmed cell death. PhD Thesis, in Hungarian, University of Szeged, Hungary, pp. 61–68.Google Scholar
  17. 17.
    Poór, P., Kovács, J., Szopkó, D., Tari, I. (2013) Ethylene signaling in salt stress- and salicylic acidinduced programmed cell death in tomato suspension cells. Protoplasma 250, 273–284.CrossRefGoogle Scholar
  18. 18.
    Poór, P., Borbély, P., Kovács, J., Papp, A., Szepesi, Á., Takács, Z., Tari, I. (2014) Opposite extremes in ethylene/nitric oxide ratio induce cell death in suspension culture and root apices of tomato exposed to salt stress. Acta Biol. Hung. 65, 428–438.CrossRefGoogle Scholar
  19. 19.
    Roberts, I. N., Caputo, C., Criado, M. V., Funk, C. (2012) Senescence-associated proteases in plants. Physiol. Plant. 145, 130–139.CrossRefGoogle Scholar
  20. 20.
    Rossano, R., Larocca, M., Riccio, P. (2011) 2-D zymographic analysis of broccoli (Brassica oleracea L. var. Italica) florets proteases: Follow up of cysteine protease isotypes in the course of post-harvest senescence. J. Plant Physiol. 168, 1517–1525.CrossRefGoogle Scholar
  21. 21.
    Sanmartín, M., Jaroszewski, L., Raikhel, N. V., Rojo, E. (2005) Caspases. Regulating death since the origin of life. Plant Physiol. 137, 841–847.CrossRefPubMedGoogle Scholar
  22. 22.
    Shindo, T., van der Horn, R. A. L. (2008) Papain-like cysteine proteases: key players at molecular battlefields employed by both plants and their invaders. Mol. Plant Pathol. 9, 119–125.Google Scholar
  23. 23.
    Shindo, T., isas-Villami, J. C., Hörger, A. J., Song, J., van der Horn, R. A. L. (2012) A role in immunity for Arabidopsis cysteine protease RD21, the ortholog of the tomato immune protease C14. PloS ONE 7(1) e29317. doi:10.1371/journal.pone.0029317CrossRefPubMedGoogle Scholar
  24. 24.
    Singh, M., Bhogal, D., Goel, A., Kumar, A. (2011) Cloning, in silico characterization and interaction of cysteine protease and cystatin for establishing their role in early blight disease in tomato. J. Plant Biochem. Biotech. 20, 110–117.CrossRefGoogle Scholar
  25. 25.
    Trobacher, C. P., Senatore, A., Greenwood, J. S. (2006) Masterminds or minions? Cysteine proteinases in plant programmed cell death. This review is one of a selection of papers published in the Special Issue on Plant Cell Biology. Botany 84, 651–667.Google Scholar
  26. 26.
    van Doorn, W. G. (2011) Classes of programmed cell death in plants, compared to those in animals. J. Exp. Bot. 62, 4749–4761.CrossRefGoogle Scholar
  27. 27.
    Woltering, E. J. (2004) Death proteases come alive. Trends Plant Sci. 9, 469–472.CrossRefGoogle Scholar
  28. 28.
    Woltering, E. J. (2010) Death proteases: alive and kicking. Trends Plant Sci. 15, 185–188.CrossRefGoogle Scholar
  29. 29.
    Yamada, K., Nishimura, M., Hara-Nishimura, I. (2004) The slow wound-response of γVPE is regulated by endogenous salicylic acid in Arabidopsis. Planta 218, 599–605.CrossRefGoogle Scholar

Copyright information

© Akadémiai Kiadó, Budapest 2016

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Judit Kovács
    • 1
  • Péter Poór
    • 1
  • Ágnes Szepesi
    • 1
  • Irma Tari
    • 1
    Email author
  1. 1.Department of Plant BiologyUniversity of SzegedSzegedHungary

Personalised recommendations