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. 1997 Sep;115(1):137–149. doi: 10.1104/pp.115.1.137

Influence of salicylic acid on H2O2 production, oxidative stress, and H2O2-metabolizing enzymes. Salicylic acid-mediated oxidative damage requires H2O2.

M V Rao 1, G Paliyath 1, D P Ormrod 1, D P Murr 1, C B Watkins 1
PMCID: PMC158469  PMID: 9306697

Abstract

We investigated how salicylic acid (SA) enhances H2O2 and the relative significance of SA-enhanced H2O2 in Arabidopsis thaliana. SA treatments enhanced H2O2 production, lipid peroxidation, and oxidative damage to proteins, and resulted in the formation of chlorophyll and carotene isomers. SA-enhanced H2O2 levels were related to increased activities of Cu,Zn-superoxide dismutase and were independent of changes in catalase and ascorbate peroxidase activities. Prolonging SA treatments inactivated catalase and ascorbate peroxidase and resulted in phytotoxic symptoms, suggesting that inactivation of H2O2-degrading enzymes serves as an indicator of hypersensitive cell death. Treatment of leaves with H2O2 alone failed to invoke SA-mediated events. Although leaves treated with H2O2 accumulated in vivo H2O2 by 2-fold compared with leaves treated with SA, the damage to membranes and proteins was significantly less, indicating that SA can cause greater damage than H2O2. However, pretreatment of leaves with dimethylthiourea, a trap for H2O2, reduced SA-induced lipid peroxidation, indicating that SA requires H2O2 to initiate oxidative damage. The relative significance of the interaction among SA, H2O2, and H2O2-metabolizing enzymes with oxidative damage and cell death is discussed.

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Selected References

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  1. Auh C. K., Murphy T. M. Plasma Membrane Redox Enzyme Is Involved in the Synthesis of O2- and H2O2 by Phytophthora Elicitor-Stimulated Rose Cells. Plant Physiol. 1995 Apr;107(4):1241–1247. doi: 10.1104/pp.107.4.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bi Y. M., Kenton P., Mur L., Darby R., Draper J. Hydrogen peroxide does not function downstream of salicylic acid in the induction of PR protein expression. Plant J. 1995 Aug;8(2):235–245. doi: 10.1046/j.1365-313x.1995.08020235.x. [DOI] [PubMed] [Google Scholar]
  3. Bolwell G. P., Butt V. S., Davies D. R., Zimmerlin A. The origin of the oxidative burst in plants. Free Radic Res. 1995 Dec;23(6):517–532. doi: 10.3109/10715769509065273. [DOI] [PubMed] [Google Scholar]
  4. 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 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Brennan T., Frenkel C. Involvement of hydrogen peroxide in the regulation of senescence in pear. Plant Physiol. 1977 Mar;59(3):411–416. doi: 10.1104/pp.59.3.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen Z., Ricigliano J. W., Klessig D. F. Purification and characterization of a soluble salicylic acid-binding protein from tobacco. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9533–9537. doi: 10.1073/pnas.90.20.9533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen Z., Silva H., Klessig D. F. Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science. 1993 Dec 17;262(5141):1883–1886. doi: 10.1126/science.8266079. [DOI] [PubMed] [Google Scholar]
  8. Conrath U., Chen Z., Ricigliano J. R., Klessig D. F. Two inducers of plant defense responses, 2,6-dichloroisonicotinec acid and salicylic acid, inhibit catalase activity in tobacco. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7143–7147. doi: 10.1073/pnas.92.16.7143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Desikan R., Hancock J. T., Coffey M. J., Neill S. J. Generation of active oxygen in elicited cells of Arabidopsis thaliana is mediated by a NADPH oxidase-like enzyme. FEBS Lett. 1996 Mar 11;382(1-2):213–217. doi: 10.1016/0014-5793(96)00177-9. [DOI] [PubMed] [Google Scholar]
  10. Durner J., Klessig D. F. Inhibition of ascorbate peroxidase by salicylic acid and 2,6-dichloroisonicotinic acid, two inducers of plant defense responses. Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11312–11316. doi: 10.1073/pnas.92.24.11312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Durner J., Klessig D. F. Salicylic acid is a modulator of tobacco and mammalian catalases. J Biol Chem. 1996 Nov 8;271(45):28492–28501. doi: 10.1074/jbc.271.45.28492. [DOI] [PubMed] [Google Scholar]
  12. Guan L., Scandalios J. G. Developmentally related responses of maize catalase genes to salicylic acid. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5930–5934. doi: 10.1073/pnas.92.13.5930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hammond-Kosack K. E., Jones J. D. Resistance gene-dependent plant defense responses. Plant Cell. 1996 Oct;8(10):1773–1791. doi: 10.1105/tpc.8.10.1773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heath R. L., Packer L. Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys. 1968 Apr;125(1):189–198. doi: 10.1016/0003-9861(68)90654-1. [DOI] [PubMed] [Google Scholar]
  15. Kauss H., Jeblick W. Pretreatment of Parsley Suspension Cultures with Salicylic Acid Enhances Spontaneous and Elicited Production of H2O2. Plant Physiol. 1995 Jul;108(3):1171–1178. doi: 10.1104/pp.108.3.1171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kauss H., Jeblick W., Ziegler J., Krabler W. Pretreatment of Parsley (Petroselinum crispum L.) Suspension Cultures with Methyl Jasmonate Enhances Elicitation of Activated Oxygen Species. Plant Physiol. 1994 May;105(1):89–94. doi: 10.1104/pp.105.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klessig D. F., Malamy J. The salicylic acid signal in plants. Plant Mol Biol. 1994 Dec;26(5):1439–1458. doi: 10.1007/BF00016484. [DOI] [PubMed] [Google Scholar]
  18. Leon J., Lawton M. A., Raskin I. Hydrogen Peroxide Stimulates Salicylic Acid Biosynthesis in Tobacco. Plant Physiol. 1995 Aug;108(4):1673–1678. doi: 10.1104/pp.108.4.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Levine A., Tenhaken R., Dixon R., Lamb C. H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell. 1994 Nov 18;79(4):583–593. doi: 10.1016/0092-8674(94)90544-4. [DOI] [PubMed] [Google Scholar]
  20. Mehdy M. C. Active Oxygen Species in Plant Defense against Pathogens. Plant Physiol. 1994 Jun;105(2):467–472. doi: 10.1104/pp.105.2.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Naton B., Hahlbrock K., Schmelzer E. Correlation of Rapid Cell Death with Metabolic Changes in Fungus-Infected, Cultured Parsley Cells. Plant Physiol. 1996 Sep;112(1):433–444. doi: 10.1104/pp.112.1.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rao M. V., Paliyath G., Ormrod D. P. Ultraviolet-B- and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol. 1996 Jan;110(1):125–136. doi: 10.1104/pp.110.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rusterucci C., Stallaert V., Milat M. L., Pugin A., Ricci P., Blein J. P. Relationship between Active Oxygen Species, Lipid Peroxidation, Necrosis, and Phytoalexin Production Induced by Elicitins in Nicotiana. Plant Physiol. 1996 Jul;111(3):885–891. doi: 10.1104/pp.111.3.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rüffer M., Steipe B., Zenk M. H. Evidence against specific binding of salicylic acid to plant catalase. FEBS Lett. 1995 Dec 18;377(2):175–180. doi: 10.1016/0014-5793(95)01334-2. [DOI] [PubMed] [Google Scholar]
  25. Sanchez-Casas P., Klessig D. F. A Salicylic Acid-Binding Activity and a Salicylic Acid-Inhibitable Catalase Activity Are Present in a Variety of Plant Species. Plant Physiol. 1994 Dec;106(4):1675–1679. doi: 10.1104/pp.106.4.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Savenkova M. L., Mueller D. M., Heinecke J. W. Tyrosyl radical generated by myeloperoxidase is a physiological catalyst for the initiation of lipid peroxidation in low density lipoprotein. J Biol Chem. 1994 Aug 12;269(32):20394–20400. [PubMed] [Google Scholar]
  27. Shah J., Tsui F., Klessig D. F. Characterization of a salicylic acid-insensitive mutant (sai1) of Arabidopsis thaliana, identified in a selective screen utilizing the SA-inducible expression of the tms2 gene. Mol Plant Microbe Interact. 1997 Jan;10(1):69–78. doi: 10.1094/MPMI.1997.10.1.69. [DOI] [PubMed] [Google Scholar]
  28. Sharma Y. K., Léon J., Raskin I., Davis K. R. Ozone-induced responses in Arabidopsis thaliana: the role of salicylic acid in the accumulation of defense-related transcripts and induced resistance. Proc Natl Acad Sci U S A. 1996 May 14;93(10):5099–5104. doi: 10.1073/pnas.93.10.5099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shirasu K., Nakajima H., Rajasekhar V. K., Dixon R. A., Lamb C. Salicylic acid potentiates an agonist-dependent gain control that amplifies pathogen signals in the activation of defense mechanisms. Plant Cell. 1997 Feb;9(2):261–270. doi: 10.1105/tpc.9.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Summermatter K., Sticher L., Metraux J. P. Systemic Responses in Arabidopsis thaliana Infected and Challenged with Pseudomonas syringae pv syringae. Plant Physiol. 1995 Aug;108(4):1379–1385. doi: 10.1104/pp.108.4.1379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vernooij B., Friedrich L., Morse A., Reist R., Kolditz-Jawhar R., Ward E., Uknes S., Kessmann H., Ryals J. Salicylic Acid Is Not the Translocated Signal Responsible for Inducing Systemic Acquired Resistance but Is Required in Signal Transduction. Plant Cell. 1994 Jul;6(7):959–965. doi: 10.1105/tpc.6.7.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Willekens H., Villarroel R., Van Montagu M., Inzé D., Van Camp W. Molecular identification of catalases from Nicotiana plumbaginifolia (L.). FEBS Lett. 1994 Sep 19;352(1):79–83. doi: 10.1016/0014-5793(94)00923-6. [DOI] [PubMed] [Google Scholar]
  33. Woodbury W., Spencer A. K., Stahman M. A. An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem. 1971 Nov;44(1):301–305. doi: 10.1016/0003-2697(71)90375-7. [DOI] [PubMed] [Google Scholar]
  34. Wu G., Shortt B. J., Lawrence E. B., Levine E. B., Fitzsimmons K. C., Shah D. M. Disease resistance conferred by expression of a gene encoding H2O2-generating glucose oxidase in transgenic potato plants. Plant Cell. 1995 Sep;7(9):1357–1368. doi: 10.1105/tpc.7.9.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Xu Y., Chang PFL., Liu D., Narasimhan M. L., Raghothama K. G., Hasegawa P. M., Bressan R. A. Plant Defense Genes Are Synergistically Induced by Ethylene and Methyl Jasmonate. Plant Cell. 1994 Aug;6(8):1077–1085. doi: 10.1105/tpc.6.8.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]

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