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Changes in the isozyme composition of antioxidant enzymes in response to aminotriazole in leaves ofArabidopsis thaliana

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Abstract

The changes in isozyme profiles of catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR) during severe deactivation of total CAT activity by aminotriazole (AT) treatment were investigated in the leaves ofArabidopsis thaliana (Columbia ecotype) in relation to H2O2-mediated oxidative stress. In spite of striking deactivation of total CAT activity by 0.1 mM AT, there were no significant differences in H2O2 levels or total leaf soluble protein contents including a Rubisco in both the control and AT-treated leaves. On the other hand, one specific protein band (molecular mass, 66 kD) was observed on the SDS-gel from leaf soluble proteins whose staining intensity was strikingly enhanced by AT treatment for 6 h. However, this band disappeared at 12 h. In the native-gel assays of CAT, POD, APX and GR isozymes, AT remarkably inhibited the expression of the CAT1 isozyme with no effects on CAT2 and CAT3, and generally had no effect on POD isozyme profiles. However, AT stimulated the intensity of activities of pre-existing APX1 and GR1 isozymes. In particular, it induced a new synthesis of one GR isozyme. Therefore, these results collectively suggest that a striking deactivation of total CAT activity by AT inA. thaliana leaves largely results from the suppression of CAT1 isozyme, and that APX1, GR1, and a newly synthesized GR isozyme could complement the role of CAT1 to metabolize H2O2 into non-toxic water.

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Literature Cited

  • Allen RD (1995) Dissection of oxidative stress tolerance using transgenic plants. Plant Physiol 107: 1049–1054

    PubMed  CAS  Google Scholar 

  • Amane M, Mae T, Ohira K (1986) Colorimetric measurement of protein stained with coomassie brilliant blue R on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by eluting with formamide. Agric Biol Chem 50: 1911–1912

    Google Scholar 

  • Anderson MD, Prasad TK, Stewart CR (1995) Changes in isozyme profiles of catalase, peroxidase, and glutathione reductase during acclimation to chilling in mesocotyls of maize seedings. Plant Physiol 109: 1247–1257

    PubMed  CAS  Google Scholar 

  • Bernt E, Bergmeyer HU (1974) Inorganic peroxides.In HU Bergmeyer, ed, Methods of Enzymatic analysis, Vol 4. Acad Press, New York, pp 2246–2248

    Google Scholar 

  • Campa A (1991) Biological roles of plant peroxidase: known and potential function.In J Everse, MB Grisham, eds, Peroxidase in Chemistry and Biology, Vol II. CRC Press, Boca Raton, FL, pp 25–50

    Google Scholar 

  • Chandlee JM, Tsaftaris AS, Scandalios JG (1983) Purification and partial characterization of three genetically defined catalase of maize. Plant Sci Lett 29: 117–131

    Article  CAS  Google Scholar 

  • Desimone M, Henke A, Waigner E (1996) Oxidative stress induces partial degradation of the large subunit of ribu-lose-1,5-bisphophate carboxylase/oxygenase in isolated chloroplast of barley. Plant Physiol 111: 789–796

    PubMed  CAS  Google Scholar 

  • Donahue Jl., Okpodu CM, Cramer CL, Grabau EA, Alscher RG (1997) Responses of antioxidants to paraquat in pea leaves. Plant Physiol 113: 249–257

    PubMed  CAS  Google Scholar 

  • Edwards EA, Enard C, Creissen GP, Mullineaux PM (1994) Synthesis and properties of glutathione reductase in stressed peas. Planta 192: 137–143

    CAS  Google Scholar 

  • Ferguson IB, Dunning Sl (1986) Effect of 3-amino-1,2,4-tri-azole, a catalase inhibitor, on peroxide content of suspension-cultured pear fruit cells. Plant Sci 43: 7–11

    Article  CAS  Google Scholar 

  • Foyer CH, Descourvieres P, Kunert KJ (1994) Protection against oxygen radicals: an important defence mechanism studied in transgenic plants. Plant Cell Environ 17: 507–523

    Article  CAS  Google Scholar 

  • Havir EA (1992) The in vivo and in vitro inhibition of catalase from leaves of Nicotiana sylvestris by 3-amino-1,2,4-triazole. Plant Physiol 99: 533–537

    Article  PubMed  CAS  Google Scholar 

  • Kang KS, Lim CJ, Han TJ, Kim JC, Jin CD (1998) Activation of ascorbate-glutathione cycle in Arabidopsis leaves in response to aminotriazole. J Plant Biol 41: 155–161

    Article  CAS  Google Scholar 

  • Klapheck S, Zimmer I, Cosse H (1990) Scavenging of hydrogen peroxide in the endosperm ofRicinus communis by ascorbate peroxidase. Plant Cell Physiol 31: 1005–1013

    CAS  Google Scholar 

  • Kunce CM, Trelaease RN (1986) Heterogenety of catalase in maturing and germinated cotten seeds. Plant Physiol 81:1134–1139

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Article  PubMed  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275

    PubMed  CAS  Google Scholar 

  • Mittler R, Zilinskas BA (1993) Detection of ascorbate peroxidase activity in native gels by inhibition of the ascor-bate-dependent reduction of nitroblue tetrazolium. Anal Biochem 212: 540–546

    Article  PubMed  CAS  Google Scholar 

  • Pacifici RE, Davis KJA (1990) Protein degradation as an index of oxidative stress. Methods Enzymol 186: 485–502

    Article  PubMed  CAS  Google Scholar 

  • Penarrubia L, Moreno J (1990) Increased susceptibility of ribulose-1,5-bisphosphate carboxylase/oxygenase to proteolytic degradation caused by oxidative treatment. Arch Bioch Biophys 281: 319–323

    Article  CAS  Google Scholar 

  • Prasad TK (1997) Role of catalase in inducing chilling tolerance in pre-emergent maize seedlings. Plant Physiol 114: 1369–1376

    PubMed  CAS  Google Scholar 

  • Prasad TK, Anderson MD, Martin BA, Stewart CR (1994) Evidence for chilling-induced oxidative stress in maize seedlings and a regulatory role for hydrogen peroxide. Plant Cell 6: 65–74

    Article  PubMed  CAS  Google Scholar 

  • Scandalios JG, Tsaftaris AS, Chandlee JM, Skadsen RW (1984) Expression of the developmentally regulated catalase (cat) genes in maize. Dev Genet 4: 281–293

    Article  CAS  Google Scholar 

  • Rao MV, Hale BA, Ormrod DP (1995) Amelioration of ozone-induced oxidative damage in wheat plants grown under high carbon dioxide. Plant Physiol 109: 421–432

    PubMed  CAS  Google Scholar 

  • Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B-and ozone-induced biochemical changes in antioxidant enzymes ofArabidopsis thaliana. Plant Physiol 110: 125–136

    Article  PubMed  CAS  Google Scholar 

  • Sen Gupta A, Webb RR, Holaday AS, Allen RD (1993) Overexpression of superoxide dismutase protects plants from oxidative stress. Plant Physiol 103: 1067–1073

    CAS  Google Scholar 

  • Stadtman ER (1990) Covalent modification reactions are marking steps in protein turnover. Biochemistry 29: 6323–6331

    Article  PubMed  CAS  Google Scholar 

  • Tsaftaris AS, Scandalios JG (1981) Genetic and biochemical characterization of a Cat2 catalase null mutant of zea mays. Mol Gen Genet 181: 158–163

    Article  CAS  Google Scholar 

  • Woodbury W, Spencer AK, Stahmann MA (1971) An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem 44: 301–305

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Chang-Duck Jin.

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Kang, KS., Lim, CJ., Han, TJ. et al. Changes in the isozyme composition of antioxidant enzymes in response to aminotriazole in leaves ofArabidopsis thaliana . J. Plant Biol. 42, 187–193 (1999). https://doi.org/10.1007/BF03030477

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  • DOI: https://doi.org/10.1007/BF03030477

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