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Regulation of ascorbate peroxidase activity in dark-grown radish cotyledons by a catalase inhibitor, 3-Amino-1,2,4-Triazole

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Abstract

The present study was performed to see the physiological role of cytosolic ascorbate peroxidase (APX) and its relationship to other enzymes involved in the H2O2 scavenging metabolism, and also to elucidate the regulation of APX expression in dark-grown radish (Raphanus sativus L. cv Taiwang) cotyledons. To do so, 3-amino-l,2,4-triazole (aminotriazole), a known specific inhibitor of catalase, was used to simulate a catalase-deficient phenomenon in cotyledons. Aminotriazole, in very low concetration (10-4 M), inhibited remarkably the development of catalase activity in cotyledons during dark germination. This inhibition of catalase by aminotriazole, however, did not result in any significant changes in the growth response and the H2O2 level of developing cotyledons. In addition, the development of guaiacol peroxidase (GPX) activity was also not significantly affected. Unlike GPX, cytosolic APX activity was induced rapidly and reached a 1.7-fold increase in aminotriazole treated cotyledons at day 7 after germination. However,in vitro incubation of cytosolic APX preparation from cotyledons with aminotriazole did not result in any significant change in activity. One cytosolic APX isozyme (APXa) band involved in this APX activation was predominantly intensified in a native polyacrylamide gel by activity staining assay. This means that this APXa isozyme seems to play a key role in the expression of cytosolic APX activity. On the other hand, 2-day-old control seedlings treated with exogenous 1 mM H2O2 for 1 h showed a significant increase of cytosolic APX acitivity even in the absence of aminotriazole. Also, 2 μM cycloheximide treatment substantially inhibited the increase of APX activity due to aminotriazole. Based on these results, we suggest that a radish cytosolic APX could probably be substituted for catalase in H2O2 removal and that the expression of APX seems to be regulated by a change of endogenous H2O2 level which couples to APX protein synthesis in a translation stage in cotyledons.

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Literature cited

  • Asada, K. 1984. Chloroplasts; Formation of active oxygen and its scavengingMethods Emzymot. 105: 422–429.

    Article  CAS  Google Scholar 

  • Asada, K. 1992. Ascorbate peroxidase-a hydrogen peroxide-scavenging enzyme in plants.Physiol. Plant. 85: 235–241.

    Article  CAS  Google Scholar 

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

    Google Scholar 

  • Bunkelmann, J.R. and R.N. Trelease. 1996. Ascorbate peroxidase-a prominent membrane protein in oil seed glyoxysomes.Plant Physiol. 110: 589–598.

    Article  PubMed  CAS  Google Scholar 

  • Chance, B. and A.C. Maehly. 1955. Assay of catalase and peroxidase.Methods Enzymol. 2: 764–775.

    Article  Google Scholar 

  • Chen, G.-X. and K. Asada. 1989. Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the difference in their enzymatic and molecular properties.Plant Cell Physiol. 30: 987–998.

    CAS  Google Scholar 

  • Feierabend, J. and B. Schubert. 1978. Comparative investigation of the action of several chlorosis-inducing herbicides on the biogenesis of chloroplasts and leaf microbodies.Plant Physiol. 61: 1017–1022.

    Article  PubMed  CAS  Google Scholar 

  • Ferguson, I.B. and S.J. Dunning. 1986. Effect of 3-amino-l, 2,4-triazole, a catalase inhibitor, on peroxide content of suspension-cultured pear fruit cells.Plant. Sci. 43: 7–11.

    Article  CAS  Google Scholar 

  • Foyer, Ch.H. and B. Halliwell. 1976. The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism.Planta 133: 21–25.

    Article  Google Scholar 

  • Havir, E.A. 1992. Thein vivo andin vitro inhibition of catalase from leaves ofNicotiana sylvestris by 3-amino-1..2.4-triazole.Plant Physiol. 99: 533–537.

    Article  PubMed  CAS  Google Scholar 

  • Heim, D.R. and I.M. Larrinua. 1989. Primary site of action of amitrole inArabidopsis thaliana involves inhibition of root elongation but not of histidine or pigment biosynthesis.Plant Physiol. 91: 1226–1231.

    Article  PubMed  CAS  Google Scholar 

  • Jablonski, P.P. and J.W. Anderson. 1982. Light-dependent reduction of hydrogen peroxide by ruptured pea chloroplasts.Plant Physiol. 69: 1407–1413.

    Article  PubMed  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227: 680–685.

    Article  PubMed  CAS  Google Scholar 

  • Lowry, O.H., M.J. Rosebrough, A.L. Farr and R.L. Randall. 1951. Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193: 265–275.

    PubMed  CAS  Google Scholar 

  • MacRae, E.A. and I.B. Ferguson. 1985. Changes in catalase activity and hydrogen peroxide concentration in plants in response to low temperature.Physiol. Plant. 65: 51–56.

    Article  CAS  Google Scholar 

  • Margoliash, E., A. Novogrodsky and A. Schejter. 1960. Irreversible reaction of 3-amino-l,2,4-triazole and related inhibitors with the protein of catalase.Biochem. J. 74: 339–348.

    PubMed  CAS  Google Scholar 

  • Mittler, R. and B.A. Zilinskas. 1991. Purification and characterization of pea cytosolic ascorbate peroxidase.Plant Physiol. 97: 962–968.

    Article  PubMed  CAS  Google Scholar 

  • Mittler, R. and B.A. Zilinskas. 1993. Detection of ascorbate peroxidase activity in native gels by inhibition of the ascorbate-dependent reduction of nitroblue tetrazolium.Anal. Biochem. 212: 540–546.

    Article  PubMed  CAS  Google Scholar 

  • Nakano, Y. and K. Asada. 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chlorolplasts.Plant Cell Physiol. 22: 867–880.

    CAS  Google Scholar 

  • Prasad, T.K., M.D. Anderson, B.A. Martin and C.R. Stewart. 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 

  • Price, A.H., A. Taylor, S.J. Ripley, A. Griffiths, A.J. Trewavas and M.R. Knight. 1994. Oxidative signals in tobacco increase cytosolic calcium.Plant Cell 6: 1301–1310.

    Article  PubMed  CAS  Google Scholar 

  • Puntarulo, S., R.A. Sanchez and A. Boveris. 1988. Hydrogen peroxide metabolism in soybean embryonic axes at the onset of germination.Plant Physiol. 86: 626–630.

    Article  PubMed  CAS  Google Scholar 

  • Rao, M.V., G. Pailyath and D.P. Ormrod. 1996. Ultraviolet-B-and ozone-induced biochemical changes in antioxidant enzymes ofArabidopsis thaliana.Plant Physiol. 110: 125–136.

    Article  PubMed  CAS  Google Scholar 

  • Singer, S.R. and C.N. McDaniel. 1982. Transport of the herbicide 3-amino-l,2,4-triazole by cultured tobacco cells and leaf protoplasts.Plant Physiol. 62: 1382–1386.

    Article  Google Scholar 

  • Smith, I.K. 1985. Stimulation of glutathione synthesis in photorespiring plants by catalase inhibitors.Plant Physiol. 79: 1044–1047.

    Article  PubMed  CAS  Google Scholar 

  • Taylorson, R.B. and S.B. Hendricks. 1977. Dormancy in seeds.Annu. Rev. Plant Physiol. 28: 331–354.

    Article  CAS  Google Scholar 

  • Tsaftaris, A.S. and J.G. Scandalios. 1981. Genetic and biochemical characterization of acat2 catalase null mutant oflea mays.Mol. Gen. Genet. 181: 158–163..

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biology, Kangwon National University, 200-701, Chuncheon, Korea

    Kyung Ah Yi & Chang-Duck Jin

  2. Department of Biology, Seoul National University, 151-742, Seoul, Korea

    Young-Nam Hong

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  1. Kyung Ah Yi
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  2. Young-Nam Hong
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Yi, K.A., Hong, YN. & Jin, CD. Regulation of ascorbate peroxidase activity in dark-grown radish cotyledons by a catalase inhibitor, 3-Amino-1,2,4-Triazole. J. Plant Biol. 40, 279–287 (1997). https://doi.org/10.1007/BF03030461

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  • DOI: https://doi.org/10.1007/BF03030461

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