Skip to main content
SpringerLink
Log in

Reactive oxygen intermediates and glutathione regulate the expression of cytosolic ascorbate peroxidase during iron-mediated oxidative stress in bean

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Excess of free iron is thought to harm plant cells by enhancing the intracellular production of reactive oxygen intermediates (ROI). Cytosolic ascorbate peroxidase (cAPX) is an iron-containing, ROI-detoxifying enzyme induced in response to iron overload or oxidative stress. We studied the expression of cAPX in leaves of de-rooted bean plants in response to iron overload. cAPX expression, i.e., mRNA and protein, was rapidly induced in response to iron overload. This induction correlated with the increase in iron content in leaves and occurred in the light as well as in the dark. Reduced glutathione (GSH), which plays an important role in activating the ROI signal transduction pathway as well as in ROI detoxification, was found to enhance the induction of APX mRNA by iron. To determine whether cAPX induction during iron overload was due to an increase in the amount of free iron, which serves as a co-factor for cAPX synthesis, or due to iron-mediated increase in ROI production, we tested the expression of APX in leaves under low oxygen pressure. This treatment, which suppresses the formation of ROI, completely abolished the induction of cAPX mRNA during iron overload, without affecting the rate of iron uptake by plants. Taken together, our results suggest that high intracellular levels of free iron in plants lead to the enhanced production of ROI, which in turn induces the expression of cAPX, possibly using GSH as an intermediate signal. We further show, using cAPX-antisense transgenic plants, that cAPX expression is essential to prevent iron-mediated tissue damage in tobacco.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Allen, R.D. 1995. Dissection of oxidative stress tolerance using transgenic plants. Plant Physiol. 107: 1049–1054.

    Google Scholar 

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

    Google Scholar 

  • Balla, G., Jacob, H.S., Balla, J., Rosenberg, M., Nath, K., Apple, F., Eaton, J.W. and Vercellotti, G.M. 1992. Ferritin: a cytoprotective antioxidant strategem of endothelium. J. Biol. Chem. 267: 18148–18153.

    Google Scholar 

  • Cadenas, E. 1989. Biochemistry of oxygen-toxicity. Annu. Rev. Biochem. 58: 79–110.

    Google Scholar 

  • Creissen, G., Firmin, J., Fryer, M., Kular, B., Leyland, N., Reynolds, H., Pastori, G., Wellburn, F., Baker, N., Wellburn, A. and Mullineaux, P. 1999. Elevated glutathione biosynthetic capacity in the chloroplasts of transgenic tobacco plants paradoxically causes increased oxidative stress. Plant Cell 11: 1277–1292.

    Google Scholar 

  • Copin, J.C., Gasche, Y., Li, Y. and Chan, P.H. 2001. Prolonged hypoxia during cell development protects mature manganese superoxide dismutase-deficient astrocytes from damage by oxidative stress. FASEB J. 15: 525–534.

    Google Scholar 

  • del-Rio, L.A., Sandalio, L.M., Palma, J.M., Bueno, P. and Corpas, F.J. 1992. Metabolism of oxygen radicals in peroxisomes and cellular implications. Free Rad. Biol. Med. 13: 577–580.

    Google Scholar 

  • Eide, D., Broderius, M., Fett, J. and Guerinot, M.L. 1996. A novel iron-regulated metal transporter from plants identified by functional expression in yeast. Proc. Natl. Acad. Sci. USA 93: 5624–5628.

    Google Scholar 

  • Foyer, C.H., Lopez-Delgado, H., Dat, J.F. and Scott, I.M. 1997. Hydrogen peroxide-and glutathione-associated mechanisms of acclimatory stress tolerance and signalling. Physiol. Plant. 100: 241–254.

    Google Scholar 

  • Foyer, C.H., Souriau, N., Perret, S., Lelandais, M., Kunert, K.J., Pruvost, C. and Jouanin, L. 1995. Overexpression of glutathione reductase but not glutathione synthetase leads to increases in antioxidant capacity and resistance to photoinhibition in poplar trees. Plant Physiol. 109: 1047–1057.

    Google Scholar 

  • Gaymard, F., Boucherez, J. and Briat, J.F. 1996. Characterization of a ferritin mRNA from Arabidopsis thaliana accumulated in response to iron through an oxidative pathway independent of abscisic acid. Biochem. J. 318: 67–73.

    Google Scholar 

  • Grant, J.J., Yun, B.W. and Loake, G.J. 2000. Oxidative burst and cognate redox signalling reported by luciferase imaging: identification of a signal network that functions independently of ethylene, SA and Me-JA but is dependent on MAPKK activity. Plant J. 24: 569–582.

    Google Scholar 

  • Halliwell, B. and Gutteridge, J. 1984. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 219: 1–14.

    Google Scholar 

  • Jacobson, M.D. and Raff, M.C. 1995. Programmed cell-death and Bcl-2 protection in very-low oxygen. Nature 374: 814–816.

    Google Scholar 

  • Jimenez, A.J. Hernandez, A., delRio, L.A. and Sevilla, F. 1997. Evidence for the presence of the ascorbate-glutathione cycle in mitochondria and peroxisomes of pea leaves. Plant Physiol. 114: 275–284.

    Google Scholar 

  • Kampfenkel, K., Van Montagu, M, and Inzé, D. 1995. Effects of iron excess on Nicotiana-plumbaginifolia plants: implications to oxidative stress. Plant Physiol. 107: 725–735.

    Google Scholar 

  • Karpinska, B., Wingsle, G. and Karpinski, S. 2000. Antagonistic effects of hydrogen peroxide and glutathione on acclimation to excess excitation energy in Arabidopsis. IUBMB Life 50: 21–26.

    Google Scholar 

  • Karpinski, S., Escobar, C., Karpinska, B., Creissen, G. and Mullineaux, P.M. 1997. Photosynthetic electron transport regulates the expression of cytosolic ascorbate peroxidase genes in Arabidopsis during excess light stress. Plant Cell 9: 627–640.

    Google Scholar 

  • Kubo, A., Saji, H., Tanaka, K. and Kondo, N. 1995. Expression of arabidopsis cytosolic ascorbate peroxidase gene in response to ozone or sulfur dioxide. Plant Mol. Biol. 29: 479–489.

    Google Scholar 

  • Lobreaux, S., Massenet, O. and Briat, J.F. 1992. Iron induces ferritin synthesis in maize plantlets. Plant Mol. Biol. 19: 563–575.

    Google Scholar 

  • Lobreaux, S., Thoiron, S. and Briat, J.F. 1995. Induction of ferritin synthesis in maize leaves by an iron-mediated oxidative stress. Plant J. 8: 443–449.

    Google Scholar 

  • Mittler, R. and Zilinskas, B.A. 1992. Molecular cloning and characterization of a gene encoding pea cytosolic ascorbate peroxidase. J. Biol. Chem. 267: 21802–21807.

    Google Scholar 

  • Mittler, R. and Zilinskas, B.A. 1994. Regulation of pea cytosolic ascorbate peroxidase and other antioxidant enzymes during the progression of drought stress and following recovery from drought. Plant J. 5: 397–405.

    Google Scholar 

  • Mittler, R., Shulaev, V., Seskar, M. and Lam, E. 1996. Inhibition of programmed cell death in tobacco plants during pathogen-induced hypersensitive response at low oxygen pressure. Plant Cell 8: 1991–2001.

    Google Scholar 

  • Mittler, R., Feng, X.Q. and Cohen, M. 1998. Post-transcriptional suppression of cytosolic ascorbate peroxidase expression during pathogen-induced programmed cell death in tobacco. Plant Cell 10: 461–473.

    Google Scholar 

  • Mittler, R., Hallak-Herr, E., Orvar, B.L., Van Camp, W., Willekens, H., Inzé, D. and Ellis, B.E. 1999a. Transgenic tobacco plants with reduced capability to detoxify reactive oxygen intermediates are hyper-responsive to pathogen infection. Proc. Natl. Acad. Sci. USA. 96: 14165–14170.

    Google Scholar 

  • Mittler, R., Lam, E., Shulaev, V. and Cohen, M. 1999b. Signals controlling the expression of cytosolic ascorbate peroxidase during pathogen-induced programmed cell death in tobacco. Plant Mol. Biol. 39: 1025–1035.

    Google Scholar 

  • Orvar, B.L. and Ellis, B.E. 1997. Transgenic tobacco plants expressing antisense RNA for cytosolic ascorbate peroxidase show increased susceptibility to ozone injury. Plant J. 11: 1297–1305.

    Google Scholar 

  • Peyret, P., Perez, P. and Alric, M. 1995. Structure, genomic organization, and expression of the Arabidopsis-thaliana aconitase gene: plant aconitase show significant homology with mammalian iron-responsive element-binding protein. J. Biol. Chem. 270: 8131–8137.

    Google Scholar 

  • Pich, A and Scholz, G. 1991. The normalizing factor for the tomato mutant chloronerva.41. Nicotianamine and the distribution of iron into apoplast and symplast of tomato (Lycopersicon esculentum Mill.) J. Exp. Bot. 42: 1517–1523.

    Google Scholar 

  • Rabinowitch, H. and Fridovich, I. 1983. Superoxide radicals, superoxide dismutases and oxygen toxicity in plants. Photochem. Photobiol. 37: 679–690.

    Google Scholar 

  • Sheinberg, O., Rubin, B., Rabinowich, H.D. and Tel-Or, E. 1996. Oxidative stress responses in beans (Phaseolus vulgaris L). Plant Physiol. 111: 246.

    Google Scholar 

  • Sheinberg, O., Rubin, B., Rabinowich, H.D., Libal, Y. and Tel-Or, E. 2000. Acclimation of beans to oxidative stress by treatment with sublethal iron levels. J. Plant Physiol. 157: 93–99.

    Google Scholar 

  • Shimizu, S., Eguchi, Y., Kosaka, H., Kamiike, W., Matsuda, H. and Tsujimoto, Y. 1995. Prevention of hypoxia-induced cell-death by Bcl-2 and Bcl-X1. Nature 374: 811–813.

    Google Scholar 

  • Shulaev, V., Leon, J. and Raskin, I., 1995. Is salicylic acid a translocated signal of systemic acquired resistance in tobacco. Plant Cell 7: 1691–1701.

    Google Scholar 

  • Sigel, A, and Sigel, H. 1998. Iron transport and storage in microorganisms, plants, and animals. Marcel Dekker, New York.

    Google Scholar 

  • Thomsen, B., Drummherrel, H. and Mohr, H. 1992. Control of the appearance of ascorbate peroxidase (EC 1.11.1.11) in mustard seedling cotyledons by phytochrome and photooxidative treatments. Planta 186: 600–608.

    Google Scholar 

  • Vansuyt, G., Lopez, F., Inzé, D., Briat, J.F. and Fourcroy, P. 1997. Iron triggers a rapid induction of ascorbate peroxidase gene expression in Brassica napus. FEBS Lett. 410: 195–200.

    Google Scholar 

  • Willekens, H., Chamnongpol, S., Davey, M., Schraudner, M., Langebartels, C. Van Montagu, M., Inzé, D. and Van Camp, W. 1997. Catalase is a sink for H2O2 and is indispensable for stress defence in C-3 plants. EMBO J. 16: 4806–1816.

    Google Scholar 

  • Wingate, V.M.P., Lawton, M.A. and Lamb, C.J. 1988. Glutathione causes a massive and selective induction of plant defense genes. Plant Physiol. 87: 206–210.

    Google Scholar 

  • Yoshimura, K., Yabuta, Y., Ishikawa, T. and Shigeoka, S. 2000. Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. Plant Physiol. 123: 223–233.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Botany, Faculty of Agriculture, Hebrew University of Jerusalem, Rehovot, 76100, Israel

    Irena Pekker & Elisha Tel-Or

  2. Department of Biology, Technion-Israel Institute of Technology, Technion City, Haifa, 32000, Israel

    Ron Mittler

Authors
  1. Irena Pekker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elisha Tel-Or
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ron Mittler
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pekker, I., Tel-Or, E. & Mittler, R. Reactive oxygen intermediates and glutathione regulate the expression of cytosolic ascorbate peroxidase during iron-mediated oxidative stress in bean. Plant Mol Biol 49, 429–438 (2002). https://doi.org/10.1023/A:1015554616358

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015554616358

Search

Navigation