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Drought induces oxidative stress in pea plants

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Abstract

Pea (Pisum sativum L. cv. Frilene) plants subjected to drought (leaf water potential of ≈-1.3 MPa) showed major reductions in photosynthesis (78‰), transpiration (83‰), and glycolate oxidase (EC 1.1.3.1) activity (44‰), and minor reductions (≈18‰) in the contents of chlorophyll a, carotenoids, and soluble protein. Water stress also led to pronounced decreases (72–85‰) in the activities of catalase (EC 1.11.1.6), dehydroascorbate reductase (EC 1.8.5.1), and glutathione reductase (EC 1.6.4.2), but resulted in the increase (32–42‰) of non-specific peroxidase (EC 1.11.1.7) and superoxide dismutase (EC 1.15.1.1). Ascorbate peroxidase (EC 1.11.1.11) and monodehydroascorbate reductase (EC 1.6.5.4) activities decreased only by 15‰ and the two enzymes acted in a cyclic manner to remove H2O2, which did not accumulate in stressed leaves. Drought had no effect on the levels of ascorbate and oxidized glutathione in leaves, but caused a 25‰ decrease in the content of reduced glutathione and a 67‰ increase in that of vitamin E. In leaves, average concentrations of catalytic Fe, i.e. Fe capable of catalyzing free-radical generation by redox cycling, were estimated as 0.7 to 7 μM (well-watered plants, depending on age) and 16 μM (water-stressed plants); those of catalytic Cu were ≈4.5 μM and 18 μM, respectively. Oxidation of lipids and proteins from leaves was enhanced two- to threefold under stress conditions and both processes were highly correlated. Fenton systems composed of the purported concentrations of ascorbate, H2O2, and catalytic metal ions in leaves produced hydroxyl radicals, peroxidized membrane lipids, and oxidized leaf proteins. It is proposed that augmented levels and decompartmentation of catalytic metals occurring during water stress are responsible for the oxidative damage observed in vivo.

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Abbreviations

ASC:

ascorbate

DW:

dry weight

DHA:

dehydroascorbate

GSH:

reduced glutathione

GSSG:

oxidized glutathione

MDHA:

monodehydroascorbate (ascorbate free radical)

SOD:

Superoxide dismutase

Ψwa:

water potential

References

  • Aebi, H. (1984) Catalase in vitro. Methods Enzymol. 105, 121–126

    Google Scholar 

  • Aruoma, O.I., Halliwell, B., Laughton, M.J., Quinlan, G.J., Gutteridge, J.M.C. (1989) The mechanism of initiation of lipid peroxidation. Evidence against a requirement for an iron (II)-iron (III) complex. Biochem. J. 258, 617–620

    Google Scholar 

  • Asada, K. (1984) Chloroplasts: formation of active oxygen and its scavenging. Methods Enzymol. 105, 422–429

    Google Scholar 

  • Asada, K., Takahashi, M. (1987) Production and scavenging of active oxygen in photosynthesis. In: Photoinhibition, pp. 227–287, Kyle, D.J., Osmond, C.B., Arntzen, C.J., eds. Elsevier, Amsterdam

    Google Scholar 

  • Babbs, C.F., Pham, J.A., Coolbaugh, R.C. (1989) Lethal hydroxyl radical production in paraquat-treated plants. Plant Physiol. 90, 1267–1270

    Google Scholar 

  • Baker, A.L., Tolbert, N.E. (1966) Glycolate oxidase (ferredoxin-containing form). Methods Enzymol. 9, 338–342

    Google Scholar 

  • Becana, M., Klucas, R.V. (1992) Transition metals in legume root nodules: iron-dependent free radical production increases during nodule senescence. Proc. Natl. Acad. Sci. USA 89, 8958–8962

    Google Scholar 

  • Becana, M., Paris, F.J., Sandalio, L.M., Del Río, L.A. (1989) Isoenzymes of superoxide dismutase in nodules of Phaseolus vulgaris L., Pisum sativum L., and Vigna unguiculata (L.) Walp. Plant Physiol. 90, 1286–1292

    Google Scholar 

  • Bielawski, W., Joy, K.W. (1986) Reduced and oxidised glutathione and glutathione-reductase activity in tissues of Pisum sativum. Planta 169, 267–272

    Google Scholar 

  • Bowler, C., Van Montagu, M., Inzé, D. (1992) Superoxide dismutase and stress tolerance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 83–116

    Google Scholar 

  • Buckland, S.M., Price, A.H., Hendry, G.A.F. (1991) The role of ascorbate in drought-treated Cochlearia atlantica Pobed. and Armeria maritima (Mill.) Willd. New Phytol. 119, 155–160

    Google Scholar 

  • Chowdhury, S.R., Choudhuri, M.A. (1985) Hydrogen peroxide metabolism as an index of water stress tolerance in jute. Physiol. Plant. 65, 476–480

    Google Scholar 

  • Dalton, D.A., Langeberg, L., Robbins, M. (1992) Purification and characterization of monodehydroascorbate reductase from soybean root nodules. Arch. Biochem. Biophys. 292, 281–286

    Google Scholar 

  • Dalton, D.A., Russell, S.A., Hanus, F.J., Pascoe, G.A., Evans, H.J. (1986) Enzymatic reactions of ascorbate and glutathione that prevent peroxide damage in soybean root nodules. Proc. Natl. Acad. Sci. USA 83, 3811–3815

    Google Scholar 

  • Del Rio, L.A., Sandalio, L.M., Palma, J.M., Bueno, P., Corpas, F.J. (1992) Metabolism of oxygen radicals in peroxisomes and cellular implications. Free Rad. Biol. Med. 13, 557–580

    Google Scholar 

  • Dhindsa, R.S. (1991) Drought stress, enzymes of glutathione metabolism, oxidation injury, and protein synthesis in Tortula ruralis. Plant Physiol. 95, 648–651

    Google Scholar 

  • Elstner, E.F. (1987) Metabolism of activated oxygen species. In: The biochemistry of plants, vol. 11, pp. 253–315, Davies, D.D., ed. Academic Press, San Diego

    Google Scholar 

  • Farquhar, G.D., Sharkey, T.D. (1982) Stomatal conductance and photosynthesis. Annu. Rev. Plant Physiol. 33, 317–345

    Google Scholar 

  • Gutteridge, J.M.C. (1991) Metalloproteins as donors of metal ions for oxygen chemistry. In: Oxidative damage and repair, pp. 355–363, Davies, K.J.A., ed. Pergamon, Oxford

    Google Scholar 

  • Gutteridge, J.M.C., Wilkins, S. (1983) Copper salt-dependent hydroxyl radical formation. Damage to proteins acting as antioxidants. Biochim. Biophys. Acta 759, 38–41

    Google Scholar 

  • Halliwell, B., Gutteridge, J.M.C. (1989) Free radicals in biology and medicine. 2nd edn. Clarendon, Oxford

    Google Scholar 

  • Irigoyen, J.J., Emerich, D.W., Sánchez-Diaz, M. (1992) Alfalfa leaf senescence induced by drought stress: photosynthesis, hydrogen peroxide metabolism, lipid peroxidation and ethylene evolution. Physiol. Plant. 84, 67–72

    Google Scholar 

  • Laidman, D.L., Gaunt, J.K., Hall, G.S., Broad, C.T. (1971) Extraction of tocopherols from plant tissues. Methods Enzymol. 18, 366–369

    Google Scholar 

  • Law, M.Y., Charles, S.A., Halliwell, B. (1983) Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. Biochem. J. 210, 899–903

    Google Scholar 

  • Levine, R.L., Garland, D., Oliver, C., Amici, A., Climent, L., Lenz, A., Ahn, B., Shaltiel, S., Stadtman, E.R. (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol. 186, 464–478

    Google Scholar 

  • Lichtenthaler, H.K. (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol. 148, 350–382

    Google Scholar 

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

    Google Scholar 

  • Miyake, C., Asada, K. (1992) Thylakoid-bound ascorbate peroxidase in spinach chloroplasts and photoreduction of its primary oxidation product monodehydroascorbate radicals in thylakoids. Plant Cell Physiol. 33, 541–553

    Google Scholar 

  • Mukherjee, S.P., Choudhuri, M.A. (1983) Implications of water stress-induced changes in the levels of endogenous ascorbic acid and hydrogen peroxide in Vigna seedlings. Physiol. Plant. 58, 166–170

    Google Scholar 

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

    Google Scholar 

  • Pacifici, R.E., Davies, K.J.A. (1990) Protein degradation as an index of oxidative stress. Methods Enzymol. 186, 485–502

    Google Scholar 

  • Price, A.H., Hendry, G.A.F. (1989) Stress and the role of activated oxygen scavengers and protective enzymes in plants subjected to drought. Biochem. Soc. Trans. 17, 493–494

    Google Scholar 

  • Price, A.H., Hendry, G.A.F. (1991) Iron-catalysed oxygen radical formation and its possible contribution to drought damage in nine native grasses and three cereals. Plant Cell Environ. 14, 477–484

    Google Scholar 

  • Pütter, J. (1974) Peroxidases. Methods Enzymatic Anal. 2, 685–690

    Google Scholar 

  • Seel, W.E., Hendry, G.A.F., Lee, J.A. (1992) Effects of desiccation on some activated oxygen processing enzymes and anti-oxidants in mosses. J. Exp. Bot. 43, 1031–1037

    Google Scholar 

  • Stadtman, E.R., Oliver, C.N. (1991) Metal-catalyzed oxidation of proteins. Physiological consequences. J. Biol. Chem. 266, 2005–2008

    Google Scholar 

  • Thompson, J.E., Legge, R.L., Barber, R.F. (1987) The role of free radicals in senescence and wounding. New Phytol. 105, 317–344

    Google Scholar 

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Author notes

  1. Silvia Frechilla & Pedro Aparicio-Tejo

    Present address: Departamento de Ciencias del Medio Natural, E.T.S. de Ingenieros Agrónomos, Universidad Pública de Navarra, E-31006, Pamplona, Spain

  2. Robert V. Klucas

    Present address: Department of Biochemistry, University of Nebraska, 68583, Lincoln, NE, USA

Authors and Affiliations

  1. Departamento de Nutrición Vegetal, Estación Experimental de Aula Dei, CSIC, Apdo. 202, E-50080, Zaragoza, Spain

    Jose F. Moran, Manuel Becana, Iñaki Iturbe-Ormaetxe, Silvia Frechilla, Robert V. Klucas & Pedro Aparicio-Tejo

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  1. Jose F. Moran
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  2. Manuel Becana
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  3. Iñaki Iturbe-Ormaetxe
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  4. Silvia Frechilla
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  5. Robert V. Klucas
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  6. Pedro Aparicio-Tejo
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Additional information

We thank Dr. R. Picorel (E.E. de Aula Dei, CSIC) for allowing us access to HPLC equipment. J.F.M., 1.1., and S.F. were the recipients of predoctoral fellowships from the Comunidades Autónomas de Aragon, Pais Vasco, and Navarra, respectively. R.V.K. thanks the U.S. Department of Agriculture (grant 91-37305-6705) for travel support. This work was financed by grants from the Comisión Interministerial de Ciencia y Tecnología (AGR-91-0857-C02 to P.A. and M.B.) and the Dirección General de Investigación Científica y Técnica (PB92-0058 to M.B) of Spain.

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Moran, J.F., Becana, M., Iturbe-Ormaetxe, I. et al. Drought induces oxidative stress in pea plants. Planta 194, 346–352 (1994). https://doi.org/10.1007/BF00197534

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  • DOI: https://doi.org/10.1007/BF00197534

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