Advertisement

Planta

, Volume 194, Issue 3, pp 346–352 | Cite as

Drought induces oxidative stress in pea plants

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

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.

Key words

Antioxidant Free radical Oxidative damage Pisum Plant senescence Water stress 

Abbreviations and Symbol

ASC

ascorbate

DW

dry weight

DHA

dehydroascorbate

GSH

reduced glutathione

GSSG

oxidized glutathione

MDHA

monodehydroascorbate (ascorbate free radical)

SOD

Superoxide dismutase

Ψwa

water potential

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aebi, H. (1984) Catalase in vitro. Methods Enzymol. 105, 121–126Google Scholar
  2. 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, 617–620Google Scholar
  3. Asada, K. (1984) Chloroplasts: formation of active oxygen and its scavenging. Methods Enzymol. 105, 422–429Google Scholar
  4. 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, AmsterdamGoogle Scholar
  5. Babbs, C.F., Pham, J.A., Coolbaugh, R.C. (1989) Lethal hydroxyl radical production in paraquat-treated plants. Plant Physiol. 90, 1267–1270Google Scholar
  6. Baker, A.L., Tolbert, N.E. (1966) Glycolate oxidase (ferredoxin-containing form). Methods Enzymol. 9, 338–342Google Scholar
  7. 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, 8958–8962Google Scholar
  8. 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, 1286–1292Google Scholar
  9. Bielawski, W., Joy, K.W. (1986) Reduced and oxidised glutathione and glutathione-reductase activity in tissues of Pisum sativum. Planta 169, 267–272Google Scholar
  10. Bowler, C., Van Montagu, M., Inzé, D. (1992) Superoxide dismutase and stress tolerance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 83–116Google Scholar
  11. 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, 155–160Google Scholar
  12. Chowdhury, S.R., Choudhuri, M.A. (1985) Hydrogen peroxide metabolism as an index of water stress tolerance in jute. Physiol. Plant. 65, 476–480Google Scholar
  13. Dalton, D.A., Langeberg, L., Robbins, M. (1992) Purification and characterization of monodehydroascorbate reductase from soybean root nodules. Arch. Biochem. Biophys. 292, 281–286Google Scholar
  14. 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, 3811–3815Google Scholar
  15. 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, 557–580Google Scholar
  16. Dhindsa, R.S. (1991) Drought stress, enzymes of glutathione metabolism, oxidation injury, and protein synthesis in Tortula ruralis. Plant Physiol. 95, 648–651Google Scholar
  17. 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 DiegoGoogle Scholar
  18. Farquhar, G.D., Sharkey, T.D. (1982) Stomatal conductance and photosynthesis. Annu. Rev. Plant Physiol. 33, 317–345Google Scholar
  19. 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, OxfordGoogle Scholar
  20. Gutteridge, J.M.C., Wilkins, S. (1983) Copper salt-dependent hydroxyl radical formation. Damage to proteins acting as antioxidants. Biochim. Biophys. Acta 759, 38–41Google Scholar
  21. Halliwell, B., Gutteridge, J.M.C. (1989) Free radicals in biology and medicine. 2nd edn. Clarendon, OxfordGoogle Scholar
  22. 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, 67–72Google Scholar
  23. Laidman, D.L., Gaunt, J.K., Hall, G.S., Broad, C.T. (1971) Extraction of tocopherols from plant tissues. Methods Enzymol. 18, 366–369Google Scholar
  24. Law, M.Y., Charles, S.A., Halliwell, B. (1983) Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. Biochem. J. 210, 899–903Google Scholar
  25. 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, 464–478Google Scholar
  26. Lichtenthaler, H.K. (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol. 148, 350–382Google Scholar
  27. 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, 51–56Google Scholar
  28. 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, 541–553Google Scholar
  29. 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, 166–170Google Scholar
  30. Nakano, Y., Asada, K. (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol. 22, 867–880Google Scholar
  31. Pacifici, R.E., Davies, K.J.A. (1990) Protein degradation as an index of oxidative stress. Methods Enzymol. 186, 485–502Google Scholar
  32. 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, 493–494Google Scholar
  33. 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, 477–484Google Scholar
  34. Pütter, J. (1974) Peroxidases. Methods Enzymatic Anal. 2, 685–690Google Scholar
  35. 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, 1031–1037Google Scholar
  36. Stadtman, E.R., Oliver, C.N. (1991) Metal-catalyzed oxidation of proteins. Physiological consequences. J. Biol. Chem. 266, 2005–2008Google Scholar
  37. Thompson, J.E., Legge, R.L., Barber, R.F. (1987) The role of free radicals in senescence and wounding. New Phytol. 105, 317–344Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Jose F. Moran
    • 1
  • Manuel Becana
    • 1
  • Iñaki Iturbe-Ormaetxe
    • 1
  • Silvia Frechilla
    • 1
  • Robert V. Klucas
    • 1
  • Pedro Aparicio-Tejo
    • 1
  1. 1.Departamento de Nutrición VegetalEstación Experimental de Aula Dei, CSICZaragozaSpain

Personalised recommendations