Abstract
A 4-h exposure of Scenedesmus sp. to Cu or Zn enhanced intracellular levels of both test metals and proline. The level of intracellular proline increased markedly up to 10 µM Cu, but higher concentrations were inhibitory. However, intracellular proline consistently increased with increasing concentration of Zn in the medium. Cu and Zn induced oxidative stress in the test alga by increasing lipid peroxidation and membrane permeability, and by reducing SH content. Pretreatment of the test alga with 1 mM proline for 30 min completely alleviated Cu-induced lipid peroxidation, minimized K+ efflux and also reduced depletion of the SH pool. But proline pretreatment could only slightly reduce Zn-induced oxidative stress. Interestingly, proline pretreatment increased the level of Cu (25–54%) and Zn (19–49%) inside the cells. It did not affect the activities of superoxide dismutase, ascorbate peroxidase or catalase, but improved glutathione reductase activity under Cu and Zn stress. A comparison of the effects of proline pretreatment on lipid peroxidation by Cu, Zn, methyl viologen and ultraviolet-B radiation suggests that proline protects cells from metal-induced oxidative stress by scavenging reactive oxygen species rather than by chelating metal ions. Pretreatment of cells with a known antioxidant (ascorbate) and a hydroxyl radical scavenger (sodium benzoate) considerably reduced metal-induced lipid peroxidation and proline accumulation. However, sodium benzoate had a very mild effect on Zn-induced lipid peroxidation and proline accumulation. The present study demonstrates that proline possibly acts by detoxifying reactive oxygen species, mainly hydroxyl radicals, rather than by improving the antioxidant defense system under metal stress.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- APOX :
-
Ascorbate peroxidase
- CAT :
-
Catalase
- GR :
-
Glutathione reductase
- MDA :
-
Malondialdehyde
- MV :
-
Methyl viologen
- ROS :
-
Reactive oxygen species
- SH :
-
Sulphydryl
- SOD :
-
Superoxide dismutase
- UV-B :
-
Ultraviolet-B radiation
References
Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126
Alia, Mohanty P, Matysik J (2001) Effect of proline on the production of singlet oxygen. Amino Acids 21:195–200
Asada K (1984) Chloroplasts: formation of active oxygen and its scavenging. Methods Enzymol 105:422–435
Bates LS, Waldren RP, Tear ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207
Bates SS, Tessier A, Campbell PGC, Buffle J (1982) Zinc adsorption and transport by Chlamydomonas variabilis and Scenedesmus subspicatus (Chlorophyceae) grown in semicontinuous culture. J Phycol 18:521–529
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 44:276–287
Briat JF (2002) Metal ion-activated oxidative stress and its control. In: Inze D, Montagu MV (eds) Oxidative stress in plants. Taylor and Francis, New York, pp 171–189
Chen THH, Murata N (2002) Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Curr Opin Plant Biol 5:250–257
Clairbone A (1985) Catalase activity. In: Greewald RA (ed) Handbook of methods for oxygen radical research. CRC Press, Boca Raton, pp 283–284
Davey MW, Montagu MV, Inze D (2002) Ascorbate metabolism and stress. In: Inze D, Montagu MV (eds) Oxidative stress in plants. Taylor and Francis, New York, pp 271–296
De Filippis LF, Pallaghy CK (1994) Heavy metals: sources and biological effects. In: Rai LC, Gaur JP, Soeder CJ (eds) Algae and water pollution. Schweizerbart’sche Verlagbuchhandlung, Stuttgart, pp 31–77
De Vos CHR, Schat H, Vooijs R, Ernst WHO (1989) Copper-induced damage to the permeability barrier in roots of Silene cucubalus. J Plant Physiol 135:164–179
Dietz KJ, Baier M, Kramer U (1999) Free radicals and reactive oxygen species as mediators of heavy metal toxicity in plants. In: Prasad MNV, Hagemeyer J (eds) Heavy metal stress in plants: from molecules to ecosystems. Springer, Berlin Heidelberg New York, pp 73–97
Farago ME, Mullen WA (1979) Plants which accumulate metals. IV. A possible copper–proline complex from the roots of Armeria maritima. Chim Acta 32:93–94
Foyer CH, Lelandias M, Kunert KJ (1994) Photooxidative stress in plants. Physiol Plant 92:696–717
Hughes EO, Gorham PR, Zehnder A (1958) Toxicity of a unialgal culture of Microcystis aeruginosa. Can J Microbiol 4:225–236
Kwon TW, Menzel DB, Olcott HS (1965) Reactivity of malondialdehyde with food constituents. J Food Sci 30:808–813
Luna CM, Gonzalez CA, Trippi VS (1994) Oxidative damage caused by an excess of copper in oat leaves. Plant Cell Physiol 35:11–15
Lutts S, Kinet JM, Bouharmont J (1996) Effects of various salts and of mannitol on ion and proline accumulation in relation to osmotic adjustment in rice (Oryza sativa L.) callus cultures. J Plant Physiol 149:186–195
Mallick N, Mohn FH (2000) Reactive oxygen species: responses of algal cells. J Plant Physiol 157:183–193
Mehta SK, Gaur JP (1999) Heavy metal-induced proline accumulation and its role in ameliorating metal toxicity in Chlorella vulgaris. New Phytol 143:253–259
Murphy AS, Eisinger WR, Shaff JE, Kochian LV, Taiz L (1999) Early copper-induced leakage of K+ from Arabidopsis seedings is mediated by ion channels and coupled to citrate efflux. Plant Physiol 121:1375–1382
Noctor G, Foyer CH (1998) Ascorbate and glutathione: kee** active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279
Schaedle M, Bassham JA (1977) Chloroplast glutathione reductase. Plant Physiol 59:1011–1012
Schickler H, Caspi H (1999) Responses of antioxidative enzymes to nickel and cadmium in hyperaccumulator plants of the genus Alyssum. Physiol Plant 105:39–44
Sedlak J, Lindsay RH (1968) Estimation of total, protein-bound and non-protein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 25:192–205
Sharma SS, Schat H, Vooijs R (1998) In vitro alleviation of heavy metal-induced enzyme inhibition by proline. Phytochemistry 49:1531–1535
Sharmila P, Pardha Saradhi P (2002) Proline accumulation in heavy metal stressed plants: an adaptive strategy. In: Prasad MNV, Strazlka K (eds) Physiology and biochemistry of metal toxicity and tolerance in plants. Kluwer, Dordrecht, pp 179–199
Siripornadulsil S, Traina S, Verma DPS, Sayre RT (2002) Molecular mechanisms of proline-mediated tolerance to toxic heavy metals in transgenic microalgae. Plant Cell 14:2837–2847
Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28:1057–1060
Smith RC, Buker KS, Hansen OH, Olson R (1980) Photoinhibition of photosynthesis in natural water. Photochem Photobiol 31:585–592
Stohs SJ, Bagchi D (1995) Oxidative mechanisms in the toxicity of metal ions. Free Radical Biol Med 18:321–336
Wu J-T, Chang SC, Chen KS (1995) Enhancement of intracellular proline levels in cells of Anacystis nidulans (cyanobacteria) exposed to deleterious concentrations of copper. J Phycol 31:376–379
Wu J-T, Hsieh MT, Kow LC (1998) Role of proline accumulation in response to toxic copper in Chlorella sp. (Chlorophyceae) cells. J Phycol 34:113–117
Yu BP (1994) Cellular defenses against damage by reactive oxygen species. Physiol Rev 74:139–162
Acknowledgments
We thank the Head, Department of Botany and Coordinator, Centre of Advanced Study in Botany, Banaras Hindu University, for providing necessary facilities, and B.N.T. thanks the Council of Scientific and Industrial Research, New Delhi, for financial support in the form of a Senior Research Fellowship.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tripathi, B.N., Gaur, J.P. Relationship between copper- and zinc-induced oxidative stress and proline accumulation in Scenedesmus sp.. Planta 219, 397–404 (2004). https://doi.org/10.1007/s00425-004-1237-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00425-004-1237-2