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Activity of antioxidant enzymes in response to cadmium in Crotalaria juncea

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Abstract

The effects of the heavy metal Cadmium (Cd) on the growth and the activities of the antioxidant enzymes, catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and glutathione reductase (GR, EC 1.6.4.2) have been investigated in Crotalaria juncea seedlings. Concentrations above 0.2 mM CdCl2 were shown to inhibit strongly the growth of roots and shoots. Cd was shown to accumulate to very high concentrations in the roots, whilst in the leaves, the maximum concentration obtained following treatment with 2 mM CdCl2, was only 6% of that determined in the roots. Although CAT activity did not exhibit any major variation in the roots following CdCl2 treatment, 2 mM CdCl2 induced a 6-fold increase in activity in the leaves when compared to the untreated control. Non-denaturing PAGE gels stained for SOD activity revealed four isoenzymes, two Mn-SOD and two Cu/Zn-SOD. The results observed for SOD were different of those observed for CAT activity, since in both, leaves and roots, no significant changes in the total activity or of the four isoenzymes were observed following the treatment with CdCl2. GR activity exhibited a similar pattern of that of CAT activity. The concentration of 2 mM CdCl2 induced a small increase in activity in the roots after 48 h of exposure, whereas in leaves a 7-fold increase in GR activity was detected after 48 hr exposure to 2 mM CdCl2. The results suggest that in C. juncea the reactive oxygen species (ROS) induced by Cd, are metabolised by CAT in the peroxisomes. In the case of GR activity, the increase observed in the leaves suggest that GR is also playing a role in the detoxification of Cd-induced ROS possibly via the glutathione-ascorbate cycle.

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References

  • Azevedo R A, Alas R M, Smith R J and Lea P J 1998 Response of antioxidant enzymes to transfer from elevated carbon dioxide to air and ozone fumigation, in the leaves and roots of wild-type and a catalase-deficient mutant of barley. Physiol. Plant. 104, 280–292.

    Google Scholar 

  • Barata R M, Chapparro A, Chabregas S M, Gonzalez R, Labate C A, Azevedo R A, Sarath G, Lea P J and Silva-Filho M C 2000 Targeting of the soybean leghemoglobin to tobacco chloroplasts: Effects on aerobic metabolism in transgenic plants. Plant Sci. 155, 193–202.

    Google Scholar 

  • Beauchamp C H and Fridovich I 1971 Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44, 276–287.

    Google Scholar 

  • Bergmann H, Machelett B, Lippmann B and Friedrich Y 2001 Influence of heavy metals on the accumulation of trimethylglycine, putrescine and spermine in food plants. Amino Acids 20, 325–329.

    Google Scholar 

  • Bhattacharjee S 1998 Membrane lipid peroxidation, free radical scavengers and ethylene evolution in Amaranthus as affected by lead and cadmium. Biol. Plant. 40, 131–135.

    Google Scholar 

  • Boussama N, Ouariti O and Ghorbal M H 1999a Changes in growth and nitrogen assimilation in barley seedlings under cadmium stress. J. Plant Nutr. 22, 731–752.

    Google Scholar 

  • Boussama N, Ouariti O, Suzuki A and Ghorbal M H 1999b Cd-stress on nitrogen assimilation. J. Plant Physiol. 155, 310–317.

    Google Scholar 

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

    Google Scholar 

  • Bradford M M 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    Google Scholar 

  • Brej T 1998 Heavy metal tolerance in Agropyron repens (L.) P. Bauv. populations from Legnica copper smelter area, Lower Silesia. Acta Soc. Pol. 67, 325–333.

    Google Scholar 

  • Bruns I, Sutter K, Menge S, Neumann D and Krauss G J 2001 Cadmium lets increase the glutathione pool in bryophytes. J. Plant Physiol. 158, 79–89.

    Google Scholar 

  • Cardoso P F, Molina S M G, Pereira G J G, Vitória A P and Azevedo R A 2002 Response of rice inbred lines to cadmium exposure. J. Plant Nutr. 25, 927–944.

    Google Scholar 

  • Chaoui A, Mazhoudi S, Ghorbal M H and El Ferjani E 1997 Cadmium and zinc induction of lipid peroxidation and effects on antioxidant enzyme activities in bean (Phaseolus vulgaris L). Plant Sci. 127, 139–147.

    Google Scholar 

  • Chaparro-Giraldo A, Barata R M, Chabregas S M, Azevedo R A and Silva-Filho M C 2000 Soybean leghemoglobin targeted to potato chloroplasts influences growth and development of transgenic plants. Plant Cell Rep. 19, 961–965.

    Google Scholar 

  • Chugh L K and Sawhney S K 1999 Effect of cadmium on activities of some enzymes of glycolysis and pentose phosphate pathway in pea. Biol. Plant. 42, 401–407.

    Google Scholar 

  • Cobbett C S 2000 Phytochelatins and their roles in heavy metal detoxification. Plant Physiol. 123, 825–832.

    Google Scholar 

  • Corpas F J, Barroso J B and del Rio L A 2001 Peroxisomes as a source of reactive oxygen species and nitric oxide signal molecules in plant cells. Trends Plant Sci. 6, 145–150.

    Google Scholar 

  • Cuypers A, Vangronsved J and Cliisters H 2000 Biphasic effect of copper on the ascorbate-glutathione pathway in primary leaves of Phaseolus vulgaris seedlings during early stages of metal assimilation. Physiol. Plant. 110, 512–517.

    Google Scholar 

  • Dalurzo H C, Sandalio L M, Gomez M and Del Rio L A 1997 Cadmium infiltration of detached pea leaves: Effect on its activated oxygen metabolism. Phyton. 37, 59–64.

    Google Scholar 

  • De Leonardis S, Dipierro N and Dipierro S 2000 Purification and characterization of an ascorbate peroxidase from potato tuber mitochondria. Plant Physiol. Biochem. 38, 773–779.

    Google Scholar 

  • Di Toppi L S, Lambardi M, Pecchioni N, Pazzagli L, Durante M and Gabbrielli R 1999 Effects of cadmium stress on hairy roots of Daucus carota. J. Plant Physiol. 154, 385–393.

    Google Scholar 

  • Fargasova A 2001 Phytotoxic effects of Cd, Zn, Pb, Cu and Fe on Sinapsis alba L. seedlings and their accumulation in roots and shoots. Biol. Plant. 44, 471–473.

    Google Scholar 

  • Ferreira R R, Fornazier R F, Vitória A P, Lea P J and Azevedo R A 2002 Changes in antioxidant enzyme activities in soybean under cadmium stress. J. Plant Nutr. 25, 327–342.

    Google Scholar 

  • Fornazier R F, Ferreira R R, Vitória A P, Molina S M G, Lea P J and Azevedo R A 2002 Effects of cadmium on antioxidant enzyme activities in sugar cane. Biol. Plant., 41, 91–97.

    Google Scholar 

  • Foyer C H and Noctor G 2000 Oxygen processing in photosynthesis: regulation and signalling. New Phytol. 146, 359–388.

    Google Scholar 

  • Gallego S M, Benavides M P and Tomaro M L 1999 Effect of cadmium ions on antioxidant defence system in sunflower cotyledons. Biol. Plant. 42, 49–55.

    Google Scholar 

  • Grant J J and Loake G J 2000 Role of reactive oxygen intermediates and cognate redox signalling in disease resistance. Plant Physiol. 124, 21–29.

    Google Scholar 

  • Groppa M D, Tomaro M L and Benavides M P 2001 Polyamines as protectors against cadmium or copper-induced oxidative damage in sunflower leaf discs. Plant Sci. 161, 481–488.

    Google Scholar 

  • Harada E, Choi Y E, Tsuchisaka A, Obata H and Sano H 2001 Transgenic tobacco plants expressing a rice cysteine synthase gene are tolerant to toxic levels of cadmium. J. Plant Physiol. 158, 655–661.

    Google Scholar 

  • Hart J J, Welch R M, Norvell W A, Sullivan L A and Kochian L V 1998 Characterization of cadmium binding, uptake and translocation in intact seedlings of bread and durum wheat cultivars. Plant Physiol. 116, 1413–1420.

    Google Scholar 

  • Hegedus A, Erdei S and Horvath G 2001 Comparative studies of H2O2 detoxifying enzymes in green and greening barley seedlings under cadmium stress. Plant Sci. 160, 1085–1093.

    Google Scholar 

  • Höfgen R, Kreft O, Willmitzer L and Hesse H 2001 Manipulation of thiol contents in plants. Amino Acids 20, 291–299.

    Google Scholar 

  • Kendall A C, Keys A J, Turner J C, Lea P J and Miflin B J 1983 The isolation and characterization of a catalase-deficient mutant of barley (Hordeum vulgare L.). Planta 159, 505–511.

    Google Scholar 

  • Khudsar T, Mahmooduzzafar and Iqbal M 2001 Cadmium-induced changes in leaf epidermes, photosynthetic rate and pigment concentration in Cajanus cajan. Biol. Plant. 44, 59–64.

    Google Scholar 

  • Kliebenstein P J, Monde R A and Last R L 1998 Superoxide dismutase in Arabidopsis: an eclectic enzyme family with disparate regulation and protein localisation. Plant Physiol. 118, 637–650.

    Google Scholar 

  • Kubota H, Sato K, Yamada T and Maitani T 2000 Phytochelatin homologs induced in hairy roots of horseradish. Phytochemistry 53, 239–245.

    Google Scholar 

  • Kumar R G and Dubey R S 1999 Glutamine synthetase isoforms from rice seedlings: Effects of stress on enzyme activity and the protective roles of osmolytes. J. Plant Physiol. 155, 118–121.

    Google Scholar 

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

    Google Scholar 

  • Lee S M and Leustek T 1999 The effect of cadmium on sulphate assimilation enzymes in Brassica juncea. Plant Sci. 141, 201–207.

    Google Scholar 

  • Leita L, Contin M and Maggioni A 1991 Distribution of cadmium and induced Cd binding proteins in roots, stems and leaves of Phaseolus vulgaris. Plant Sci. 77, 139–147.

    Google Scholar 

  • Mullineaux P, Ball L, Escobar C, Karpinska B, Creissen G and Karpinski S 2000 Are diverse signalling pathways in the regulation of Arabidopsis antioxidant defence gene expression in response to excess excitation energy. Phil. Trans. Roy. Soc. Lond. B. 355, 1531–1540.

    Google Scholar 

  • Noctor G and Foyer C H 1998 Ascorbate and glutathione: Keeping active oxygen under control. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49, 249–279.

    Google Scholar 

  • Pilon-Smits E A H, Zhu Y L, Sears T and Terry N 2000 Over-expression of glutathione reductase in Brassica juncea: Effects on cadmium accumulation and tolerance. Physiol. Plant. 110, 455–460.

    Google Scholar 

  • Polidoros A N and Scandalios J G 1999 Role of hydrogen peroxide and different classes of antioxidants in the regulation of catalase and glutathione S-transferase gene expression in maize (Zea mays L.). Physiol. Plant. 106, 112–120.

    Google Scholar 

  • Rauser W E 2000 Roots of maize seedlings retain most of their cadmium through two complexes. J. Plant Physiol. 156, 545–551.

    Google Scholar 

  • Scandalios J G 1994 Regulation and properties of plant catalases. In Causes of Photo-oxidative Stress and Amelioration of Defense Systems in Plants. Eds C H Foyer and P M Mullineaux. pp 275–316. CRC Press, Boca Raton, FL.

    Google Scholar 

  • Schickler H and Caspi H 1999 Response of antioxidative enzymes to nickel and cadmium stress in hyperaccumulator plants of the genus Alyssum. Physiol. Plant. 105, 39–44.

    Google Scholar 

  • Shaw B P 1995 Effects of mercury and cadmium on the activities of antioxidative enzymes in the seedlings of Phaseolus aureus. Biol. Plant. 37, 587–596.

    Google Scholar 

  • Siedlecka A and Krupa Z 1999 Cd/Fe interaction in higher plants-its consequences for the photosynthetic apparatus. Photosynthetica 36, 321–331.

    Google Scholar 

  • Somashekaraiah B V, Padmaja K and Prasad A R K 1992 Phytotoxicity of cadmium ions on germinating seedlings of mung bean (Phaseoulus vulgaris)-involvement of lipid peroxides in chlorophyll degradation. Physiol. Plant. 85, 85–89.

    Google Scholar 

  • Stevens R G, Creissen G P and Mullineaux P M 1997 Cloning and characterisation of a cytosolic glutathione reductase cDNA from pea (Pisum sativum L.) and its expression in response to stress. Plant Mol. Biol. 35, 641–654.

    Google Scholar 

  • Stroinski A, Kubis J and Zielezinska M 1999 Effect of cadmium on glutathione reductase in potato tubers. Acta Physiol. Plant. 21, 201–207.

    Google Scholar 

  • Van Assche F and Clijsters H 1990 Effects of metals on enzyme activity in plants. Plant Cell Environ. 13, 195–206.

    Google Scholar 

  • Verma S and Dubey R S 2001 Effect of cadmium on soluble sugars and enzymes of their metabolism in rice. Biol. Plant. 44, 117–123.

    Google Scholar 

  • Vitória A P, Lea P J and Azevedo R A 2001 Antioxidant enzymes responses to cadmium in radish tissues. Phytochemistry 57, 710–710.

    Google Scholar 

  • Willekens H, Langebartels C, Tire D, Van Montagu M, Inzé D and Van Camp W 1994 Differential expression of catalase genes in Nicotiana plumbaginifolia (L.). Proc. Natl Acad. Sci. USA 91, 10450–10454.

    Google Scholar 

  • Woodbury W, Spencer A K and Stahmann M A 1971 An improved procedure using ferricyanide for detecting catalase isoenzymes. Anal. Biochem. 44, 301–305.

    Google Scholar 

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Pereira, G., Molina, S., Lea, P. et al. Activity of antioxidant enzymes in response to cadmium in Crotalaria juncea . Plant and Soil 239, 123–132 (2002). https://doi.org/10.1023/A:1014951524286

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