Involvement of heme oxygenase as antioxidant defense in soybean nodules

Objective: We have previously demonstrated that the inducible form of heme oxygenase plays a critical role in protecting against oxidative stress in mammals. To gain further insight into the functions of this enzyme in plants, we have tested its activity and expression in soybean nodules subjected to cadmium (Cd) stress. Materials and methods: Four-weeks-old soybean nodulated plants were treated with different cadmium chloride concentrations (0, 50 and 200 μM) during 48 h. Oxidative stress parameters such as TBARS content, GSH levels and antioxidant enzyme activities were measured as well as heme oxygenase activity and expression. Besides, the effect of biliverdin and Zn-protophorphyrin IX were analized. Results: Treatment with 200 μM Cd during 48 h caused a 67% increase in TBARS content, whereas GSH decreased 44%, and total superoxide dismutase, gluthatione reductase and guaiacol peroxidase were also inhibited 54, 20 and 60%, respectively. A total of 200 μM Cd produced the overexpression of heme oxygenase-1, as well as a 10-fold enhancement of its activity. Co-administration of biliverdin (10 μM) completely prevented the effects caused by Cd. Treatment with Zn protoporphyrin IX, a strong inhibitor of heme oxygenase, expectedly decreased heme oxygenase-1 activity to half. When the inhibitor was given together with Cd, completely prevented the enzyme induction and oxidative stress parameters were significantly enhanced. Conclusion: Taking together, these results are indicating that heme oxygenase plays a protective role against oxidative cell damage in soybean nodules.

[1]  A. Puppo,et al.  A Medicago sativa haem oxygenase gene is preferentially expressed in root nodules. , 2003, Journal of experimental botany.

[2]  A. M. del Batlle,et al.  Bilirubin is highly effective in preventing in vivo delta-aminolevulinic acid-induced oxidative cell damage. , 2003, Biochimica et biophysica acta.

[3]  M. Benavides,et al.  Effect of cadmium stress on nitrogen metabolism in nodules and roots of soybean plants. , 2003, Functional plant biology : FPB.

[4]  T. Kohchi,et al.  Expression and Biochemical Properties of a Ferredoxin-Dependent Heme Oxygenase Required for Phytochrome Chromophore Synthesis1 , 2002, Plant Physiology.

[5]  M. Tomaro,et al.  Bilirubin and ferritin as protectors against hemin-induced oxidative stress in rat liver. , 2002, Cellular and molecular biology.

[6]  A. Batlle,et al.  Bilirubin: its role in cytoprotection against oxidative stress. , 2002, The international journal of biochemistry & cell biology.

[7]  A. M. del C. Batlle,et al.  Paraquat-generated Oxidative Stress in Rat Liver Induces Heme Oxygenase-1 and Aminolevulinic Acid Synthase , 2002, Free radical research.

[8]  T. Kohchi,et al.  Making light of it: the role of plant haem oxygenases in phytochrome chromophore synthesis. , 2001, Biochemical Society transactions.

[9]  S. Gallego,et al.  Response of antioxidant defence system in soybean nodules and roots subjected to cadmium stress , 2001 .

[10]  R. Vierstra,et al.  The heme-oxygenase family required for phytochrome chromophore biosynthesis is necessary for proper photomorphogenesis in higher plants. , 2001, Plant physiology.

[11]  R. Foresti,et al.  Dynamics of haem oxygenase-1 expression and bilirubin production in cellular protection against oxidative stress. , 2000, The Biochemical journal.

[12]  S. Ryter,et al.  The heme synthesis and degradation pathways: role in oxidant sensitivity. Heme oxygenase has both pro- and antioxidant properties. , 2000, Free radical biology & medicine.

[13]  R. Vierstra,et al.  The Arabidopsis thaliana HY1 locus, required for phytochrome-chromophore biosynthesis, encodes a protein related to heme oxygenases. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[14]  L. Toppi,et al.  Response to cadmium in higher plants , 1999 .

[15]  I. Hwang,et al.  The Arabidopsis Photomorphogenic Mutant hy1 Is Deficient in Phytochrome Chromophore Biosynthesis as a Result of a Mutation in a Plastid Heme Oxygenase , 1999, Plant Cell.

[16]  R. Willows,et al.  Phytobilin biosynthesis: cloning and expression of a gene encoding soluble ferredoxin-dependent heme oxygenase from Synechocystis sp. PCC 6803. , 1998, The Plant journal : for cell and molecular biology.

[17]  M. Tomaro,et al.  Heme oxygenase induction by UVA radiation. A response to oxidative stress in rat liver. , 1998, The international journal of biochemistry & cell biology.

[18]  M. Tomaro,et al.  Heme oxygenase induction by cadmium chloride: evidence for oxidative stress involvement. , 1995, Toxicology.

[19]  G. Rhie,et al.  Phycobilin biosynthesis: reductant requirements and product identification for heme oxygenase from Cyanidium caldarium. , 1995, Archives of biochemistry and biophysics.

[20]  M L Tomaro,et al.  Heme oxygenase and oxidative stress. Evidence of involvement of bilirubin as physiological protector against oxidative damage. , 1994, Biochimica et biophysica acta.

[21]  J. Frydman,et al.  Heme oxygenase induction by CoCl2, Co-protoporphyrin IX, phenylhydrazine, and diamide: evidence for oxidative stress involvement. , 1991, Archives of biochemistry and biophysics.

[22]  M. Maines.  Heme oxygenase: function, multiplicity, regulatory mechanisms, and clinical applications , 1988, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[23]  J. Irigoyen,et al.  Some Enzymes of Hydrogen Peroxide Metabolism in Leaves and Root Nodules of Medicago sativa. , 1986, Plant physiology.

[24]  G. Kikuchi,et al.  Features of the reaction of heme degradation catalyzed by the reconstituted microsomal heme oxygenase system. , 1978, The Journal of biological chemistry.

[25]  J. .. Bassham,et al.  Chloroplast glutathione reductase. , 1977, Plant physiology.

[26]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[27]  A. Kappas,et al.  Studies on the mechanism of induction of haem oxygenase by cobalt and other metal ions. , 1976, The Biochemical journal.

[28]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[29]  H. Marver,et al.  The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. , 1968, Proceedings of the National Academy of Sciences of the United States of America.

[30]  L. Packer,et al.  Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. , 1968, Archives of biochemistry and biophysics.

[31]  Paul R. Ortiz De Montellanoa,et al.  75 – Heme Oxygenase Structure and Mechanism , 2003 .

[32]  A. Batlle,et al.  Paraquat-generated oxidative stress in rat liver induces heme oxygenase-1 and aminolevulinic acid synthase. , 2002, Free radical research.

[33]  P. Das,et al.  Studies on cadmium toxicity in plants: a review. , 1997, Environmental pollution.

[34]  A. Puppo,et al.  Direct detection of a globin-derived radical in leghaemoglobin treated with peroxides. , 1992, The Biochemical journal.

[35]  R. Stocker,et al.  Induction of haem oxygenase as a defence against oxidative stress. , 1990, Free radical research communications.

[36]  R. Tyrrell,et al.  Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[37]  M. Anderson,et al.  Determination of glutathione and glutathione disulfide in biological samples. , 1985, Methods in enzymology.