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Plant Cell Reports

, Volume 26, Issue 11, pp 2027–2038 | Cite as

Involvement of oxidative stress and role of antioxidative defense system in growing rice seedlings exposed to toxic concentrations of aluminum

  • Pallavi Sharma
  • R. S. DubeyEmail author
Biotic and Abiotic Stress

Abstract

When seedlings of rice (Oryza sativa L.) cultivar Pant-12 were raised in sand cultures containing 80 and 160 μM Al3+ in the medium for 5–20 days, a regular increase in Al3+ uptake with a concomitant decrease in the length of roots as well as shoots was observed. Al3+ treatment of 160 μM resulted in increased generation of superoxide anion (O2 ) and hydrogen peroxide (H2O2), elevated amount of malondialdehyde, soluble protein and oxidized glutathione and decline in the concentrations of thiols (-SH) and ascorbic acid. Among antioxidative enzymes, activities of superoxide dismutase (SOD EC 1.15.1.1), guaiacol peroxidase (Guaiacol POX EC 1.11.1.7), ascorbate peroxidase (APX EC 1.11.1.11), monodehydroascorbate reductase (MDHAR EC 1.6.5.4), dehydroascorbate reductase (EC 1.8.5.1) and glutathione reductase (EC 1.6.4.2) increased significantly, whereas the activities of catalase (EC EC 1.11.1.6) and chloroplastic APX declined in 160 μM Al3+ stressed seedlings as compared to control seedlings. The results suggest that Al3+ toxicity is associated with induction of oxidative stress in rice plants and among antioxidative enzymes SOD, Guaiacol POX and cytosolic APX appear to serve as important components of an antioxidative defense mechanism under Al3+ toxicity. PAGE analysis confirmed the increased activity as well as appearance of new isoenzymes of APX in Al3+ stressed seedlings. Immunoblot analysis revealed that changes in the activities of APX are due to changes in the amounts of enzyme protein. Similar findings were obtained when the experiments were repeated using another popular rice cv. Malviya-36.

Keywords

Aluminum stress Antioxidative enzymes Lipid peroxidation Oryza sativa Oxidative stress 

Abbreviations

chl-APX

Chloroplastic ascorbate peroxidase

APX

Ascorbate peroxidase

AsA

Ascorbic acid

CAT

Catalase

DHAR

Dehydroascorbate reductase

Guaiacol POX

Guaiacol peroxidase

GR

Glutathione reductase

GSH

Glutathione

MDA

Malondialdehyde

MDHAR

Monodehydroascorbate reductase

SOD

Superoxide dismutase

Notes

Acknowledgments

We gratefully acknowledge Dr Ishikawa of Faculty of Life and Environmental Science, Shimane University, Japan for the gift of APX antibody. We are thankful to Dr Yoko Yamamoto of Research Institute for Bioresources, Kurashiki, Okayama University, Japan for providing the facilities related to image analysis of our photographs using image analyzer model LAS-1000 Plus Fuji Photo film Co. Ltd., Japan and to Hirotoshi Motoda for assisting in the analysis of images. PS is thankful to the Council of Scientific and Industrial Research (CSIR), New Delhi for providing a Senior Research Fellowship.

References

  1. Aebi HE (1983) Catalase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Verlag Chemie, Weinhern, pp 273–286Google Scholar
  2. Arroyo-Serralta GA, Kú-González A, Hernández-Sotomayor SMT, Aguilar JJZ (2005) Exposure to toxic concentrations of aluminum activates a MAPK-like protein in cell suspension cultures of Coffea Arabica. Plant Physiol Biochem 43:27–35PubMedCrossRefGoogle Scholar
  3. Asada K (1994) Production and action of active oxygen species in photosynthetic tissues. In: Foyer CH, Mullineaux PM (eds) Causes of photooxidative stress and amelioration of defense systems in plants. CRC Press, Boca Raton, pp 77–104Google Scholar
  4. Atal N, Saradhi PP, Mohanty P (1991) Inhibition of the chloroplast photochemical reactions by treatment of wheat seedlings with low concentrations of cadmium: analysis of electron transport activities and changes in fluorescence yield. Plant Cell Physiol 32:943–951Google Scholar
  5. Boscolo PRS, Menossi M, Jorge RA (2003) Aluminium induced oxidative stress in maize. Phytochem 62:181–189CrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  7. Cakmak I, Horst WJ (1991) Effect of aluminium on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycine max). Physiol Plant 83:463–468CrossRefGoogle Scholar
  8. Cuypers A, Vangronsveld J, Clijsters H (2002) Peroxidases in roots and primary leaves of Phaseolus vulgaris, copper and zinc phytotoxicity: a comparison. J Plant Physiol 159:869–876CrossRefGoogle Scholar
  9. De Gara L, Tullio MDE, Paciolla C, Liso R, Arrigoni O (1993) Cytosolic ascorbate peroxidase in angiosperms and the different expression of its isoforms in maize embryo during germination. In: Welinder KG, Rasmussen SK, Penel C, Greppin H (eds) Plant peroxidases: biochemistry and physiology. University of Geneva, Geneva, pp 251–255Google Scholar
  10. deKok LJ, Kuiper PJC (1986) Effect of short term dark incubation with chloride and selenate on the glutathione content of spinach leaf discs. Physiol Plant 68:477–482CrossRefGoogle Scholar
  11. Dipierro N, Mondelli D, Paciolla C, Brunetti G, Dipierro S (2005) Changes in the ascorbate system in the response of pumpkin root to aluminium stress. Plant Physiol 162:529–536CrossRefGoogle Scholar
  12. Dobermann A, Fairhurst T (2000) Rice: nutrient disorders and nutrient management. Potash and Phosphate Institute (PPI), Potash and Phosphate Institute of Canada (PPIC) and International Rice Research Institute (IRRI), Philippines, pp 135–138Google Scholar
  13. Doulis AG, Debian N, Kingston-Smith AH, Foyer CH (1997) Differential localization of antioxidants in maize leaves. Plant Physiol 116:1031–1037Google Scholar
  14. Edjolo A, Laffray D, Guerrier G (2001) The ascorbate-glutathione cycle in the cytosolic and chloroplastic fractions of drought-tolerant and drought-sensitive poplars. J Plant Physiol 158:1511–1517CrossRefGoogle Scholar
  15. Egley GH, Paul RN, Vaughn KC, Duke SO (1983) Role of peroxidase in the development of water impermeable seed coats in Sida spinosa L. Planta 157:224–232CrossRefGoogle Scholar
  16. Ezaki B, Gardner RC, Ezaki Y, Matsumoto H (2000) Expression of aluminium-induced genes in transgenic Arabidopsis plants can ameliorate aluminium stress and and/or oxidative stress. Plant Physiol 122:657–665PubMedCrossRefGoogle Scholar
  17. Foyer CH, Halliwell B (1976) The presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21–25CrossRefGoogle Scholar
  18. Giannopolitis CN, Ries SK (1972) Superoxide dismutase I. Occurrence in higher plants. Plant Physiol 59:309–314Google Scholar
  19. Griffith O (1980) Determination of glutathione and glutathione disulphide using glutathione reductase and 2- vinyl pyridine. Anal Biochem 106:207–212PubMedCrossRefGoogle Scholar
  20. Guo Z, Tan H, Zhu Z, Lu S, Zhou B (2005) Effect of intermediates on ascorbic acid and oxalate biosynthesis of rice and in relation to its stress resistance. Plant Physiol Biochem 43:955–963PubMedCrossRefGoogle Scholar
  21. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198PubMedCrossRefGoogle Scholar
  22. Hossain MA, Nakano Y, Asada K (1984) Monodehydroascorbate reductase in spinach chloroplasts and its participation in regeneration of ascorbate for scavenging hydrogen peroxide. Plant Cell Physiol 25:385–395Google Scholar
  23. Hsu PH (1963) Effect of pH, phosphate and silicate on the determination of aluminium with aluminon. Soil Sci 96:230–238CrossRefGoogle Scholar
  24. Ishikawa T, Takeda T, Kohno H, Shigeoka S (1996) Molecular characterization of Euglena ascorbate peroxidase using monoclonal antibody. Biochim Biophys Acta 1290:69–75PubMedGoogle Scholar
  25. Jana S, Chaudhuri A (1981) Glycolate metabolism of three submerged aquatic angiosperms during aging. Aquat Bot 12:345–354CrossRefGoogle Scholar
  26. Jones DL, Blancaflor EB, Kochian LV, Gilroy S (2006) Spatial coordination of aluminium uptake, production of reactive oxygen species, callose production and wall rigidification in maize roots. Plant Cell Environ 29:1309–1318PubMedCrossRefGoogle Scholar
  27. Kinraide TR, Ryan PR, Kochian LV (1992) Interactive effects of Al3+, H+, and other cations on root elongation considered in terms of cell surface electrical potential. Plant Physiol 99:1461–1468PubMedCrossRefGoogle Scholar
  28. Kochian LV (1995) Cellular mechanisms of aluminum toxicity and resistance in plants. Annu Rev Plant Physiol Plant Mol Biol 46:237–260CrossRefGoogle Scholar
  29. Kuniak E, Sklodowska M (2005) Compartment-specific role of the ascorbate-glutathione cycle in the response of tomato leaf cells to Botrytis cinerea infection. J Exp Bot 56:921–933CrossRefGoogle Scholar
  30. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  31. Larsen PB, Kochian LV, Howell SH (1997) Al inhibits both shoot development and root growth in als3, an Al-sensitive Arabidopsis mutant. Plant Physiol 114:1207–1214PubMedGoogle Scholar
  32. Law MY, Charles SA, Halliwell B (1983) Glutathione and ascorbic acid in spinach (Spinacea oleracea) chloroplasts. The effect of hydrogen peroxide and of paraquat. Biochem J 210:899–903PubMedGoogle Scholar
  33. Ma JF, Nagao S, Sato K, Ito H, Furukawa J, Tekeda K (2004) Molecular mapping of a gene responsible for Al-activated secretion of citrate in barley. J Exp Bot 55:1335–1341PubMedCrossRefGoogle Scholar
  34. Meriga B, Reddy BK, Rao KR, Kishor PBK (2003) Aluminium-induced production of oxygen radicals, lipid peroxidation and DNA damage in seedlings of rice (Oryza sativa). J Plant Physiol 161:63–68CrossRefGoogle Scholar
  35. Mishra HP, Fridovich I (1972) The role of superoxide anion in auto-oxidation of the epinephrine and sample assay for SOD. J Biol Chem 247:3170–3175Google Scholar
  36. Mittler R, Zilinskas BA (1994) Regulation of pea cytosolic ascorbate peroxidase and other antioxidant enzymes during the progression of drought stress and following recovery from drought. Plant J 5:397–405PubMedCrossRefGoogle Scholar
  37. Nakano Y, Asada K (1987) Purification of ascorbate peroxidase in spinach chloroplasts; its inactivation in ascorbate depleted medium and reactivation by monodehydro ascorbate radical. Plant Cell Physiol 28:131–140Google Scholar
  38. Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279PubMedCrossRefGoogle Scholar
  39. Pereira LM, Tabaldi LA, Gonçalves JF, Jucoski GO, Pauletto MM, Weis SN, Nicoloso FT, Borher D, Rocha JBT, Schetinger MRC (2006) Effect of aluminum on δ-aminolevulinic acid dehydratase (ALA-D) and the development of cucumber (Cucumis sativus). Environ Exp Bot 57:106–115CrossRefGoogle Scholar
  40. Richharia A, Shah K, Dubey RS (1997) Nitrate reductase from rice seedlings: partial purification, characterization and the effects of in situ and in vitro NaCl salinity. J Plant Physiol 151:316–322Google Scholar
  41. Sharma P, Dubey RS (2004) Ascorbate peroxidase from rice seedlings: properties of enzyme isoforms, effects of stresses and protective roles of osmolytes. Plant Sci 167:541–550CrossRefGoogle Scholar
  42. Simonovicova M, Tamas L, Huttova J, Mistrík I (2004) Effect of aluminium on oxidative stress related enzymes activities in barley roots. Biol Plant 48:261–266CrossRefGoogle Scholar
  43. Sivaguru M, Baluska F, Volkmann D, Felle HH, Horst WJ (1999) Impacts of aluminum on the cytoskeleton of the maize root apex. Short-term effects on the distal part of the transition zone. Plant Physiol 119:1073–1082PubMedCrossRefGoogle Scholar
  44. Sivaguru M, Yamamoto Y, Rengel Z, Ahn SJ, Matsumoto H (2005) Early events responsible for aluminum toxicity symptoms in suspension-cultured tobacco cells. New Phytol 165:99–109PubMedCrossRefGoogle Scholar
  45. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A, 76:4350–4354PubMedCrossRefGoogle Scholar
  46. Ushimaru T, Kanematsu S, Shibasaka M, Tsuji H (1999) Effect of hypoxia on antioxidant enzymes in aerobically grown rice (Oryza sativa) seedlings. Physiol Plant 107:181–187CrossRefGoogle Scholar
  47. Verma S, Dubey RS (2003) Lead toxicity induces lipid peroxidation and alters the activities of antioxidant enzymes in growing rice plants. Plant Sci 164:645–655CrossRefGoogle Scholar
  48. Wang JW, Kao CH (2006) Aluminum-inhibited root growth of rice seedlings is mediated through putrescine accumulation. Plant Soil 288:373–381CrossRefGoogle Scholar
  49. Wang J, Zhang H, Allen RD (1999) Overexpression of an Arabidopsis peroxisomal ascorbate peroxidase gene in tobacco increases protection against oxidative stress. Plant Cell Physiol 40:725–732PubMedGoogle Scholar
  50. Watt DA (2004) Aluminium-responsive genes in sugarcane: identification and analysis of expression under oxidative stress. J Exp Bot 385:1163–1174Google Scholar
  51. Yamamoto Y, Rikiishi S, Chang YC, Ono K, Kasai M, Matsumoto H (1994) Quantitative estimation of aluminum toxicity in cultured tobacco cells: correlation between aluminum uptake and growth inhibition. Plant Cell Physiol 35:575–583Google Scholar
  52. Yamamoto Y, Kobayashi Y, Matsumoto H (2001) Lipid peroxidation is an early symptom triggered by aluminium, but not the primary cause of elongation inhibition in pea roots. Plant Physiol 125:199–208PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.Department of Biochemistry, Faculty of ScienceBanaras Hindu UniversityVaranasiIndia
  2. 2.Department of PhysicsUniversity of ArkansasFayettevilleUSA

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