Plant Growth Regulation

, Volume 67, Issue 1, pp 83–92 | Cite as

Response of antioxidant systems in oxygen deprived suspension cultures of rice (Oryza sativa L.)

  • Revandy Iskandar Damanik
  • Mohd Razi Ismail
  • Zulkifli Shamsuddin
  • Sariam Othman
  • Abd Mohd Zain
  • Mahmood Maziah
Original paper


The effect of oxygen deprivation (anoxia) on the antioxidant system in suspension culture of anoxia-intolerant Malaysian rice mutants cells was examined. Abiotic stresses have been reported to adversely affect cell division, damage cellular and organelle membranes. The signaling defense mechanisms, such as molecular and biochemical aspects responding to stress have been proven to be very complex, and still largely untapped. The objective of this study was to determine the potential involvement of activated oxygen species, such as superoxide dismutase, catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase which occur in cells of rice plants exposed to anoxia stress in two Malaysian rice mutants, MR219-4 and MR219-9, and rice cultivar FR13A which is known to be tolerant to anoxia stress during 5–30 days of exposure. The antioxidative enzymes were decreased for MR219-4 and MR219-9 mutants for CAT and APX activities, and increased in FR13A cultivar starting at 20 days in suspension culture compared to that of control. CAT and APX activities were maintained higher in anoxia condition for all mutants and cultivar. These findings suggested that anoxia stress in suspension cultures induced the level of H2O2 to toxic levels.


Antioxidant enzymes Suspension culture Cultivars Periods of stress Rice Anoxia stress 



This research was supported by Graduate Research Fund (GRF) of Universiti Putra Malaysia (UPM).


  1. Ahmed S, Nawata E, Hosokawa M, Domae Y, Sakuratani T (2002) Alterations in photosynthesis and some antioxidant enzymatic activities of mungbean subjected to waterlogging. Plant Sci 163:117–123CrossRefGoogle Scholar
  2. Anbazhagan M, Rajendran R, Kalpana M, Natarajan V, Panneerselvam R (2009) Agrobacterium mediated transformation of rice var. Pusa Basmati-1. J Ecol Biotech 1(1):7–11Google Scholar
  3. Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50:601–639PubMedCrossRefGoogle Scholar
  4. Babcock GT (1999) How oxygen is activated and reduced in respiration. Proc Natl Acad Sci USA 96:12971–12973PubMedCrossRefGoogle Scholar
  5. Bergmeyer N (1970) Methoden der enzymatischen, analyse, vol 1. Akademie Verlag, Berlin, pp 636–647Google Scholar
  6. Blokhina O, Eija V, Kurt VF (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot 91:179–194PubMedCrossRefGoogle Scholar
  7. Boscolo PRS, Menossi M, Jorge RA (2003) Aluminium induced oxidative stress in maize. Phytochemistry 62:181–189PubMedCrossRefGoogle Scholar
  8. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  9. Bray EA, Bailey-Serres J, Weretilnyk E (2000) Responses to abiotic stress. In: Buchanan BB, Gruissem W, Jones RL (eds) Biochemistry and molecular biology of plants. American Society of Plant Biologists, Waldorf, pp 1158–1203Google Scholar
  10. Cotter TG, Al-Rubeai M (1995) Cell death (apoptosis) in cell culture systems. Trends Biotechnol 13:150–155PubMedCrossRefGoogle Scholar
  11. Damanik RI, Maziah M, Ismail MR, Syahida A, Zain AM (2010) Responses of antioxidative enzymes in Malaysian rice (Oryza sativa L.) cultivars under submergence condition. Acta Physiol Plant 32:739–747CrossRefGoogle Scholar
  12. Dempsey DA, Shah J, Klessig DF (1999) Salicylic acid and disease resistance in plants. Crit Rev Plant Sci 18:547–575CrossRefGoogle Scholar
  13. Dewir YH, Chakrabarty D, Ali MB, Hahn EJ, Paek KY (2006) Lipid peroxidation and antioxidant enzyme activities of Euphorbia millii hyperhydric shoots. Environ Exp Bot 58:93–99CrossRefGoogle Scholar
  14. Dionisio-Sese ML, Tobita S (1998) Antioxidant responses of rice seedlings to salinity stress. Plant Sci 135:1–9CrossRefGoogle Scholar
  15. Elstner EF, Osswald W (1994) Mechanisms of oxygen activation during plant stress. Proc Roy Soc Edinb 102B:131–154Google Scholar
  16. Foyer C, Descourvieres P, Kunert KJ (1994) Protection against oxygen radicals: an important defense mechanism studied in transgenic plant. Plant Cell Environ 17:507–523CrossRefGoogle Scholar
  17. Fukao T, Bailey-Serres J (2008) Ethylene—A key regulator of submergence responses in rice. Plant Sci 175:43–51CrossRefGoogle Scholar
  18. Goldberg DM, Spooner RJ (1983) Glutathione reductase. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 3. Basel, pp 258–265Google Scholar
  19. Henzler T, Steudle E (2000) Transport and metabolic degradation of hydrogen peroxide in Chara coralline: model calculations and measurements with the pressure probe suggest transport of H2O2 across water channels. J Exp Bot 353:2053–2066CrossRefGoogle Scholar
  20. Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611CrossRefGoogle Scholar
  21. Htwe NN, Maziah M, Ho CL, Zaman FQ, Zain AM (2011) Responses of some selected Malaysian rice genotypes to callus induction under in vitro salt stress. Afr J Biotech 10(3):350–362Google Scholar
  22. Hung SH, Yu CW, Lin CH (2005) Hydrogen peroxide functions as a stress signal in plants. Bot Bull Acad Sin 46:1–10Google Scholar
  23. IRRI (1988) Standard evaluation system for rice, 3rd edn. International Rice Research Institute, Manila, p 54Google Scholar
  24. Jain M, Mathur G, Koul S, Sarin NB (2001) Ameliorative effects of proline on salt stress-induced lipid peroxidation in cell lines of groundnut (Arachis hypogea L.). Plant Cell Rep 20:463–468CrossRefGoogle Scholar
  25. Kocsy G, Galiba G, Brunold C (2001) Role of glutathione in adaptation and signaling during chilling and cold acclimation in plants. Physiol Plant 113:158–164PubMedCrossRefGoogle Scholar
  26. Kocsy G, Szalai G, Galiba G (2004) Effect of osmotic stress on glutathione and hydroxymethylglutathione accumulation in wheat. J Plant Physiol 161:785–794PubMedCrossRefGoogle Scholar
  27. Lafitte HR, Yongsheng G, Yan S, Li ZK (2007) Whole plant responses, key processes, and adaptation to drought stress: the case of rice. J Exp Bot 58:169–175PubMedCrossRefGoogle Scholar
  28. Lee SH, Ahsan N, Lee KW, Kim DH, Lee DG, Kwak SS, Kwon SY, Kim TH, Lee BH (2007) Simultaneous overexpression of both CuZn superoxide dismutase and ascorbate peroxidase in transgenic tall fescue plants confers increased tolerance to a wide range of abiotic stresses. J Plant Physiol 64:1626–1638CrossRefGoogle Scholar
  29. Lin CC, Kao CH (2000) Effect of NaCl stress on H2O2 metabolism in rice leaves. Plant Growth Regul 30:151–155CrossRefGoogle Scholar
  30. Małecka A, Derba-Maceluch M, Kaczorowska K, Piechalak A, Tomaszewska B (2009) Reactive oxygen species production and antioxidative defense system in pea root tissues treated with lead ions: mitochondrial and peroxisomal level. Acta Physiol Plant 31:1065–1075CrossRefGoogle Scholar
  31. Matés JM (2000) Effects of antioxidant enzymes in the molecular control of reactive oxygen species. Toxicology 153:83–104PubMedCrossRefGoogle Scholar
  32. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–479CrossRefGoogle Scholar
  33. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-spesific peroxidase in spinach chloroplast. Plant Cell Physiol 22:867–880Google Scholar
  34. Parida AK, Das AB, Mohanty P (2004) Defense potentials to NaCl in a mangrove, Bruguiera parviflora: differential changes of isoforms of some antioxidative enzymes. J Plant Physiol 161:531–542PubMedCrossRefGoogle Scholar
  35. Park SW, Jeon JH, Kim HS, Park YM, Aswath C, Joung H (2004) Effect of sealed and vented gaseous microenvironments on the hyperhydricity of potato shoots in vitro. Sci Hortic 99:199–205CrossRefGoogle Scholar
  36. Peng M, Kuc J (1992) Peroxidase—generated hydrogen peroxide as a source of antifungal activity in vitro and on tobacco leaf discs. Phytopathology 82:696–699CrossRefGoogle Scholar
  37. Perata P, Geshi N, Yamaguchi J, Akazawa T (1993) Effect of anoxia on the induction of α-amylase in cereal seeds. Planta 191:402–408CrossRefGoogle Scholar
  38. Perata P, Guglielminetti L, Alpi A (1996) Anaerobic carbohydrate metabolism in wheat and barley, two anoxia-intolerant cereal seeds. J Exp Bot 47:999–1006CrossRefGoogle Scholar
  39. Ryals JA, Neuenschwander UH, Willits MG, Molina A, Steiner H-Y, Hunt MD (1996) Systemic acquired resistance. Plant Cell 8:1809–1819PubMedCrossRefGoogle Scholar
  40. Saglio PH (1985) Effect of path or sink anoxia on sugar translocation in roots of maize seedlings. Plant Physiol 77:285–290PubMedCrossRefGoogle Scholar
  41. Saher S, Fernández-García N, Piqueras A, Hellin E, Olmos E (2005) Reducing properties, energy efficiency and carbohydrate metabolism in hyperhydric and normal carnation shoots cultured in vitro: a hypoxia stress? Plant Physiol Biochem 43:573–582PubMedCrossRefGoogle Scholar
  42. Samarajeewa PK, Barrero RA, Umeda-Hara C, Kawai M, Uchimiya H (1999) Cortical cell death, cell proliferation, macromolecular movements and rTip1 expression pattern in roots of rice (Oryza sativa L.) under NaCl stress. Planta 207:354–361CrossRefGoogle Scholar
  43. Shalata A, Mittova V, Volokita M, Guy M, Tal M (2001) Response of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii to salt-dependent oxidative stress: the root antioxidative system. Physiol Plant 112:487–494PubMedCrossRefGoogle Scholar
  44. Shanker AK, Djanaguiraman M, Sudhagar R, Chandrashekar CN, Pathmanabhan G (2004) Differential antioxidative response of ascorbate glutathione pathway enzymes and metabolites to chromium speciation stress in green gram (Vigna radiate (L.) R.Wilczek. cv CO4) roots. Plant Sci 166:1035–1043CrossRefGoogle Scholar
  45. Stewart RC, Bewley JD (1980) Lipid peroxidation associated with accelerated aging of soybean axes. Plant Physiol 65:245–248PubMedCrossRefGoogle Scholar
  46. Sticher L, Mauch-Mani B, Métrauxs J-P (1997) Systemic acquired resistance. Annu Rev Phytopath 35:235–270CrossRefGoogle Scholar
  47. Ushimaru T, Kanematsu S, Shibasaka M, Tsuji H (1999) Effect of hypoxia on the antioxidative enzymes in aerobically grown rice (Oryza sativa) seedlings. Physiol Plant 107:181–187CrossRefGoogle Scholar
  48. Vartapetian BB, Andreeva IN, Kozlova GI, Agapova LP (1977) Mitochondrial ultrastructure of roots of mesophytes and hydrophytes at anoxia and after glucose feeding. Protoplasma 91:243–256CrossRefGoogle Scholar
  49. Wang Y, Xue Y, Jiayang LJ (2005) Toward molecular breeding and improvement of rice in China. Trends Plant Sci 10(12):610–614PubMedCrossRefGoogle Scholar
  50. Webb T, Armstrong W (1983) The effects of anoxia and carbohydrates on the growth and viability of rice, pea and pumpkin roots. J Exp Bot 34:579–603CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Revandy Iskandar Damanik
    • 1
  • Mohd Razi Ismail
    • 2
  • Zulkifli Shamsuddin
    • 3
  • Sariam Othman
    • 4
  • Abd Mohd Zain
    • 5
  • Mahmood Maziah
    • 1
  1. 1.Department of Biochemistry, Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra MalaysiaSerdangMalaysia
  2. 2.Institute of Tropicial AgricultureUniversiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of Land Management, Faculty of AgricultureUniversiti Putra MalaysiaSerdangMalaysia
  4. 4.Rice and Industrial Crop Research Centre MARDI Head QuartersKuala LumpurMalaysia
  5. 5.Department of Agrotechnology, Faculty of Agrotechnology and Food ScienceUniversiti Malaysia TerengganuKuala TerengganuMalaysia

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