Journal of Plant Biology

, Volume 55, Issue 6, pp 469–480 | Cite as

Dehydroascorbate reductase and glutathione reductase play an important role in scavenging hydrogen peroxide during natural and artificial dehydration of Jatropha curcas seeds

  • Samar A. Omar
  • Nabil I. Elsheery
  • Hazem M. Kalaji
  • Zeng-Fu Xu
  • Song Song-Quan
  • Robert Carpentier
  • Choon-Hwan Lee
  • Suleyman I. Allakhverdiev
Original Article


Changes in H2O2 and the main antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR), in endospermic and embryonic tissues were studied in developing and artificially dried Jatropha curcas seeds. Immature seeds were desiccation-tolerant at 80 days after flowering, as they were able to germinate fully after artificial drying on silica gel had reduced their water content to 10–12% of fresh weight. In both endospermic and embryonic tissues, H2O2 level and, consequently, lipid peroxide content, decreased during seed development as well as after artificial dehydration of developing seeds. All examined antioxidant enzymes except DHAR showed a decrease in total activity in mature stages as compared with early stages. Expression analysis of SOD genes revealed that the decrease in total SOD activities was related to the decrease in Cu/Zn-SOD expression, while the continuous activity of SOD during maturation was related to an increase in Mn-SOD expression. Artificial drying resulted in increased SOD and DHAR activity, irrespective of the developmental stage. Our results revealed weak participation of CAT and APX in H2O2 scavenging, as well as no significant alterations in GR activities either during maturation or after artificial drying. Changes in SOD and GR isoenzyme patterns occurred during maturation-related drying, but not after artificial drying. These results highlight the role of ascorbate-glutathione cycle enzymes (DHAR and GR) in H2O2 scavenging during maturation or after artificial drying of developing J. curcas seeds.

Key words

dehydroascorbate reductase glutathione reductase hydrogen peroxide Jatropha curcas scavenging 


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  1. Aebi HE (1983) Catalase, In HU Bergmeyer, eds, Methods of Enzymatic Analysis. Verlage Chemie, Weinheim, Germany, pp 123–456Google Scholar
  2. Alscher R, Erturk N, Heath L (2002) Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. J Exp Bot 53:1331–1341PubMedCrossRefGoogle Scholar
  3. Alscher RG, Hess JL (1993) Antioxidants in higher plants, In CH Foyer, PM Mullineaux, eds, Causes of photooxidative streaa and Amelioration of defense system in plants. CRC Press, Boca Raton, USA, pp 59–87Google Scholar
  4. Allakhverdiev SI, Murata N (2004) Environmental stress inhibits the synthesis de novo of proteins involved in the photodamage-repair cycle of Photosystem II in Synechocystis sp. PCC 6803. Biochim Biophys Acta 1657:23–32PubMedCrossRefGoogle Scholar
  5. Allakhverdiev SI, Murata N (2008) Salt stress inhibits photosystems II and I in cyanobacteria. Photosynth Res 98:529–539PubMedCrossRefGoogle Scholar
  6. Allakhverdiev SI, Kreslavski VD, Klimov VV, Los DA, Carpentier R, Mohanty P (2008) Heat stress: An overview of molecular responses in photosynthesis. Photosynth Res 98:541–550PubMedCrossRefGoogle Scholar
  7. Asada K, Takahashi M (1987) Production, scavenging and action of active oxygen. In: DJ Kyle, CB Osmond, CJ Arntzen, eds, Photosynthesis. Elsevier, Amsterdam, pp 227–287Google Scholar
  8. Bailly C, Leymarie J, Lehner A, Rousseau S, Come D, Corbineau F (2004) Catalase activity and expression in developing sunflower seeds as related to drying. J Exp Bot 55:475–483PubMedCrossRefGoogle Scholar
  9. Beauchamp CO, Fridovich I (1971) Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287PubMedCrossRefGoogle Scholar
  10. Bewley JD, Black M (1983) Development, germination and growth. In: JD Bewley, M Black, eds, Physiology and Biochemistry of Seeds: In Relation to Germination. Springer-Verlag, New York, pp 119–207Google Scholar
  11. Bewley JD, Black M (1994) Seeds: physiology and germination. Plenum Press, New York, 445Google Scholar
  12. Bhattacharjee S (2005) Reactive oxygen species and oxidative burst: roles in stress, senescence and signal transduction in plants. Curr Sci India 89:1113–1121Google Scholar
  13. Borraccino G, Mastropasqua L, de Leonardis S, Dipierro S (1994) The role of the ascorbic acid system in delaying the senesce of oat (Avena sative L.) leaf segments. J Plant Physiol 144:161–166CrossRefGoogle Scholar
  14. Bowler C, Slooten L, Vandenbranden S, de Rycke R, Botterman J, Sybesma C, van Montagu M, Inzé D (1991) Manganese superoxide dismutase can reduce cellular damage mediated by oxygen radicals in transgenic plants. EMBO J 10:1723–1732PubMedGoogle Scholar
  15. Bowler C, van Camp W, van Montagu M, Inzé D. 1994. Superoxide dismutase in plants. Plant Sci 13:199–218Google Scholar
  16. Bradford MM (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
  17. Buchanan BB, Balmer Y (2005) Redox regulation: A broadening horizon. Ann Rev Plant Biol 56:187–220.CrossRefGoogle Scholar
  18. Bukhov NG, Govindachary S, Egorova EA, Carpentier R (2004) Recovery of photosystem I and II activities during re-hydration of lichen Hypogymnia physodes thalli. Planta 219:110–120PubMedCrossRefGoogle Scholar
  19. Cakmak I, Strbac D, Marschner H (1993) Activities of hydrogen peroxide-scavenging enzymes in germinating wheat seeds. J Exp Bot 44:127–132CrossRefGoogle Scholar
  20. Creissen GP, Edwards EA, Mullineaux PM (1994) Glutathione reductase and ascorbate peroxidase. In: CH Foyer, PM Mullineaux, eds, Causes of Photooxidative Stress and Amelioration of Defense System in Plants. CRC Press, Boca Raton, FL, USA, pp 343–364Google Scholar
  21. de Gara L, de Pinto MC, Paciolla C, Cappetti V, Arrigoni O (1996) Is Ascorbate peroxidase only a scavenger of hydrogen peroxide? In C Obinger, U Burner, R Ederman, C Penel, H Greppen, eds, Plant peroxidases biochemistry and physiology. University of Geneve, Geneve, Switzerland, pp 157–162Google Scholar
  22. de Gara L, de Pinto MC, Moliterni VM, d’Egidio MG (2003) Redox regulation and storage processes during maturation in kernels of Triticum durum. J Exp Bot 54:249–258PubMedCrossRefGoogle Scholar
  23. de Gara L, de Pinto MC. Arrigoni O (1997) Ascorbate synthesis and ascorbate peroxidase activity during the early stage of wheat germination. Physiol Plantarum 100:894–900CrossRefGoogle Scholar
  24. de Pinto MC, Tommasi F, de Gara L (2000) Enzymes of the ascorbate biosynthesis and ascorbate-glutathione cycle in cultured cells of tobacco Bright Yellow 2. Plant Physiol Biochem 38:541–550CrossRefGoogle Scholar
  25. Ding LW, Sun QY, Wang ZY, Sun YB, Xu ZF (2008) Using silica particles to isolate total RNA from plant tissues recalcitrant to extraction in guanidine thiocyanate. Anal Biochem 374:426–428PubMedCrossRefGoogle Scholar
  26. Dizengremel P, Le Thiec D, Hasenfratz-Sauder MP, Vaultier MN, Bagard M, Jolivet Y (2009) Metabolic-dependent changes in plant cell redox power after ozone exposure. Plant Biol 11:35–42PubMedCrossRefGoogle Scholar
  27. Donahue JL, Okpodu CM, Cramer CL, Grabau EA, Alscher RG (1997) Responses of antioxidants to paraquat in pea leaves (Relationships to resistance). Plant Physiol 113:249PubMedGoogle Scholar
  28. Finch-Savage WE, Grang RI, Hendry GAF, Atherton NM (1993) Embryo water status and loss of viability during desiccation in the recalcitrant species Quercus robur L. In D Côme, F Corbineau, (eds), Fourth International Workshope on Seeds: Basic and Applied Aspects of Seed Biology. ASFIS, Paris, pp 723–730Google Scholar
  29. Foyer C, Lelandais M, Galap C, Kunert KJ (1991) Effects of elevated cytosolic glutathione reductase activity on the cellular glutathione pool and photosynthesis in leaves undernormal and stress conditions. Plant Physiol 97:863–872PubMedCrossRefGoogle Scholar
  30. Foyer CH, Descourvieres P, Kunert KJ (1994) Protection against oxygen radicals: an important defence mechanism studied in transgenic plants. Plant Cell Environ 17:507–523CrossRefGoogle Scholar
  31. Foyer CH, Lopez-Delgado H, Dat JF, Scott IM (1997) Hydrogen peroxide-and glutathione-associated mechanisms of acclimatory stress tolerance and signalling. Physiol Plantarum 100:241–254CrossRefGoogle Scholar
  32. Garnczarska M, Bednarski W, Jancelewicz M (2009) Ability of lupine seeds to germinate and to tolerate desiccation as related to changes in free radical level and antioxidants in freshly harvested seeds. Plant Physiol Biochem 47:56–62PubMedCrossRefGoogle Scholar
  33. Garnczarska M, Wojtyla L (2008) Differential response of antioxidative enzymes in embryonic axes and cotyledons of germinating lupine seeds. Acta Physiol Plantarum 30:427–432CrossRefGoogle Scholar
  34. Goh C-H, Ko S-M, Koh S, Kim Y-J, Bae H-J (2012) Photosynthesis and environments: photoinhibition and repair mechanisms in plants. J Plant Biol 55:93–101CrossRefGoogle Scholar
  35. Gruissem W, Lee C-H, Oliver M, Pogson B (2012) The global plant council: Increasing the impact of plant research to meet global challenges. J Plant Biol 55:343–348CrossRefGoogle Scholar
  36. Haberer K, Herbinger K, Alexou M, Tausz M, Rennenberg H (2009) Antioxidative defence of old growth beech (Fagus sylvatica) under double ambient O3 concentrations in a free-air exposure system. Plant Biol 9:215–226CrossRefGoogle Scholar
  37. Halliwell B, Foyer CH (1978) Properties and physical function of glutathione reductase purified from spinach leaves by affinity chromatography. Planta 139:7–9CrossRefGoogle Scholar
  38. Heath RL, Packer L (1986) Photo-peroxidation in isolated chloroplasts. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefGoogle Scholar
  39. Hendry GAF, Finch-Savage WE, Thorpe PC, Atherton NM, Buckland SM, Nilsson KA, Seel WE (1992) Free radical processes and loss of seed viability during desiccation in the recalcitrant species Quercus robur L. New Phytol 122:273–279CrossRefGoogle Scholar
  40. Hendry GAF, Thorpe PC, Merzlyak MN (1993) Stress indicators: lipid peroxidation. In GAF Hendrey, JP Grime, eds, Methods in comparative plant ecology. Chapman & Hall, London, UK, pp 154–156CrossRefGoogle Scholar
  41. Henning RK (2003) Jatropha curcas L. in Africa. Baganì, Weissenberg, GermanyGoogle Scholar
  42. Hossain MA, Asada K (1984) Purification of dehydroascorbate reductase from spinach and its characterization as a thiol enzyme. Plant Cell Physiol 25:85–92Google Scholar
  43. Huang H, Song SQ, Wu XJ (2008) Response of Chinese Wampee Axes and Maize Embryos to Dehydration at Different Rates. J Integ Plant Biol 51:67–74CrossRefGoogle Scholar
  44. Kaniuga Z (2008) Chilling response of plants: importance of galactolipase, free fatty acids and free radicals. Plant Biol 10: 171–184PubMedCrossRefGoogle Scholar
  45. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  46. Lee DH, Lee CB (2000) Chilling stress-induced changes of antioxidant enzymes in the leaves of cucumber: in gel enzyme activity assays. Plant Sci 59:75–85CrossRefGoogle Scholar
  47. Lehner A, Bailly C, Flechel B, Poels P, Come D, Corbineau F (2006) Changes in wheat seed germination ability, soluble carbohydrate contents and antioxidant enzyme activities in the embryo during the desiccation phase of maturation. J Cereal Sci 43:175–182CrossRefGoogle Scholar
  48. Leprince O, Hendry GAF, McKersie BD (1993) The mechanisms of desiccation tolerance in developing seeds. Seed Sci Res 3:231–231CrossRefGoogle Scholar
  49. Leprince O, Hoekstra FA, Harren FJM (2000) Unravelling the responses of metabolism to dehydration points to a role for cytoplasmic viscosity in desiccation tolerance. In M Black, KJ Bradford, J Vasquez-Ramos, eds, Seed Biology: Advances and Application. CABI, New York, USA, pp 57–66Google Scholar
  50. Li C, Sun WQ (1999) Desiccation sensitivity and activities of free radical-scavenging enzymes in recalcitrant Theobroma cacao seeds. Seed Sci Res 9:209–217Google Scholar
  51. Li W, Qi L, Lin X, Chen H, Ma Z, Wu K, Huang S (2009) The expression of manganese superoxide dismutase gene from Nelumbo nucifera responds strongly to chilling and oxidative stresses. J Integ Plant Biol 51:279–286CrossRefGoogle Scholar
  52. Mattana E, Pritchard HW, Porceddu M, Stuppy WH, Bacchetta G (2012) Interchangeable effects of gibberellic acid and temperature on embryo growth, seed germination and epicotyl emergence in Ribes multiflorum ssp. sandalioticum (Grossulariaceae). Plant Biol 14:77–87PubMedGoogle Scholar
  53. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410PubMedCrossRefGoogle Scholar
  54. Murata N, Takahashi S, Nishiyama Y, Allakhverdiev SI (2007) Photoinhibition of photosystem II under environmental stress. Biochim Biophys Acta 1767:414–421PubMedCrossRefGoogle Scholar
  55. Murata N, Allakhverdiev SI, Nishiyama Y (2012) The mechanism of photoinhibition in vivo: Re-evaluation of the roles of catalase, α-tocopherol, non-photochemical quenching, and electron transport. Biochim Biophys Acta 1817:1127–1133PubMedCrossRefGoogle Scholar
  56. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  57. Nishiyama Y, Allakhverdiev SI, Murata N (2006) A new paradigm for the action of reactive oxygen species in the photoinhibition of photosystem II. Biochim Biophys Acta 1757:742–749PubMedCrossRefGoogle Scholar
  58. Nishiyama Y, Allakhverdiev SI, Murata N (2011) Protein synthesis is the primary target of reactive oxygen species in the photoinhibition of photosystem II. Physiol Plantarum 142:35–46CrossRefGoogle Scholar
  59. Navarri-Izzo F, Quartacci MF, Pinzino C, Vecchia FD, Sgherri CLM (1998) Thylakoid-bound and stromal antioxidative enzymes in wheat treated with excess copper. Physiol Plantarum 104:630–638CrossRefGoogle Scholar
  60. Neill S, Desikan R, Hancock J (2002) Hydrogen peroxide signaling. Curr Opin Plant Biol 5:388–395.PubMedCrossRefGoogle Scholar
  61. Openshaw K (2000) A review of Jatropha curcas: an oil plant of unfulfilled promise. Biomass Bioenerg 19:1–19CrossRefGoogle Scholar
  62. Pammenter NW, Berjak P (1999) A review of recalcitrant seed physiology in relation to desiccation-tolerance mechanisms. Seed Sci Res 9: 13–37CrossRefGoogle Scholar
  63. Parrish DJ, Leopold AC (1978) On the Mechanism of Aging in Soybean Seeds 1. Plant Physiol 61:365–368PubMedCrossRefGoogle Scholar
  64. Patterson BD, Macrae EA, Ferguson IB (1984) Estimation of hydrogen peroxide in plant extracts using titanium (IV). Anal Biochem 139:487–492PubMedCrossRefGoogle Scholar
  65. Posch S, Bennett LT (2009) Photosynthesis, photochemistry and antioxidative defence in response to two drought severities and with re-watering in Allocasuarina luehmannii. Plant Biol 11:83–93PubMedCrossRefGoogle Scholar
  66. Pukacka S, Hoffmann SK, Goslar J, Pukacki PM, Wójkiewicz E (2003) Water and lipid relations in beech (Fagus sylvatica L.) seeds and its effect on storage behaviour. Biophys Biochim Acta 1621:48–56CrossRefGoogle Scholar
  67. Pukacka S, Ratajczak E (2007) Ascorbate and glutathione metabolism during development and desiccation of orthodox and recalcitrant seeds of the genus Acer. Funct Plant Biol 34:601–613CrossRefGoogle Scholar
  68. Smirnoff N (1993) The role of active oxygen in the response of plants to water deficit and desiccation. New Phytol 125:27–58CrossRefGoogle Scholar
  69. Srikanta DKG, Hatti KS, Ravikumar P, Kush A (2011) Structural and functional analyses of a saturated acyl ACP thioesterase, type B from immature seed tissue of Jatropha curcas. Plant Biol 13: 453–461CrossRefGoogle Scholar
  70. Tommasi F, Paciolla C, Arrigoni O (1999) The ascorbate system in recalcitrant and orthodox seeds. Physiol Plantarum 105:193–198CrossRefGoogle Scholar
  71. Vertucci CW, Farrant JM (1995) Acquisition and loss of desiccation tolerance. In J Kigel, G Galili, eds, Seed Development and Germination. Marcel Dekker, New York, USA, pp 237–271Google Scholar
  72. Willekens H, Inze D, van Montagu M, van Camp W (1995) Catalases in plants. Mol Breeding 1:207–228CrossRefGoogle Scholar
  73. Woodbury W, Spencer AK, Stahmann MA (1971) An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem 44:301–305PubMedCrossRefGoogle Scholar
  74. Yang QH, Yin SH, Song SQ, Ye WH (2004) Development of desiccation tolerance and germination physiology of Crotalaria pallida Ait seeds. Seed Sci Technol 32:99–111Google Scholar
  75. Zulfugarov I, Tovuu A, Kim J-H, Lee C-H (2011) Detection of reactive oxygen species in higher plants. J Plant Biol 54:351–357CrossRefGoogle Scholar

Copyright information

© Korean Society of Plant Biologists and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Samar A. Omar
    • 1
    • 2
  • Nabil I. Elsheery
    • 3
  • Hazem M. Kalaji
    • 4
  • Zeng-Fu Xu
    • 1
    • 2
  • Song Song-Quan
    • 5
  • Robert Carpentier
    • 6
  • Choon-Hwan Lee
    • 7
  • Suleyman I. Allakhverdiev
    • 8
    • 9
  1. 1.Xishuangbanna Tropical Botanical GardenChinese Academy of SciencesKunmingChina
  2. 2.Graduate School of the Chinese Academy of SciencesBeijingChina
  3. 3.Agricultural Botany Department, Faculty of AgricultureTanta UniversityTantaEgypt
  4. 4.Department of Plant PhysiologyWarsaw University of Life Sciences SGGWWarsawPoland
  5. 5.Institute of BotanyChinese Academy of SciencesBeijingChina
  6. 6.Groupe de Recherche en Biologie Végétale (GRBV)Université du Québec à Trois-RivièresTrois-RivièresCanada
  7. 7.Department of Molecular BiologyPusan National UniversityBusanKorea
  8. 8.Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia
  9. 9.Institute of Basic Biological ProblemsRussian Academy of SciencesPushchino, Moscow RegionRussia

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