Protoplasma

, Volume 250, Issue 2, pp 585–600 | Cite as

Reactive oxygen species, ascorbate–glutathione pool, and enzymes of their metabolism in drought-sensitive and tolerant indica rice (Oryza sativa L.) seedlings subjected to progressing levels of water deficit

Original Article

Abstract

Water deficit for rice is a worldwide concern, and to produce drought-tolerant varieties, it is essential to elucidate molecular mechanisms associated with water deficit tolerance. In the present study, we investigated the differential responses of nonenzymatic antioxidants ascorbate (AsA), glutathione (GSH), and their redox pool as well as activity levels of enzymes of ascorbate–glutathione cycle in seedlings of drought-sensitive rice (Oryza sativa L.) cv. Malviya-36 and drought-tolerant cv. Brown Gora subjected to water deficit treatment of −1.0 and −2.1 MPa for 24–72 h using PEG-6000 in sand cultures. Water deficit caused increased production of reactive oxygen species such as O2, H2O2, and HO⋅ in the tissues, and the level of production was higher in the sensitive than the tolerant cultivar. Water deficit caused reduction in AsA and GSH and decline in their redox ratios (AsA/DHA and GSH/GSSG) with lesser decline in tolerant than the sensitive seedlings. With progressive level of water deficit, the activities of monodehydroascorbate reductase, dehydroascorbate reductase, ascorbate peroxidase (APX), and glutathione transferase increased in the seedlings of both rice cultivars, but the increased activity levels were higher in the seedlings of drought-tolerant cv. Brown Gora compared to the sensitive cv. Malviya-36. Greater accumulation of proline was observed in stressed seedlings of tolerant than the sensitive cultivar. In-gel activity staining of APX revealed varying numbers of their isoforms and their differential expression in sensitive and tolerant seedlings under water deficit. Results suggest that an enhanced oxidative stress tolerance by a well-coordinated cellular redox state of ascorbate and glutathione in reduced forms and induction of antioxidant defense system by elevated activity levels of enzymes of ascorbate–glutathione cycle is associated with water deficit tolerance in rice.

Keywords

Ascorbate Glutathione Proline Water deficit Rice Oryza sativa L. 

References

  1. Asada K (1999) The water cycle in chloroplast: scavenging of active oxygens and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50:601–639PubMedCrossRefGoogle Scholar
  2. Ashraf M, Foolad MR (2007) Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot 59:206–216CrossRefGoogle Scholar
  3. Azevedo-Neto AD, Prisco JT, Eneas-Filho J, de Abrau CEB, Gomez-Filho E (2006) Effect of salt stress on antioxidative enzymes and lipid peroxidation in leaves and roots of salt-tolerant and salt-sensitive maize genotypes. Environ Exp Bot 56:87–94CrossRefGoogle Scholar
  4. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  6. De Gara L, Paciolla C, Tullio MDE, Liso R, Arrigoni O (1993) Cytosolic ascorbste peroxidase in angiosperms and the different expression of its isoforms in maize embryo during germination. In: Welinder KG, Rasmussen SK, Panel C, Greppin H (eds) Plant peroxidases: biochemistry and physiology. University of Geneva, Geneva, pp 251–255Google Scholar
  7. Demiral T, Turkan I (2005) Comparative lipid peroxidation, antioxidants defense systems and proline content in roots of two rice cultivars differing in salt tolerance. Environ Exp Bot 53:247–257CrossRefGoogle Scholar
  8. Doulis A, Debian N, Kingston-Smith A, Foyer CH (1997) Characterization of chilling sensitivity in maize: differential localization of antioxidants in maize leaves. Plant Physiol 114:1031–1037PubMedGoogle Scholar
  9. Eltayeb AE, Kawano N, Badawi GH, Kaminaka H, Sanekata T, Shibahara T, Inanaga S, Tanaka K (2007) Overexpression of monodehydroascorbate reductase in transgenic tobacco confers enhanced tolerance to ozone, salt and polyethylene glycol stresses. Planta 225:1255–1264PubMedCrossRefGoogle Scholar
  10. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875PubMedCrossRefGoogle Scholar
  11. Frahry G, Schopfer P (2001) NADH-stimulated, cyanide-restistant superoxide production in maize coleoptiles analysed with a tetrazolium-based assay. Planta 212:175–183PubMedCrossRefGoogle Scholar
  12. Frova C (2003) The glutathione transferase gene family: genomic structure, functions, expression and evolution. Physiol Plant 119:469–479CrossRefGoogle Scholar
  13. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930PubMedCrossRefGoogle Scholar
  14. Griffith OW (1980) Determination of glutathione disulphide using glutathione reductase and 2-vinylpyridine. Anal Biochem 6:207–212CrossRefGoogle Scholar
  15. Halliwell B, Gutteridge JMC (1981) Formation of a thiobarbituric-acid-reactive substance from deoxyribose in the presence of iron salts. FEBS Lett 128:347–352PubMedCrossRefGoogle Scholar
  16. Haluskova L, Valentovicova K, Huttova J, Mistrik I, Tamas L (2009) Effect of abiotic stresses on glutathione peroxidase and glutathione S-transferase activity in barley root tips. Plant Physiol Biochem 47:1069–1074PubMedCrossRefGoogle Scholar
  17. Hare PD, Cress WA (1997) Metabolic implications of stress-induced proline accumulation in plants. Plant Growth Regul 21:79–102CrossRefGoogle Scholar
  18. 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
  19. Jogeswar G, Pallela R, Jakka NM, Reddy PS, Venkateswara Rao J, Sreenivasulu N, Kavi Kishor PB (2006) Antioxidative response in different sorghum species under short-term salinity stress. Acta Physiol Plant 28:465–475CrossRefGoogle Scholar
  20. Law MY, Charles SA, Halliwell B (1983) Glutathione and ascorbic acid and spinach (Spinacea oleracea) chloroplasts: the effect of hydrogen peroxide and paraquat. Biochem J 210:899–903PubMedGoogle Scholar
  21. Li ZS, Zhen RG, Rea PA (1995) 1-Chloro-2,4-dinitrobenzene-elicited increase in vacuolar glutathione-S-conjugate transport activity. Plant Physiol 109:177–185PubMedGoogle Scholar
  22. Maheshwari R, Dubey RS (2009) Nickel-induced oxidative stress and the role of antioxidative defense in rice seedlings. Plant Growth Regul 59:37–49CrossRefGoogle Scholar
  23. Michel BE, Kaufmann MR (1973) The osmotic potential of polyethylene glycol 6000. Plant Physiol 51:914–916PubMedCrossRefGoogle Scholar
  24. Mishra P, Bhoomika K, Dubey RS (2012) Differential responses of antioxidative defense system to prolonged salinity stress in salt-tolerant and salt-sensitive Indica rice (Oryza sativa L.) seedlings. Protoplasma. doi:10.1007/s00709-011-0365-3
  25. Moradi F, Ismail AM (2007) Responses of photosynthesis, chlorophyll fluorescence and ROS-scavenging systems to salt stress during seedling and reproductive stages in rice. Ann Bot 99:1161–1173PubMedCrossRefGoogle Scholar
  26. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  27. Noctor G (2006) Metabolic signalling in defence and stress: the central roles of soluble redox couples. Plant Cell Environ 29:409–425PubMedCrossRefGoogle Scholar
  28. Noctor G, Gomez L, Vanacker H, Foyer CH (2002) Interactions between biosynthesis, compartmentation, and transport in the control of glutathione homeostasis and signaling. J Exp Bot 53:1283–1304PubMedCrossRefGoogle Scholar
  29. Phung T, Jung H, Park J, Kim J-G, Back J, Jung S (2011) Porphyrin biosynthesis control under water stress: sustained porphyrin status correlates with drought tolerance in transgenic rice. Plant Physiol 157:1746–1764PubMedCrossRefGoogle Scholar
  30. Polle A (2001) Dissecting the superoxide dismutase–ascorbate–glutathione-pathway in chloroplasts by metabolic modeling. Computer simulations as a step towards flux analysis. Plant Physiol 126:445–462PubMedCrossRefGoogle Scholar
  31. Pourcel L, Routaboul JM, Cheynier V, Lepiniec L, Debeaujon I (2007) Flavonoid oxidation in plants: from biochemical properties to physiological functions. Trends Plant Sci 12:29–36PubMedCrossRefGoogle Scholar
  32. Price AH, Atherton N, Hendry GAF (1989) Plants under drought stress generated activated oxygen. Free Radic Res Commun 8:61–66PubMedCrossRefGoogle Scholar
  33. Schaedle M, Bassham JA (1977) Chloroplast glutathione reductase. Plant Physiol 59:1011–1012PubMedCrossRefGoogle Scholar
  34. Schutzendübel A, Schwanz P, Teichmann T, Gross K, Langenfeld-Heyser R, Godbold DL, Polle A (2001) Cadmium-induced changes in antioxidative systems, H2O2 content and differentiation in pine (Pinus sylvestris) roots. Plant Physiol 127:887–898PubMedCrossRefGoogle Scholar
  35. 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
  36. Sharma P, Dubey RS (2005) Drought induces oxidative stress and enhances the activities of antioxidant enzymes in growing rice seedlings. Plant Growth Regul 46:209–221CrossRefGoogle Scholar
  37. Sharma P, Jha AB, Dubey RS (2010) Oxidative stress and antioxidative defense system in plants growing under abiotic stresses. In: Pessarakli M (ed) Handbook of plant and crop stress, 3rd edn. Taylor & Francis, Florida, pp 89–138CrossRefGoogle Scholar
  38. Sharma P, Jha AB, Dubey RS, Pessarakli, M (2012) Reactive oxygen species, oxidative damage and antioxidative defense mechanism in plants under stressful conditions. J Bot. article ID 217037, 26 pages, doi:10.1155/2012/217037.
  39. Smith AP, DeRidder BP, Guo WJ, Seeley EH, Regnier FE, Goldsbrough PB (2004) Proteomic analysis of Arabidopsis glutathione S-transferases from benoxacor- and copper-treated seedlings. J Biol Chem 279:26098–26104PubMedCrossRefGoogle Scholar
  40. Stevens R, Page D, Gouble B, Garchery C, Zamir D, Causse M (2008) Tomato fruit ascorbic acid content is linked with monodehydroascorbate reductase activity and tolerance to chilling stress. Plant Cell Environ 31:1086–1096PubMedCrossRefGoogle Scholar
  41. Szalai G, Kell T, Galiba G, Kocsy G (2009) Glutathione as an antioxidant and regulatory molecule in plants under abiotic stress conditions. J Plant Growth Regul 28:66–80CrossRefGoogle Scholar
  42. Tausz M, Sircelj H, Grill D (2004) The glutathione system as a stress marker in plant ecophysiology: is a stress-response concept valid? J Exp Bot 55:1955–1962PubMedCrossRefGoogle Scholar
  43. Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants, H2O2 accumulation in papillae and hypersensitive response during barley powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  44. Trovato M, Mattioli R, Costanstion P (2008) Multiple roles of proline in plant stress tolerance and development. Rend Lincei Sci Fis Nat 19:325–346CrossRefGoogle Scholar
  45. Wang F, Wang Q, Kwon S, Kwak S, Su W (2005) Enhanced drought tolerance of transgenic rice plants expressing a pea manganese superoxide dismutase. J Plant Physiol 162:465–472PubMedCrossRefGoogle Scholar
  46. Wang Z, Xiao Y, Chen W, Tang K, Zhang L (2010) Increased vitamin C content accompanied by an enhanced recycling pathway confers oxidative stress tolerance in Arabidopsis. J Integr Plant Biol 52:400–409PubMedCrossRefGoogle Scholar
  47. Yin L, Wang S, Eltayeb AE, Uddin MI, Yamamoto Y, Tsuji W, Takeuchi Y, Anaka K (2010) Overexpression of dehydroascorbate reductase, but not monodehydroascorbate reductase, confers tolerance to aluminium stress in transgenic tobacco. Planta 231:609–621PubMedCrossRefGoogle Scholar
  48. Yoshida S, Forno DA, Cock JH, Gomez KA (1976) Laboratory manual for physiological studies of rice. IRRI, Los BañosGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Samantha Pyngrope
    • 1
  • Kumari Bhoomika
    • 1
  • R. S. Dubey
    • 1
  1. 1.Department of Biochemistry, Faculty of ScienceBanaras Hindu UniversityVaranasiIndia

Personalised recommendations