Advertisement

Protoplasma

, Volume 245, Issue 1–4, pp 85–96 | Cite as

Redox homeostasis, antioxidant defense, and methylglyoxal detoxification as markers for salt tolerance in Pokkali rice

  • Hattem El-Shabrawi
  • Bhumesh Kumar
  • Tanushri Kaul
  • Malireddy K. Reddy
  • Sneh L. Singla-Pareek
  • Sudhir K. SoporyEmail author
Original Article

Abstract

To identify biochemical markers for salt tolerance, two contrasting cultivars of rice (Oryza sativa L.) differing in salt tolerance were analyzed for various parameters. Pokkali, a salt-tolerant cultivar, showed considerably lower level of H2O2 as compared to IR64, a sensitive cultivar, and such a physiology may be ascribed to the higher activity of enzymes in Pokkali, which either directly or indirectly are involved in the detoxification of H2O2. Enzyme activities and the isoenzyme pattern of antioxidant enzymes also showed higher activity of different types and forms in Pokkali as compared to IR64, suggesting that Pokkali possesses a more efficient antioxidant defense system to cope up with salt-induced oxidative stress. Further, Pokkali exhibited a higher GSH/GSSG ratio along with a higher ratio of reduced ascorbate/oxidized ascorbate as compared to IR64 under NaCl stress. In addition, the activity of methylglyoxal detoxification system (glyoxalase I and II) was significantly higher in Pokkali as compared to IR64. As reduced glutathione is involved in the ascorbate–glutathione pathway as well as in the methylglyoxal detoxification pathway, it may be a point of interaction between these two. Our results suggest that both ascorbate and glutathione homeostasis, modulated also via glyoxalase enzymes, can be considered as biomarkers for salt tolerance in Pokkali rice. In addition, status of reactive oxygen species and oxidative DNA damage can serve as a quick and sensitive biomarker for screening against salt and other abiotic stresses in crop plants.

Keywords

Antioxidative defense ROS Methylglyoxal Rice Salt stress 

Notes

Acknowledgements

H.E.-S. is thankful to ICGEB for pre-doctoral fellowship and B.K. is grateful for salary support from a Young Scientist grant from the Department of Science and Technology, Government of India. The research was partly supported by a grant from the Department of Biotechnology, Government of India, and the internal grants from ICGEB.

Conflict of interest

The authors declare that they have no conflict of interest. There is no financial or other relationship that might be perceived as leading to a conflict of interest.

References

  1. Aebi H (1983) Catalase. In: Bergmeies H (ed) Methods of enzyme analysis. Chemie Wenhein, Verlag, pp 273–277Google Scholar
  2. Beyer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal Biochem 161:559–566CrossRefPubMedGoogle Scholar
  3. Bhattacharjee S, Mukherjee AK (1997) Role of free radicals in membrane deterioration in three rice (Oryza sativa L.) cultivars under NaCl salinity at early germination stage. Indian J Exp Bot 35:1365–1369Google Scholar
  4. 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 72:248–254CrossRefPubMedGoogle Scholar
  5. Casano LM, Martin M, Sabater B (1994) Sensitivity of superoxide dismutase transcript levels and activities to oxidative stress is lower in mature-senescent than in young barley leaves. Plant Physiol 106:1033–1039PubMedGoogle Scholar
  6. Comba ME, Benavides MP, Tomaro ML (1998) Effect of salt stress on antioxidant defence system in soybean root nodules. Aust J Plant Physiol 25:665–671CrossRefGoogle Scholar
  7. Corpas F, Gomez JM, Hernandez JA, Del-Rio JA (1993) Metabolism of activated oxygen in leaf peroxisomes from two Pisum sativum L. cultivars with different sensitivity to sodium chloride. J Plant Physiol 141:160–165Google Scholar
  8. De Gara L, de Pinto MC, Arrigoni O (1997) Ascorbate synthesis and ascorbate peroxidase activity during the early stage of wheat germination. Physiol Plant 100:894–900CrossRefGoogle Scholar
  9. Deswal R, Sopory SK (1999) Glyoxalase I from Brassica juncea is a calmodulin stimulated protein. 1450:460–467Google Scholar
  10. Dionisio-Sese ML, Tobita S (1998) Antioxidant responses of rice seedlings to salinity stress. Plant Sci 135:1–9CrossRefGoogle Scholar
  11. Dutilleul C, Garmier M, Noctor G, Mathieu C, Chetrit P, Foyer CH, de Paepe R (2003) Leaf mitochondria modulate whole cell redox homeostasis, set antioxidant capacity, and determine stress resistance through altered signaling and diurnal regulation. Plant Cell 15:1212–1226CrossRefPubMedGoogle Scholar
  12. Evans WR, Fuchsman DE, Calveri HE, Pyati PV, Alter GM, Subha Rao NS (1999) Bacteriochlorophyll and photosynthetic reaction centers in Rhizobium strain BTAII. Appl Environ Microbiol 56:3445–3449Google Scholar
  13. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: A metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875CrossRefPubMedGoogle Scholar
  14. Gossett DR, Banks SW, Millhollon EP, Lucas MC (1996) Antioxidant response to NaCl stress in control and NaCl-tolerant cotton cell line grown in the presence of paraquat, buthionine sulfoximine and exogenous glutathione. Plant Physiol 112:803–809PubMedGoogle Scholar
  15. Griffith OW (1981) The role of glutathione turnover in the apparent renal secretion of cystine. J Biol Chem 256:12263–12268PubMedGoogle Scholar
  16. Halliwell B, Gutteridge JMC (1989) Free radicals in biology and medicine. Oxford, Clarendan Press, pp 446–489Google Scholar
  17. Hernandez JA, Jimenez A, Mullineaux P, Sevilla F (2000) Tolerance of pea (Pisum sativum L.) to long-term salt stress is associated with induction of antioxidant defences. Plant Cell Environ 23:853–862CrossRefGoogle Scholar
  18. Hoque MA, Banu MN, Nakamura Y, Shimoishi Y, Murata Y (2008) Proline and glycinebetaine enhance antioxidant and methylglyoxal detoxification systems and reduce NaCl-induced damage in cultured tobacco cells. J Plant Physiol 165:813–824CrossRefPubMedGoogle Scholar
  19. Hung SH, Yu CW, Lin CH (2005) Hydrogen peroxide functions as a stress signal in plants. Bot Bull Acad Sin 46:1–10Google Scholar
  20. Inoue Y, Matsuda T, Sugiyama K, Izawa S, Kimura A (1999) Genetic analysis of glutathione peroxidase in oxidative stress response of Saccharomyces cerevisiae. J Biol Chem 274:27002–27009CrossRefPubMedGoogle Scholar
  21. Jiménez-Bremont JF, Becerra-Flora A, Hernández-Lucero E, Rodríguez-Kessler M, Acosta-Gallegos JA, Ramírez-Pimentel JG (2006) Proline accumulation in two bean cultivars under salt stress and the effect of polyamines and ornithine. Biol Plant 50:763–766CrossRefGoogle Scholar
  22. Kayupova GA, Klyshev LK (1984) Superoxide dismutase of pea roots under the influence of high NaCl concentrations. Fiziol Rast 31:555–559Google Scholar
  23. Kazuya Y, Kazuhiro M, Ahmed G, Toru T, Haruo K, Hitoshi M, Shigeru S (2004) Enhancement of stress tolerance in transgenic tobacco plants overexpressing Chlamydomonas glutathione peroxidase in chloroplasts or cytosol. Plant J 37:21–33CrossRefGoogle Scholar
  24. Khan MA, Shirazi MU, Khan MA, Mujtaba SM, Islam E, Mumtaz S, Shereen A, Ansari RU, Ashraf MY (2009) Role of proline, K/Na ratio and chlorophyll content in salt tolerance of wheat (Triticum aestivum L.). Pak J Bot 41:633–638Google Scholar
  25. Kho CW, Park SG, Lee DH, Cho S, Oh GT, Kang S, Park BC (2004) Activity staining of glutathione peroxidase after two-dimensional gel electrophoresis. Mol Cell 18:369–373Google Scholar
  26. Laemmli UK (1970) Cleavage of structural proteins during the assembly of head of the bacteriophage T4. Nature 277:680–685Google Scholar
  27. Larkindale J, Huang B (2004a) Thermotolerance and antioxidant systems in Agrostis stolonifera: Involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene. J Plant Physiol 161:405–413CrossRefPubMedGoogle Scholar
  28. Larkindale J, Huang B (2004b) Thermotolerance and antioxidant systems in Agrostis stolonifera: Involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene. J Plant Physiol 161:405–413CrossRefPubMedGoogle Scholar
  29. Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–595CrossRefPubMedGoogle Scholar
  30. Maiti MK, Krishnasamy S, Owen HA, Makaroff CA (1997) Molecular characterization of glyoxalase II from Arabidopsis thaliana. Plant Mol Biol 35:471–481CrossRefPubMedGoogle Scholar
  31. Mittova V, Tal M, Volokita M, Guy M (2003) Up-regulation of the leaf mitochondrial and peroxisomal antioxidative systems in response to salt-induced oxidative stress in the wild salt-tolerant tomato species Lycopersicon pennellii. Plant Cell Environ 26:845–856CrossRefPubMedGoogle Scholar
  32. Munns R (2002) Salinity, growth and phytohormones. In: Läuchli A, Lüttge U (eds) Salinity: environment—plants—molecules. Kluwer, Dordrecht, pp 271–290Google Scholar
  33. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  34. Noctor G, Foyer CH (1998) Ascorbate and glutathione: Keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279CrossRefPubMedGoogle Scholar
  35. Noctor G, Gomez L, Vanacker H, Foyer CH (2002) Interactions between biosynthesis, compartmentation, and transport in the control of glutathione homeostasis and signalling. J Exp Bot 53:1283–1304CrossRefPubMedGoogle Scholar
  36. Ogawa K (2005) Glutathione-associated regulation of plant growth and stress responses. Antioxid Redox Signal 7:973–981CrossRefPubMedGoogle Scholar
  37. Prasad TK, Anderson MD, Stewart LR (1994) Acclimation, hydrogen peroxide and abscisic acid protect mitochondria against irreversible chilling injury in maize seedlings. Plant Physiol 105:619–627PubMedGoogle Scholar
  38. Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol 110:125–136CrossRefPubMedGoogle Scholar
  39. Ray PKS, Islam MA (2008) Genetic analysis of salinity tolerance in rice. Bangladesh J Agric Res 33:519–529Google Scholar
  40. Roy SD, Saxena M, Bhomkar PS, Pooggin M, Hohn T, Neera B-S (2008) Generation of marker free salt tolerant transgenic plants of Arabidopsis thaliana using the gly I gene and cre gene under inducible promoters. Plant Cell Tissue Organ Culture 95:1–11CrossRefGoogle Scholar
  41. Rucinska R, Waplak S, Gwozdz E (1999) Free radical formation and activity of antioxidant enzymes in lupin roots exposed to lead. Plant Physiol Biochem 37:187–194CrossRefGoogle Scholar
  42. Sabu A, Sheeja TE, Nambisan P (1995) Comparison of proline accumulation in callus and seedlings of two cultivars of Oryza sativa L. differing in salt tolerance. Indian J Exp Biol 33:139–141Google Scholar
  43. Sagi M, Fluhr R (2006) Production of reactive oxygen species by plant NADPH oxidases. Plant Physiol 141:336–340CrossRefPubMedGoogle Scholar
  44. Singla-Pareek SL, Reddy MK, Sopory SK (2003) Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance. Proc Natl Acad Sci USA 100:14672–14677CrossRefPubMedGoogle Scholar
  45. Singla-Pareek SL, Yadav SK, Pareek A, Reddy MK, Sopory SK (2006) Transgenic tobacco overexpressing glyoxalase pathway enzymes grow and set viable seeds in zinc-spiked soils. Plant Physiol 140:613–623CrossRefPubMedGoogle Scholar
  46. Singla-Pareek SL, Yadav SK, Pareek A, Reddy MK, Sopory SK (2008) Enhancing salt tolerance in a crop plant by overexpression of glyoxalase II. Transgenic Res 17(2):171–180CrossRefPubMedGoogle Scholar
  47. Smith IK, Vierheller TL, Thorne CA (1988) Assay of glutathione reductase in crude tissue homogenates using 5, 5' dithiobis (2-nitrobenzoic acid). Anal Biochem 175:408–413CrossRefPubMedGoogle Scholar
  48. Sreenivasulu N, Grimm B, Wobus U, Weschke W (2000) Differential response of antioxidant compounds to salinity stress in salt-tolerant and salt-sensitive seedlings of foxtail millet (Setaria italica). Physiol Plant 109:435–442CrossRefGoogle Scholar
  49. Sreenivasulu N, Miranda M, Prakash HS, Wobus U, Weschke W (2004) Transcriptome changes in foxtail millet genotypes at high salinity: Identification and characterization of a PHGPX gene specifically up-regulated by NaCl in a salt-tolerant line. J Plant Physiol 161:467–477CrossRefPubMedGoogle Scholar
  50. Suzuki N, Mittler R (2006) Reactive oxygen species and temperature stresses: A delicate balance between signaling and destruction. Physiol Plant 126:45–51CrossRefGoogle Scholar
  51. 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 55:1955–1962CrossRefPubMedGoogle Scholar
  52. Trotel P, Bouchereau A, Niogret MF, Larher F (1996) The fate of osmo-accumulated proline in leaf disc of rape (Brassica napus L) incubated in a medium of low osmolarity. Plant Sci 118:31–45CrossRefGoogle Scholar
  53. Uddin MI, Rashid MH, Khan N, Perveen MF, Tai TH, Tanaka K (2007) Selection of promising salt tolerant rice mutants derived from cultivar ‘drew’ and their antioxidant enzymes activity under salt stress. SABRAO J Breed Genet 39:89–98Google Scholar
  54. Vaidyanathan H, Sivakumar P, Chakrabarty R, Thomas G (2003) Scavenging of reactive oxygen species in NaCl-stressed rice (Oryza sativa L.) differential response in salt-tolerant and sensitive varieties. Plant Sci 165:1411–1418CrossRefGoogle Scholar
  55. Yadav SK, Singla-Pareek SL, Reddy MK, Sopory SK (2005) Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress. FEBS Lett 579:6265–6271CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Hattem El-Shabrawi
    • 1
  • Bhumesh Kumar
    • 1
  • Tanushri Kaul
    • 1
  • Malireddy K. Reddy
    • 1
  • Sneh L. Singla-Pareek
    • 1
  • Sudhir K. Sopory
    • 1
    Email author
  1. 1.Plant Molecular BiologyInternational Centre for Genetic Engineering and Biotechnology (ICGEB)New DelhiIndia

Personalised recommendations