Skip to main content
Log in

Expression of SbGSTU (tau class glutathione S-transferase) gene isolated from Salicornia brachiata in tobacco for salt tolerance

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Tau class glutathione transferases (GSTU) genes are plant specific, induced by different abiotic stress, and important for protecting plants against oxidative damage. GST gene was isolated using 5′ RACE from an extreme halophyte Salicorniabrachiata, cloned, sequenced and its protein structure was predicted. Transcript profiling of SbGST gene expression was studied under different abiotic stress conditions and plant growth regulator treatments, viz. salt, cold, drought, ABA and salicylic acid, with time period point and concentration point. The expression of SbGST gene was up-regulated in all stress conditions, except SA treatment. Seed germination percentage, GST enzyme assay, fresh weight and other growth parameters (root length, shoot length and leaf area) were studied and results indicate that over-expression of SbGST gene in transgenic tobacco leads to enhanced seed germination and growth under salt stress. Transgenic lines were evaluated for their performance under salt stress and tobacco plants over-expressing SbGST showed higher seed germination and survival compared to wild type. These results confirm that expression of SbGST gene is up-regulated by different stresses and over-expression of tau class SbGST gene in transgenic tobacco plays a vital role in abiotic stress tolerance. SbGST gene expressed conspicuously under salt stress leading to enhance seed germination and better growth. Furthermore, GST is a potential candidate gene to be used in genetic engineering for enhancing abiotic stress tolerance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Edwards R, Dixon DP (2000) The role of glutathione transferases in herbicide metabolism. In: Cobb AH, Kirkwood RC (eds) Herbicides and their mechanisms of action. Sheffield Academic Press, Sheffield, England, pp 38–71

    Google Scholar 

  2. Seppänen MM, Cardi T, Hyökki MB, Pehu E (2000) Characterization and expression of cold-induced glutathione S-transferase in freezing tolerant Solanum commersonii, sensitive S. tuberosum and their interspecific somatic hybrids. Plant Sci 153:125–133

    Article  PubMed  Google Scholar 

  3. Moons A (2003) OsGSTU3 and OsGTU4, encoding tau class glutathione S-transferases, are heavy metal and hypoxic stress-induced and differentially salt stress-responsive in rice roots. FEBS Lett 553:427–432

    Article  PubMed  CAS  Google Scholar 

  4. Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K (1993) Characterization of two cDNAs (ERD11 and ERD13) for dehydration inducible genes that encode putative glutathione-S-transferases in Arabidopsis thaliana L. FEBS Lett 335:189–192

    Article  PubMed  CAS  Google Scholar 

  5. Bianchi MW, Roux C, Vartanian N (2002) Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase. Physiol Plant 116:96–105

    Article  PubMed  CAS  Google Scholar 

  6. Vollenweider S, Weber H, Stolz S, Chetelat A, Farmer EE (2000) Fatty acid ketodienes and fatty acid ketotrienes: Michael addition acceptors that accumulate in wounded and diseased Arabidopsis leaves. Plant J 24:467–476

    Article  PubMed  CAS  Google Scholar 

  7. Mauch F, Dudler R (1993) Differential induction of distinct glutathione transferases of wheat by xenobiotics and by pathogen attack. Plant Physiol 102:1193–1201

    Article  PubMed  CAS  Google Scholar 

  8. Roxas VP, RKJr Smith, Allen ER, Allen RD (1997) Overexpression of glutathione S-transferase/glutathione peroxidase enhances the growth of transgenic tobacco seedlings during stress. Nat Biotechnol 15:988–991

    Article  PubMed  CAS  Google Scholar 

  9. Zhou J, Goldsbrough PB (1993) An Arabidopsis gene with homology to glutathione S-transferase is regulated by ethylene. Plant Mol Biol 22:517–523

    Article  PubMed  CAS  Google Scholar 

  10. Chen W, Singh KB (1999) The auxin, hydrogen peroxide and salicylic acid induced expression of the Arabidopsis GST6 promoter is mediated in part by an ocs element. Plant J 19:667–677

    Article  PubMed  CAS  Google Scholar 

  11. Marrs KA (1996) The functions and regulation of glutathione S-transferases in plants. Ann Rev Plant Physiol Plant Mol Biol 47:127–158

    Article  CAS  Google Scholar 

  12. Wagner U, Edwards R, Dixon DP, Mauch F (2002) Probing the diversity of the Arabidopsis glutathione S-transferase gene family. Plant Mol Biol 49:515–532

    Article  PubMed  CAS  Google Scholar 

  13. Edwards R, Dixon DP, Walbot V (2000) Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci 5:193–198

    Article  PubMed  CAS  Google Scholar 

  14. Dixon DP, Davis BG, Edwards E (2002) Functional divergence in the glutathione transferase superfamily in plants. J Biol Chem 277:30859–30869

    Article  PubMed  CAS  Google Scholar 

  15. Frova C (2003) The plant glutathione transferase gene family: genomic structure, functions, expression and evolution. Physiol Plant 119:469–479

    Article  CAS  Google Scholar 

  16. Moons A (2005) Regulatory and functional interactions of plant growth regulators and plant glutathione-S-transferases (GSTs). Vitam Horm (New York) 72:155–202

    CAS  Google Scholar 

  17. Axarli IA, Rigden DJ, Labrou NE (2004) Characterization of the ligand in site of maize glutathione transferase. Biochem J 382:885–893

    Article  PubMed  CAS  Google Scholar 

  18. LoPiero AR, Puglisi I, Petrone G (2006) Gene isolation, analysis of expression and in vitro synthesis of a glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]. J Agric Food Chem 54:9227–9233

    Article  CAS  Google Scholar 

  19. Dixon DP, Lapthorn A, Madesis P, Mudd EA, Day A, Edwards R (2008) Binding and glutathione conjugation of porphyrinogens by plant glutathione transferases. J Biol Chem 283:20268–20276

    Article  PubMed  CAS  Google Scholar 

  20. Edwards R, Dixon DP (2005) Plant glutathione transferases. Meth Enzymol 401:169–186

    Article  PubMed  CAS  Google Scholar 

  21. Bhatnagar-Mathur P, Vadez V, Sharma K (2008) Transgenic approaches for abiotic stress tolerance in plants: retrospect and prospects. Plant Cell Rep 27:411–424

    Article  PubMed  CAS  Google Scholar 

  22. Jha B, Agarwal PK, Reddy PS, Lal S, Sopory SK, Reddy MK (2009) Identification of salt -induced genes from Salicornia brachiata, an extreme halophyte through expressed sequence tags analysis. Genes Genet Sys 84:111–120

    Article  CAS  Google Scholar 

  23. Agarwal PK, Gupta K, Jha B (2010) Molecular characterization of the Salicornia brachiata SbMAPKK gene and its expression by abiotic stress. Mol Biol Rep 37:981–986

    Article  PubMed  CAS  Google Scholar 

  24. Jha A, Joshi M, Yadav NS, Agarwal PK, Jha B (2010) Cloning and characterization of the Salicornia brachiata Na+/H+ antiporter gene SbNHX1 and its expression by abiotic stress. Mol Biol Rep Online doi:10.1007/s11033-010-0318-5

  25. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:431–497

    Article  Google Scholar 

  26. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159

    Article  PubMed  CAS  Google Scholar 

  27. Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the expasy server. In: Walker JM (ed) The proteomics protocols handbook. Humana Press, Totowa, USA, pp 571–607

    Chapter  Google Scholar 

  28. McGuffin LJ, Bryson K, Jones DT (2000) The PSIPRED protein structure prediction server. Bioinformatics 16:404–405

    Article  PubMed  CAS  Google Scholar 

  29. Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schäffer AA, Yu YK (2005) Protein database searches using compositionally adjusted substitution matrices. FEBS J 272:5101–5109

    Article  PubMed  CAS  Google Scholar 

  30. Marchler-Bauer A, Anderson JB, Chitsaz F, Derbyshire MK, DeWeese-Scott C, Fong JH, Geer LY, Geer RC, Gonzales NR, Gwadz M, He S, Hurwitz DI, Jackson JD, Ke Z, Lanczycki CL, Liebert CA, Liu C, Lu F, Lu S, Marchler GH, Mullokandov M, Song JS, Tasneem A, Thanki N, Yamashita RA, Zhang D, Zhang N, Bryant SH (2009) CDD: specific functional annotation with the conserved domain database. Nucleic Acids Res 37:205–210

    Article  Google Scholar 

  31. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Farley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  32. Jefferson RA (1987) Assaying chimeric genes in plants: the GUS fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  33. Dean VJ, Gronwald JW, Eberlein CV (1990) Induction of glutathione S-transferase isozyme in sorghum by herbicide antidotes. Plant Physiol 92:267–273

    Article  Google Scholar 

  34. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  35. Sokal RR, Rohlf FJ (1995) Biometry, the principles and practice of statistics in biological research, 3rd edn. WH Freeman and Company, New York, pp 321–356

    Google Scholar 

  36. Droog FNJ (1997) Plant glutathione S-transferase, a tale of theta and tau. J Plant Growth Reg 16:95–107

    Article  CAS  Google Scholar 

  37. Thom R, Cummins I, Dixon DP, Edwards R, Cole DJ, Lapthorn AJ (2002) Structure of a tau class glutathione S-transferase from wheat active in herbicide detoxification. Biochemistry 41:7008–7020

    Article  PubMed  CAS  Google Scholar 

  38. Axarli I, Dhavala P, Papageorgiou AC, Labrou NE (2009) Crystallographic and functional characterization of the fluorodifen-inducible glutathione transferase from Glycine max reveals an active site topography suited for diphenylether herbicides and a novel L-site. J Mol Biol 385:984–1002

    Article  PubMed  CAS  Google Scholar 

  39. Reinemer P, Prade L, Hof P, Neuefeind T, Huber R, Zettl R, Palme K, Schell J, Koelnn I, Bartunik HD, Bieseler B (1996) Three dimensional structure of glutathione S-transferase from Arabidopsis thaliana at 2.2 Å resolution: structural characterization of herbicide-conjugating plant glutathione S-transferases and a novel active site architecture. J Mol Biol 255:289–309

    Article  PubMed  CAS  Google Scholar 

  40. Board PG, Coggan M, Chelvanayagam G, Easteal S, Jermiin LS, Schulte GK, Danley DE, Hoth LR, Griffor MC, Kamath AV, Rosner MH, Chrunyk BA, Perregaux DE, Gabel CA, Geoghegan KF, Pandit J (2000) Identification, characterization, and crystal structure of the omega class glutathione transferases. J Biol Chem 275:24798–24806

    Article  PubMed  CAS  Google Scholar 

  41. Ji XH, Vonrosenvinge EC, Johnson WW, Tomarev SI, Piatigorsky J, Armstrong RN, Gilliland GL (1995) Three-dimensional structure, catalytic properties, and evolution of a sigma class glutathione transferase from squid, a progenitor of the lens S-crystallins of cephalopods. Biochemistry 34:5317–5328

    Article  PubMed  CAS  Google Scholar 

  42. Wilce MCJ, Board PG, Feil SC, Parker MW (1995) Crystal structure of a theta-class glutathione transferase. EMBO J 14:2132–2143

    Google Scholar 

  43. Yang KY, Kim EY, Kim CS, Guh JO, Kim KC, Cho BH (1998) Characterization of a glutathione S-transferase gene ATGST1 in Arabidopsis thaliana. Plant Cell Rep 17:700–704

    Article  CAS  Google Scholar 

  44. Gong HY, Jiao WH, Pua EC (2005) Expression of glutathione S-transferase and its role in plant growth and development in vivo and shoot morphogenesis in vitro. Plant Mol Biol 57:53–66

    Article  PubMed  CAS  Google Scholar 

  45. Roxas VP, Lodhi SA, Garrett DK, Mahan JR, Allen RD (2000) Stress tolerance in transgenic tobacco seedlings that overexpress glutathione S-transferase/glutathione peroxidase. Plant Cell Physiol 41:1229–1234

    Article  PubMed  CAS  Google Scholar 

  46. Takesawa T, Ito M, Kanzaki H, Kameya N, Nakamura I (2002) Over-expression of glutathione S-transferase in transgenic rice enhances germination and growth at low temperature. Mol Breed 9:93–101

    Article  CAS  Google Scholar 

  47. Anderson JV, Davis DG (2004) Abiotic stress alters transcript profiles and activity of glutathione S-transferase, glutathione peroxidase, and glutathione reductase in Euphorbia esula. Physiol Plant 120:421–433

    Article  PubMed  CAS  Google Scholar 

  48. Liu X, He BW, Zhang YZ (2009) Stress-responsive expression analysis of glutathione-S-transferase gene of Ipomoea batatas (L.) Lam. Yi Chuan/Hereditas (Beijing) 8:859–864

    Article  Google Scholar 

  49. Gallé Á, Csiszár J, Secenji M, Guóth A, Cseuz L, Tari I, Györgyey J, Erdei L (2009) Glutathione transferase activity and expression patterns during grain filling in flag leaves of wheat genotypes differing in drought tolerance: Response to water deficit. J Plant Physiol 116:1878–1891

    Article  Google Scholar 

  50. Dixon DP, Cummins I, Cole DJ, Edwards R (1998) Glutathione-mediated detoxification systems in plants. Curr Opin Plant Biol 1:258–266

    Article  PubMed  CAS  Google Scholar 

  51. Sappl PG, Carroll AJ, Clifton R, Lister R, Whelan J, Millar AH, Singh KB (2009) The Arabidopsis glutathione transferase gene family displays complex stress regulation and co-silencing multiple genes results in altered metabolic sensitivity to oxidative stress. Plant J 58:53–58

    Article  CAS  Google Scholar 

  52. Ryu HY, Kim SY, Park HM, You JY, Kim BH, Lee JS, Nama KH (2009) Modulations of AtGSTF10 expression induce stress tolerance and BAK1-mediated cell death. Biochem Biophys Res Commun 379:417–422

    Article  PubMed  CAS  Google Scholar 

  53. Cummins I, Cole DJ, Edwards R (1999) A role for glutathione transferases functioning as glutathione peroxidises in resistance to multiple herbicides in black-grass. Plant J 18:285–292

    Article  PubMed  CAS  Google Scholar 

  54. Lim JD, Hahn SJC, Yu Y, Chung M (2005) Expression of the glutathione S-transferase gene (NT107) in transgenic Dianthus superbus. Plant Cell Tissue Organ Cult 80:277–286

    Article  CAS  Google Scholar 

Download references

Acknowledgment

The financial support of CSIR Network Project (NWP-020), Govt. of India for carrying out this study is thankfully acknowledged.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Bhavanath Jha.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 1527 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jha, B., Sharma, A. & Mishra, A. Expression of SbGSTU (tau class glutathione S-transferase) gene isolated from Salicornia brachiata in tobacco for salt tolerance. Mol Biol Rep 38, 4823–4832 (2011). https://doi.org/10.1007/s11033-010-0625-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-010-0625-x

Keywords

Navigation