Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Cloning and characterisation of glutathione reductase cDNAs and identification of two genes encoding the tobacco enzyme


We have isolated 4 cDNA clones (GRT1-4) encoding glutathione reductase (GR) from a tobacco (Nicotiana tabacum L.) leaf cDNA library. The cDNAs were almost identical: GRT1, GRT3 and GRT4 represented the same gene, differing only in that GRT4 contained an intron within the C-terminal part of the coding sequence. Failure to splice out this intron resulted in a substitution of the final 13 amino acids of the deduced amino acid sequence. A second gene was represented by GRT2. Southern blots indicated that there were two related GR genes in tobacco. The presence of multiple isoforms of GR in tobacco may be explained in part by the expression of a small gene family. In addition, alternative isoforms may result from translation of different mRNAs derived from the same gene by intron skipping during the splicing of nascent GR mRNAs.

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



base pairs


glutathione reductase


rapid amplification of cDNA ends


  1. Arondel V, Tchang F, Baillet B, Bignols F, Grellet F, Delseny M, Kader J-C, Puigdomenech P (1991) Multiple mRNAs coding for phospholipid transfer protein from Zea mays arise from alternative splicing. Gene 99: 133–136

  2. Creissen G, Edwards EA, Enard C, Wellburn A, Mullineaux P (1992) Molecular characterization of glutathione reductase cDNAs from pea (Pisum sativum L.). Plant J 2: 129–131

  3. Creissen GP, Broadbent P, Kular B, Reynolds H, Wellburn AR, Mullineaux PM (1994) Manipulation of glutathione reductase in transgenic plants: implications for plants' responses to environmental stress. Proc R Soc Edin 102B:167–175

  4. Creissen G, Reynolds H, Xue Y, Mullineaux P (1995) Simultaneous targeting of pea glutathione reductase and of a bacterial fusion protein to chloroplasts and mitochondria in transgenic tobacco. Plant J 8:167–175

  5. Dronn M, Clouse SD, Dixon RA, Lawton MA, Lamb CJ (1988) Glutathione and fungal elicitor regulation of a plant defense gene promoter in electroporated protoplasts. Proc Natl Acad Sci USA 85:6738–6742

  6. Dalton DA, Russell SA, Hanus FJ, Pascoe GA, Evans HJ (1986) Enzymatic reactions of ascorbate and glutathione that prevent peroxide damage in soybean root nodules. Proc Natl Acad Sci USA 83:3811–3815

  7. Edwards EA, Rawsthorne S, Mullineaux PM (1990) Subcellular distribution of multiple forms of glutathione reductase in leaves of pea (Pisum sativum L.). Planta 180:278–284

  8. Edwards EA, Enard C, Creissen GP, Mullineaux PM (1994) Synthesis and properties of glutathione reductase in stressed peas. Planta 192:137–143

  9. Feinberg AP Vogelstein B (1983) Atechnique for radiolabelling DNA restriction endonuclease fragments to high specificity. Anal Biochem 132:6–13

  10. Foyer CH, Halliwell B (1976) Presence of glutathione and glutathione reductase in chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133: 21–25

  11. Foyer CH, Lelandais M, Galap C, Kunert KJ (1991) Effects of elevated cytosolic glutathione reductase activity on the cellular glutathione pool and photosynthesis in leaves under normal and stress conditions. Plant Physiol 97: 863–872

  12. Frohman MA, Dush MK, Martin GR (1988) Rapid amplification of full-length cDNAs from rare transcripts: Amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002

  13. Gavel Y, Heinje G von (1990) A conserved cleavage-site motif in chloroplast transit peptides. FEBS Lett 261:455–458

  14. Kubo A, Sano T, Saji H, Tanaka K, Kondo N, Tanaka K (1993) Primary structure and properties of glutathione reductase from Arabidopsis thaliana. Plant Cell Physiol 34:1259–1266

  15. Madamanchi NR, Anderson JV, Alscher RG, Cramer CL, Hess JL (1992) Purification of multiple forms of glutathione reductase from pea (Pisum sativum L.) seedlings and enzyme levels in ozone-fumigated pea leaves. Plant Physiol 100:138–145

  16. Nakano Y, Asada K (1980) Spinach chloroplasts scavenge hydrogen peroxide on illumination. Plant Cell Physiol 21:1295–1307

  17. Ortiz DF, Strommer JN (1990) The MuI maize transposable element induces tissue-specific aberrant splicing and polyadenylation in two AdhI mutants. Mol Cell Biol 10:1039–1049

  18. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

  19. Smith HH (1968) Recent cytogenetic studies in the genus Nicotiana. Adv Genet 14:1–54

  20. Sharp PA, Konarska MM, Grabowski AI, Lamond AI, Marciniak R, Seiler SR (1987) Splicing of messenger RNA precursors. Cold Spring Harbor Symp Quant Biol, vol LII, 277–285

  21. Wingate VPM, Lawton MA, Lamb CJ (1988) Glutathione elicits a massive and selective induction of plant defense genes. Plant Physiol 87:206–210

  22. Winter J, Wright R, Duck N, Gasser C, Frahley R, Shah D (1988)The inhibition of petunia hsp70 mRNA processing during CdCl2 stress. Mol Gen Genet 211:325–319

  23. Ziegler DM (1985) Role of reversible oxidation-reduction of enzyme thiol-disulphides in metabolic regulation. Annu Rev Biochem 54:305–329

Download references

Author information

Correspondence to Gary P. Creissen.

Additional information

We thank Dr. Rod Casey for critical reading of the manuscript. This work was supported by the Biotechnology and Biological Sciences Research Council through a grant-in-aid to the John Innes Centre and by the Commission of the European Union Frame-work III AIR programme (contract number AIR1-CT92-0205)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Creissen, G.P., Mullineaux, P.M. Cloning and characterisation of glutathione reductase cDNAs and identification of two genes encoding the tobacco enzyme. Planta 197, 422–425 (1995).

Download citation

Key words

  • Glutathione reductase (cDNAs)
  • Intron skipping
  • Nicotiana