Abstract
Glutathione S-transferases (GST) are multifunctional proteins encoded by a large gene family, divided on the basis of sequence identity into phi, tau, theta, zeta and lambda classes. The phi and tau classes are present only in plants. GSTs appear to be ubiquitous in plants and are involved in herbicide detoxification and stress response, but little is known about the precise role of GSTs in normal plant physiology and during biotic and abiotic stress response. Two cDNAs representing the two plant classes tau and phi, AtGSTF9 and AtGSTU26, were expressed in vitro and the corresponding proteins were analysed. Both GSTs were able to catalyse a glutathione conjugation to 1-chloro-2,4-dinitrobenzene (CDNB), but they were inactive as transferases towards p-nitrobenzylchloride (pNBC). AtGSTF9 showed activity towards benzyl isothiocyanate (BITC) and an activity as glutathione peroxidase with cumene hydroperoxide (CumHPO). AtGSTU26 was not active as glutathione peroxidase and towards BITC. RT-PCR analysis was used to evaluate the expression of the two genes in response to treatment with herbicides and safeners, chemicals, low and high temperature. Our results reveal that AtGSTU26 is induced by the chloroacetanilide herbicides alachlor and metolachlor and the safener benoxacor, and after exposure to low temperatures. In contrast, AtGSTF9 seems not to be influenced by the treatments employed.
Similar content being viewed by others
Abbreviations
- AGI:
-
Arabidopsis Genome Initiative
- BITC:
-
Benzyl isothiocyanate
- CDNB:
-
1-Chloro-2,4-dinitrobenzene
- CumHPO:
-
Cumene hydroperoxide
- DTT:
-
Dithiothreitol
- GSH:
-
Glutathione
- GST:
-
Glutathione S-transferase
- MJ:
-
Methyl jasmonic acid
- PCIB:
-
2-(p-Chlorophenoxy)-2-methylpropionic acid
- pNBC:
-
p-Nitrobenzylchloride
- RT-PCR:
-
Reverse transcript polymerase chain reaction
- RTS:
-
Rapid Translation System
- SA:
-
Salicylic acid
- SAR:
-
Systemic acquired resistance
- SDS-PAGE:
-
Sodium dodecyl sulphate polyacrylamide gel electrophoresis
- 2,4,6-T:
-
2,4,6-Trichlorophenol
References
Davies J, Caseley JC (1999) Herbicide safeners: a review. Pestic Sci 55:1043–1058
DeRidder BP, Dixon DP, Beussman DJ, Edwards R, Goldsbrough PB (2002) Induction of glutathione S-transferase in Arabidopsis by herbicides safeners. Plant Physiol 130:1497–1505
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
Ezaki B, Suzuki M, Motoda H, Kawamura M, Nakashima S, Matsumoto H (2004) Mechanism of gene expression of Arabidopsis glutathione S-transferase, AtGST1, and AtGST11 in response to aluminium stress. Plant Physiol 134:1–11
Flohè L, Günzler WA (1984) Assay of glutathione peroxidase. Methods Enzymol 105:114–121
Jepson I, Holt DC, Roussel V, Wright SY, Greenland AJ (1997) Transgenic plant analysis as a tool for the study of maize glutathione S-transferases. In: Hatzios KK (ed) Regulation of enzymatic systems detoxifying xenobiotics in plants. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 313–323
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
Kolm RH, Danielson UH, Zhang Y, Talalay P, Mannervik B (1995) Isothiocyanates as substrates for human glutathione transferases: structure–activity studies. Biochem J 311:453–459
Lamoureux GL, Rusness DG (1993) Glutathione in the metabolism and detoxification of xenobiotics in plants. In de Kok LJ, Stulen I, Rennenberg H, Brunold C, Rauser WE (eds) Sulfur nutrition and assimilation in higher plants. SPB Academic Publishing, The Hague, The Netherlands, pp. 221–237
Mannervik B, Guthenberg C (1981) Glutathione transferase (human placenta). Methods Enzymol 77:231–235
Marrs KA (1996) The functions and regulation of glutathione S-transferases in plants. Annu Rev Plant Physiol Plant Mol Biol 47:127–158
Martinoia E, Grill E, Tommasini R, Kreuz R, Amrhein N (1993) ATP-dependent glutathione S-conjugate ‘export’ pump in the vacuolar membrane of plants. Nature 364:247–249
Neuefeind T, Reinemer P, Bieseler B (1997) Plant glutathione S-transferases and herbicide detoxification. Biol Chem 378:199–205
Rea PA (1999) MRP subfamily ABC transporters from plants and yeast. J Exp Bot 50:895–913
Reinemer P, Prade L, Hof P, Neuefeind T, Huber R, Zettl R, Palme R, Schell K, 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
Sappl PG, Oñate-Sanchez L, Singh KB, Millar AH (2004) Proteomic analysis of glutathione S-transferases of Arabidopsis thaliana reveals differential salicylic acid-induced expression of the plant-specific phi and tau class. Plant Mol Biol 54:205–219
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 279:26098–26104
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
Zimmermann P, Hirsh-Hoffmann M, Hennig L, Gruissem W (2004) GENEVESTIGATOR: Arabidopsis thaliana microarray database and analysis toolbox. Plant Physiol 136:2621–2632
Zimmermann P, Hennig L, Gruissem W (2005) Gene-expression analysis and network discovery using Genevestigator. Trends Plant Sci 10:407–409
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by P. Puigdoménech
Rights and permissions
About this article
Cite this article
Nutricati, E., Miceli, A., Blando, F. et al. Characterization of two Arabidopsis thaliana glutathione S-transferases. Plant Cell Rep 25, 997–1005 (2006). https://doi.org/10.1007/s00299-006-0146-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-006-0146-1