The Arabidopsis glutathione transferase gene family displays complex stress regulation and co-silencing multiple genes results in altered metabolic sensitivity to oxidative stress.

Plant glutathione transferases (GSTs) are induced by diverse biotic and abiotic stimuli, and are important for protecting plants against oxidative damage. We have studied the primary transcriptional stress response of the entire Arabidopsis GST family to seven stresses, including both biotic and abiotic stimuli, with a focus on early changes in gene expression. Our results indicate that individual GST genes are highly specific in their induction patterns. Furthermore, we have been able to link individual GSTs to particular stress stimuli. Using RNAi, we successfully co-silenced a group of four phi GSTs that represent some of the most highly expressed GST genes. Despite a marked reduction in total phi GST protein levels, the transgenic plants showed no reduction in GST activity as measured using the model substrate 1-chloro-2,4-dinitrobenzene (CDNB), and appeared to have surprisingly robust physical phenotypes during stress. However, analysis of metabolite pools showed oxidation of the glutathione pool in the RNAi lines, and we observed alterations in carbon and nitrogen compounds following salicylic acid and hydrogen peroxide stress treatments, indicative of oxidative modification of primary metabolism. Thus, there appears to be a high degree of functional redundancy within the Arabidopsis GST family, with extensive disruption being required to reveal the roles of phi GSTs in protection against oxidative stress.

[1]  R. Lister,et al.  Highly Integrated Single-Base Resolution Maps of the Epigenome in Arabidopsis , 2008, Cell.

[2]  S. Nelson,et al.  Shotgun bisulphite sequencing of the Arabidopsis genome reveals DNA methylation patterning , 2008, Nature.

[3]  Joachim Selbig,et al.  The Metabolic Response of Heterotrophic Arabidopsis Cells to Oxidative Stress1[W] , 2006, Plant Physiology.

[4]  A. McDonald,et al.  Changes in plant mitochondrial electron transport alter cellular levels of reactive oxygen species and susceptibility to cell death signaling molecules. , 2006, Plant & cell physiology.

[5]  A. Millar,et al.  Desensitization of GSTF8 Induction by a Prior Chemical Treatment Is Long Lasting and Operates in a Tissue-Dependent Manner1[W] , 2006, Plant Physiology.

[6]  Nicolas L Taylor,et al.  Differential Impact of Environmental Stresses on the Pea Mitochondrial Proteome*S , 2005, Molecular & Cellular Proteomics.

[7]  C. Foyer,et al.  Redox Homeostasis and Antioxidant Signaling: A Metabolic Interface between Stress Perception and Physiological Responses , 2005, The Plant Cell Online.

[8]  K. Torii,et al.  Two callose synthases, GSL1 and GSL5, play an essential and redundant role in plant and pollen development and in fertility , 2005, Plant Molecular Biology.

[9]  A. Harvey Millar,et al.  Stress-induced co-expression of alternative respiratory chain components in Arabidopsis thaliana , 2005, Plant Molecular Biology.

[10]  R. Allen,et al.  Transgenic cotton (Gossypium hirsutum L.) seedlings expressing a tobacco glutathione S-transferase fail to provide improved stress tolerance , 2005, Planta.

[11]  P. Böger,et al.  A Ligand Function of Glutathione S-Transferase , 2005, Zeitschrift fur Naturforschung. C, Journal of biosciences.

[12]  G. Angenent,et al.  ASYMMETRIC LEAVES2-LIKE1gene, a member of the AS2/LOB family, controls proximal–distal patterning in Arabidopsis petals , 2005, Plant Molecular Biology.

[13]  A. Moons.  Regulatory and functional interactions of plant growth regulators and plant glutathione S-transferases (GSTs). , 2005, Vitamins and hormones.

[14]  P. Christou,et al.  Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[15]  S. Tachibana,et al.  Overexpression of spermidine synthase enhances tolerance to multiple environmental stresses and up-regulates the expression of various stress-regulated genes in transgenic Arabidopsis thaliana. , 2004, Plant & cell physiology.

[16]  A. Millar,et al.  Untangling multi-gene families in plants by integrating proteomics into functional genomics. , 2004, Phytochemistry.

[17]  Jonathan D. G. Jones,et al.  The Transcriptional Innate Immune Response to flg22. Interplay and Overlap with Avr Gene-Dependent Defense Responses and Bacterial Pathogenesis1[w] , 2004, Plant Physiology.

[18]  Marta Matvienko,et al.  Arabidopsis MPSS. An Online Resource for Quantitative Expression Analysis1[w] , 2004, Plant Physiology.

[19]  B. Halkier,et al.  Functional Analysis of the Tandem-Duplicated P450 Genes SPS/BUS/CYP79F1 and CYP79F2 in Glucosinolate Biosynthesis and Plant Development by Ds Transposition-Generated Double Mutants1 , 2004, Plant Physiology.

[20]  Jonathan D. G. Jones,et al.  Bacterial disease resistance in Arabidopsis through flagellin perception , 2004, Nature.

[21]  Klaus F. X. Mayer,et al.  Crosstalk and differential response to abiotic and biotic stressors reflected at the transcriptional level of effector genes from secondary metabolism , 2004, Plant Molecular Biology.

[22]  A. Millar,et al.  Salicylic Acid Is an Uncoupler and Inhibitor of Mitochondrial Electron Transport1 , 2004, Plant Physiology.

[23]  Ulrich Wagner,et al.  Probing the Diversity of the Arabidopsis glutathione S-Transferase Gene Family , 2002, Plant Molecular Biology.

[24]  F. Brun,et al.  Photoaffinity labelling with the cytokinin agonist azido-CPPU of a 34 kDa peptide of the intracellular pathogenesis-related protein family in the moss Physcomitrella patens , 2001, Plant Molecular Biology.

[25]  K. Dixon,et al.  Acetyl salicylic acid (Aspirin) and salicylic acid induce multiple stress tolerance in bean and tomato plants , 2000, Plant Growth Regulation.

[26]  P. Goldsbrough,et al.  An Arabidopsis gene with homology to glutathione S-transferases is regulated by ethylene , 1993, Plant Molecular Biology.

[27]  A. Millar,et al.  Proteomic Analysis of Glutathione S-Transferases of Arabidopsis thaliana Reveals Differential Salicylic Acid-Induced Expression of the Plant-Specific Phi and Tau Classes , 2004, Plant Molecular Biology.

[28]  S. Kitamura,et al.  TRANSPARENT TESTA 19 is involved in the accumulation of both anthocyanins and proanthocyanidins in Arabidopsis. , 2004, The Plant journal : for cell and molecular biology.

[29]  P. Goldsbrough,et al.  Arabidopsis AtGSTF2 is regulated by ethylene and auxin, and encodes a glutathione S-transferase that interacts with flavonoids. , 2003, The Plant journal : for cell and molecular biology.

[30]  A. Moons.  Osgstu3 and osgtu4, encoding tau class glutathione S‐transferases, are heavy metal‐ and hypoxic stress‐induced and differentially salt stress‐responsive in rice roots 1 , 2003, FEBS letters.

[31]  Lothar Willmitzer,et al.  De Novo Amino Acid Biosynthesis in Potato Tubers Is Regulated by Sucrose Levels1[w] , 2003, Plant Physiology.

[32]  P. Waterhouse,et al.  Constructs and methods for high-throughput gene silencing in plants. , 2003, Methods.

[33]  P. Böger,et al.  Binding and protection of porphyrins by glutathione S-transferases of Zea mays L. , 2003, Biochimica et biophysica acta.

[34]  C. Foyer,et al.  Leaf Mitochondria Modulate Whole Cell Redox Homeostasis, Set Antioxidant Capacity, and Determine Stress Resistance through Altered Signaling and Diurnal Regulation Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.009464. , 2003, The Plant Cell Online.

[35]  Ying Zhu,et al.  Transgenic tobacco plants overexpressing cotton glutathione S-transferase (GST) show enhanced resistance to methyl viologen. , 2003, Journal of plant physiology.

[36]  M. Bianchi,et al.  Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase. , 2002, Physiologia plantarum.

[37]  B. G. Davis,et al.  Functional Divergence in the Glutathione Transferase Superfamily in Plants , 2002, The Journal of Biological Chemistry.

[38]  A. Lapthorn,et al.  Plant glutathione transferases , 2002, Genome Biology.

[39]  C. Helliwell,et al.  The Arabidopsis AMP1 Gene Encodes a Putative Glutamate Carboxypeptidase , 2001, The Plant Cell Online.

[40]  J. Hancock,et al.  Regulation of the Arabidopsis transcriptome by oxidative stress. , 2001, Plant physiology.

[41]  O. Borsani,et al.  Evidence for a role of salicylic acid in the oxidative damage generated by NaCl and osmotic stress in Arabidopsis seedlings. , 2001, Plant physiology.

[42]  R. Allen,et al.  Stress tolerance in transgenic tobacco seedlings that overexpress glutathione S-transferase/glutathione peroxidase. , 2000, Plant & cell physiology.

[43]  S. Stolz,et al.  Fatty acid ketodienes and fatty acid ketotrienes: Michael addition acceptors that accumulate in wounded and diseased Arabidopsis leaves. , 2000, The Plant journal : for cell and molecular biology.

[44]  S. J. Keeler,et al.  A genomics approach to the comprehensive analysis of the glutathione S-transferase gene family in soybean and maize. , 2000, Plant physiology.

[45]  V. Walbot,et al.  AN9, a petunia glutathione S-transferase required for anthocyanin sequestration, is a flavonoid-binding protein. , 2000, Plant physiology.

[46]  R Edwards,et al.  Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. , 2000, Trends in plant science.

[47]  Seppänen,et al.  Characterization and expression of cold-induced glutathione S-transferase in freezing tolerant Solanum commersonii, sensitive S. tuberosum and their interspecific somatic hybrids. , 2000, Plant science : an international journal of experimental plant biology.

[48]  J. Hancock,et al.  Hydrogen peroxide-induced gene expression in Arabidopsis thaliana. , 2000, Free radical biology & medicine.

[49]  K B Singh,et al.  The auxin, hydrogen peroxide and salicylic acid induced expression of the Arabidopsis GST6 promoter is mediated in part by an ocs element. , 1999, The Plant journal : for cell and molecular biology.

[50]  Jyoti Shah,et al.  Salicylic acid and disease resistance in plants. , 1999 .

[51]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[52]  C. Foyer,et al.  Simultaneous measurement of foliar glutathione, gamma-glutamylcysteine, and amino acids by high-performance liquid chromatography: comparison with two other assay methods for glutathione. , 1998, Analytical biochemistry.

[53]  D. Inzé,et al.  Polyamines and paraquat toxicity in Arabidopsis thaliana. , 1998, Plant & cell physiology.

[54]  J. W. Gronwald,et al.  Isolation and characterization of glutathione S-transferase isozymes from sorghum. , 1998, Plant physiology.

[55]  V. Walbot,et al.  Functional Complementation of Anthocyanin Sequestration in the Vacuole by Widely Divergent Glutathione S-Transferases , 1998, Plant Cell.

[56]  H. H. Muller,et al.  Constitutively Elevated Levels of Putrescine and Putrescine-Generating Enzymes Correlated with Oxidant Stress Resistance in Conyza bonariensis and Wheat , 1997, Plant physiology.

[57]  R. Allen,et al.  Engineering stress tolerance in transgenic plants , 1997, Acta Physiologiae Plantarum.

[58]  E. R. Allen,et al.  Overexpression of glutathione S-transferase/glutathioneperoxidase enhances the growth of transgenic tobacco seedlings during stress , 1997, Nature Biotechnology.

[59]  F. Droog.  Plant Glutathione S-Transferases, a Tale of Theta and Tau , 1997, Journal of Plant Growth Regulation.

[60]  J. Coleman,et al.  Detoxification of xenobiotics by plants: chemical modification and vacuolar compartmentation , 1997 .

[61]  Z. Mei.  Regulation of Glutathione S-Transferase Gene Expression , 1997 .

[62]  G. Chao,et al.  The promoter of a H2O2-inducible, Arabidopsis glutathione S-transferase gene contains closely linked OBF- and OBP1-binding sites. , 1996, The Plant journal : for cell and molecular biology.

[63]  K. Marrs.  THE FUNCTIONS AND REGULATION OF GLUTATHIONE S-TRANSFERASES IN PLANTS. , 1996, Annual review of plant physiology and plant molecular biology.

[64]  I. Raskin,et al.  Ozone-induced responses in Arabidopsis thaliana: the role of salicylic acid in the accumulation of defense-related transcripts and induced resistance. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[65]  A. Sturm,et al.  Cloning and Characterization of a Glutathione S-Transferase That Can Be Photolabeled with 5-Azido-indole-3-acetic Acid , 1995, Plant physiology.

[66]  V. Walbot,et al.  A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2 , 1995, Nature.

[67]  K. Palme,et al.  Photoaffinity labeling of Arabidopsis thaliana plasma membrane vesicles by 5-azido-[7-3H]indole-3-acetic acid: identification of a glutathione S-transferase. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[68]  K. Shinozaki,et al.  Characterization of two cDNAs (ERD11 and ERD13) for dehydration‐inducible genes that encode putative glutathione S‐transferases in Arabidopsis thaliana L , 1993, FEBS letters.

[69]  F. Mauch,et al.  Differential Induction of Distinct Glutathione-S-Transferases of Wheat by Xenobiotics and by Pathogen Attack , 1993, Plant physiology.

[70]  Yasunori Nakamura,et al.  Carbohydrate Metabolism in the Developing Endosperm of Rice Grains , 1989 .

[71]  W B Jakoby,et al.  Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. , 1974, The Journal of biological chemistry.