Changes in glutathione and ascorbate content, hydrogen peroxide and abscisic acid were studied in developing leaves in wild-type (Col-0) and glutathione-deficient mutant pad 2-1 lines of Arabidopsis thaliana over a time period of 9 days of drought followed by re-watering. Glutathione deficient mutant ( pad 2-1) presents mechanisms of acclimation to water stress through the reduction of plant biomass and increase in endogenous concentrations of ascorbate and glutathione. These acclimation responses to stress appeared along with the first symptoms of stress and we suggest here that they are regulated by ABA and H 2 O 2 . Normally, wild type plants under conditions of stress require an initial response phase in which a decrease in antioxidants is observed before reaching acclimation through the increase in levels of antioxidants. pad 2-1 is more sensitive to stress and reacts to it ; however, it did not suffer more oxidative stress than Col-0 plants, even though pad 2-1 had higher levels of endogenous H 2 O 2 relative to wild-type. In both water stressed Col-0 and pad 2-1 plants increases in ABA were observed , however, more sharply in wild-type stressed plants. Low levels of glutathione together with high levels of H 2 O 2 may regulate endogenous ABA concentrations and could be related to the slow growth rates which were observed during the experiment. The results highlighted the double function of glutathione as an antioxidant and signal molecule and also, the different response patterns of wild-type and pad 2-1 when faced with drought stress. The results bring new insights to the responses of pad 2-1 under conditions of water stress.
Apel, K. and Hirt, H. (2004) Reactive Oxygen Species: Metabolism, Oxidative Stress, and Signal Transduction. Annual Review of Plant Biology, 55, 373-379. http://dx.doi.org/10.1146/annurev.arplant.55.031903.141701
Brossa, R., López-Carbonell, M., Jubany-Marí, T. and Alegre, L. (2011) Interplay between Abscisic Acid and Jasmonic Acid and Its Role in Water-Oxidative Stress in Wild-Type, ABA Deficient, JA-Deficient, and Ascorbate Deficient Arabidopsis Plants. Journal of Plant Regulation, 30, 322-333. http://dx.doi.org/10.1007/s00344-011-9194-z
Foyer, C.H. and Noctor, G. (2011) Ascorbate and Glutathione: The Hearth of the Redox Hub. Plant Physiology, 155, 2-18. http://dx.doi.org/10.1104/pp.110.167569
Noctor, G. (2006) Metabolic Signaling in Defence and Stress. The Central Roles of Soluble Redox Couples. Plant Cell and Environment, 29, 409-425. http://dx.doi.org/10.1111/j.1365-3040.2005.01476.x
Noctor, G., Mhamdi, A., Chaouch, S., Han, Y., Neukerman, J., Marquez-Garcia, B., et al. (2012) Glutathione Functions in Plants: An Integrated Overwiew. Plant Cell and Environment, 35, 454-484. http://dx.doi.org/10.1111/j.1365-3040.2011.02400.x
Noctor, G., Mhamdi, A., Queval, G. and Foyer, C.H. (2013) Regulating the Redox Gatekeeper: Vacuolar Sequestration Puts Glutathione Disulfide in Its Place. Plant Physiology, 163, 665-671. http://dx.doi.org/10.1104/pp.113.223545
Potters, G., De Gara, L., Asard, H. and Horemans, N. (2002) Ascorbate and Glutathione Guardians of the Cell Cycle, Partners in Crime? Plant Physiol.ogy and Biochemistry, 40, 537-548. http://dx.doi.org/10.1016/S0981-9428(02)01414-6
May, M.J., Vernoux, T., Leaver, C., Van Montagu, M. and Inzé, D. (1998) Glutathione Homeostasis in Plants: Implications for environmental Sensing and Plant Development. Journal of Experimental Botany, 49, 649-667.
Mullineaux, P.M. and Raush, T. (2005) Glutathione, Photosynthesis and the Redox Regulation of Stress-Responsive Gene Expression. Photosyntesis Research, 86, 459-474. http://dx.doi.org/10.1007/s11120-005-8811-8
Bick, J.A., Setterdahl, A.T., Knaff, D.B., Chen. Y., Pitcher, L.H., Zilinkas, B., et al. (2001) Regulation of the Plant-Type 5’-Adenylyl Sulfate Reductase by Oxidative Stress. Biochemistry, 40, 9040-9048. http://dx.doi.org/10.1021/bi010518v
Gomez, L., Vanacker, H., Buchner, P., Noctor, G. and Foyer, C.H. (2004) The Intercellular Distribution of Glutathione Synthesis and Its Response to Chilling in Maize. Plant Physiology, 134, 1662-1671. http://dx.doi.org/10.1104/pp.103.033027
Holler, K., Kiraly, L., Künstler, A., Müller, M., Gullner, G., Fatinger, M., et al. (2010) Enhanced Glutathione Metabolism Is Correlated with Sulfur-Induced Resistance in Tobaco Mosaic Virus-Infected Genetically Susceptible Nicotiana tabacum Plants. Molecular Plant-Microbe Interactions, 23, 1448-1459. http://dx.doi.org/10.1094/MPMI-05-10-0117
Ball, L., Accotto, G.P., Bechtold, U., Creissen, G., Funck, D., Jimenez, A., et al. (2004) Evidence for a Direct Link between Glutathione Biosynthesis and Stress Defense Gene Expression in Arabidopsis. Plant Cell, 16, 2448-2462. http://dx.doi.org/10.1105/tpc.104.022608
Cobbett, C.S., May, M.J., Howden, R. and Rolls, B. (1998) The Glutathione-Deficient Cadmium-Sensitive Mutant cad2-1, of Arabidopsis thaliana Is Deficient in Gamma-Glutamylcysteine Synthase. Plant Journal, 16, 73-78. http://dx.doi.org/10.1046/j.1365-313x.1998.00262.x
Howden, R., Andersen, C.R., Goldsbrough, P.B. and Cobbett, C.S. (1995) A Cadmium-Sensitive, Glutathione-Deficient Mutant of Arabidopsis thaliana. Plant Physiology, 107, 1067-1073. http://dx.doi.org/10.1104/pp.107.4.1067
Parisy, V., Poinssot, B., Owsianowki, A., Glazebrook, J. and Mauch, F. (2006) Identification of pad2 as a γ-Glutamylcisteine Synthatase Highlights the Importance og Gluatahione Disease Resistance of Arabidopsis. The Plant Journal, 49, 159-172. http://dx.doi.org/10.1111/j.1365-313X.2006.02938.x
Dubreuil-Maurizi, C., Vitecek, J., Marty, L., Branciard, L., Frettinger, P., Wendehenne, D., et al. (2011) Glutathione Deficiency od the Arabidopsis Mutant pad2-1 Affects Oxidative Stress-Related Events, Defense Gene Expression and the Hypersensitive Response. Plant Physiology, 157, 2000-2012. http://dx.doi.org/10.1104/pp.111.182667
Jubany-Marí, T., Munné-Bosch, S., López-Carbonell, M. and Alegre, L. (2009) Hydrogen Peroxide Is Involved in the Acclimation of the Mediterranean Shrub Cistus albidus L., to Summer Drought. Journal of Experimental Botany, 60, 107-120. http://dx.doi.org/10.1093/jxb/ern274
Jubany-Marí, T., Prinsen, E., Munné-Bosch, S. and Alegre, L. (2010) The Timing of Methyl Jasmonate, Hydrogen Peroxide and Ascorbate Accumulation during Water Deficit and Subsequent Recovery in the Mediterranean Shrub Cistus albidus L. Environmental and Experimental Botany, 69, 47-55. http://dx.doi.org/10.1016/j.envexpbot.2010.02.003
Asada, K. (1999) The Water-Water Cycle in Chloroplasts: Scavenging of Active Oxygens and Dissipation of Excess Photons. Annual Review of Plant Physiology and Molecular Biology, 50, 601-639. http://dx.doi.org/10.1146/annurev.arplant.50.1.601
Mhamdi, A., Hager, J., Chaoux, S., Queval, G., Han, Y., Taconnat, L., Saindrenan, P., et al. (2010) Arabidopsis GLUTATHIONE REDUCTASE1 Plays a Crucial Role in Leaf Responses to Intracellular Hydrogen Peroxide and in Ensuring Appropriate Gene Expression Signaling Pathways. Plant Physiology, 153, 1144-1160. http://dx.doi.org/10.1104/pp.110.153767
Queval, G., Issakidis-Bourguet, E., Hoeberischts, F.A., Vandorpe, M., Gakière, B., Vanaker, H., et al. (2007) Conditional Oxidative Stress Responses in the Arabidopsis Photorespiratory Mutant cat2 Demonstrate That Redox Status Is a Key Modulator of Day-Length-Dependent Gene Expression and Define Photoperiod as a Crucial Factor in the Regulation of H2O2 Induced Cell Death. The Plant Journal, 52, 640-657. http://dx.doi.org/10.1111/j.1365-313X.2007.03263.x
Queval, G., Thominet, D., Vanacker, H., Miginiac-Maslow, M., Gakière, B. and Noctor, G. (2009) H2O2-Activated Up-Regulation of Glutathione in Arabidopsis Involves Induction of Genes Encoding Enzymes Involved in Cysteine Synthesis in the Chloroplasts. Molecular Plant, 2, 344-356. http://dx.doi.org/10.1093/mp/ssp002
Smith, I.K., Kendall, A.C., Keys, A.J., Turner, J.C. and Lea, P.J. (1984) Increased Levels of Glutathione in Catalase-Deficient Mutant of Barley (Hordeum vulgare L.). Plant Science Letters, 37, 29-33. http://dx.doi.org/10.1016/0304-4211(84)90198-6
Willekens, H., Chamnongpol, S., Davey, M., Schraudner, M., Langebartels, C., Van Montagu, M., et al. (1997) Catalase Is a Sink for H2O2 and Is Indispensable for Stress Defense in C3 Plants. The EMBO Journal, 16, 4806-4816. http://dx.doi.org/10.1093/emboj/16.16.4806
Tognetti, V.B., Van Aken, O., Morreel, K., Vandnbroucke, K., Van de Cotte, B., De Clercq I., et al. (2010) Perturbation of Indole-3-Butyric Acid Homeostasis by the UDP-Glucosyltrasferase UGT74E2 Modulates Arabidopsis Architecture and Water Stress Tolerance. The Plant Cell, 22, 2660-2679. http://dx.doi.org/10.1105/tpc.109.071316
Queval, G. and Noctor, G. (2007) A Plate Reader Method for the Measurement of NAD, NADP, Glutathione, and Ascorbate in the Tissue Extracts: Application to Redox Profiling during Arabidopsis Rosette Development. Analytical Biochemistry, 363, 58-69. http://dx.doi.org/10.1016/j.ab.2007.01.005
Foyer, C., Rowell, J. and Walker, D. (1983) Measurement of the Ascorbate Content of Spinach Leaf Protoplasts during Illumination. Planta, 157, 239-244. http://dx.doi.org/10.1007/BF00405188
Tietze, F. (1969) Enzymic Method for Quantitative Determination of Nanogram Amounts of Total and Oxidized Glutathione Applications to Mammalian Blood and Other Tissues. Analytical Biochemistry, 27, 502-522. http://dx.doi.org/10.1016/0003-2697(69)90064-5
López-Carbonell, M., Gabassa, M. and Jáuregui, O. (2009) Enhanced Determination of Abscisic Acid (ABA) and Abscisic Acid Glucose Ester (ABA-GE) in Cistus albidus Plants by Liquid Chromatography Mass Spectrometry in Tandem Mode. Plant Physiology and Biochemistry, 47, 256-261. http://dx.doi.org/10.1016/j.plaphy.2008.12.016
Brossa, R., Pintó-Marijuan, M., Jiang, K., Alegre, L. and Feldman, L. (2013) Assessing the Regulation of Leaf Redox Status under Water Stress Conditions in Arabidopsis thaliana. Plant Signaling & Behavior, 8, e24781. http://dx.doi.org/10.4161/psb.24781
Reicheld, J.P., Khafif, M., Riondet, C., Droux, M., Bonnard, G. and Meyer, Y. (2007) Inactivation of Thioredoxin Reductases Reveals a Complex Interplay between Thioredoxin and Glutathione Pathways in Arabidopsis Development. The Plant Cell, 19, 1851-1865. http://dx.doi.org/10.1105/tpc.107.050849
Vernoux, T., Wilson, R.C., Seeley, K.A., Reicheld, J.P., Murot, S., Brown, S., et al. (2000) The ROOMERISTEMLESS1/CADMIUM SENSITIVE2 Gene Defines a Glutathione-Dependent Pathway Involved in Initiation and Maintenance of Cell Division during Postembryonic Root Development. The Plant Cell, 12, 97-110.
Koprivova, A., Mugford, S.T. and Kopriva, S. (2010) Arabidopsis Root Growth Dependence on Glutathione Is Linked to Auxin Transport. Plant Cell Reports, 29, 1157-1167. http://dx.doi.org/10.1007/s00299-010-0902-0
García-Giménez, J.L., Markovic, J., Dasí, F., Queval, G., Schanaubelt, D., Foyer, C.H., et al. (2013) Nuclear Glutathione. Biochimica et Biophysica Acta (BBA)-General Subjects, 1830, 3304-3316. http://dx.doi.org/10.1016/j.bbagen.2012.10.005
Bashandy, T., Guilleminot, J., Vernoux, T., Caparros-Ruiz, D., Ljung, K., Meyer, T., et al. (2010) Interplay between the NADP-Linked Thioredoxin and Glutathione Systems in Arabidopsis Auxin Signaling. The Plant Cell, 22, 376-391. http://dx.doi.org/10.1105/tpc.109.071225
Schnaubelt, D., Queval, G., Dong, Y., Diaz-Vivancos, P., Makgopa, M.E., Howell, G., et al. (2015) Low Glutathione Regulates Gene Expression and the Redox Potentials of the Nucleus and Cytosol in Arabidopsis thaliana. Plant, Cell & Environment, 38, 266-279. http://dx.doi.org/10.1111/pce.12252
Claeys, H. and Inzé, D. (2013) The Agony of Choice: How Plants Balance Growth and Survival under Water-Limiting Conditions. Plant Physiology, 162, 1768-1779. http://dx.doi.org/10.1104/pp.113.220921
Clauw, P., Coppens, F., De Beuf, K., Dhondt, S., Van Daele, T., Maleux, K., et al. (2015) Leaf Responses to Mild Drought Stress in Natural Variants of Arabidopsis. Plant Physiology, 167, 800-816. http://dx.doi.org/10.1104/pp.114.254284
Tardieu, F., Parent, B. and Simonneau, T. (2010) Control of Leaf Growth by Abscisic Acid Hydraulic or Non-Hydraulic Processes? Plant, Cell & Environment, 33, 636-647. http://dx.doi.org/10.1111/j.1365-3040.2009.02091.x
Heyneke, E., Lushi-Ebengreuth, N., Krajcer, V., Müller, M. and Zechmann, B. (2013) Dynamic Compartment Specific Changes in Glutathione and Ascorbate Levels in Arabidopsis Plants Exposed to Different Light Intensities. BMC Plant Biology, 13, 104. http://dx.doi.org/10.1186/1471-2229-13-104
Tausz, M., Sircelj, H. and Grill, D. (2004) The Glutathione System as a Stress Markers in Plant Ecophysiology: Is a Stress-Response Concept Valid? Journal of Experimental Botany, 55, 1955-1962. http://dx.doi.org/10.1093/jxb/erh194
Schachtman, D.P. and Goodger, Q.D. (2008) Chemical Root to Shoot Signaling under Drought. Trends in Plant Science, 13, 281-287. http://dx.doi.org/10.1016/j.tplants.2008.04.003
Koffler, B.E., Luschin-Ebengreuth, N.L., Stabentheir, E., Müller, M. and Zechmann, B. (2014) Compartment Specific Response of Antioxidants to Drought Stress in Arabidopsis. Plant Science, 227, 133-144. http://dx.doi.org/10.1016/j.plantsci.2014.08.002
Koffler, B.E., Luschin-Ebengreuth, N.L. and Zechmann, B. (2015) Compartment Specific Changes of the Antioxidative Status in Arabidopsis thaliana during Salt Stress. Journal of. Plant Biology, 58, 8-16. http://dx.doi.org/10.1007/s12374-014-0264-1
Finkelstein, R.B. and Rock, C.D. (2002) Abscisic Acid Biosynthesis and Response. The Arabidopsis Book, 1, e0058. http://dx.doi.org/10.1199/tab.0058
Yamaguchi-Shinozaki, K. and Shinozaki, K. (2006) Transcriptional Regulatory Networks in Cellular Response and Tolerance to Dehydration and Cold Stresses. Annual Review of Plant Biology, 57, 781-803. http://dx.doi.org/10.1146/annurev.arplant.57.032905.105444
Chan, K.X., Wirtz, M., Phua, S.Y., Estavillo, G.M. and Pogson, B.J. (2012) Balancing Metabolites in Drought: The Sulfur Assimilation Conundrum. Trends in Plant Science, 18, 18-29. http://dx.doi.org/10.1016/j.tplants.2012.07.005
Jiang, M. and Zhang, J. (2002) Water Stress-Induced Abscisic Acid Accumulation Triggers the Increased Generation of Reactive Oxygen Species and Up-Regulates the Activities of Antioxidant Enzymes in Maize Leaves. Journal of Experimental Botany, 53, 2401-2410. http://dx.doi.org/10.1093/jxb/erf090
Han, Y., Chaouch, S., Mhamdi, A., Queval, G., Zechmann, B. and Noctor, G. (2013) Functional Analysis of Arabidopsis Mutants Points to Novel Roles for Glutathione in Coupling H2O2 to Activation of Salicylic Accumulation and Signaling. Antioxidants & Redox Signaling, 18, 2106-2121. http://dx.doi.org/10.1089/ars.2012.5052
Niu, Y., Wang, Y., Li, P., Zhang, F., Liu, H. and Zheng, G. (2013) Drought Stress Induces Oxidative Stress and the Antioxidant Defense System in Ascorbate-Deficient vtc1 Mutants of Arabidopsis thaliana. Acta Physiologiae Plantarum, 35, 1189-1200. http://dx.doi.org/10.1007/s11738-012-1158-9
Petrov, V.D. and Van Breusegem, F. (2012) Hydrogen Peroxide—A Central Hub for Information Flow in Plants. AoB Plants, 2012, pls014. http://dx.doi.org/10.1093/aobpla/pls014