Antioxidant enzyme activities and gene expression patterns in peanut nodules during a drought and rehydration cycle
Ana Laura Furlan A B C , Eliana Bianucci A , María del Carmen Tordable A , Stella Castro A and Karl-Josef Dietz B
+ Author Affiliations
- Author Affiliations
A Departamento de Ciencias Naturales, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto. Ruta 36, Km 601, 5800 Río Cuarto, Córdoba, Argentina.
B Biochemistry and Physiology of Plants, Bielefeld University, D-33501 Bielefeld, Germany.
C Corresponding author. Email: afurlan@exa.unrc.edu.ar
Functional Plant Biology 41(7) 704-713 https://doi.org/10.1071/FP13311
Submitted: 24 October 2013 Accepted: 29 January 2014 Published: 26 March 2014
Abstract
Drought stress is one of the most important environmental factors that affect plant growth and limit biomass production. Most studies focus on drought stress development but the reversibility of the effects receives less attention. Therefore, the present work aims to explore the biological nitrogen fixation (BNF) of the symbiotic association between peanut (Arachis hypogaea L.) and Bradyrhizobium sp. during a drought–recovery cycle with a focus on the response of enzyme activity and gene expression of the antioxidant system. Peanuts exposed to drought stress had impaired BNF, as indicated by lower nitrogenase activity, and decreased leghaemoglobin content; the latter was reversed to control values upon rehydration. Previous results demonstrated that reactive oxygen species (O2·− and H2O2) were accumulated as a consequence of drought stress, suggesting that nodules experience oxidative stress. In addition, marker transcripts responsive to drought, abscisic acid and H2O2 were upregulated. Increased transcript levels of glutathione reductase were associated with an increased enzyme activity but superoxide dismutase and glutathione S-transferase activities were unchanged, despite upregulated gene transcription. In contrast, increased activity of ascorbate peroxidase (APX) was unrelated with changes in cytosolic APX transcript levels suggesting isogene specificity. In conclusion, the work exemplarily demonstrates the efficient and dynamic regulation of antioxidant enzymes and marker compounds during drought cycling, which is likely to be a prerequisite for functional optimisation of nodule metabolism.
Additional keywords: antioxidant system, Arachis hypogaea, biological nitrogen fixation, oxidative stress, reactive oxygen species.
References
Aebi H (1984) Catalase in vitro. Methods in Enzymology 105, 121–126.| Catalase in vitro.Crossref | GoogleScholarGoogle Scholar | 6727660PubMed |
Amako K, Chen GX, Asada K (1994) Separate assays specific for ascorbate peroxidase and guaiacol peroxidase and for the chloroplastic and cytosolic isozymes of ascorbate peroxidase in plants. Plant & Cell Physiology 35, 497–504.
Anderson ME (1985) Determination of glutathione and glutathione disulfide in biological samples. Methods in Enzymology 113, 548–555.
| Determination of glutathione and glutathione disulfide in biological samples.Crossref | GoogleScholarGoogle Scholar | 4088074PubMed |
Appleby C, Bergersen F (1980) Preparation and experimental use of leghemoglobin. In ‘Methods for evaluating biological nitrogen fixation’. (Ed FJ Bergersen) pp. 315–335. (John Wiley: Chichester)
Arrese-Igor C, González EM, Gordon AJ, Minchin FR, Gálvez L, Royuela M, Cabrerizo PM, Aparicio-Tejo PM (1999) Sucrose synthase and nodule nitrogen fixation under drought and other environmental stresses. Symbiosis 27, 189–212.
Arrese-Igor C, González EM, Marino D, Ladrera R, Larrainzar E, Gil-Quintana E (2011) Physiological responses of legume nodules to drought. In ‘Plant nutrition and abiotic stress tolerance III. Plant stress 5 (Special issue 1)’. (Eds NA Anjum, F Lopez-Lauri) pp. 24–31. (Global Science Books: Ikenobe, Japan)
Asensio AC, Marino D, James EK, Ariz I, Arrese-Igor C, Aparicio-Tejo PM, Arredondo-Peter R, Moran JF (2011) Expression and localization of a Rhizobium-derived cambialistic superoxide dismutase in pea (Pisum sativum) nodules subjected to oxidative stress. Molecular Plant-Microbe Interactions 24, 1247–1257.
| Expression and localization of a Rhizobium-derived cambialistic superoxide dismutase in pea (Pisum sativum) nodules subjected to oxidative stress.Crossref | GoogleScholarGoogle Scholar | 21774575PubMed |
Beauchamp CO, Fridovich I (1973) Isoenzymes of SOD from wheat germ. Biochimica et Biophysica Acta 317, 50–64.
| Isoenzymes of SOD from wheat germ.Crossref | GoogleScholarGoogle Scholar | 4723247PubMed |
Becana M, Matamoros MA, Udvardi M, Dalton DA (2010) Recent insights into antioxidant defenses of legume root nodules. New Phytologist 188, 960–976.
| Recent insights into antioxidant defenses of legume root nodules.Crossref | GoogleScholarGoogle Scholar | 21039567PubMed |
Bensmihen S, Rippa S, Lambert G, Jublot D, Pautot V, Granier F, Giraudat J, Parcy F (2002) The homologous ABI5 and EEL transcription factors function antagonistically to fine-tune gene expression during late embryogenesis. The Plant Cell 14, 1391–1403.
| The homologous ABI5 and EEL transcription factors function antagonistically to fine-tune gene expression during late embryogenesis.Crossref | GoogleScholarGoogle Scholar | 12084834PubMed |
Bian S, Jiang Y (2009) Reactive oxygen species, antioxidant enzyme activities and gene expression patterns in leaves and roots of Kentucky bluegrass in response to drought stress and recovery. Scientia Horticulturae 120, 264–270.
| Reactive oxygen species, antioxidant enzyme activities and gene expression patterns in leaves and roots of Kentucky bluegrass in response to drought stress and recovery.Crossref | GoogleScholarGoogle Scholar |
Boote K (1982) Growth stages of peanut (Arachis hypogaea L.). Peanut Science 9, 35–40.
| Growth stages of peanut (Arachis hypogaea L.).Crossref | GoogleScholarGoogle Scholar |
Bradford M (1976) A rapid sensitive method for the quantification the microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.
| A rapid sensitive method for the quantification the microgram quantities of protein utilizing the principle of protein-dye binding.Crossref | GoogleScholarGoogle Scholar | 942051PubMed |
Bright J, Desikan R, Tancock JT, Weir IS, Neill SJ (2006) ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis. The Plant Journal 45, 113–122.
| ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H2O2 synthesis.Crossref | GoogleScholarGoogle Scholar | 16367958PubMed |
Burk R (1996) ‘Soil survey laboratory methods manual, soil survey investigations report 42, version 3.0.’ (National Soil Survey Center: Lincoln, NE, USA).
Collino DJ, Dardanelli JL, Sereno R, Racca RW (2001) Physiological responses of Argentine peanut varieties to water stress. Light interception, radiation use efficiency and partitioning of assimilates. Field Crops Research 70, 177–184.
| Physiological responses of Argentine peanut varieties to water stress. Light interception, radiation use efficiency and partitioning of assimilates.Crossref | GoogleScholarGoogle Scholar |
Dalton DA (1995) Antioxidant defenses of plants and fungi. In ‘Oxidative stress and antioxidant defenses in biology’. (Ed S Ahmad) pp. 298–355. (Chapman and Hall: New York)
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. Proceedings of the National Academy of Sciences of the United States of America 83, 3811–3815.
| Enzymatic reactions of ascorbate and glutathione that prevent peroxide damage in soybean root nodules.Crossref | GoogleScholarGoogle Scholar | 16593704PubMed |
Finkelstein RR, Gampala SS, Rock CD (2002) Abscisic acid signaling in seeds and seedlings. The Plant Cell 14, S15–S45.
Finkemeier I, Goodman M, Lamkemeyer P, Kandlbinder A, Sweetlove LJ, Dietz KJ (2005) The mitochondrial type II peroxiredoxin F is essential for redox homeostasis and root growth of Arabidopsis thaliana under stress. The Journal of Biological Chemistry 280, 12 168–12 180.
| The mitochondrial type II peroxiredoxin F is essential for redox homeostasis and root growth of Arabidopsis thaliana under stress.Crossref | GoogleScholarGoogle Scholar |
Flohé L, Günzler W (1984) Assays of glutathione peroxidase. Methods in Enzymology 105, 114–120.
| Assays of glutathione peroxidase.Crossref | GoogleScholarGoogle Scholar | 6727659PubMed |
Foyer CH, Noctor G (2009) Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxidants & Redox Signalling 11, 861–905.
| Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications.Crossref | GoogleScholarGoogle Scholar |
Furlan A Llanes A Luna V Castro S 2012
Furlan A, Llanes A, Luna V, Castro S (2013) Abscisic acid mediation in hydrogen peroxide production in peanut under water stress. Biologia Plantarum 57, 555–558.
| Abscisic acid mediation in hydrogen peroxide production in peanut under water stress.Crossref | GoogleScholarGoogle Scholar |
Gogorcena Y, Iturbe-Ormaetxe I, Escuredo PR, Becana M (1995) Antioxidant defenses against activated oxygen in pea nodules subjected to water stress. Plant Physiology 108, 753–759.
Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases: the first enzymatic step in mercaptric acid formation. The Journal of Biological Chemistry 249, 7130–7139.
Halliwell B, Gutteridge JMC (1999) ‘Free radicals in biology and medicine.’ 3rd edn. (Oxford University Press, Oxford)
Hardy R, Burns R, Holsten T (1973) Application of the acetylene reduction assay for the measurement of nitrogen fixation. Soil Biology & Biochemistry 5, 47–81.
| Application of the acetylene reduction assay for the measurement of nitrogen fixation.Crossref | GoogleScholarGoogle Scholar |
Hoagland D, Arnon D (1950) The water culture method for growing plants without soil. California Agricultural Experiment Station Bulletin 347, 1–39.
Hong L, Hu B, Liu X, He CY, Yao Y, Li XL, Li L (2013) Molecular cloning and expression analysis of a new stress-related AREB gene from Arachis hypogaea. Biologia Plantarum 57, 56–62.
| Molecular cloning and expression analysis of a new stress-related AREB gene from Arachis hypogaea.Crossref | GoogleScholarGoogle Scholar |
Irigoyen JJ, Emerich DW, Sánchez-Díaz M (1992) Phosphoenolpyruvate carboxylase, malate and alcohol dehydrogenase activities in alfalfa (Medicago sativa) nodules under water stress. Physiologia Plantarum 84, 61–66.
| Phosphoenolpyruvate carboxylase, malate and alcohol dehydrogenase activities in alfalfa (Medicago sativa) nodules under water stress.Crossref | GoogleScholarGoogle Scholar |
Jacquot JP, Dietz KJ, Rouhier N, Meux E, Lallement PA, Selles B, Hecker A (2013) Redox regulation in plants: glutathione and “redoxin” related families. In ‘Oxidative stress and redox regulation’. (Eds U Jakob, D Reichmann) pp. 213–231. (Springer Science Business Media: Dordrecht)
Law M, Charles S, Halliwell B (1983) Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of Paraquat. Biochemical Journal 210, 899–903.
Lu S, Su W, Li H, Guo Z (2009) Abscisic acid improves drought tolerance of triploid bermudagrass and involves H2O2- and NO-induced antioxidant enzyme activities. Plant Physiology and Biochemistry 47, 132–138.
| Abscisic acid improves drought tolerance of triploid bermudagrass and involves H2O2- and NO-induced antioxidant enzyme activities.Crossref | GoogleScholarGoogle Scholar | 19042137PubMed |
Luo M, Liang XQ, Dang P, Holbrook CC, Bausher MG, Lee RD, Guo BZ (2005) Microarray-based screening of differentially expressed genes in peanut in response to Aspergillus parasiticus infection and drought stress. Plant Science 169, 695–703.
| Microarray-based screening of differentially expressed genes in peanut in response to Aspergillus parasiticus infection and drought stress.Crossref | GoogleScholarGoogle Scholar |
Marino D, Gonzalez EM, Arrese-Igor C (2006) Drought effects on carbon and nitrogen metabolism of pea nodules can be mimicked by Paraquat: evidence for the occurrence of two regulation pathways under oxidative stresses. Journal of Experimental Botany 57, 665–673.
| Drought effects on carbon and nitrogen metabolism of pea nodules can be mimicked by Paraquat: evidence for the occurrence of two regulation pathways under oxidative stresses.Crossref | GoogleScholarGoogle Scholar | 16415332PubMed |
Marino D, Pucciariello C, Puppo P, Frendo P (2009) The redox state, a referee of the legume–rhizobia symbiotic game. Advances in Botanical Research 52, 115–151.
| The redox state, a referee of the legume–rhizobia symbiotic game.Crossref | GoogleScholarGoogle Scholar |
Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends in Plant Science 9, 490–498.
| Reactive oxygen gene network of plants.Crossref | GoogleScholarGoogle Scholar | 15465684PubMed |
Morgante CV, Guimarães PM, Martins ACQ, Araújo ACG, Leal-Bertioli SCM, Bertioli DJ, Brasileiro ACM (2011) Reference genes for quantitative reverse transcription-polymerase chain reaction expression studies in wild and cultivated peanut. BMC Research Notes 4, 339
| Reference genes for quantitative reverse transcription-polymerase chain reaction expression studies in wild and cultivated peanut.Crossref | GoogleScholarGoogle Scholar | 21906295PubMed |
Nakano Y, Asada K (1987) Purification of ascorbate peroxidase in spinach chloroplasts; its inactivation in ascorbate-depleted medium and reactivation by monodehydroascorbate radical. Plant & Cell Physiology 28, 131–140.
Navrot N, Collin V, Gualberto J, Gelhaye E, Hirasawa M, Rey P, Knaff DB, Issakidis E, Jacquot JP, Rouhier N (2006) Plant glutathione peroxidases are functional peroxiredoxins distributed in several subcellular compartments and regulated during biotic and abiotic stresses. Plant Physiology 142, 1364–1379.
| Plant glutathione peroxidases are functional peroxiredoxins distributed in several subcellular compartments and regulated during biotic and abiotic stresses.Crossref | GoogleScholarGoogle Scholar | 17071643PubMed |
Naya L, Ladrera R, Ramos J, Gonzalez EM, Arrese-Igor C, Minchin FR, Becana M (2007) The response of carbon metabolism and antioxidant defenses of alfalfa nodules to drought stress and to the subsequent recovery of plants. Plant Physiology 144, 1104–1114.
| The response of carbon metabolism and antioxidant defenses of alfalfa nodules to drought stress and to the subsequent recovery of plants.Crossref | GoogleScholarGoogle Scholar | 17468213PubMed |
Oelze ML, Vogel MO, Alsharafa K, Kahmann U, Viehhauser A, Maurino VG, Dietz KJ (2012) Efficient acclimation of the chloroplast antioxidant defence of Arabidopsis thaliana leaves in response to a 10- or 100-fold light increment and the possible involvement of retrograde signals. Journal of Experimental Botany 63, 1297–1313.
| Efficient acclimation of the chloroplast antioxidant defence of Arabidopsis thaliana leaves in response to a 10- or 100-fold light increment and the possible involvement of retrograde signals.Crossref | GoogleScholarGoogle Scholar | 22131159PubMed |
op den Camp RGL, Przybyla D, Ochsenbein C, Laloi C, Kim C, Danon A, Wagner D, Hideg E, Göbel C, Feussner I, Nater M, Apel K (2003) Rapid induction of distinct stress responses after the release of singlet oxygen in Arabidopsis. Plant Cell 15, 2320–2332.
| Rapid induction of distinct stress responses after the release of singlet oxygen in Arabidopsis.Crossref | GoogleScholarGoogle Scholar |
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research 29, e45
| A new mathematical model for relative quantification in real-time RT-PCR.Crossref | GoogleScholarGoogle Scholar | 11328886PubMed |
Pnueli L, Hallak-Herr E, Rozenberg M, Cohen M, Goloubinoff P, Kaplan A, Mittler R (2002) Molecular and biochemical mechanisms associated with dormancy and drought tolerance in the desert legume Retama raetam. The Plant Journal 31, 319–330.
| Molecular and biochemical mechanisms associated with dormancy and drought tolerance in the desert legume Retama raetam.Crossref | GoogleScholarGoogle Scholar | 12164811PubMed |
Porcel R, Barea JM, Ruiz-Lozano JM (2003) Antioxidant activities in mycorrhizal soybean plants under drought stress and their possible relationship to the process of nodule senescence. New Phytologist 157, 135–143.
| Antioxidant activities in mycorrhizal soybean plants under drought stress and their possible relationship to the process of nodule senescence.Crossref | GoogleScholarGoogle Scholar |
Puppo A, Rigaud J, Job D (1981) Role of superoxide anion in leghemoglobin autoxidation. Plant Science Letters 22, 353–360.
| Role of superoxide anion in leghemoglobin autoxidation.Crossref | GoogleScholarGoogle Scholar |
Rao MV, Hale BA, Ormrod DP (1995) Amelioration of ozone-induced oxidative damage in wheat plants grown under high carbon dioxide. Role of antioxidant enzymes. Plant Physiology 109, 421–432.
Reid DE, Ferguson BJ, Hayashi S, Lin YH, Gresshoff PM (2011) Molecular mechanisms controlling legume autoregulation of nodulation. Annals of Botany 108, 789–795.
| Molecular mechanisms controlling legume autoregulation of nodulation.Crossref | GoogleScholarGoogle Scholar | 21856632PubMed |
Ruijter JM, Ramakers C, Hoogaars W, Bakker O, van den Hoff MJB, Karlen Y, Moorman AFM (2009) Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Research 37, e45
| Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data.Crossref | GoogleScholarGoogle Scholar | 19237396PubMed |
Sassi S, González EM, Aydi S, Arrese-Igor C, Abdely C (2008) Tolerance of common bean to long-term osmotic stress is related to nodule carbon flux and antioxidant defenses, evidence from two cultivars with contrasting tolerance. Plant and Soil 312, 39–48.
| Tolerance of common bean to long-term osmotic stress is related to nodule carbon flux and antioxidant defenses, evidence from two cultivars with contrasting tolerance.Crossref | GoogleScholarGoogle Scholar |
Seki M, Umezawa T, Urano K, Shinozaki K (2007) Regulatory metabolic networks in drought stress responses. Current Opinion in Plant Biology 10, 296–302.
| Regulatory metabolic networks in drought stress responses.Crossref | GoogleScholarGoogle Scholar | 17468040PubMed |
Shaedle M, Bassham J (1977) Chloroplast glutathione reductase. Plant Physiology 59, 1011–1012.
| Chloroplast glutathione reductase.Crossref | GoogleScholarGoogle Scholar |
Vincent J (1970) ‘A manual for the practical study of root nodule bacteria, IBP handbook no 15.’ (Blackwell Scientific Publication, Oxford, UK)
Wormuth D, Baier M, Kandlbinder A, Scheibe R, Hartung W, Dietz KJ (2006) Regulation of gene expression by photosynthetic signals triggered through modified CO2 availability. BMC Plant Biology 6, 15
| Regulation of gene expression by photosynthetic signals triggered through modified CO2 availability.Crossref | GoogleScholarGoogle Scholar | 16916444PubMed |
Zabalza A, Gálvez L, Marino D, Royuela M, Arrese-Igor C, González EM (2008) Effects of ascorbate and its immediate precursor, galactono-1,4-lactone on the response of nitrogen-fixing pea nodules to water stress. Journal of Plant Physiology 165, 805–812.
| Effects of ascorbate and its immediate precursor, galactono-1,4-lactone on the response of nitrogen-fixing pea nodules to water stress.Crossref | GoogleScholarGoogle Scholar | 17931744PubMed |
Zhang A, Jiang M, Zhang J, Ding H, Xu S, Hu X, Tan M (2007) Nitric oxide induced by hydrogen peroxide mediates abscisic acid-induced activation of the mitogen-activated protein kinase cascade involved in antioxidant defense in maize leaves. New Phytologist 175, 36–50.
| Nitric oxide induced by hydrogen peroxide mediates abscisic acid-induced activation of the mitogen-activated protein kinase cascade involved in antioxidant defense in maize leaves.Crossref | GoogleScholarGoogle Scholar | 17547665PubMed |
Zhou B, Guo Z, Xing J, Huang B (2005) Nitric oxide is involved in abscisic acid induced antioxidant activities in Stylosanthes guianensis. Journal of Experimental Botany 56, 3223–3228.
| Nitric oxide is involved in abscisic acid induced antioxidant activities in Stylosanthes guianensis.Crossref | GoogleScholarGoogle Scholar | 16263901PubMed |
