Plant Cell Reports

, Volume 29, Issue 1, pp 37–50

Transcriptional differences in gene families of the ascorbate–glutathione cycle in wheat during mild water deficit

  • Maria Sečenji
  • Éva Hideg
  • Attila Bebes
  • János Györgyey
Original Paper

Abstract

When comparing the responses of two wheat (Triticum aestivum L.) genotypes, the drought-tolerant Plainsman V and the drought-sensitive Cappelle Desprez, to reduced amounts of irrigation water, we found differences in ascorbate metabolism: both ascorbate oxidation and transcription levels of enzymes processing ascorbate were changed. Relative transcript levels of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) isoenzymes, predicted to localize in distinct subcellular organelles, showed different transcriptional changes in the two genotypes. Among APX coding mRNAs, expression levels of two cytosolic (cAPX I, II) and a thylakoid-bound (tAPX) variants increased significantly in Plainsman V while a cytosolic (cAPX I) and a stromal (sAPX II) APX coding transcripts were found to be higher in Cappelle Desprez after a 4-week-long water-deficit stress. Examining the MDARs, two cytosolic isoforms (cMDAR I, II) displayed significant up-regulation of mRNA levels in the sensitive genotype, whereas only one of them (cMDAR II) did in the tolerant cultivar. We found an up-regulated chloroplastic DHAR (chlDHAR) mRNA only in the sensitive Cappelle Desprez. However, increased expression levels of a cytosolic GR (cGR) and a chloroplastic GR (chlGR) were detected only in the tolerant Plainsman V. After 4 weeks of reduced irrigation, a significantly lower ascorbate/dehydroascorbate ratio was detected in leaves of the sensitive Cappelle Desprez than in the tolerant Plainsman V. Our results indicate that more robust transcription of ascorbate-based detoxification machinery may prevent an adverse shift of the cellular redox balance.

Keywords

Ascorbate peroxidase Monodehydroascorbate reductase Dehydroascorbate reductase Glutathione reductase Transcriptional regulation Oxidative stress Triticum aestivum L. 

Abbreviations

APX

Ascorbate peroxidase

AsA

Ascorbic acid

DHAR

Dehydroascorbate reductase

DHAsA

Dehydroascorbic acid

GR

Glutathione reductase

GSH

Glutathione

MDAR

Monodehydroascorbate reductase

ORF

Open reading frame

P5CS

l1-pyrroline-5-carboxylate synthetase

TA

Transcript assembly

TBARS

Thiobarbituric acid reacting substances

References

  1. 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 Physiol 35:497–504Google Scholar
  2. Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygen and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50:601–639CrossRefPubMedGoogle Scholar
  3. Baek K-H, Skinner DZ (2003) Alteration of antioxidant enzyme gene expression during cold acclimation of near-isogenic wheat lines. Plant Sci 165:1221–1227CrossRefGoogle Scholar
  4. Bannai H, Tamada Y, Maruyama O, Nakai K, Miyano S (2002) Extensive feature detection of N-terminal protein sorting signals. Bioinformatics 18:298–305CrossRefPubMedGoogle Scholar
  5. Bartoli CG, Yu J, Gómez F, Fernández L, McIntosh L, Foyer CH (2006) Inter-relationships between light and respiration in the control of ascorbic acid synthesis and accumulation in Arabidopsis thaliana leaves. J Exp Bot 57:1621–1631CrossRefPubMedGoogle Scholar
  6. Bashir K, Nagasaka S, Itai RN, Kobayashi T, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2007) Expression and enzyme activity of glutathione reductase is upregulated by Fe-deficiency in graminaceous plants. Plant Mol Biol 65:277–284CrossRefPubMedGoogle Scholar
  7. Bendtsen JD, Nielsen H, von Heijne G, Brunak S (2004) Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340:783–795CrossRefPubMedGoogle Scholar
  8. Bray EA (2004) Genes commonly regulated by water-deficit stress in Arabidopsis thaliana. J Exp Bot 55:2331–2341CrossRefPubMedGoogle Scholar
  9. Burritt DJ, Larkindale J, Hurd CL (2002) Antioxidant metabolism in the intertidal red seaweed Stictosiphonia arbuscula following desiccation. Planta 215:829–838CrossRefPubMedGoogle Scholar
  10. Chen Z, Young TE, Ling J, Chang SC, Gallie DR (2003) Increasing vitamin C content of plants through enhanced ascorbate recycling. Proc Natl Acad Sci USA 100:3525–3530CrossRefPubMedGoogle Scholar
  11. Chen Y, Wang H, Wang X, Cao A, Chen P (2006) Cloning and expression of peroxisomal ascorbate peroxidase gene from wheat. Mol Biol Rep 33:207–213CrossRefPubMedGoogle Scholar
  12. Chen JB, Wang SM, Jing RL, Mao XG (2009) Cloning the PvP5CS gene from common bean (Phaseolus vulgaris) and its expression patterns under abiotic stresses. J Plant Physiol 166:12–19CrossRefPubMedGoogle Scholar
  13. Chew O, Whelan J, Millar AH (2003) Molecular definition of the ascorbate–glutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants. J Biol Chem 278:46869–46877CrossRefPubMedGoogle Scholar
  14. Childs KL, Hamilton JP, Zhu W, Ly E, Cheung F, Wu H, Rabinowicz PD, Town CD, Buell CR, Chan AP (2007) The TIGR Plant Transcript Assemblies database. Nucleic Acids Res 35:D846–D851CrossRefPubMedGoogle Scholar
  15. D’Arcy-Lameta A, Ferrari-Iliou R, Contour-Ansel D, Pham-Thi AT, Zuily-Fodil Y (2006) Isolation and characterization of four ascorbate peroxidase cDNAs responsive to water deficit in cowpea leaves. Ann Bot (Lond) 97:133–140CrossRefGoogle Scholar
  16. Dalton DA, Baird LM, Langeberg L, Taugher CY, Anyan WR, Vance CP, Sarath G (1993) Subcellular localization of oxygen defense enzymes in soybean (Glycine max [L.] Merr.) root nodules. Plant Physiol 102:481–489PubMedGoogle Scholar
  17. Davletova S, Rizhsky L, Liang H, Shengqiang Z, Oliver DJ, Coutu J, Shulaev V, Schlauch K, Mittler R (2005) Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell 17:268–281CrossRefPubMedGoogle Scholar
  18. De Leonardis S, De Lorenzo G, Borraccino G, Dipierro S (1995) A specific ascorbate free radical reductase isozyme participates in the regeneration of ascorbate for scavenging toxic oxygen species in potato tuber mitochondria. Plant Physiol 109:847–851PubMedGoogle Scholar
  19. Edjolo A, Laffray D, Guerrier G (2001) The ascorbate–glutathione cycle in the cytosolic and chloroplastic fractions of drought-tolerant and drought-sensitive poplars. J Plant Physiol 158:1511–1517CrossRefGoogle Scholar
  20. Eltayeb AE, Kawano N, Badawi GH, Kaminaka H, Sanekata T, Shibahara T, Inanaga S, Tanaka K (2007) Overexpression of monodehydroascorbate reductase in transgenic tobacco confers enhanced tolerance to ozone, salt and polyethylene glycol stresses. Planta 225:1255–1264CrossRefPubMedGoogle Scholar
  21. Emanuelsson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016CrossRefPubMedGoogle Scholar
  22. Foyer CH, Noctor G (2009) Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxid Redox Signal 11:861–905CrossRefPubMedGoogle Scholar
  23. Foyer CH, Lelandais M, Kunert KJ (1994) Photooxidative stress in plants. Physiol Plantarum 92:696–717CrossRefGoogle Scholar
  24. Guo Z, Ou W, Lu S, Zhong Q (2006) Differential responses of antioxidative system to chilling and drought in four rice cultivars differing in sensitivity. Plant Physiol Biochem 44:828–836CrossRefPubMedGoogle Scholar
  25. Guóth A, Tari I, Gallé Á, Csiszár J, Pécsváradi A, Cseuz L, Erdei L (2009) Comparison of the drought stress responses of tolerant and sensitive wheat cultivars during grain filling: changes in flag leaf photosynthetic activity, ABA levels, and grain yield. J Plant Growth Regul 28:167–176CrossRefGoogle Scholar
  26. Hong C-Y, Hsu YT, Tsai Y-C, Kao CH (2007) Expression of ascorbate peroxidase 8 in roots of rice (Oryza sativa L.) seedlings in response to NaCl. J Exp Bot 58:3273–3283CrossRefPubMedGoogle Scholar
  27. Janda T, Szalai G, Lesko K, Yordanova R, Apostol S, Popova LP (2007) Factors contributing to enhanced freezing tolerance in wheat during frost hardening in the light. Phytochemistry 68:1674–1682CrossRefPubMedGoogle Scholar
  28. Kranner I, Beckett RP, Wornik S, Zorn M, Pfeifhofer HW (2002) Revival of a resurrection plant correlates with its antioxidant status. Plant J 31:13–24CrossRefPubMedGoogle Scholar
  29. Lai Q-X, Bao Z-Y, Zhu Z-J, Qian Q-Q, Mao B-Z (2007) Effects of osmotic stress on antioxidant enzymes activities in leaf discs of PSAG12-IPT modified gerbera. J Zhejiang Univ Sci B 8:458–464CrossRefPubMedGoogle Scholar
  30. Lascano HR, Antonicelli GE, Luna CM, Melchiorre MN, Gómez LD, Racca RW, Trippi VS, Casano LM (2001) Antioxidant system response of different wheat cultivars under drought: field and in vitro studies. Aust J Plant Physiol 28:1095–1102Google Scholar
  31. Lascano HR, Melchiorre MN, Luna CM, Trippi VS (2003) Effect of photooxidative stress induced by paraquat in two wheat cultivars with differential tolerance to water stress. Plant Sci 164:841–848CrossRefGoogle Scholar
  32. Leterrier M, Corpas FJ, Barroso JB, Sandalio LM, del Rio LA (2005) Peroxisomal monodehydroascorbate reductase. Genomic clone characterization and functional analysis under environmental stress conditions. Plant Physiol 138:2111–2123CrossRefPubMedGoogle Scholar
  33. Lin R, Wang X, Luo Y, Du W, Guo H, Yin D (2007) Effects of soil cadmium on growth, oxidative stress and antioxidant system in wheat seedlings (Triticum aestivum L.). Chemosphere 69:89–98CrossRefPubMedGoogle Scholar
  34. Lisenbee CS, Lingard MJ, Trelease RN (2005) Arabidopsis peroxisomes possess functionally redundant membrane and matrix isoforms of monodehydroascorbate reductase. Plant J 43:900–914CrossRefPubMedGoogle Scholar
  35. Livak KJ, Schmittgen TD (2001) Analysis of relative expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  36. Lunde C, Baumann U, Shirley NJ, Drew DP, Fincher GB (2006) Gene structure and expression pattern analysis of three monodehydroascorbate reductase (Mdhar) genes in Physcomitrella patens: implications for the evolution of the MDHAR family in plants. Plant Mol Biol 60:259–275CrossRefPubMedGoogle Scholar
  37. Miller G, Suzuki N, Rizhsky L, Hegie A, Koussevitzky S, Mittler R (2007) Double mutants deficient in cytosolic and thylakoid ascorbate peroxidase reveal a complex mode of interaction between reactive oxygen species, plant development, and response to abiotic stresses. Plant Physiol 144:1777–1785CrossRefPubMedGoogle Scholar
  38. Mittova V, Volokita M, Guy M, Tal M (2000) Activities of SOD and the ascorbate–glutathione cycle enzymes in subcellular compartments in leaves and roots of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii. Physiol Plantarum 110:42–51CrossRefGoogle Scholar
  39. Moradi F, Ismail AM (2007) Responses of photosynthesis, chlorophyll fluorescence and ROS-scavenging systems to salt stress during seedling and reproductive stages in rice. Ann Bot (Lond) 99:1161–1173CrossRefGoogle Scholar
  40. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  41. Noctor G (2006) Metabolic signalling in defence and stress: the central roles of soluble redox couples. Plant Cell Environ 29:409–425CrossRefPubMedGoogle Scholar
  42. Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279CrossRefPubMedGoogle Scholar
  43. Obara K, Sumi K, Fukuda H (2002) The use of multiple transcription starts causes the dual targeting of Arabidopsis putative monodehydroascorbate reductase to both mitochondria and chloroplasts. Plant Cell Physiol 43:697–705CrossRefPubMedGoogle Scholar
  44. Ozturk ZN, Talame V, Deyholos M, Michalowski CB, Galbraith DW, Gozukirmizi N, Tuberosa R, Bohnert HJ (2002) Monitoring large-scale changes in transcript abundance in drought-and salt-stressed barley. Plant Mol Biol 48:551–573CrossRefGoogle Scholar
  45. Panchuk II, Volkov RA, Schöffl F (2002) Heat stress- and heat shock transcription factor-dependent expression and activity of ascorbate peroxidase in Arabidopsis. Plant Physiol 129:838–853CrossRefPubMedGoogle Scholar
  46. Passioura JB, Condon AG, Richards RA (1993) Water deficits, the development of leaf area crop productivity. In: Smith JAC, Griffiths H (eds) Water deficits. Bios Scientific Publishers, Oxford, pp 253–263Google Scholar
  47. Pierleoni A, Martelli PL, Fariselli P, Casadio R (2006) BaCelLo: a balanced subcellular localization predictor. Bioinformatics 22:e408–e416CrossRefPubMedGoogle Scholar
  48. Polle A (2001) Dissecting the superoxide dismutase–ascorbate–glutathione pathway in chloroplasts by metabolic modeling. Computer simulations as a step towards flux analysis. Plant Physiol 126:445–462CrossRefPubMedGoogle Scholar
  49. Pukacka S, Ratajczak E (2006) Antioxidative response of ascorbate–glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds. J Plant Physiol 163:1259–1266CrossRefPubMedGoogle Scholar
  50. Qiu Z-B, Liu X, Tian X-J, Yue M (2008) Effects of CO2 laser pretreatment on drought stress resistance in wheat. J Photochem Photobiol B 90:17–25PubMedGoogle Scholar
  51. Rossel JB, Walter PB, Hendrickson L, Chow WS, Poole A, Mullineaux PM, Pogson BJ (2006) A mutation affecting ascorbate peroxidase 2 gene expression reveals a link between responses to high light and drought tolerance. Plant Cell Environ 29:269–281CrossRefPubMedGoogle Scholar
  52. Saitou N, Nei N (1987) A neighbor-joining method: a new method for constructing phylogenetic tree. Mol Biol Evol 44:406–425Google Scholar
  53. Sano S, Tao S, Endo Y, Inaba T, Hossain MA, Miyake C, Matsuo M, Aoki H, Asada K, Saito K (2005) Purification and cDNA cloning of chloroplastic monodehydroascorbate reductase from spinach. Biosci Biotechnol Biochem 69:762–772CrossRefPubMedGoogle Scholar
  54. Scandalios JG (1993) Oxygen stress and superoxide dismutases. Plant Physiol 101:7–12PubMedGoogle Scholar
  55. Shao HB, Liang ZS, Shao MA, Sun Q (2005) Dynamic changes of anti-oxidative enzymes of 10 wheat genotypes at soil water deficits. Colloids Surf B Biointerfaces 42:187–195CrossRefPubMedGoogle Scholar
  56. Sharma P, Dubey RS (2007) Involvement of oxidative stress and role of antioxidative defense system in growing rice seedlings exposed to toxic concentrations of aluminum. Plant Cell Rep 26:2027–2038CrossRefPubMedGoogle Scholar
  57. Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Takeda T, Yabuta Y, Yoshimura K (2002) Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53:1305–1319CrossRefPubMedGoogle Scholar
  58. Shimaoka T, Yokota A, Miyake C (2000) Purification and characterization of chloroplast dehydroascorbate reductase from spinach leaves. Plant Cell Physiol 41:1110–1118CrossRefPubMedGoogle Scholar
  59. Small I, Peeters N, Legeai F, Lurin C (2004) Predotar: a tool for rapidly screening proteomes for N-terminal targeting sequences. Proteomics 4:1581–1590CrossRefPubMedGoogle Scholar
  60. Székely G, Ábrahám E, Cséplő Á, Rigó G, Zsigmond L, Csiszár J, Ayaydin F, Strizhov N, Jásik J, Schmelzer E, Koncz C, Szabados L (2008) Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. Plant J 53:11–28CrossRefPubMedGoogle Scholar
  61. Takahama U, Oniki T (1992) Regulation of peroxidase-dependent oxidation of phenolics in the apoplast of spinach leaves by ascorbate. Plant Cell Physiol 33:379–387Google Scholar
  62. Teixeira FK, Menezes-Benavente L, Galvao VC, Margis R, Margis-Pinheiro M (2006) Rice ascorbate peroxidase gene family encodes functionally diverse isoforms localized in different subcellular compartments. Planta 224:300–314CrossRefPubMedGoogle Scholar
  63. Yagi K (1976) A simple fluorometric assay for lipoperoxide in blood plasma. Biochem Med 15:212–216CrossRefPubMedGoogle Scholar
  64. Yoon H-S, Lee H, Lee I-A, Kim K-Y, Jo J (2004) Molecular cloning of the monodehydroascorbate reductase gene from Brassica campestris and analysis of its mRNA level in response to oxidative stress. Biochim Biophys Acta 1658:181–186CrossRefPubMedGoogle Scholar
  65. Zhang J, Kirkham MB (1996) Enzymatic responses of the ascorbate–glutathione cycle to drought in sorghum and sunflower plants. Plant Sci 113:139–147CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Maria Sečenji
    • 1
  • Éva Hideg
    • 1
  • Attila Bebes
    • 1
  • János Györgyey
    • 1
  1. 1.Biological Research CenterHungarian Academy of Sciences (BRC)SzegedHungary

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