Skip to main content
SpringerLink
Log in

Antioxidant defences of the apoplast

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

The apoplast of barley and oat leaves contained superoxide dismutase (SOD), catalase, ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, and glutathione reductase activities. The activities of these enzymes in the apoplastic extracts were greatly modified 24 h after inoculation with the biotrophic fungal pathogenBlumeria graminis. The quantum efficiency of photosystem II, which is related to photosynthetic electron transport flux, was comparable in inoculated and healthy leaves during this period. Apoplastic soluble acid invertase activity was also modified in inoculated leaves. Inoculation-dependent increases in apoplastic SOD activity were observed in all lines. Major bands of SOD activity, observed in apoplastic protein extracts by activity staining of gels following isoelectric focusing, were similar to those observed in whole leaves but two additional minor bands were found in the apoplastic fraction. The apoplastic extracts contained substantial amounts of dehydroascorbate (DHA) but little or no glutathione (GSH). Biotic stress decreased apoplastic ascorbate and DHA but increased apoplastic GSH in resistant lines. The antioxidant cycle enzymes may function to remove apoplastic H2O2 with ascorbate and GSH derived from the cytoplasm. DHA and oxidized glutathione may be reduced in the apoplast or returned to the cytosol for rereduction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

AA:

reduced ascorbate

APX:

ascorbate peroxidase

DHA:

dehydroascorbate (oxidised ascorbate)

DHAR:

dehydroascorbate reductase

G6PDH:

glucose-6-phosphate dehydrogenase

GSH:

reduced glutathione

GSSG:

glutathione disulphide

GR:

glutathione reductase

MDHA:

monodehydroascorbate

MDHAR:

monodehydroascorbate reductase

SOD:

superoxide dismutase

References

  • Asard H, Horemans N, Caubergs RJ (1995) Involvement of ascorbic acid and ab-type cytochrome in plant plasmamembrane redox reactions. Protoplasma 184: 36–41

    Google Scholar 

  • Beauchamp CO, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44: 276–287

    Google Scholar 

  • Bérczi A, Asard K (1995) NAD(P)H-utilising oxidoreductases of the plasmamembrane: an overview of presently purified proteins. Protoplasma 184: 140–144

    Google Scholar 

  • Bolwell GP, Butt VS, Davies DR, Zimmerlin A (1995) The origin of the oxidative burst in plants. Free Radie Res 23: 517–532

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    Google Scholar 

  • Carver TLW, Robbins MP, Zeyen RJ (1991) Effects of two PAL inhibitors on the susceptibility and localized autofluorescent host cell responses of oat leaves attacked byErysiphe graminis D.C. Physiol Mol Plant Pathol 39: 269–287

    Google Scholar 

  • Castillo FJ, Greppin H (1988) Extracellular ascorbic acid and enzyme activities related to ascorbic acid metabolism inSedum album L. leaves after ozone exposure. Environ Exp Bot 28: 231–238

    Google Scholar 

  • Dietz K-J (1997) Functions and responses of the leaf apoplast under stress. Prog Bot 58: 221–254

    Google Scholar 

  • Draper J (1997) Salicylate, superoxide synthesis and cell suicide in plant defence. Trends Plant Sci 2: 162–165

    Google Scholar 

  • Edwards R, Blount JW, Dixon RA (1991) Glutathione and elicitation of the phytoalexin response in legume cell cultures. Planta 184: 403–409

    Google Scholar 

  • El-Zahaby HM, Gullner G, Kiràly Z (1995) Effects of powdery mildew infection of barley on the ascorbate-glutathione cycle and other antioxidants in different host-pathogen interactions. Phytopathology 85: 1225–1230

    Google Scholar 

  • Felix G, Regenass M, Boller T (1993) Specific perception of subnanomolar concentrations of chitin fragments by tomato cells: induction of extracellular alkanisation, changes in protein phosphorylation, and establishment of a refractory state. Plant J 4: 307–316

    Google Scholar 

  • Fodor J, Gullner G, Adam AL, Barna B, Kömives T, Kiràly Z (1997) Local and systemic responses of antioxidants to tobacco mosaic virus infection and to salicylic acid in tobacco. Plant Physiol 114: 1443–1451

    Google Scholar 

  • Foyer CH, Lelandais M (1996) A comparison of the relative rates of transport of ascorbate and glucose across the thylakoid, chloroplast and plasmalemma membranes of pea leaf mesophyll cells. J Plant Physiol 148: 391–398

    Google Scholar 

  • —, Lopez-Delgado H, Dat JF, Scott IM (1997) Hydrogen peroxide- and glutathione-associated mechanisms of acclimatory stress tolerance and signalling. Physiol Plant 100: 241–254

    Google Scholar 

  • Gamalei YV (1991) Phloem loading and its development related to plant evolution from trees to herbs. Trees 5: 50–64

    Google Scholar 

  • Genty B, Meyer S (1994) Quantitative mapping of leaf photosynthesis using chlorophyll fluorescence imaging. Aust J Plant Physiol 22: 277–284

    Google Scholar 

  • —, Briantais JM, Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching chlorophyll fluorescence. Biochim Biophys Acta 990: 87–92

    Google Scholar 

  • Gönner MV, Schlösser E (1993) Oxidative stress in interactions betweenAvena sativa L. andDrechslera spp. Physiol Mol Plant Pathol 42: 221–234

    Google Scholar 

  • Grignon C, Sentenac H (1991) pH and ionic conditions in the apoplast. Annu Rev Plant Physiol Plant Mol Biol 42: 103–128

    Google Scholar 

  • Gross GG, Vanse C, Elstner EF (1997) Involvement of malate, monophenols, of the superoxide radical in H2O2 formation by isolated cell walls from horseradish. Planta 136: 271–276

    Google Scholar 

  • Guo Z-J, Nakagawara S, Sumitani K, Ohta Y (1993) Effect of intracellular glutathione level on the production of 6-methoxymellein in cultured carrot (Daucus carota) cells. Plant Physiol 102: 45–51

    Google Scholar 

  • Halliwell B (1978) Lignin synthesis: the generation of H2O2 and superoxide by horseradish peroxidase and its stimulation by manganese (II) and phenols. Planta 140: 81–88

    Google Scholar 

  • Hippe-Sanwald S, Hermanns M, Somerville SC (1992) Ultrastructural comparison of incompatible and compatible interactions in the barley powdery mildew disease. Protoplasma 168: 27–40

    Google Scholar 

  • Horemans N, Asard H, Caubergs RJ (1996) Transport of ascorbate into plasma membrane vesicles ofPhaseolus vulgaris L. Protoplasma 194: 177–185

    Google Scholar 

  • — — — (1997) Transport of dehydroascorbate into purified plasma membrane vesicles ofPhaseolus vulgaris L. Plant Physiol 114: 1247–1253

    Google Scholar 

  • Jabs T, Dietrich RA, Dangl JL (1996) Extracellular superoxide is necessary and sufficient for runaway cell-death in an arabidopsis mutant. Plant Physiol 111: 276

    Google Scholar 

  • Jamaï A, Tommasini R, Martinoia E, Delrot S (1996) Characterization of glutathione uptake in broad bean leaf protoplasts. Plant Physiol 111: 1145–1152

    Google Scholar 

  • Johnson LEB, Bushnell WR, Zeyen RJ (1979) Binary pathways for analysis of primary infection and host response in populations of powdery mildew fungi. Can J Bot 57: 497–511

    Google Scholar 

  • Jones IT (1982) Transgressive segregation for enhanced level of adult plant resistance to mildew in the oat cross Mostyn X Maldwyn. Euphytica 32: 499–503

    Google Scholar 

  • — (1986) Inheritance of adult plant resistance to mildew in oats. Ann Appl Biol 109: 187–192

    Google Scholar 

  • —, Hayes JD (1971) The effects of sowing date on adult plant resistance toErysiphe graminis f. sp.avenae in oats. Ann Appl Biol 68: 31–39

    Google Scholar 

  • —, Roderick HW (1986) Sources of resistance in oats to mildew. Welsh Plant Breed Stat Aberystwyth Annu Rep 1985: 113–114

    Google Scholar 

  • Koga H, Zeyen RJ, Bushnell WR, Ahlstrand GG (1988) Hypersensitive cell-death, autofluorescence, and insoluble silicon accumulation in barley leaf epidermal-cells under attack byErysiphe graminis f. sp.hordei. Physiol Mol Plant Pathol 32: 395–409

    Google Scholar 

  • —, Bushnell WR, Zeyen RJ (1990) Specificity of cell type and timing of events associated with papilla formation and the hypersensitive reaction in leaves ofHordeum vulgare attacked byErysiphe graminis f. sp.hordei. Can J Bot 68: 2344–2352

    Google Scholar 

  • Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79: 583–593

    Google Scholar 

  • Luster DG, Buckhout TJ (1988) Characterization and partial-purification of multiple electron-transport activities in plasma-membranes from maize (Zea mays) roots. Physiol Plant 73: 339–347

    Google Scholar 

  • Luwe M (1996) Antioxidants in the apoplast and symplast of beech (Fagus sylvatica L.) leaves, seasonal variations and responses to changing ozone concentrations in air. Plant Cell Environ 19: 321–328

    Google Scholar 

  • —, Heber U (1995) Ozone detoxification in the apoplasm and symplasm of spinach, broad bean and beech leaves at ambient and elevated concentrations of ozone in air. Planta 197: 448–455

    Google Scholar 

  • —, Takahama U, Heber U (1993) Role of ascorbate in detoxifying ozone in the apoplast of spinach (Spinacia oleracea L.) leaves. Plant Physiol 101: 969–976

    Google Scholar 

  • Mathieu Y, Kurkdjian A, Xia H, Guern J, Koller A, Spiro MD, O'Neill M, Albersheim P, Darvill A (1991) Membrane responses induced by oligogalacturonides in suspension-cultured tobacco cells. Plant J 1: 333–343

    Google Scholar 

  • Mehdy MC, Sharma YK, Sathasivan K, Bays NW (1996) The role of activated oxygen species in plant disease resistance. Physiol Plant 98: 365–374

    Google Scholar 

  • Mittler R, Feng X, Cohen M (1998) Post-transcriptional suppression of cytosolic ascorbate peroxidase expression during pathogeninduced programmed cell death in tobacco. Plant Cell 10: 461–473

    Google Scholar 

  • Mozafar A, Oertli JJ (1993) Vitamin C (ascorbic acid): uptake and metabolism by soybean. J Plant Physiol 141: 316–321

    Google Scholar 

  • Ogawa K, Kanematsu S, Asada K (1996) Intra- and extra-cellular localization of “cytosolic” CuZn-superoxide dismutase in spinach leaf and hypocotyl. Plant Cell Physiol 37: 790–799

    Google Scholar 

  • Oxborough K, Baker NR (1997) An instrument capable of imaging chlorophylla fluorescence from intact leaves at very low irradiance and at cellular and subcellular levels of organisation. Plant Cell Environ 20: 1473–1483

    Google Scholar 

  • Polle A, Chakrabarti K, Schürmann W, Rennenberg H (1990) Composition and properties of hydrogen peroxide decomposing systems in apoplastic and total extracts from needles of Norway spruce (Picea abies L. Karst.). Plant Physiol 94: 312–319

    Google Scholar 

  • Ranieri A, Durso G, Nali C, Lorenzini G, Soldatini GF (1996) Ozone stimulates apoplastic antioxidant systems in pumpkin leaves. Physiol Plant 97: 381–387

    Google Scholar 

  • Rautenkranz AAF, Li L, Mächler F, Martinoia E, Oertli JJ (1994) Transport of ascorbic and dehydroascorbic acids across protoplast and vacuole membranes -isolated from barley (Hordeum vulgare L., cv. Gerbel) leaves. Plant Physiol 106: 187–193

    Google Scholar 

  • Schrenzel J, Serrander L, Bánfi B, Nübe O, Foyouzi R, Lew DP, Demaurex N, Krause K-H (1998) Electron currents generated by the human phagocyte NADPH oxidase. Nature 392: 734–737

    Google Scholar 

  • Segal AW, Abo A (1993) The biochemical basis of the NADPH oxidase of phagocytes. Trends Biochem Sci 18: 43–47

    Google Scholar 

  • Skøt L, Gordon AJ, Timms E, James CL, Webb KJ, Mizen S (1997) Down-regulation of sucrose synthase expression and activity in transgenic hairy roots ofLotus japonicus. Symbiosis 22: 241–254

    Google Scholar 

  • Streller S, Wingsle G (1994)Pinus sylvestris L. needles contain extracellular CuZn superoxide dismutase. Planta 192: 195–201

    Google Scholar 

  • Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localisation of H2O2 in plants, H2O2 accumulation in papillae and hypersensitive response during barley-powdery mildew interaction. Plant J 11: 1187–1194

    Google Scholar 

  • Vanacker H, Carver TLW, Foyer CH (1998) Pathogen-induced changes in the antioxidant status of the apoplast in barley leaves. Plant Physiol 117: 1103–1114

    Google Scholar 

  • - Foyer CH, Carver TLW (1999) Changes in apoplastic antioxidants induced by powdery mildew attack in oat genotypes with race non-specific resistance. Planta (in press)

  • Van Bel AJE, Gamalai YV (1991) Multiprogrammed phloem loading. In: Bonnemain J-L, Delrot S, Lucas WJ, Dainty J (eds) Recent advances in phloem transport and assimilate compartmentation. Nantes, Ouest Edition, pp 128–139

  • Wingsle G, Gardestrom P, Hallgren JE, Karpinski S (1991) Isolation, purification and sub-cellular localization of isozymes of superoxide-dismutase from Scots pine (Pinus sylvestris L.) needles. Plant Physiol 95: 21–28

    Google Scholar 

  • Winter H, Robinson DG, Heldt HW (1993) Subcellular volumes and metabolite concentrations in barley leaves. Planta 191: 180–190

    Google Scholar 

  • —, Robinson DG, Heldt HW (1994) Subcellular volumes and metabolites in spinach leaves. Planta 193: 530–535

    Google Scholar 

  • Zhang L, Robbins MP, Carver TLW, Zeyen RJ (1997) Induction of phenylpropanoid gene transcripts in oat attacked byErysiphe graminis at 20 °C and 10 °C. Physiol Mol Plant Pathol 51: 15–33

    Google Scholar 

  • Zeyen RJ, Bushnell WR (1979) Papillae response of barley epidermal cells caused byErysiphe graminis: rate and method of deposition determined by microcinematography and transmission electron microscopy. Can J Bot 57: 898–913

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Environmental Biology, IGER, SY23 3EB, Aberystwyth, UK

    H. Vanacker & T. L. W. Carver

  2. DLO-Agrotechnological Research Institute, Wageningen

    J. Harbinson & J. Ruisch

  3. Department of Biochemistry and Physiology, Institute of Arable Crops Research-Rothamstedt, Harpenden

    C. H. Foyer

Authors
  1. H. Vanacker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Harbinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Ruisch
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. L. W. Carver
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C. H. Foyer
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vanacker, H., Harbinson, J., Ruisch, J. et al. Antioxidant defences of the apoplast. Protoplasma 205, 129–140 (1998). https://doi.org/10.1007/BF01279303

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01279303

Keywords

Search

Navigation