Skip to main content
SpringerLink
Log in

Role of hydrogen peroxide and apoplastic peroxidase in tomato — Botrytis cinerea interaction

  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

The aim of the research was to estimate the sensitivity of tomato tissue and spore from necrotrophic isolate of B. cinerea on H2O2. The influence of exogenic H2O2 and B. cinerea on plant tissue and on the activity of peroxidases (PO), catalase (CAT) and superoxide dismutase (SOD) in apoplastic tomato leaves fraction were investigated.

It was proved that 40 mM H2O2 damaged the cells of a host, and inhibited in vitro germination of B.cinerea spores. Complete inhibition of germination was observed after the use 100 mM H2O2. In the presence of spores H2O2 was decomposed to H2O and O2. Trace activity of catalase was observed in a solution of spores used for inoculation. Necrosis which appeared on the leaves after 40 mM H2O2 treatment resembled hypersensitive response. On the leaves pretreated at this concentration the development of infection was observed. The H2O2 concentration harmful for the tissues, stimulated the PO activity measured with NADH — responsible for generation of ·O −2 , as well as with syringaldazine (S) and ferulic acid (FA), substrates characteristics of forms lignifying and strengthening the cell wall. Clear increase in CAT activity, resulting from infection and early pretreatment with H2O2 was observed in apoplast. No effect on SOD activity was observed.

A hypothesis may be put forward, that germinating spores produce enzymes which allow them to decompose H2O2 generated in apoplast, so there is little likelihood that B. cinerea can be directly inhibited by reactive oxygen forms (ROS) during initial stages of infection. Necrotic lesions resembling HR generated by exogenous H2O2 as well as induction of activity of apoplastic plant enzymes, particularly PO connected with strengthening and lignification of cell wall, were not sufficient factors to inhibit fungal expansion.

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

ROS:

reactive oxygen species

CAT:

catalase

PO:

peroxidase

SOD:

superoxide dismutase

TMB:

tetramethylbenzidine

FA:

ferulic acid

S:

syringaldazine

MBTH:

3-methylbenzothiazoline hydrazone

References

  • Apostol I., Heinstein P.F., Low P.S. 1989. Rapid stimulation an oxidative burst during elicitation of cultured plant cells. Role in defence and signal transduction. Plant Physiol. 90: 109–116.

    PubMed  CAS  Google Scholar 

  • Baker C.J., Orlandi E.W. 1995. Active oxygen in plant pathogenesis. Annu. Rev. Phytopathol. 33: 299–321.

    Article  CAS  PubMed  Google Scholar 

  • Barcelo R.A. 1998. The generation of H2O2 in the xylem of Zinnia elegans is mediated by an NADPH oxidase-like enzyme. Planta 207: 207–216.

    Article  CAS  Google Scholar 

  • Bestwick C.S., Brown I.R., Mansfield J.W. 1998. Localized changes in peroxidase activity accompany hydrogen peroxide generation during the development of a nonhost hypersensitive reaction in lettuce. Plant Physiol. 118: 1067–1078.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Bowell G.P., Davies D.R., Gerrish C., Auh C-K., Murphy T.M. 1998. Comparative biochemistry of the oxidative burst produced by rose and French bean cells reveals two distinct mechanisms. Plant Physiol. 116: 1379–1385.

    Article  Google Scholar 

  • Bradford M.M. 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.

    Article  PubMed  CAS  Google Scholar 

  • Bradley D.J., Kjellbom P., Lamb C.J. 1992. Elicitor-and wound-induced oxidative cross-linking of a proline-rich plant cell wall protein: a novel, rapid defense response. Cell 70: 21–30.

    Article  PubMed  CAS  Google Scholar 

  • Capaldi D.J., Taylor K.E. 1983. A new peroxidase color reaction: oxidative coupling of 3-methyl-2-benzothiazolinone hydrazone (MBTH) with its formaldehyde azine application to glucose and choline oxidases. Anal. Biochem. 129: 329–336.

    Article  PubMed  CAS  Google Scholar 

  • Creissen G., Firmin J., Freyer M., Kular B., Leyland N., Reynolds H., Pastori G., Wellburn F., Baker N., Wellburn A., Mullineaux P. 1999. Elevated glutathione biosynthetic capacity in the chloroplasts of transgenic tobacco plants paradoxically causes increased oxidative stress. Plant Cell 11: 1277–1292.

    Article  PubMed  CAS  Google Scholar 

  • Doke N. 1983. Involvement of superoxide anion generation in the hypersensitive response of potato tuber tissues to infection with an incompatible race of Phytophtora infestans and to the hyphal wall components. Physiol. Plant Pathol. 23: 345–357.

    CAS  Google Scholar 

  • Dhindsa R.S., Plumb-Dhindsa P., Thorpe T.A. 1981. Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decrease levels of superoxide dismutase and catalase. J. Exp. Bot. 32: 93–101.

    Article  CAS  Google Scholar 

  • Durrant W.E., Rowland O., Piedras P., Hammond-Kosack K.E., Jones J.D.G. 2000. cDNA-AFLP reveals a striking overlap in race-specific resistance and wound response gene expression profiles. Plant Cell 12: 963–977.

    Article  PubMed  CAS  Google Scholar 

  • Figueroa-Spinoza M.C., Rouau X. 1998. Oxidative cross-linking of pentosans by a fungal laccase and horseradish peroxidase: Mechanism of linking between feruloylated arabinoxylans. Cereal Chem. 75: 259–265.

    Article  Google Scholar 

  • Frahry G., Schopfer P. 1998. Hydrogen peroxide production by roots and its stimulation by exogenous NADH. Physiol. Plantarum 103: 395–404.

    Article  CAS  Google Scholar 

  • Fry S.C. 1998. Oxidative scission of plant cell wall polysacchardes by ascorbate-induced hydroxy radicals. Biochem. J. 332: 507–515.

    PubMed  CAS  Google Scholar 

  • Gil-ad N.L., Bar-Nun N., Noy T., Mayer A.M. 2000. Enzymes of Botrytis cinerea capable of breaking down hydrogen peroxide. FEMS Microb. Letters 190: 121–126.

    Article  CAS  Google Scholar 

  • Govrin E.M., Levine A. 2000. The hypersensitive response facilitates plant infection by the nectrotrophic pathogen Botrytis cinerea. Current Biology 10: 751–757.

    Article  PubMed  CAS  Google Scholar 

  • Halliwell B., Guttering J.M.C. 1990. Role of free radicals and catalytic metal ions in human disease: An overview. Methods in Enzymology 186: 1–85.

    Article  PubMed  CAS  Google Scholar 

  • Hckelhoven R., Kogel K-H. 2003. Reactive oxygen intermediates in plant-microbe interactions: Who is who in powdery mildew resistance? Planta 216: 891–902.

    Google Scholar 

  • Imberty A., Goldberg R., Catesson A.M. 1985. Isolation and characterization of Populus isoperoxidases involved in the last step of lignification. Planta 164: 221–226.

    Article  CAS  Google Scholar 

  • Ishida A., Ookubu K., Ono K. 1987. Formation of hydrogen peroxide by NAD(P)H oxidation with isolated cell wall-associated peroxidase from cultured liverwort cells, Marchantia polymorpha L. Plant Cell Physiol. 28(4): 723–726.

    CAS  Google Scholar 

  • Lamb C.J., Brisson L., Bradley D.J., Kjellborn P. 1993. Stimulus-dependent oxidative cross-linking of a proline-rich plant cell wall protein: a novel, rapid defence response and control point in cellular maturation. In: Friting B, Legrand M, ed. Mechanisms of Plant Defence Responses. Kluwar Academic Publisher, 250–256.

  • Lamb C., Dixon R.A. 1997. The oxidative burst in plant disease resistance. Annual Rev. Plant Physiol. Mol. Biol. 48: 251–275.

    Article  CAS  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.

    Article  PubMed  CAS  Google Scholar 

  • Lu H., Higgins V.J. 1999. The effect of hydrogen peroxide on the viability of tomato cells and of the fungal pathogen Cladosporium fulvum. Physiol. Mol. Plant Pathol. 54: 131–143.

    Article  CAS  Google Scholar 

  • Lüthje S., Böttger M., Döring O. 2000. Are plants stacked neutrophils? Comparison of pathogen-induced oxidative burst in plants and mammals. Progress in Botany 61: 187–222.

    Google Scholar 

  • Malolepsza U., Urbanek H. 2000. The oxidants and antioxidant enzymes in tomato leaves treated with o-hydroxyethylorutin and infected with Botrytis cinerea. Eur. J. Plant Pathol. 106: 657–665.

    Article  CAS  Google Scholar 

  • Mayer A. M., Staples R.C., Gil-ad N.L. 2001. Mechanisms of survival fungal plant pathogens in hosts expressing the hypersensitive response. Phytochemistry 58: 33–41.

    Article  PubMed  CAS  Google Scholar 

  • Minibayeva F.V., Gordon L.K., Kolesnikov O.P., Chasov A.V. 2001. Role of extracellular peroxidase in the superoxide production by wheat root cells. Protoplasma 217: 125–128.

    Article  PubMed  CAS  Google Scholar 

  • Milosevic N., Slusarenko A.J. 1996. Activity oxygen metabolism and lignification in the hypersensitive response in bean. Physiol. Mol. Plant Pathol. 49: 143–158.

    Article  Google Scholar 

  • Moerschbacher B.M., Noll U., Gorrichon I., Reisener H-J. 1990. Specific inhibition of lignification breaks hypersensitive resistance to wheat stem rust. Plant Physiology 93: 465–470.

    Article  PubMed  CAS  Google Scholar 

  • Ogawa K., Kanematsu S., Asada K. 1997. Generation of superoxide anion and localization of CuZn-superoxide dismutase in the vascular tissue of spinach hypocotyls: their association with lignification. Plant Cell Physiol. 38: 1118–1126.

    PubMed  CAS  Google Scholar 

  • Patykowski J., Urbanek H. 2003. Activity of enzymes related to H2O2 generation and metabolism in leaf apoplastic fraction of tomato leaves infected with Botrytis cinerea. J. Phytopathology 151: 153–161.

    Article  CAS  Google Scholar 

  • Peng M., Ku J. 1992. Peroxidase-generated hydrogen peroxide as a source of antifungal activity in vitro and tobacco leaf discs. Phytopathology 64: 696–699.

    Google Scholar 

  • Rij R.E., Forney C.F. 1995. Phytotoxicity of vapour phase hydrogen peroxide to Thompson Seedlees grapes and Botrytis cinerea spores. Crop Protection 14: 131–135.

    Article  CAS  Google Scholar 

  • Schouten A., Tenberge K.B., Vermeer J., Stewart J., Wagemaker L., Williamson B., van Kan J.A.L. 2002. Functional analysis of an extracellular catalase of Botrytis cinerea. Mol. Plant Pathol. 3: 227–238.

    Article  CAS  Google Scholar 

  • Schweikert C., Liszkay A., Schopfer P. 2000. Scission of polysaccharide by peroxidase-generated hydroxyl radicals. Phytochemistry 53: 565–570.

    Article  PubMed  CAS  Google Scholar 

  • Takahama U. 1995. Oxidation of hydroxycinnamic acid and hydroxycinnamyl alcohol derivatives by laccase and peroxidase. Interactions among p-hydroxyphenyl, guaiacyl and syringyl groups during the oxidation reactions. Physiol. Plant. 93: 61–68.

    Article  CAS  Google Scholar 

  • Tiedemann A.v. 1997. Evidence for a primary role of active oxygen species in induction of host cell death during infection of bean leaves with Botrytis cinerea. Physiol. Mol. Plant Pathol. 50: 151–166.

    Article  CAS  Google Scholar 

  • Wojtaszek P. 1997. Oxidative burst: an early plant response to pathogen infection. Biochem. J. 322: 681–692.

    PubMed  CAS  Google Scholar 

  • Zimmerlin A., Wojtaszek P., Bolwell G.P. 1994. Synthesis of dehydrogenation polymers of ferulic acid with high specificity by a purified cell-wall peroxidase from French bean (Phaseolus vulgaris L.). Biochem. J. 299: 747–753.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Physiology and Biochemistry, University of Łód, Banacha 12/16 90-237, Łód, Poland

    Jacek Patykowski

Authors
  1. Jacek Patykowski
    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

Patykowski, J. Role of hydrogen peroxide and apoplastic peroxidase in tomato — Botrytis cinerea interaction. Acta Physiol Plant 28, 589–598 (2006). https://doi.org/10.1007/s11738-006-0054-6

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-006-0054-6

Key words

Search

Navigation