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, Volume 26, Issue 1, pp 131–139 | Cite as

Effects of apple (Malus × domestica Borkh.) phenolic compounds on proteins and cell wall-degrading enzymes of Venturia inaequalis

  • Beate Golba
  • Dieter TreutterEmail author
  • Andreas Kollar
Original Paper

Abstract

Cell wall-degrading enzymes of Venturia inaequalis are supposed to be fungal virulence factors whereas phenolic compounds of the host plant may be involved in defence. Since phenolic structures are predestined for an interaction with proteins we studied the effects on enzymes and proteins in course of in vitro culture and with preparations from culture filtrates and mycelia, respectively. The native compounds epicatechin, catechin, phloridzin, chlorogenic acid, caffeic acid, p-coumaric acid and phloridzin tested under non-oxidizing conditions had no or weak effects on enzyme activities. A significant inhibition of pectinase was only detected with the highest concentrations of procyanidins and phloretin. Aerobe conditions resulted in a fast oxidation of most phenolics which was enhanced by fungal phenoloxidases. Generally, no inhibition of fungal growth occurred in vitro but distinct irreversible effects on proteins and enzymes were detected with oxidized phenolics in course of in vitro-cultures as well as with the corresponding preparations. Efficacy of inhibitory activity in in vitro-cultures depended on media, culture technique and time course. Direct treatment of enzyme preparations with the oxidized phenolics resulted in a distinct inhibition of cellulolytic and especially pectinolytic activity. Apart from cellulase pattern altered by phenolics, in vitro-culture zymograms revealed a non specific reduction of enzymatic activities, whereas action on total culture filtrate proteins resulted in specific effects due to phenolic compounds and incubation time. An attempt was made to characterize the oxidation products of epicatechin. Chromatographic fractionation revealed a non-resolvable complex of inhibitory compounds which were not consistent with the typical yellow oxidation products.

Keywords

Apple scab Cellulase Pectinase Oxidized phenolics Enzyme activity 

Abbreviations

BSA

Bovine serum albumin

CC

Column chromatography

CMC

Carboxymethylcellulose

IEC

Ion exchange chromatography

IEF

Isoelectric focusing

Mes

2-(N-morpholino)ethanesulfonic acid

PDB

Potato dextrose broth

PGA

Polygalacturonic acid

SPE

Solid phase extraction

Notes

Acknowledgments

The authors thank the Deutsche Forschungsgemeinschaft for financial support of the project.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Julius Kuehn-Institute, Federal Research Centre for Cultivated PlantsInstitute for Plant Protection in Fruit Crops and ViticultureDossenheimGermany
  2. 2.Institute of Fruit SciencesTechnische Universität MünchenFreising-WeihenstephanGermany

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