Fungal Elicitors Induce a Transient Release of Active Oxygen Species from Cultured Spruce Cells That is Dependent on Ca(2+) and Protein-kinase Activity

Cell-wall components from the ectomycorrhizal fungi Amanita muscaria and Hebeloma crustuliniforme and from the spruce pathogen Heterobasidion annosum elicited a transient release of active oxygen species from cultured spruce cells (Picea abies (L.) Karst.). Since the detection of active oxygen was suppressed by catalase, H2O2 was assumed to be the prevailing O2 species. On the other hand, superoxide dismutase enhanced the concentration of detectable H2O2 indicating that the superoxide anion was formed before dismutating to H2O2. The elicitors induced the formation of active oxygen in a dose-dependent manner. Interestingly, elicitors from mycorrhizal fungi had a lower H2O2-inducing activity than equal amounts of cell-wall preparations from the pathogen H. annosum. In Ca(2+)-depleted medium the production of active oxygen by elicitor-treated spruce cells was suppressed. Additionally, the ionophore A 23187 induced active oxygen formation in a medium with Ca(2+) but not in a Ca(2+)-depleted medium. Furthermore, the protein-kinase inhibitor staurosporine inhibited the oxidative burst. At a concentration of 34 nM the effect was diminished to 50%. From these results it is suggested that the release of active oxygen species from cultured spruce cells triggered by cell-wall-derived fungal elicitors depends on external Ca(2+) and a protein-kinase activity. In these respects the effect shows similarities with the well-studied respiratory burst of mammalian neutrophils.