Some Properties of the H2O2/O2−Generating System from the Lignifying Xylem ofZinnia elegans

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Abstract
The properties of the enzymatic system responsible for generating H2O2/O2- in the lignifying xylem of Zinnia elegans were studied using the starch/KI method for monitoring H2O2 production and the nitroblue tetrazolium method for monitoring superoxide anion production. The results showed that H2O2/O2- production by lignifying xylem tissues was insensitive to inhibitors of peroxidase and poly(di)amine oxidases. However, H2O2/O2 production in the xylem of Z. elegans was sensitive to the inhibitors of phagocytic plasma membrane NADPH oxidase, pyridine, imidazole, quinacrine and diphenylene iodonium. The sensitivity of H2O2/O2- production to the respective inhibitors of calmodulin (R-24571), phospholipase C (neomycin sulfate), and protein kinase (staurosporine), and its reversion by an inhibitor of protein phosphatases (cantharidin); pointed to the analogies existing between the mechanism of H2O2/O2- production in the lignifying xylem of Z. elegans and the oxidative burst observed during the hypersensitive plant cell response. These results suggest the existence of a metabolic cascade involving calmodulin, IP3 and protein phosphorylation in the activation of the enzymatic system responsible for H2O2/O2- production in the lignifying xylem of Z. elegans.