A Noninvasive Technique for Monitoring Peroxidative and H2O2-Scavenging Activities during Interactions between Bacterial Plant Pathogens and Suspension Cells

Stimulation of active oxygen metabolism occurs during the early stages of interactions involving bacteria and plant cell suspensions. Although many cellular processes are known to affect active oxygen metabolism in plants, it is not known which of these factors affect active oxygen levels during plant-bacteria interactions. Extracellular peroxidases have been shown to participate in both the production and utilization of active oxygen species such as H2O2 and superoxide. Catalase and other scavenging mechanisms also affect the overall level of active oxygen. In this study the luminol-dependent chemiluminescent reaction previously used to measure H2O2 levels in suspension cells was modified to allow the assay of both peroxidase and H2O2-scavenging activity. The early stages of the interactions between tobacco (Nicotiana tabacum) and Pseudomonas syringae pv syringae, as well as between soybean (Glycine max) and P. syringae pv glycinea, were investigated. This method of monitoring peroxidase and H2O2-scavenging activity proved to be rapid, sensitive, and nonintrusive, allowing the processing of multiple samples using intact cells or cell-free preparations. The results from the study demonstrate that the scavenging activities can be significant and must be considered when studying active oxygen production in biological interactions.