ROS Detection and Quantification in Plant–Nematode Interactions

Reactive oxygen species (ROS) accumulation is one of the earliest hallmarks upon successful pathogen recognition in plants. H2O2 is considered the most important ROS in plant defense considering its relatively high stability and capacity to cross long distances in the plant. However, ROS also play roles in cell development and could hence facilitate nematode feeding site development. Several methods to analyze the cellular redox state exist, among which ROS detection and quantification and the evaluation of ROS scavenging enzyme activity (peroxidase activity, catalase activity, etc.). Here, we describe DAB staining, which is used to detect and localize ROS in planta upon an external trigger. Furthermore, ROS quantification using the FOX assay is described. Both methods have been used extensively in research and yield repeatable results in various plants.