Live Whole-Plant Detection of Rapidly Accumulating Reactive Oxygen Species Following Applied Stress in Arabidopsis thaliana

Abstract

Reactive Oxygen Species (ROS) waves serve as key systemic signals within plants. Following the initial sensation of a stress, auto-propagation of ROS (the ROS wave) begins and rapidly spreads to distant, systemic tissues of the plant and invokes important physiological responses. Highly sensitive methods capable of imaging this systemic signal at the whole-plant level have long been desired for the study of ROS signaling. Here, we describe a straightforward and highly sensitive method for the detection and quantification of ROS in planta at the whole-plant level in Arabidopsis thaliana with the In Vivo Imaging System (IVIS) Lumina S5 imaging platform and the fluorescent probe 2′,7′-dichlorofluorescin diacetate (H2DCFDA). This method can be used for high-throughput screening of the ROS Wave within Arabidopsis plants, with up to 16 plants capable of being imaged approximately every half hour.