Crosstalk between Calcium and ROS Signaling during Flg22-Triggered Immune Response in Arabidopsis Leaves

Calcium and reactive oxygen species (ROS) are two of the earliest second messengers in response to environmental stresses in plants. The rise and sequestration of these messengers in the cytosol and apoplast are formed by various channels, transporters, and enzymes that are required for proper defense responses. It remains unclear how calcium and ROS signals regulate each other during pattern-triggered immunity (PTI). In the present study, we examined the effects of perturbing one signal on the other in Arabidopsis leaves upon the addition of flg22, a well-studied microbe-associated molecular pattern (MAMP). To this end, a variety of pharmacological agents were used to suppress either calcium or ROS signaling. Our data suggest that cytosolic calcium elevation is required to initiate and regulate apoplastic ROS production generated by respiratory burst oxidase homologs (RBOHs). In contrast, ROS has no effect on the initiation of the calcium signal, but is required for forming a sufficient amplitude of the calcium signal. This finding using pharmacological agents is corroborated by the result of using a genetic double mutant, rbohd rbohf. Our study provides an insight into the mutual interplay of calcium and ROS signals during the MAMP-induced PTI response in plants.