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Nitric oxide (NO) is a key regulator of plant development, growth, and responses to the environment. Together with hydrogen peroxide (H₂O₂), NO modifies the structure and function of proteins, controlling redox signaling. Although NO has been studied extensively at the cellular and subcellular levels, very little is known about changes in NO content at the whole‐plant level.Here, we report on the development of an aboveground whole‐plant live imaging method for NO. Using mutants with altered NO levels, as well as an NO donor/scavenger, we demonstrate the specificity of the detection method for NO.Arabidopsis thalianaplants were found to produce a basal level of NO under control conditions. NO levels accumulated enzymatically in plants following heat stress applied to the entire plant, as well as in a systemic manner following different locally applied stimuli. Similar or opposing accumulation patterns were also found for NO and H₂O₂during the response of plants to different stimuli.Our findings reveal that NO accumulates during the systemic response of plants to a local stimulus. In addition, they shed new light on the intricate relationships between NO and H₂O₂. The new method reported opens the way for multiple future studies of NO's role in plant biology. 

Abstract Reactive oxygen species (ROS) play a critical role in plant development and stress responses, acting as key components in rapid signalling pathways. The ‘ROS wave’ triggers essential acclimation processes, ultimately ensuring plant survival under diverse challenges. This review explores recent advances in understanding the composition and functionality of the ROS wave within plant cells. During their initiation and propagation, ROS waves interact with other rapid signalling pathways, hormones and various molecular compounds. Recent research sheds light on the intriguing lack of a rigid hierarchy governing these interactions, highlighting a complex interplay between diverse signals. Notably, ROS waves culminate in systemic acclimation, a crucial outcome for enhanced stress tolerance. This review emphasizes the versatility of ROS, which act as flexible players within a network of short‐ and long‐term factors contributing to plant stress resilience. Unveiling the intricacies of these interactions between ROS and various signalling molecules holds immense potential for developing strategies to augment plant stress tolerance, contributing to improved agricultural practices and overall ecosystem well‐being. 

Systemic signaling and systemic acquired acclimation (SAA) are key to the survival of plants during episodes of abiotic stress. These processes depend on a continuous chain of cell-to-cell signaling events that extends from the initial tissue that senses the stress (the local tissue) to the entire plant (systemic tissues). Reactive oxygen species (ROS) and Ca 2+ are key signaling molecules thought to be involved in this cell-to-cell mechanism. Here, we report that the systemic response of Arabidopsis thaliana to a local treatment of high light stress, which resulted in local ROS accumulation, required ROS generated by respiratory burst oxidase homolog D (RBOHD). ROS increased cell-to-cell transport and plasmodesmata (PD) pore size in a manner dependent on PD-localized protein 1 (PDLP1) and PDLP5, and this process was required for the propagation of the systemic ROS signals and SAA. Furthermore, aquaporins and several Ca 2+ -permeable channels in the glutamate receptor–like (GLR), mechanosensitive small conductance–like (MSL), and cyclic nucleotide–gated (CNGC) families were involved in this systemic signaling process. However, we determined that these channels were required primarily to amplify the systemic signal in each cell along the path of the systemic ROS wave, as well as to establish local and systemic acclimation. Thus, PD and RBOHD-generated ROS orchestrate light stress–induced rapid cell-to-cell spread of systemic signals in Arabidopsis .