MAcro Plant Projection Imaging (MAPPI): An open, scalable platform for whole-plant fluorescence real-time imaging

Understanding how plants perceive and respond to environmental and developmental cues requires tools capable of monitoring molecular signals in vivo, across whole tissues, and in real time. Genetically encoded fluorescent indicators, coupled with fluorescence microscopy, have transformed plant biology, but their application remains largely confined to small model organisms and specialized microscopy instrumentation. Here, we present MAcro Plant Projection Imaging (MAPPI), an open-source, low-cost, and modular fluorescence imaging platform for soil-grown plants beyond the model organism or seedling stage. MAPPI enables wide field-of-view, dual-projection imaging of fluorescent reporters, supporting real-time visualization of systemic signals under near-physiological conditions. We validate MAPPI by tracking calcium and l-glutamate dynamics in adult Nicotiana benthamiana plants, revealing developmentally regulated long-distance calcium waves triggered by wounding, burning, or submergence, including bidirectional shoot-to-root and root-to-shoot signaling. By democratizing access to whole-plant functional imaging, MAPPI provides a scalable tool for dissecting signal propagation, stress adaptation, and systemic communication in both model and nonmodel species.