Live Imaging of Abscisic Acid Dynamics Using Genetically Encoded Fluorescence Resonance Energy Transfer (FRET )-Based...

The phytohormone abscisic acid (ABA) regulates various aspects of plant physiology, growth, and development to maintain a balanced plant water status. Cellular ABA levels are regulated through the combined activities of biosynthesis, catabolism, and transport proteins and depend on the developmental stage, the cell-type and on environmental conditions. Genetically encoded Förster (fluorescence) Resonance Energy Transfer (FRET)-based ABA-responsive biosensors enable the direct monitoring of ABA dynamics in intact plants. Thus, ABA biosensor-based in vivo imaging can provide novel insights about the spatiotemporal patterns of biosynthesis- and transport-dependent ABA dynamics that are required for the regulation of seed dormancy and germination, root growth and hydrotropism, and stomatal closure under water limiting conditions. Here, I describe a protocol for the in vivo analysis of ABA in 5-day-old Arabidopsis seedlings (roots) expressing the FRET-based ABA biosensor ABAleonSD1-3L21.