Phosphatidic acid (PA) is a structurally simplest membrane phospholipid,which plays key roles in membrane trafficking,stress responses and cytoskeletal dynamics.However it remains unknown about spatio-temporal distribution of PA produced in living plant cells.Here we generated PA biosensor PPB (Plant Phosphatidic acid Biosensor) to determine PA changes at plasma membrane using F(o)rster resonance energy transfer (FRET) approaches.A PA-binding domain of RbohD (0-250 amino acids) was sandwiched with the cyan fluorescent protein and yellow fluorescent protein and was tagged with the plasma membrane-targeting sequence of K-Ras4B-CT.The transgenic lines with PPB vectors were generated for both wild-type and pldα1 Arabidopsis.The cell imaging of PA dynamics in transgenic Arabidopsis plants was performed using confocal laser scanning microscopy.Using these toolkits,we identified temporally distinct PA responses to different lipids,NaCl,and ABA.The PPB biosensor also revealed the different accumulation of PA between WT and pldα1 in these processes.In conclusion,we have set a method to detect spatial and temporal characteristics of PA signatures in plant cells.