The Toolbox to Study Protein–Protein Interactions in Plants

The ability to detect the interaction partners of a protein and how those interactions occur are central to the study of biology. Indeed, such knowledge can provide important clues to unravel the functional context of proteins in specific pathways. This review provides an overview of the different techniques available to plant researchers to study protein–protein interactions (PPIs), and briefly summarizes key benefits and disadvantages of each technique. We discuss the most commonly used in vitro-based techniques (i.e., co-immunoprecipitation and microscale thermophoresis), yeast-based screening methods (i.e., yeast two-hybrid and the split-ubiquitin system), as well as imaging and FRET-based approaches (i.e., bimolecular fluorescence complementation, modification of intracellular localisation, protein mobility measurements, acceptor photobleaching, fluorescence lifetime imaging microscopy, and fluorescence polarization/anisotropy) that are used in the field of plant biology. Finally, we offer suggestions that may provide future directions for how to best use and combine these techniques.