Phase separation in plants: mechanisms, functions, and a simplified workflow for investigation

Liquid-liquid phase separation (LLPS) has emerged as a fundamental mechanism organizing membrane-less compartments within cells, driving crucial biological processes. In plants, LLPS facilitates the spatiotemporal regulation of diverse functions, from growth and development, such as hormone signaling, photo-perception, and floral transition, to adaptive responses against abiotic and biotic stresses. This review outlines the properties and major driving forces governing LLPS, emphasizing multivalent interactions mediated by intrinsically disordered regions, repeated domains, and nucleic acids. Key influencing factors, including concentration, temperature, and ionic conditions, are discussed. We further describe a streamlined experimental workflow for studying plant LLPS, encompassing prediction, assessment, and validation. Understanding LLPS dynamics offers profound insights into plant adaptation and resilience, positioning phase separation as a pivotal regulatory paradigm in plant biology.