With the significant advances in "-omics" technologies, various biological approaches and analytical platforms have been developed to uncover the functions and involved biological processes of proteins in plants. However, in-depth characterization of protein functions at the molecular level, such as protein activities, interactions and post-translational modifications (PTMs), remains one of the most challenging tasks in plant science. As an alternative, chemical proteomics has emerged as a precise and dynamic analytical method that employs chemical tools to explore the functional states of proteins within complex proteomes. This review highlights the contributions of chemical tools in plant functional proteomics studies, with focus on their applications in evaluating enzyme activity, identifying receptors or targets of bioactive small molecules, elucidating biosynthetic pathways of plant natural products, and profiling PTMs in plant-derived biological systems.