Cytochrome P450 enzymes (CYPs), a class of heme monooxygenases, are widely distributed across bacteria, archaea, viruses, as well as in higher plants and animals. They regulate the metabolism of various endogenous substances and exogenous compounds, accounting for approximately 1% of the protein-coding genes in plants. These enzymes play a crucial role in the biosynthesis of plant bioactive metabolites, growth regulation and stress adaptation. Currently, over 300,000 CYPs are recorded in databases, yet only a small fraction (less than 0.2 %) has undergone functional characterization. This review will summarize the latest research progress from two perspectives: the involvement of CYPs in the biosynthesis of important bioactive compounds in plants, and their significant physiological functions in plant growth, development, and stress adaptation. The aim is to enhance our understanding and provide examples for the functional exploration of CYPs in plants, thereby paving the way for innovative applications in the field of biotechnological breeding and synthetic biology.