Mushrooms are widely recognized for their rich nutritional profile and diverse array of bioactive compounds, including proteins, polysaccharides, and secondary metabolites, which contribute to various health benefits. Among these compounds, phospholipids stand out due to their essential role in cellular processes and their potential applications in food, nutraceuticals, and pharmaceuticals. This review comprehensively examines the phospholipid composition in mushrooms, highlighting the methods of analysis, and delving into their functional applications. By comparing lipid profiles across different stages of growth and between various species, this review highlights the significant role of phospholipids in mushroom cellular structure and function. The review also discusses the biosynthetic pathways and extraction methods employed for this class of compounds. By shedding light on this underexplored aspect of mushroom chemistry, this work aims to spur further research and innovation in the utilization of mushrooms in food science and medicine.