Abstract: Shiitake mushrooms (Lentinus edodes) are highly favored by consumers worldwide and hold a high market share on the global consumption of mushrooms. The stems of L. edodes are often discarded because of their coarse fiber and poor taste, resulting in many environmental problems and waste of effective resources. In recent years, growing research interests in the comprehensive utilization of L. edodes stems have attracted extensive attention. This paper systematically summarizes the various nutritional components and flavor substances of L. edodes stems, coupling with their hypoglycemic, hypolipidemic, antioxidant and antitumor activities. The comprehensive utilization of L. edodes stems in the fields of food, agriculture and biotechnology are summarized, including utilizations in food ingredients, texture modifiers, flavor enhancers, culture media substrates, biomass energy materials, and environmental remediation agents, as well as applications in improving crop traits, postharvest quality enhancement of fruits and vegetables, and functional feed production. This review aimed to provide theoretical basis and reference for the high value utilization of L. edodes stems, as well as the quality and efficiency improvement of edible mushroom industry and the development of green economy.