One of the most important functions of plant roots is to uptake nutrients from the soil and upwards them to the aerial parts. Water and nutrients are transported across epidermis, cortex and endodermis of the concentric root cell layers before reaching the central vasculature for aboveground metabolism. The endodermis undergoes two stages of differentiation with forming two diffusion barriers, Casparian strips and suberin lamellae, which play important roles in controlling nutrients acquisition and loss. This review summarizes the research progress of endodermal differentiation and barrier function in the past few years, and put forward the future research directions, so as to provide a theoretical basis for the further exploring the role of endodermal barrier in plant growth and development, as well as stress adaptation, and opens up a new thought for plant breeding in turn.