Heterosis utilization is the main way to significantly improve crop yield， which helps to solve the contradiction between the increasing population and limited arable land. Soybean， which identified as an important grain， oil and feed crop in the world， the utilization of heterosis in it has been used for more than 30 years. Among them， the three-line hybrid breeding system based on cytoplasmic male sterility is the main way to utilize the heterosis in soybean. At present， more than 40 hybrid soybean varieties have been approved and popularized in production. Furthermore， hybrid soybean is in the stage of advancing from pilot to industrialization. Therefore， this paper reviewed the genetic basis and breeding application of cytoplasmic male sterility in soybean. The discovery and utilization of various cytoplasmic male sterility lines， the genetic and molecular mechanism of sterility traits， the map-based clone of restorer-of-fertility gene and inhibitor of fertility restorer genes were systematically described. Based on the description and analysis of current situation， the challenges and solutions of three-line hybrid soybean breeding were put forward， and the innovation of three-line hybrid soybean technology was prospected， aiming to provide new methods and new ideas for the molecular basic and application research of soybean heterosis.