The development of new business subjects is an important force for the implementation of rural revitalization strategy, which is of great significance for the realization of agricultural modernization. This paper constructs a dynamic game model of the tripartite business subjects of the government, enterprises, and farmers, focuses on the evolutionary game process of the new business subjects to promote the development of rural revitalization, carries out the dynamic game analysis of the model at the macro level, micro level, macro and micro level and multi-level, and performs numerical simulation on it. Then, taking the development of new management subjects in a certain place as an example, the game model is used for example verification, and the optimization path of new management subjects to promote the development of rural revitalization is further proposed. The results show that among the eight evolutionary paths (1, 1, 1) is the optimal choice for new management subjects to promote rural revitalization and development, and when the government is highly supportive, the enterprises actively cooperate, and the farmers participate in generating the operation, the interests among the subjects can be maximized. The benefits of new business subjects at the equilibrium benefit point in the example analysis are close to the actual situation, reflecting the effectiveness of the game model. The role of new business subjects in promoting the development of rural revitalization should be optimized by gathering modern production factors, improving the mechanism of interest relations, establishing new urban-rural relations, guiding large-scale operations, and cultivating talents.