Creating a knowledge of alliance between the urban and rural cold chain logistics enterprises in Yunnan is an important content of coordinated and efficient operation of the entire cold chain system. The study shows that the information disclosure in the knowledge alliance and enterprises lacks of knowledge sharing motivation. The lack of professional and technical personnel of enterprises factors restricts the establishment of urban and rural areas of cold chain logistics enterprise knowledge alliance. Measures (establish security and incentives in a knowledge-based alliance, develop a plan to nurture talent and establish a high level of trust between knowledge sharing) have to be taken to solve the problems in knowledge alliance building process.