This study aims to provide scientific basis and practical guidance for the optimization and improvement of blueberry substrate cultivation in Yunnan Province. 12 blueberry substrate cultivation bases with the same planting variety and tree age in Yunnan Province were selected. Substrate samples were collected during the full fruit period, and indicators such as bulk density, EC value, pH, air porosity, water holding capacity, steam-water ratio, ≤1 mm weight, >1 mm weight, total nitrogen, available phosphorus, and available potassium were measured and evaluated using the membership function comprehensive analysis method. The results showed that there were extremely significant differences in various indicators of the substrates in different bases. The order of membership degree from large to small was Yanshan> Lanping> Mengzi> Shilin> Yulong> Chengjiang> Jingdong> Mile> Jianshui 2> Jianshui 1> Qilin> Qiubei. The Yanshan base had the highest comprehensive score, and its substrate ratio was coconut husk: peat: perlite= 2:1:1; the Lanping base was second, with a substrate ratio of coconut husk: peat: perlite= 4:3:3; the Mengzi base ranked third, with a substrate ratio of coconut husk: coconut block: perlite= 2:1:2. The research analysis found that the addition of peat improved the substrate performance, but there is relatively less high-quality peat on the market. It is recommended to prioritize coconut husk as the main component of the substrate in production and appropriately add perlite to improve the physical properties of the substrate. The substrate formula can be adjusted according to the actual situation to achieve the best cultivation effect.