In order to analyze the impact of mixing different tree species and Pinus massoniana on the physiological characteristics of Pinus massoniana seedlings, and reveal the physiological mechanism of mixing Pinus massoniana with different tree species, indoor simulation experiments were conducted to select 6 local tree species[Robinia pseudoacacia L., Alnus cremastogyne Burk., Liquidambar formosana Hance, Cinnamomum camphora (Linn) Presl, Schima superba Gardn. et Champ., Cunninghamia lanceolata (Lamb.) Hook.] and Pinus massoniana for mixed planting in a 1∶1 ratio, denoted as CH-M, QM-M, FX-M, XZ-M, MH-M, SM-M, and Pinus massoniana separately planted as the control, denoted as M (CK). The chlorophyll (chlorophyll a, chlorophyll b, and total chlorophyll), antioxidant enzyme activity [superoxide dismutase (SOD), peroxidase (POD)], osmoregulation substances (soluble protein and proline content) changes in plant invertase activity [nitrate reductase (NR), acid phosphatase (ACP)], and malondialdehyde (MDA) content were analyzed, and the short-term comprehensive physiological effects after mixing were also analyzed using Fuzzy comprehensive evaluation method. The results show that the chlorophyll content (except CH-M), antioxidant enzyme activity (SOD, POD) and soluble protein content of Pinus massoniana seedlings under different treatments are higher than in the control, and the effect of FX-M and MH-M treatments are obvious. The proline content in FX-M treatment is the highest, and that in SM-M and MH-M treatment is lower than in the control. The NR and ACP activities under MH-M treatment are higher than in the control. The MDA content in FX-M and QM-M treatments is significantly higher than in control and other treatments, and the MDA content in MH-M treatment is the lowest. Using Fuzzy membership function to comprehensively evaluate the physiological effects, it is concluded that the comprehensive physiological effects of Pinus massoniana seedlings under different treatments are MH-M> FX-M>QM-M>XZ-M >CH-M >SM-M > M (CK). The comprehensive physiological effect of Pinus massoniana mixed with other tree species is higher than in the control, and the comprehensive evaluation value of mixed needle and broadleaf is higher than that of mixed needle and leaf (SM-M). In terms of comprehensive physiological effects, the mixed effect of MH-M treatment is the best, the next is FX-M and QM-M treatment. concluded that the comprehensive physiological effects of Pinus massoniana seedlings under different treatments are MH-M> FX-M>QM-M>XZ-M >CH-M >SM-M > M (CK). The comprehensive physiological effect of Pinus massoniana mixed with other tree species is higher than in the control, and the comprehensive evaluation value of mixed needle and broadleaf is higher than that of mixed needle and leaf (SM-M). In terms of comprehensive physiological effects, the mixed effect of MH-M treatment is the best, the next is FX-M and QM-M treatment.