Quadratic rotation-orthogonal composite experimental design was applied in this experiment to analyze the ultraweak luminescence property of osmanthus tree seedings affected by multilateral growth environmental factors. The influence models of four factors on luminescence were established including illumination, temperature, humidity, and moisture. The ANOVA (analysis of variance) analysis showed that different factors and their interaction had different effect on luminescence property, among which the linear effects of four variables, the quadratic effects of illumination, relative humidity and moisture, and the interaction effect of illumination × moisture and temperature × relative humidity all have significant effect on the luminous value of osmanthus (p ≤ 0.01). The maximum luminous value was obtained when ten stick of lamp tube (intensity of illumination value 4229 Lx) was used, mean temperature, humidity and water were 35 ℃, 85% and 90 mL/d respectively. On the base of that, linear effects and interactions that affected ultraweak luminescence were further studied.