Abstract: To reduce peat consumption and select suitable substituted matrices for tobacco seedling culturing in main tobacco-planting areas of Sichuan, tobacco seedling culturing experiments were conducted in floating systems with alternative matrices (yellow sands from Liangshan, Panzhihua and Luzhou and powdered stone from Yibin mixed with 75%, 50% and 25% of a commercial peat matrix). The effects of substituted matrices and their proportions in floating seedling system on the growing, physiological and biochemical indexes, seedling stage and seedling rate were studied. The results showed that the substituted matrices mixing commercial peat matrix by yellow sands or powdered stone from the four areas met the needs of germination and growth of tobacco seedlings. The optimal proportion for substituted matrices was 50% yellow sand/powdered stone+50% commercial peat matrix, and the physicochemical properties of matrices with the optimal proportion satisfied the requirements in floating seedling system. The emergence rate, seedling rate and seedling stage of tobacco seedlings cultured in the matrices of 50% yellow sand/powdered stone+50% commercial peat matrix were above 90%, 82%-84% and 62-64 days, respectively. Comparing with control, 50% yellow sand/powdered stone+50% commercial peat matrix were significantly better or did not significantly differ in terms of plant height, stalk circumference, leaf number, the number of the first lateral roots, root vigor, the activities of superoxide dismutase (SOD)and peroxidase (POD)and the content of chlorophyll; and the cost of substituted matrices decreased by 46.70%. The proportion of 50% yellow sand/powdered stone with 50% commercial peat matrix is suitable for Sichuan tobacco-planting areas.