Abstract: In order to investigate the feasibility of using waste tobacco stem as the matrix material in tobacco floating seedling system, seedling culture experiments were carried out with the matrix containing prepared tobacco stem peat (TSP). The results showed that the TSP 1 (50% tobacco stem peat + 50% natural peat), TSP 2 (50% tobacco stem peat + 50% coconut husk) and TSP 3 (50% tobacco stem peat + 25% natural peat + 25% coconut husk) could be the alternatives to natural peat in tobacco floating seedling system. The tobacco seedlings in TSP 3 presented better comprehensive growth vigor than, while those in TSP 1 and TSP 2 were comparable to, those in natural peat matrix. Therefore, it is feasible to use waste stems as a substitute for natural peat in tobacco floating seedling system.