Abstract: The research aims to verify the feasibility of using edible fungus residue as a seedling substrate to replace non- renewable peat resources, and promote the sustainable development of the industry and environment. Effects of five different ratios of seedling substrates, commercial seedling substrate(A), grass charcoal: high-quality loess: perlite= 1:1:1(B), edible fungus residue: perlite: high-quality loess= 1:1:1(C), edible fungus residue: high-quality loess: perlite: compound fertilizer= 1:1:1:0.005(D), and high-quality loess: fire-ash ash= 1:1(E), and three seedling culture modes, plug seedling, floating seedling, and tidal seedlings, on seedling rate, seedling height and stem diameter, seedling uniformity, seedling true leaf number, seedling root and shoot fresh weight ratio, seedling index, and recover period after planting were investigated. Results indicated that seedling substrate A was the best one. Except for substrate E, the other substrates could meet the production demand. The differences between substrate C, D and B were not significant, but the production management was more complicated. The seedling substrate E had the greatest impact on seedling rate and seedling index. Comparing the results from different seedling culture modes, the tidal seedling cultivation was better than the other two seedling culture modes. The floating seedling had a longer recover period after planting, and there was also partial yellowing phenomenon during seedling cultivation. The plug seedling culture mode had the problems of small number of sowing plants per unit area, large difference between plants, and complicated management. Edible fungus residue has certain feasibility as a seedling substrate, but corresponding measures are needed to tackle the problems to reduce the disadvantages.