Abstract: In order to discuss the localization application of cotton seedling using substrate, the author used cotton seed hulls residue after the production of edible fungi, river sand and peat in different proportion formula to conduct cotton seedling test in field and laboratory. The results showed that: dry mass per plant and leaf area per plant of formula 3 (bulk ratio of seed hulls residue, river sand and peat was 6:3:1) were higher than Jiangxi cotton seedling nursery substrate for 48.4% and 73.5%, respectively; the rate of forming plantlets, survival rate of transplant and the unginned cotton yield had no obvious difference with the other matrix seedling and transplanting; nursery substrate was returned to field as fertilizer to reduce environmental pollution and achieve comprehensive and cyclic utilization. Therefore, being a cotton seedling substrate material, cotton seed hulls residue after the production of edible fungi is easily-obtained, cost-effective and environment-friendly and has a good application prospect.