Abstract: Suitable of submerged culture conditions of Pleurotus nebrodensis were studied by flask shaking. The results showed that the suitable carbon source and nitrogen source are glucose and peptone. The mycelium grows best when glucose concentration is 3.0% and C/N is 20:1. The optimum culture conditions were as below: inoculation loading 10%,medium capacity 100mL／25OmL flask,250mLflask,initial pH7.0, temperature 30℃.