Abstract: Pleurotus nebrodensis is a specially rare mushroom for food and medicine which can regulate physiological balance and strengthen the immune function. In this paper, we used pleurotus nebrodensis as material to study the effects of different temperature, pH vale, carbon source, nitrogen source and ventilation conditions on the mycelium growth. The results showed that: The appropriate growth temperature range of mycelia growth is from 22 ° C to 30 ° C, the optimal temperature is 25 ° C; the appropriate pH vale range is from 5.0 to 8.0, the optimal pH vale is 7.5; the optimal carbon source is sugar; the optimal nitrogen source is peptone; adequate oxygen is conducive to mycelial growth.