Abstract: Pleurotus eryngii is a promising edible fungus. In this study， a Pleurotus eryngii strain was screened for rapid growth. The best protoplast preparation conditions were obtained via optimization. The optimum conditions for the preparation of protoplasts are: snailase of 3% and cellulase of 1% pH 55， enzymatic hydrolysis temperature of 37℃， cell age of 3 days， and mannitol of 6 mol·L-1 as osmotic stabilizer. The results presented here lay an important foundation of UV mutation and fusion protoplast of Pleurotus eryngii strains， which has importantly practical values for the breeding of Pleurotus eryngii strains.

Key words: Pleurotus eryngii, screening, protoplast preparation, conditions optimization