Abstract: The factors affecting the preparation of Agrocybe aegerita protoplasts from and the fusion between the protoplasts were studied. The results showed that enzyme type, enzymolysis time, enzymolysis temperature, osmotic pressure stabilizer and pH significantly affect the preparation of protoplasts, and enzymolysis temperature and pH among them are the most important influence factors. The best preparation conditions are as follows: digesting for 4h with 1% cellulase and 1% snailase (1:1) at 28 ℃ and taking 0.6 mol/L mannitol solution at pH 6.0 as osmotic pressure stabilizer. Protoplast fusion is made by chemical method with 30% PEG (MW 4000) and 0.2 mol/L CaCl2 (pH 10.5).

Key words: Agrocybe aegerita, protoplast, preparation, fusion