The effects of some factors on the production of protoplasts of flammulina velutipes, such as enzyme dosage, culture age, osmotic stabilizer, hydrolytic temperature and time, were investigated and the optimum conditions for preparation of protoplasts were also investigated. After 4-day liquid culture and 10-day solid culture, the Flammulina velutipes Mycelia was hydrolyzed by 1% cellulase and 1% lysozyme for 1.5h at 25℃ with 0.5M KCl as osmotic stabilizer. The maximum yield was achieved to be above (05~7)×107/ml .
