Abstract:The protoplasts isolated enzymically from monokaryotic mycelia of Lentinus edodes, Pleurotus sapidus and Pleurotus ostreatus cultured in liquid medium were electrofused by means of supplying electric field of sine wave 0.5MHz, 500PV / cm and square pulse 50us, 6000PV / cm. The colonies grew up from the fusants or protoplasts plated on solid medium af-ter electrofusion. The strains from fusants were selected by discovering clamp connection of fusant mycelia under a microscope. Frequence of regenerating strain from fusant was 61.53% between Lentinus edodes and Pleurotus sapidus, 32.58% between Lentinus edodes and Pleurotus ostreatus. It was demonstrated that fusant strains grew from the heterkaryon according to an-tagonistic action of fusant strains and parents and difference of their electrophoretogram of esterase and peroxidase isoenzyme from those of their parents. While the electrofusion method and results were discussed.