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This experiment was undertaken to investigate proper conditions for protoplast isolation from strains of Agaricus bisporus, Flammulina velutipes, Lentinus edodes, Pleurotus florida, Pleurotus ostreatus, Pleurotus sajor-caju and Volvariella volvacea. Combination of β-D-glucanase, novozym 234 and snail enzyme with 0.6M MgSO₄ in 0.05M Na-maleate buffer, pH 5.8 was the most effective for isolation of protoplasts. High yields of protoplasts were obtained from 4∼5 days old mycelia of P. florida and P. ostreatus on mushroom complete agar medium. Protoplasts were also obtained in good yield from other fungi but A. bisporus and V. volvacea. In the latter two cases protoplast release was observed; however, the yield was much lower than those of the other fungi.
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