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KMID : 0379119860140020141
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Korean Journal of Mycology
1986 Volume.14 No. 2 p.141 ~ p.148
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Studies on Protoplast Isolation of Pleurotus cornucopiae
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박용환/Park, Yong Hwan
민경희/이연희/유영복/Min, Kyung Hee/Lee, Yeon Hee/Yoo, Young Bok
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Abstract
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The optimal conditions for high yields of mycelial protoplasts from P. cornucopiae were established. The concentraion of enzyme system containing Novozym 234, β-D-glucanase and β-glucuronidase was 5㎎ ㎖^(-1) each. The osmotic stabilizer most effective for protoplast isolation was 0.6 M sucrose. The optimal reaction time of mycelium with the lptic mixture was 90 thin is a shaking condition at 120 strokes min^(-1). When the myelium of P. cornucopiae was cultured for 4 days on mushroom complete medium at 28℃, the formation of protoplast was effective. When the pH of the digestion mizture with 0.6 M sucrose as stabilizer vazied between pH 4.0 and 7.0. the production of protoplasts was effective in phosphate buffer (pH 6.2) and Na-maleate buffer (pH 5.0). Generally, phosphate buffer was more effective for protoplast isolation than Na-maleate buffer, but 0.6 M sucrose osmotic stabilizer without adjusting pH was most effective. Using these conditions, protoplasts from P. cornucopiae were obtained at a ratio 1×10^7 ㎖^(-1).
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