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KMID : 0613820030130020143
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Journal of Life Science
2003 Volume.13 No. 2 p.143 ~ p.149
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Studies on Protoplast Isolation and Regeneration of Lyophyllum ulmarium
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Choi Hye-Jin
Kim Byung-Kak
Hyun Jin-Won
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Abstract
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This experiment was undertaken to investigate proper conditions for protoplast isolation and regeneration from the mycelia of Lyophyllum ulmarium. Protoplast isolation and regeneration are influenced by a variety of factors such as enzyme, osmotic stabilizer, reaction time and age of mycelia. A combination of Novozyme 234 (10 mg/ml) and cellulase Onozuka R-10 (10 mg/ml) with 0.6 M MgSO4 was most effective for isolation of the protoplasts. The optimum reaction time of the mycelia with the lytic enzymes was 3.5～4 hours at 28℃ in shaking condition at 120 strokes per min. High yield of the protoplasts were obtained from its 4～5 days old mycelia on complete agar media. Its protoplasts were regenerated to normal hyphae. Regeneration media with 0.6 M sucrose were proper for regeneration of the protoplasts. Their regeneration frequency on complete agar media was 2.3～2.7%.
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KEYWORD
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Basidiomycetes, Lyophyllum ulmarium, Protoplast isolation, Protoplast regeneration, Osmotic stabilizer
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