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KMID : 0379119940220020130
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Korean Journal of Mycology
1994 Volume.22 No. 2 p.130 ~ p.137
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Studies on Protoplast Formation and Regeneration of Lyophyllum decastes
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Choi Eung-Chil
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Abstract
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This experiment was carried out to investigate proper conditions for protoplast isolation and regeneration from mycelia of Lyophyllum decastes. Novozym 234(10 ㎎/㎖) with 0.6 M MgSO₄ in phosphate buffer(pH 4.0) was proper for protoplast isolation. The optimal reaction time of the myceliun with the lytic enzyme was four hours in shaking condition at 120 strokes per min. When the mycelium of L. decastes was cultured at 24℃ for 5 days, the formation of protoplasts was effective. The liquid medium was more effective for protoplast isolation than the solid medium. In the liquid medium, high yields of protoplasts were obtained from 0.6 M MgSO₄, osmotic stabilizer. Protoplasts of L. decastes were regenerated to normal hyphal growth snd the regeneration frequency of the protoplasts in the complete agar medium containing Triton X-100(0.0025%) was 5.94∼8.32%. The regeneration medium stabilized with 0.6 M sucrose was the best for regeneration of the protoplasts. In contrast to protoplast formation, regeneration was inhibited by the inorganic salts used as osmotic stabilizer.
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