• Korean Journal of Microbiology
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• v.19 no.4
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• pp.192-198
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• 1981

Protoplasts from Trichoderma koningii were fractioned at varing digestion periods. There were distinct differences between the fractionated protoplasts in CMCase activity and protein content ; namely, protein contents and CMCase activities in the early-protoplasts were higher than the late-produced protoplasts. Reversion frequencies of the early-produced protoplasts. Reversion frequencies of the early-produced protoplasts (0-1hr.) and the late-produced protoplasts (1-2.5 hr.)were 1.25% and 0.56%, respectively. From these results it was assumed that the early-produced protoplasts were more active in physiological metabolism. Meanwhile, in osmotically stabilized liquid medium two distinct reversion patterns of these protoplasts were found. In the first, normal germ tube was developed from the opposite side of the protoplast after the production of an aberrent tube. In the second, reversion occurred through germination of protoplast, but this patterns was uncommon. It is suggested that the the physiological and morphological heterogeneities in protoplast reversion are related to the heterogeneous origins of protoplasts from highly differentiated mycelium.

• The Korean Journal of Mycology
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• v.13 no.2
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• pp.79-82
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• 1985

The experiment of protoplast regeneration and reversion were undertaken to provide a basic techniques for protoplast manipulation. Protoplasts of Pleurotus florida and Pleurotus ostreatus were reverted to normal hyphal growth and the reversion frequency of both fungal protoplasts were $0.24{\sim}3.19%$. Reversion medium stabilized with 0.6 M potassium chloride and sucrose was better than the other stabilized one. The protoplast reversion frequency was increased when various amino acid and vitamin compounds were added to the hypertonic mushroom complete agar medium.

• The Korean Journal of Mycology
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• v.15 no.1
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• pp.38-41
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• 1987

Neohaplonts were obtained to improve mushroom strain from dikaryon strains of Flammulina Velutips, Ganoderma lucidum, Lyophyllum ulmarium, Pleurotus cornucopiae and Pleurotus spodoleucus by protoplast reversion. The noehaplont frequency from reversion colonies of protoplasts was 4.47-47.7%. Four more types of morphological characteristics were detected from neohaplonts of protoplast reversion in L. ulmarium, P. cornucopiae and P. spodoleucus, but one or two types were neohaplonts in F.velutipes and G. lucidum. Growth rate of neohaplonts was slow compared with dikaryon strains.

• Korean Journal of Agricultural Science
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• v.19 no.1
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• pp.33-39
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• 1992

Effects of buprofezin, a chitin synthesis inhibitor of insects, on mycelial growth, protoplast formation and reversion of Coriolus versicolor, Ganoderma applanatum and G. lucidum were investigated. The mycelial growth of C. versicolor, G, applanatum and G, lucidum was severely inhibited by buprofezin treatment, and the inhibition rate severely as the concentration of the buprofezin increased. Aerial mycelia and oidia formation of the mushrooms were increased by buprofezin treatment, but mycelial morphology was not changed. The rate of protoplast formation and reversion of G, applanatum was greatly increased by the treatment of the buprofezin, while that of G. lucidium was decreased. The rate of protoplast formation of C. versicolor was also increased when buprofezin was added to the medium, but the rate of protoplast reversion was not affected by the treatment, From the results obtained in this experiment, we found that the rate of protoplast. formation and reversion was increased by the treatment of the buprofezin in the mushrooms such as C, versicolor and G, applanatum, whose mycelial growth was fast on the medium, while that of G. lucidum, whose mycelial growth was relatively slow, was decreased by the treatment.

• The Korean Journal of Mycology
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• v.15 no.1
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• pp.14-18
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• 1987

This experiment was undertaken to investigate proper conditions for protoplast formation from Lyophyllum ulmarium. Combination of Novozym 234, ${\beta}-Glucuronidase$ and ${\beta}-D-Glucanase$ with 0.6 M Sucrose was the most effective for isolation of protoplasts. The optimal reaction time of mycelium with the lytic mixture was 3 hrs in shaking condition at 120 strokes $min-^1$. When the mycelium of L. ulmarium was cultured for 6 days on yeast glucose agar medium at $25^{\circ}C$, the formation of protoplasts was effective. The yeast glucose agar medium stabilized with 0.6 M sucrose was the most effective for reversion of protoplasts.

• The Korean Journal of Mycology
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• v.16 no.4
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• pp.214-219
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• 1988

Factors affecting protoplast formation and reversion were investigated in Pleurotus sapidus kalchbr. For release of protoplast, enzyme mixture of Novozyme 234, ${\beta}-D-glucanase$ and ${\beta}-glucuronidase$ was most effective, when mycelium of 0.6 M sucrose solution as osmotic stabilizer without addition of buffer solution. The yield of protoplast was highest with mycelium cultured for 4 days on mushroom complete agar medium at ${30}^{\circ}C$. Protoplasts of Pleurotus sapidus were reverted to normal hyphal growth with maximum reversion frequency of 2% on Mushroom complete agar medium stabilized with 0.6 M sucrose solution and covered by 0.75% agar layer.

• The Korean Journal of Mycology
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• v.14 no.3
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• pp.215-223
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• 1986

Protoplasts of P. cornucopiae were reverted to normal hyphal growth and reversion frequency was $0.04{\sim}19%$. The complete medium stabilized with 0.6 M sucrose was most effective for regeneration of protoplasts. When hypertonic mushroom complete medium not containing agar was overlaid, regeneration frequency of protoplasts was the highest rate among the others of topagar. The protoplast reversion frequency and mycelial growth of P. cornucopiae were increased when various amino acids, nucleic acid components and vitamin compound were added to the hypertonic minimal medium. The relation between sources increasing reversion frequency and sources accelerating mycelial growth was similar in amino acids and nucleic acid components but it was different in vitamins. The protoplast reversion frequency showed the highest rate when all sources were added to the regeneration minimal medium. Microscopically, regeneration patterns of protoplasts showed formation of a bud-like structure, direct germination, yeast-like cell chain of the protoplast, and the production of both direct germ tube and yeast-like cell chain from a protoplast.

• Korean Journal of Agricultural Science
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• v.17 no.2
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• pp.77-81
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• 1990

Effects of buprofezin, an inhibitor of chitin synthesis, on mycelial growth, protoplast formation and reversion of Pleurotus ostreatus and P. sajor-caju were investigated. The mycelial growth of Pleurotus ostreatus and P. sajor-caju was the inhibited by buprofezin treatment, and the inhibition rate was severer as the concentration of the buprofezin increased. Aerial mycelium formation was increased by buprofezin treatment, but mycelial morphology was not changed. Protoplast formation of Pleurotus ostreatus and P. sajor-caju. was significantly increased when buprofezin was added to the culture medium at the concentration of 200~500 ppm and the protoplast reversion of the mushrooms was also increased by the treatment of the buprofezin.

• The Korean Journal of Mycology
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• v.16 no.1
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• pp.21-25
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• 1988

This experiment was carried out to investigate proper conditions for protoplast isolation and reversion from Ganoderma lucidum and Gctnoderma sp.. In G. lucidum, 10 mg. $ml^{-1}$ Novozyme 234 with 0.6 M sucrose was proper for protoplast isolation. The optimal reaction time of mycelium with lytic enzyme was five hrs. Protoplast isolation from four-day-old mycelium was the most effective. Protoplast isolation from four-day-old mycelium in G. sp. was optimum in the combination of N ovozyme 234 and ${\beta}-glucuronidase$ with 0.6 M sucrose. MCM was suitable for reversion in G. lucidum while SCM was good for G. sp.. The most effective osmoticum stabilizer for protoplast reversion in G. lucidum and G. sp. was 0.6 M sucrose.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.514-518
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• 1992

The present study has been perromed to investigate the optimal conditions for protoplast formation and regeneration of oleandomycin-producing Streptomyces antibioticus (S. antibioticus) KCTC 1081. Mycelia were grown in YME medium containing 0.2% (w/v) glycine and converted into the protoplast by incubating at 35.deg.C for 60 minutes in protoplast buffer (P buffer) containing 4 mg/ml lysozyme. The reversion of protoplasts to the normal filamentous state was examined by the growth on various synthetic agar media. A high reversion rate was obtained by incubating the protoplasts on a hypertonic agar medium containing 20 mM $Mg^{++}$, 5 mM $Ca^{++}$ and 0.3 M sucrose at 28.deg.C for 5 days. From these experiments, we established the improved regeneration medium and a protocol which supports higher and more consistent levels of regeneration of S. antibioticus protoplasts. The regenerant showed an increased antimicrobial activity compared with that of the initial strain.n.n.