• 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.

• Journal of Mushroom
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• v.8 no.4
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• pp.161-164
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• 2010

For the development of a sporeless strain of P. ostreatus we used sporeless strain ASI 2069. We have recovered both nuclear types of strain ASI 2069 as monokaryons of nh9, nh15, nh26 and nh36 (here after referred to as neohaplonts) by protoplasting the mycelium. Crosses between neohaplonts and SSI's(single spore isolates) obtained from a sporulating commercial strain ASI 2180. Five excellent strains are selected from 30 bred strains by quality of fruitbodies and spore number. To development of molecular markers linked to sporeless strain of P. ostreatus, we are screened helicase, recombinase(DMC1) and topoisomerase(Spo11) genes related meiosis by PCR and sequencing. Among three genes, Spo11 gene was identified into molecular marker of sporeless from neohaplonts and bred strains of P. ostreatus.

• Journal of Mushroom
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• v.2 no.1
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• pp.33-37
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• 2004

To develop neohaplonts for Ganoderma breeding, protoplasts were isolated from dikaryotic mycelium and regenerated. Selection rate of neohaplonts varied between ASI7074, ASI7091, ASI7094, ASI7100 and ASI7115, showing 5.24% on the average. Auxotropic mutants from Ganoderma monokarions were recovered by UV irradiation on protoplasts. Protoplast survival rates were 1.9% ASI 7074, 0.17% ASI 7091, and zero percent ASI 7100 using 300 second irradiation. Four auxotrophic strains were recovered from 1,536 colonies screened that will be further utilized for protoplast fusion and transformation.

• Journal of Mushroom
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• v.4 no.2
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• pp.53-56
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• 2006

The enormous production of spores by the fruitbodies in the cultivation of oyster mushrooms (Pleurotus ostreatus) is develop an allergy with symptoms similar to an "extrinsic allergic alveolitis (EEA)". the sporeless strain would noy only benefit health of mushroom workers but also reduce the risk of viral infections on the mushroom farms. For the development of a sporeless strain of P. ostreatus we used strain ASI 2069. This non-commercial strain is completely nonsporulating. We have recovered both nuclear types of strain ASI 2069 as monokaryons (hereafter referred to as neohaplonts) by protoplasting the mycelium. Crosses between neohaplonts and SSI's(single spore isolates) obtained from a sporulating commercial strain ASI 2180 yielded fruitbodies that isolated 128 strains. 13 excellent strains are selected from 30 bred strains by quality of fruitbodies and spore number. Among 13 excellent strains, G192 strain is chose finally to high yield and sporelessness.

• Journal of Mushroom
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• v.13 no.3
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• pp.270-273
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• 2015

Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.