• Mycobiology
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• v.50 no.5
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• pp.374-381
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• 2022

In the mating of filamentous basidiomycetes, dikaryotic mycelia are generated through the reciprocal movement of nuclei to a monokaryotic cytoplasm where a nucleus of compatible mating type resides, resulting in the establishment of two different dikaryotic strains having the same nuclei but different mitochondria. To better understand the role of mitochondria in mushrooms, we created four sets of dikaryotic strains of Lentinula edodes, including B2×E13 (B2 side) and B2×E13 (E13 side), B5×E13 (B5 side) and B5×E13 (E13 side), E8×H3 (E8 side) and E8×H3 (H3 side), and K3×H3 (K3 side) and K3×H3 (H3 side). The karyotypes and mitochondrial types of the dikaryotic strains were successfully identified by the A mating type markers and the mitochondrial variable length tandem repeat markers, respectively. Comparative analyses of the dikaryotic strains on the mycelial growth, substrate browning, fruiting characteristics, and mitochondrial gene expression revealed that certain mitochondria are more effective in the mycelial growth and the production of fruiting body, possibly through the activated energy metabolism. Our findings indicate that mitochondria affect the physiology of dikaryotic strains having the same nuclear information and therefore a selection strategy aimed at mitochondrial function is needed in the development of new mushroom strain.

• Journal of Mushroom
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• v.13 no.3
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• pp.212-216
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• 2015

A new commercial strain "Mongdol" of oyster mushroom was developed by hyphal anastomosis. It was improved with hybridization between monokaryotic strain derived from Pleurotus ostreatus ASI 0627 and dikaryotic strain derived from P. ostreatus ASI 2929. The optimum temperature of mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $12{\sim}18^{\circ}C$, respectively. When two different media including PDA (potato dextrose agar medium) and MCM (mushroom complete medium) were compared, the mycelial growth of this mushroom was faster in MCM than in PDA. Similar result was observed with the control strain P. ostreatus ASI 2504. Analysis of the genetic characteristics of the new cultivar "Mongdol" showed a different DNA profile as that of the control strain ASI 2504, when RAPD (Random Amplified Polymorphic DNA) primer URP3 and URP6 were used. Fruiting body production per bottle was about 106 g using demonstration farms. The color of pileus was blackish gray and the stipe was long. Therefore, we expect that this new strain "Mongdol" will satisfy the consumer's demand for high quality mushrooms.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.53-53
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• 2015

Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

• Journal of Mushroom
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• v.14 no.2
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• pp.59-63
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• 2016

A new commercial strain of oyster mushroom (was developed by hyphal anastomosis, and was improved byhybridization between a monokaryotic strain derived from Pleurotus ostreatus ASI 0635 (Gonji 7ho) and a dikaryotic strain derived from P. ostreatus ASI 0666 (Mongdol). The optimum temperatures for mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $12{\sim}18^{\circ}C$, respectively. When PDA (potato dextrose agar medium) and MCM (mushroom complete medium) were compared, mycelial growth was faster in MCM. Similar results were observed with the control strain P. ostreatus ASI 2504 (Suhan 1ho). Analysis of the genetic characteristics of the new cultivar ('Gosol') showed a different DNA profile from that of the control ASI 2504 strain, when RAPD (raurpDNA) primers URP1, 2, 3, and 7 were used. Fruiting body production per bottle was approximately116 g based on a production performance test. In addition, yields from a farm field trial were stably achieved in an inadequate production enviro. The color of the pileus was blackish gray, and the stipe was long and thick. Therefore, we expect that this new strain will satisfy consumer demand for high quality mushrooms.

• Korean Journal of Agricultural Science
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• v.15 no.1
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• pp.15-22
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• 1988

Factors affecting the regeneration, reversion of protoplasts from mycelium of F. velutipes were investigated and the selection of auxotrophic mutants from protoplasts of F. velutipes was performed. PDP medium stabilized with 0.6M sucrose was suitable for the regeneration of protoplasts, and regeneration frequency was 0.47-1.32. The regeneration frequency of protoplasts was increased when nutrients were added to the regeneration medium. Especially, yeast extract was the most effective to regeneration of protoplasts. Regeneration pattern of protoplasts was formation of germ tubes from bud-like cells. 13-18% of monokaryotic strains was appeared from reverted protoplasts. Five of auxotrophic mutants were isolated from strains showed survival frequency of 1.9-16.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.39-43
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• 2011

A new cultivar "Dan Bi" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2312 and KNR2596. The parental strains, KNR2312 and KNR2596, are characterized by the property of high quality and a small number of primordia formation, respectively. The optimum temperature of mycelial growth was 25 and that of fruiting body development was $15{\sim}16^{\circ}C$. The period of harvesting including primordia formation was 0.7~1.3 days later than that of control strain Knneutari No. 3 in the culling cultivation. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has a scale like as cobweb. The yield was $93{\pm}9.7$ g per 850 cc of plastic bottle. Analysis of the genetic characteristics of the new commercial variety "Dan Bi" showed a different profile as that of the control strain, Knneutari No. 3, when RAPD (Random Amplified Polymorphic DNA) primer #8005 was used. This new variety "Dan Bi" of Pleurotus eryngii is characterized by a small number of primordia formation after scratching.

• Journal of Mushroom
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• v.5 no.1
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• pp.7-12
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• 2007

All intra-specific hybrids among ASI $2172{\times}2104$, $2172{\times}2307$, $2186{\times}2172$ and $2186{\times}2307$ in P. salmoneostramineus produced pink fruiting bodies as like wild parental types. However, three hybrids of them between ASI 2186 and 2104 were white fruiting bodies. A new commercial strain "Noeul" of Pink Oyster mushroom was developed by intra-specific hyphal anastomosis. It was improved with hybridization between monokaryotic strain derived from ASI 2172 and ASI 2104. The optimum temperature of mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $19{\sim}24^{\circ}C$, respectively. The pileus was bright reddish pink. Commercial strain "Noeul" was as prolific as the more commonly cultivated Pleurotus ostreatus in the conversion of substrate mass to mushrooms using bottle cultivation. Mushroom cultivator can save money for mushroom growing on summer in Korea. Mushrooms should be picked when moderately young, and handled carefully so as to not bruise the brilliantly colored gills. This pink color makes marketing an interesting challenge depending upon the market niche.

• Journal of Mushroom
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• v.6 no.2
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• pp.47-51
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• 2008

The Oyster mushroom is saprophytic fungus. The pileus is stemmed at the side and later depressed. It grows to 5-15 cm, and is of grey, grey-lilac, blackish-grey, steel grey, grey-brown, and blue-blackish. Various kinds of Oyster mushrooms such as golden, pink, brown, grey, white, and blue make marketing an interesting challenge depending upon the market niche. A new commercial strain "Chung" of oyster mushroom was developed by hyphal anastomosis. It was improved with hybridization between monokaryotic strain derived from Pleurotus ostreatus ASI 2194 and ASI 2487. The pileus of parental strain ASI 2194 and ASI 2487 was grey and light blue-grey, respectively. Most of intra-specific hybrids between strain ASI 2194 and ASI 2487 were showed similar pileus color of parents. By the way, the pileus color of variety 'Chung' was blue to bluish grey. The optimum temperatures of mycelial growth and fruiting body development was $25{\sim}30^{\circ}C$ and $12{\sim}16^{\circ}C$, respectively.

• The Korean Journal of Mycology
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• v.27 no.5 s.92
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• pp.322-327
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• 1999

Five white fruitbodies of Ganoderma lucidum found from two different mushroom farms, and the characteristics of atypical fruiting structure formation of these strains were described. The white fruitbodies were spontaneously generated on Quercus-log during the cultivation. They did not differentiate to the normal fruitbodies with pileus, hymenium, stipe and coloration, and fruitbodies remained non-laccateed even after 3 months. Dikaryotic mycelia isolated from the five white fruitbodies differed from wild-type strains in the mycelial growth rate, colony color, and the capacity of atypical fruiting structure (AFS) formation on agar media. These white mutants readily induced brown colored AFSs on the colonies under ventilation and illumination conditions. Both isolates Gl-010 and Gl-011 that were obtained from a normal and white fruitbody, respectively, did not form AFSs in the dark and/or under black light blue (BLB) light illumination, but induced under the visible light. They required dim light for the AFS formation, and the AFS formation was inhibited up to $0.5{\mu}mol\;m^{-2}\;S^{-1}$ in light intensity. However, the other four isolates induced AFSs even in the dark and BLB illumination, although their parent strain, isolate Gl-030, did not form AFSs under any light conditions. The monokaryotic mycelia derived from basidiospores of the AFSs of the white mutants were compatible with the original culture (dikaryon) on a dual culture.

• Journal of Mushroom
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• v.15 no.1
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• pp.25-30
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• 2017

'Baekseung', a new variety of Flammulina velutipes, was bred by mating two monokaryotic strains isolated from KMCC 4210 and KMCC 4216 in the Mushroom Research Division, Baekseung ARES in 2016. The Baekseung and Uri1ho strains showed fast mycelial growth and mycelial density on malt extract agar media after 7 days of incubation. The spawn running period on the sawdust substrate required a cultivation period and temperature of 30 days and $25^{\circ}C$, respectively, for primordia formation where in fruiting body development occurred from $11{\pm}1days$ at $14^{\circ}C$ and $14{\pm}1days$ at $7^{\circ}C$. The length of the pilei and stipes of the Baekseung harvested in optimal stag were $11.3{\pm}0.4$ and $89.2{\pm}7.1mm$, respectively, whereas the values for Uri1ho were $10.7{\pm}1.0$ and $91.3{\pm}20.8mm$, respectively. The yield of the Baekseung and Uri1ho strain grown on the sawdust substrate was $153.7{\pm}12.5$ and $139.8{\pm}17.8g$, respectively, per 850 ml in bottle cultivation. The inferred tree exhibited a phylogenetic relationship between the Korean white fruiting body strains of Baekseung, Uri1ho and Fv-14-a-38, Fv-14-a-51, and the Japanese white fruiting body-forming strains of KMCC 4226, and these were confirmed to be genetically related.