• Mycobiology
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• v.51 no.1
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• pp.26-35
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• 2023

The diversity of A mating type in wild strains of Lentinula edodes was extensively analyzed to characterize and utilize them for developing new cultivars. One hundred twenty-three A mating type alleles, including 67 newly discovered alleles, were identified from 106 wild strains collected for the past four decades in Korea. Based on previous studies and current findings, a total of 130 A mating type alleles have been found, 124 of which were discovered from wild strains, indicating the hyper-variability of A mating type alleles of L. edodes. About half of the A mating type alleles in wild strains were found in more than two strains, whereas the other half of the alleles were found in only one strain. About 90% of A mating type combinations in dikaryotic wild strains showed a single occurrence. Geographically, diverse A mating type alleles were intensively located in the central region of the Korean peninsula, whereas only allele A17 was observed throughout Korea. We also found the conservation of the TCCCAC motif in addition to the previously reported motifs, including ATTGT, ACAAT, and GCGGAG, in the intergenic regions of A mating loci. Sequence comparison among some alleles indicated that accumulated mutation and recombination would contribute to the diversification of A mating type alleles in L. edodes. Our data support the rapid evolution of A mating locus in L. edodes, and would help to understand the characteristics of A mating loci of wild strains in Korea and help to utilize them for developing new cultivars.

• Journal of Mushroom
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• v.12 no.1
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• pp.58-62
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• 2014

To develop a new cultivar of King oyster mushroom white variety (Pleurotus ferulae), GW10-68 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2798-24 derived from ASI 2798 and dikaryotic strain ASI 2803. The GW10-95(ASI 2803 ${\times}$ ASI 2798-24) was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Beesan No.2'. The 'Beesan No.2' was formed incompatibility line distinctly in the confrontation growth of parental strains ASI 2803, ASI 2798 and Beesan No.1. Analysis of the genetic characteristics of the new cultivar 'Beesan No.2' showed a different DNA profile as that of the control strains, ASI 2803, ASI 2798 and 'Beesan No.1', when RAPD(Random Amplified Polymorphic DNA) primer URP4 was used. The optimum temperature and pH arrange for mycelial growth were $25^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about 123.3 g. And also the stipe was long. This new cultivar 'Beesan No.2' of Pleurotus ferulae was characterized white-like variety of King oyster mushroom in the color of stipe, that was long and low yield compared to that of other cultuvar 'Beesan No.1'. We therefore expect that this new strain will increase of export and substitute cultivar of King oyster mushroom.

• Journal of Mushroom
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• v.12 no.1
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• pp.52-57
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• 2014

To develop a new cultivar of King oyster mushroom white variety (Pleurotus ferulae), GW10-95 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2850-24 derived from ASI 2850 and dikaryotic strain ASI 2803. The GW10-95(ASI 2803 ${\times}$ ASI 2850-24) was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Beesan No.1'. The 'Beesan No.1' was formed incompatibility line distinctly in the confrontation growth of parental strains ASI 2803 and ASI 2850. Analysis of the genetic characteristics of the new cultivar 'Beesan No.1' showed a different DNA profile as that of the control strains, ASI 2803 and ASI 2850, when RAPD(Random Amplified Polymorphic DNA) primer URP6 was used. The optimum temperature and pH arrange for mycelial growth were $25^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about 245 g using demonstration farms. And also the stipe was thick and long. This new cultivar 'Beesan No.1' of Pleurotus ferulae was characterized by fast fruitbody formation, the stipe was thick and long and high quality yield compared to that of other cultuvars. We therefore expect that this new strain will increase of farmer's income by construction of stabilized production system.

• Journal of Mushroom
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• v.11 no.2
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• pp.77-81
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• 2013

To develop a new cultivar of king oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2824-21 derived from ASI 2824(Keunneutari No. 2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-53(G09-21-10 x ASI 2844-9) was shown the best cultural characteristics, selected to be a new cultivar and designated as 'Seolsong'. The 'Seolsong' was distinctly formed incompatibility line in the confrontation growth of parental strains Keunneutari No. 2, Aeryni 3 and ASI 2844. Analysis of the genetic characteristics of the new cultivar 'Seolsong' showed a different DNA profile as that of the control strains, Keunneutari No. 2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primer URP4 was used. The optimum temperature and pH arrange for mycelial growth were $25{\sim}30^{\circ}C$ and pH 5~8, respectively. This new cultivar 'Seolsong' of fruiting body production per bottle was about $131.4{{\pm}}43.1$ g which is about 102% quantity compared to that of other cultivar Keunneutari No. 2. And also the stipe is thick and long, but the number of available stipe is few. Particularly, it was tolerant of high moisture above 90% during the growth period after primodia formation. We therefore expect that this new strain will save of labor and cost of cultivation by without culling work.

• Journal of Mushroom
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• v.13 no.2
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• pp.129-134
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• 2015

Hybrid strains was selected by crossing between dikaryotic strain ASI 0628 (Dagul) bred from ASI 2596 (Suhan No.3) and ASI 2782(Black pileus mutant) and monokaryotic strain ASI 2344-84. The strain 84 that shown the best cultural characteristics was selected to be a new cytoplasmic hybrid variety and named as 'Chunhwashim'. The 'Chunhwashim' shown incompatibility among them forming the confrontation growth line against parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho). Fruiting body produced about $161.4{\pm}4.8g$ per bottle. And also the individual generation of fruit body is multiple than chunchu2ho as 40.9. The 'Chunhwashim' was cytoplasmic hybrid included hybrid DNA bands of parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho) by Random Amplified Polymorphic DNA (RAPD) primer, and mitochondria DNA band of monokaryotic ASI 2344 (chunchu 2ho) using mitochondria microsatellite DNA marker. This new cytoplasmic hybrid variety 'Chunhwashim' of oyster mushroom is characterized by multiple of individual generation and deeply grey color of pileus.

• The Korean Journal of Mycology
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• v.42 no.1
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• pp.86-90
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• 2014

Button mushroom (Agaricus bisporus) strains from diverse origins were compared in this study to obtain basic information on their growth and biochemical properties that are helpful for breeding. Among 31 dikaryotic strains tested, most strains showed better mycelial growth on oatmeal agar than on MEA and PDA. Mycelia of the mushroom strains revealed ${\beta}$-glucosidase activity the most clearly among the seven extracellular enzymes tested. All the strains showed protease activity, but ${\beta}$-glucosidase activity was found in 27 strains and xylanase activity was found in 30 strains. The activity of avicelase, CM-cellulase, amylase, and pectinase was detected in less than 20 strains. These results implied that enzymatic characteristics could be used as a criterion of strain selection for breeding study.

• Journal of Mushroom
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• v.13 no.2
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• pp.135-138
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• 2015

'Daejang 3ho' is a new cultivar of oyster mushroom for high productivity than that of Chunchu 2ho. This cultivar was bred by Di-mon mating between monokaryotic strains isolated from 'Daejang 1ho' and dikaryotic strain 'Jangan 5ho'. The color of pileus and the shape of pileus of new cultivar were dark gray and deeply funnel shape, respectively. The length and thickness of stipe were longer and thicker than those of Chunchu 2ho. It took 4~5 days to formation of primordia, that was a similar to Chunchu 2ho. The optical temperature of fruit body was $14-17^{\circ}C$. Yield of 'Daejang 3ho' was 13.9% higher than that of Chunchu 2ho. Analysis of RAPD by URP-primer of #03, #08, #10 and #11 showed that different band pattern those of other strains. There were no significant differences between the texture properties of 'Daejang 3ho' and Chunchu 2ho.

• Mycobiology
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• v.32 no.2
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• pp.88-94
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• 2004

Protoplasts of the wild type strain of Pleurotus osteatus were mutagenized with UV light, and 3,000 colonies were examined for abnormal mycelial and fruiting phenotypes. Forty one strains displayed variant phenotypes in mycelia and fruiting processes. The variant phenotypes were classified into 6 groups: (1) auxotrophic strains, which are incapable of growing on minimal media and can only grow when provided with their specific requirements; (2) abnormal vegetative strains, which grow very slowly on minimal and complete media; (3) primordiumless strains, which fail to develop to the formation of primordia; (4) maturationless strains, which form primordia, but do not form mature fruiting bodies; (5) specifically colored strains, which have Specific bluish grey or bluish white pileus; (6) poorly spored strains, which fail to produce basidiospore or which produce few spores. These variant strains may be useful in genetic breeding programs and for the studies of fungal development and genetics.

• Mycobiology
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• v.43 no.1
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• pp.81-86
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• 2015

To promote the selection of promising monokaryotic strains of button mushroom (Agaricus bisporus) during breeding, 61 progeny strains derived from basidiospores of two different lines of dikaryotic parental strains, ASI1038 and ASI1346, were analyzed by nucleotide sequencing of the intergenic spacer I (IGS I) region in their rDNA and by extracellular enzyme assays. Nineteen different sizes of IGS I, which ranged from 1,301 to 1,348 bp, were present among twenty ASI1346-derived progeny strains, while 15 different sizes of IGS I, which ranged from 700 to 1,347 bp, were present among twenty ASI1038-derived progeny strains. Phylogenetic analysis of the IGS sequences revealed that different clades were present in both the ASI10388- and ASI1346-derived progeny strains. Plating assays of seven kinds of extracellular enzymes (${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease) also revealed apparent variation in the ability to produce extracellular enzymes among the 40 tested progeny strains from both parental A. bisporus strains. Overall, this study demonstrates that characterization of IGS I regions and extracellular enzymes is useful for the assessment of the substrate-degrading ability and heterogenicity of A. bisporus monokaryotic strains.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.35-35
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• 2015

Pleurotus eryngii has recently become a major cultivated mushroom; it uses tetrapolar heterothallism as a part of its reproductive process. Sexual development progresses only when the A and B mating types are compatible. Such mating incompatibility occasionally limits the efficiency of breeding programs in which crossing within loci-shared strains or backcrossing strategies are employed. Therefore, understanding the mating system in edible mushroom fungi will help provide a short cut in the development of new strains. We isolated and identified pheromone and receptor genes in the B3 locus of P. eryngii and performed a functional analysis of the genes in the mating process by transformation. A genomic DNA library was constructed to map the entire mating-type locus. The B3 locus was found to contain four pheromone precursor genes and four receptor genes. Remarkably, receptor PESTE3.3.1 has just 34 amino acid residues in its C-terminal cytoplasmic region; therefore, it seems likely to be a receptor-like gene. Real-time quantitative RT-PCR (real-time qRT-PCR) revealed that most pheromone and receptor genes showed significantly higher expression in monokaryotic cells than dikaryotic cells. The pheromone genes PEphb3.1 and PEphb3.3 and the receptor gene PESTE3.3.1 were transformed into P5 (A3B4). The transformants were mated with a tester strain (A4B4), and the progeny showed clamp connections and a normal fruiting body, which indicates the proposed role of these genes in mating and fruiting processes. This result also confirms that PESTE3.3.1 is a receptor gene. In this study, we identified pheromone and receptor genes in the B3 locus of P. eryngii and found that some of those genes appear to play a role in the mating and fruiting processes. These results might help elucidate the mechanism of fruiting differentiation and improve breeding efficiency.