• 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.

• The Korean Journal of Mycology
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• v.14 no.1
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• pp.17-23
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• 1986

The mating system of monokaryotic isolates in Pleurotus sajor-caju was controlled by two incompatibility factors A and B of tetrapola mating system. The mycelia of dikaryotic isolates grew faster than those of their component monokaryons, but no correlation between dikaryotic and their component monokaryotic isolates was found. The primodia formed well on the potato dextrose agar (PDA) media not only in the dikaryotic isolates but also in the monokaryons under irradiated conditions. The dikaryotic isolates produced normal sporophores; however, the monokaryotic isolates produced abnormal sporophores when they were cultivated with sawdust substrates. Some dikaryotic isolates derived by mating between monokaryotic isolates showed high yields of sporophores more than those of parental strains. Both the dikaryotic mycelial growth rate and primodia formation number on the PDA plate showed significant correlation with its sporophore products on sawdust substrates.

• Journal of Mushroom
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• v.10 no.2
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• pp.49-56
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• 2012

Two strains which have good traits in quality and speedy growing were selected based on previous report(Ryu, 2007) to breed a new strain carrying the traits in it. KNR2322 carrying high quality and KNR2503 carrying a speedy growing trait were cultivated to harvest monokaryons. The monokaryotic mycelia were characterized in growth rate, color, and stability. Selfing of monokaryons from KNR2322 and crossing between KNR2322- and KNR2503-derived monokaryons were performed. Two parental lines, $KNR2322-4{\times}37-6{\times}12(B)$ showing thremmatological values in weight and quality, 68.5g and 6.5 and $KNR2322-30{\times}KNR2503-60(A)$ was harvested in 13.5 days after scraping old medium were selected. Crossing between A and B resulted in two high quality and speedy growing strain, A8B8 and A8B10. Their days for harvest after scraping, weight, and quality were 14.0 days, 88.8g and 92.7g, 7.0 and 7.1. Originality tests were conducted with genomic DNAs of A8B8 and Keuneutari 3ho(commercial strain) by DNA fingerprinting a confrontation culture. The results showed polymorphism and a inhibition line between them. It suggested that the new strain have originality from the commercial strain. A8B8 have been registered in Korea Seed and Variety Service with as "Saesongi 1ho".

• Journal of Mushroom
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• v.20 no.4
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• pp.199-207
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• 2022

The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

• Journal of Mushroom
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• v.7 no.4
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• pp.141-149
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• 2009

The sexual incompatibility system was investigated in the new strain Pleurotus ostreatus 'Miso'. Matings among monosporous progeny collected from fruiting bodies revealed that the new oyster mushroom is heterothallic and has a tetrapolar mating system. Single basidiospore isolates that did not have clamp connections were determined to be monokaryotic. Compatible parings of monokaryons were distinguishable macroscopically by the rapid growth and gross morphology to some extent but they were distinguished obviously by the presence of clamp connections from incompatible pairings under microscopic examination. Rarely dikaryotization was unidirectional with clamp connections only at the margin of one side of mating. In tetrapolarity of matings, four kinds of patterns, A1B1, A1B2, A2B1, and A2B2, were decided depending on compatibility tests in a parental strain with a mating factor of A1A2B1B2. The mating tests with the tester strains were performed for the detection of mating types on the rest of selected monokaryons.

• Journal of Mushroom
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• v.21 no.3
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• pp.179-184
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• 2023

The development of automated bottle cultivation systems has facilitated the large-scale production of Pleurotus ostreatus, a commonly cultivated oyster mushroom species in South Korea. However, as the consumption of this product is decreasing and production quantities are exceeding demand, farmers are seeking various other mushroom types and cultivars. In response to this, we have developed a new oyster mushroom cultivar named 'Sena'. This high-yielding cultivar has a white pileus and excellent quality. The white oyster mushroom cultivars 'Goni' and 'Miso' were selected as parental strains from the genetic resources of the National Institute of Horticultural and Herbal Science's Mushroom Division. By crossing their monokaryons, hybrids were developed and subjected to cultivation trials and characteristic evaluations to select the superior cultivar. The optimal temperature for 'Sena' mycelial growth is 25-30℃, with inhibition occurring at temperatures above 30℃, whereas the temperature for mushroom growth is 14-18℃. The mushrooms grow in clusters, with the white pileus having a shallow funnel shape. Optimal mycelial growth occurs in malt extract agar medium. When cultivated in 1,100 cc bottles, the 'Sena' cultivar had 35 available individuals, surpassing the number 16 available from the control cultivar 'Goni'. The yield per bottle also increased by approximately 157 g, a 24% increase over the control cultivar amount. When 300 g samples of harvested mushrooms were packed and stored at 4℃ in a cold storage facility for 28 days, the weight loss rate of 'Sena' was approximately 4.22%, lower than that of 'Goni'. Moreover, the changes in pileus and stipe whiteness (measuring 6.99 and 8.33, respectively) were also lower than those of the control cultivar. Since the appearance of a white cap is crucial for quality assessment, the 'Sena' cultivar is superior to the 'Goni' cultivar in terms of both weight and quality after undergoing low-temperature storage.

• Journal of Mushroom
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• v.17 no.4
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• pp.224-229
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• 2019

The two most common mushroom species grown in Korea are pearl oyster mushroom (Pleurotus ostreatus) and king oyster mushroom (P. eryngii). In recent years, the production of king oyster mushroom greatly increased due to the automation of the cultivation facilities, and it became a major export mushroom owing to its excellent shelf life. However, the increase in the production of king oyster mushroom led to a decline in its market price; thus, necessitating the development of new mushroom species that could replace king oyster mushroom, to diversify the mushroom market for the benefit of both, the producers and the consumers. The Mushroom division at the National Institute of Horticultural & Herbal Science (NIHHS) reported the development of a new interspecific hybrid between P. ferulae and P. tuoliensis, referred to as 'Creamy.' Two parental strains KMCC00430 (Bisan2ho, P. ferulae) and KMCC00461 (P. tuoliensis) were selected based on the results of genetic resource analysis, and their monokaryons were collected. About 1,000 Mon-Mon crosses were performed and 73 of them were selected. Following repeated cultivation tests and strain analyses, we selected strain 7773, which had a bright creamy pileus and a thick straight stipe, and named it 'Creamy.' Optimum temperature for mycelial growth of Creamy was 25-30℃, and that for fruiting body growth was 16℃. The pileus, which had a brighter creamy color, was small in size with a diameter of 61.2 mm. Although it was cultivated in suboptimal conditions, such as low temperature and high CO₂ concentration, Creamy was characterized by its straight and smooth stipe. Field production tests and further analyses indicated that the yield of Creamy was 5% higher than that of Baekhwang. It is expected that Creamy, the new interspecific hybrid with a bright creamy pileus and a pleasant flavor, will help create new opportunities for mushroom farmers and diversify the mushroom market.