• Journal of Mushroom
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• v.11 no.1
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• pp.1-8
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• 2013

In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morphological characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in $K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, and $K4{\times}K6$. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better morphological characteristics through a special mating of L. nuda.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.187-193
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• 1998

The effects of medium, incubation temperature, incubation period, pH of medium and $CO_2$ ondition during mycelial growth were investigated to study the factors associated with the formation of oidia in Flammulina velutipes. Oidia formation was increased when mycelial growth was poor, while oidia formation was inhibited in optimum condition of mycelial growth. Mating type of oidia was investigated to examine the effect of oidia formation on dikaryotic strain. Di-mon matings between oidia strains and original dikaryotic strain were carried out. Monokaryotic strains derived from oidia showed only one genotype. Seventy percent among Dimon mating strains showed slow mycelial growth and low yield of fruit-body, but others showed similar or high mycelial growth and yields in comparision with original dikaryon strain. One strain from di-mon mating demonstrated some differences in isozyme band pattern.

• Journal of Mushroom
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• v.13 no.3
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• pp.228-232
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• 2015

'Nongjin-go' is a new breed strain of Lentinula edodes, saw-dust bag variety. It is a cross combination of dikaryon Lentinula edodes ASI 3305mut and monokaryon L5-16 of L. edodes ASI 3305(Sanjo701ho). We crossbred them by 2011 and verified productive capacity from 2012 to 2013 in Rural Development Administration. Optimum temperature of mycelial growth is $30^{\circ}C$ and it of fruit-body primodium formation is range from 15 to $23^{\circ}C$. Nongjigo is agricultured at mid-high temperature well. Fruiting body is platy-hemisphere, light brown and centralizing. And bast is formed around edge of pileus. Yield productions per period is regular than 'Sanjo701'. Plastic bag culture medium is 1.5 kilogram and culture periods are 90~100 days. As its browning of pileus in culture is a little slow, Light and ventilation is needed a lot in light-culturing. Humidity is controlled properly for its color in fruit-body growing. Tested culture medium is consisted of 80% Oak-Tree saw-dust and 20% rice-bran.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.249-253
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• 2016

The inky cap, Coprinellus congregatus, produces several laccase isozymes during its life cycle: both hyphal tip laccase and sclerotial laccase are involved in the fungal development. When this fungus was transferred to an acid liquid medium (pH 4.0-4.5), a new laccase was synthesized and secreted into the culture supernatant. In order to examine its regulation by external pH, green fluorescent protein gene was ligated at the downstream of the promoters having different lengths. These expression vectors having different promoter lengths were inserted into the fungal transformation vector, pBARGEM7-1. These expression vectors were introduced to the mating type a1 and a2 monokaryons, and the transformants were selected by the phosphinothricin resistance. Transformant a1 (a1TF) and transformant a2 (a2TF) were mated with each other to generate homozygotic dikaryon transformants. All these transformants were grown in neutral liquid medium for 5 days, and then the whole cell homogenates were transferred to the acidic liquid medium (pH 4.1). After 36 h incubation at $25^{\circ}C$, cells were harvested for the analysis of GFP expression. GFP expression was detected in the transformant having full-length promoter (2.0 kb), but other transformants having shorter length promoter (shorter than 1.29 kb) failed to show the fluorescence. Therefore, the acid-responsive element in the laccase promoter should be localized between -2.0 kb ~ -1.29 kb region.

• The Korean Journal of Mycology
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• v.45 no.2
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• pp.114-120
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• 2017

Lentinula edodes is an edible mushroom that is mainly cultivated in Asian countries. Recently, new cultivars of this mushroom have been developed in Korea; variety protection is very important, so the development of efficient molecular markers that can distinguish each variety is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanmaru 1ho and Chunjang 3ho). These markers were developed from whole genomic sequencing data from L. edodes monokaryon strain B17 and resequencing data from 10 dikaryon strains. A single nucleotide polymorphism changed in scaffold 9 POS 1630048 in Sanmaru 1ho($G{\rightarrow}T$), and in scaffold 13 POS 920681 in Chunjang 3ho ($G{\rightarrow}A$). The restriction enzymes TspR I and Xho I distinguished Sanmaru 1ho and Chunjang 3ho, respectively, from other strains. Thus, we developed 2 CAPS markers for the identification of the L. edodes cultivars Sanmaru 1ho and Chunjang 3ho.

• Journal of Mushroom
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• v.10 no.3
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• pp.109-114
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• 2012

To develop new variety of oyster mushroom, 63 intra-specific hybrids between the strain Suhan and #Nongi201 were developed using hyphal anastomosis technique in 2004. The Po2008-275 hybrid between the dikaryon strain 04-154(Suhan x #Nongi201) and the monokaryon strain derived from ASI2487 were developed using hyphal anastomosis in 2008. The Po2008-275 was shown the best cultural characteristics, selected to be a new variety and named as 'Guseol'. The new commercial strain, 'Guseol' had dark grey pilei and grows well under spring and autumn conditions in Korea. The fruiting bodies of 'Guseol' were of an excellent quality in that not only the stipe was thick and long but also the pileus was small and hard. The optimum temperatures for mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $10{\sim}16^{\circ}C$, respectively. Time period required for the initiation of the first fruiting body was about 3 to 5 days depending on the temperatures. The shape of fruiting body was thin funnel shape. Fruiting body production per box($43{\times}43{\times}12cm$) was about $1545{\pm}400.9g$ which was almost 137% quantity compared to that of parental strain 04-154. Relatively low temperature incubation ($11^{\circ}C$) resulted in the development of better quality of 'Guseol' mushrooms. When two different media including potato dextrose medium and mushroom complete medium were compared, the mycelial growth of this mushroom were much faster in mushroom complete medium. Similar results were observed with other variety '#Chunchu2'. Analysis of the genetic characteristics of the new commercial strain 'Guseol' showed a major DNA profile as that of the parental 04-154 when primer URP 1, primer URP 2 and primer URP 5 were used, but different to '#Chunchu2' that was used as a control. This new variety of the dark grey oyster mushroom had smart and high quality image that corresponds well to "health food". We therefore expect that this new strain will satisfy the consumers demand for variety and excellent mushrooms.