• Korean Journal of Microbiology
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• v.43 no.2
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• pp.147-149
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• 2007

White-rot fungi which degrade lignin can also degrade diverse recalcitrant compounds such as polymeric dyes, explosives, pesticides, and endocrine disrupting chemicals. Lignin degrading enzymes are involved in the degradation reactions, and introduction of foreign genes into a white-rot fungus is required in order to increase the degrading capacity. Genetic transformation experiment has been carried out in Irpex lacteus and Phlebia tremellosa to an antibiotic resistance. The transformation yields were 50-70 transformants/${\mu}g$ DNA and 15-25 transformants/${\mu}g$ DNA in I. lacteus and P. tremellosa, respectively. The stable replication of the plasmid was confirmed by PCR using the plasmid-specific primers, and many mutants were generated during this integration in both fungi.

• The Korean Journal of Mycology
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• v.49 no.1
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• pp.33-44
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• 2021

Lentinula edodes (Berk.) Pegler, the most produced mushroom in the world, is an edible mushroom with very high nutritional and pharmacological value. Currently, interest in the protection of genetic resources is increasing worldwide, and securing the distinction between new cultivars is very important. Therefore, the development of efficient molecular markers that can discriminate between L. edodes cultivars is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanbaekhyang and Sulbaekhyang). These markers were developed from whole genome sequencing data from L. edodes monokaryon strain B17 and resequencing data from 40 cultivars. A nucleotide deletion existed in scaffold 19 POS 214449 in Sanbaekhyang (GT→G), and a single nucleotide polymorphism changed in scaffold 7 POS 215801 in Sulbaekhyang (G→A). The restriction enzymes Hha I and HpyCH4IV distinguished Sanbaekhyang and Sulbaekhyang, respectively, from other cultivars. Thus, we developed two CAPS markers for the identification of the L. edodes cultivars Sanbaekhyang and Sulbaekhyang.

• Journal of Mushroom
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• v.12 no.3
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• pp.171-180
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• 2014

The primary objective of the present study is the characterization of the hybrids of monokaryon- monokaryon (mono-mono) crosses in mushroom breeding. We employed this technique for developing superior strains from Pleurotus species strains with 85 mono-mono intraspecific hybrids of 7 combinations between six Pleurotus ostreatus strains and one Pleurotus florida strain. In this study, the results of analysis on hybridization rate, nuclear DNA patterns, and colors and yields of fruit-bodies, are presented as follows. The crossability between mono-mono crossing ranges between 50 and 93.75%. The results of the analysis on the nuclear DNA patterns of 85 hybrid strains of mono-mono crosses share the nuclei of both parents, but their genetic similarities were predominated by either parent. The hybrid strain between P. florida and P. ostreatus showed patterns more similar to P. florida, while the hybrid strain between P. ostreatus and P. ostreatus either had patterns predominated by either parent strain. The fruiting body colors of the mono-mono crosses mostly had combined colors of both parents but showed the tendency of being more similar to that of either parent. 82% of the hybrid strain indicated similar fruiting body yields compared to both parent strains, while 0% was higher and 18% were lower than both parents. The present study was able to find out and suggest superior hybrid trains by identifying the nuclear DNA patterns of hybrids between Pleurotus species as well as the characteristics of their fruiting bodies. This study expects that the advantages of the mono-mono crossing are needs to be fully utilized in mushroom breeding and it is better to develop superior strains of Pleurotus species strains together with the mono-mono crossing.

• 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.

• Journal of Mushroom
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• v.5 no.2
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• pp.86-90
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• 2007

Jeakdan', a new variety of pink oyster mushroom was bred in Mushroom Research Institute, Gyeonggido Agricultural Research & Extension Services in 2006. This pink oyster mushroom was selected one which cross with monokaryon collected from strain KME20063 and KME20193. The major characteristics of the mushroom were showed a lot of pinheadings, the pink-colored and infundibuliform pileus. The optimum temperature for the mycelial growth was around $26{\sim}30^{\circ}C$ and that for the primordia and growth of fruitbody was around $18{\sim}23^{\circ}C$. Primordia and young fruiting bodies were bright pink but become less intensely colored while the mushroom were mature. Incubation period was required about 20 days in bottle culture and 21 days in bag culture. Number of valid stipes was 31.5/850cc bottle and 47.0/2kg bag. The fruiting body yield was 134.5g/850cc bottle and 350.0g/2kg bag.

• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.574-582
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• 1998

Five basidiocarps of Pholiota species have been collected from the areas of BubJu Temple for last two years, and identified to those of P. adiposa or Pholiota species. The taxonomy of these basidiocarps with the morphological aspects was compared with the analysis obtained from the polymorphisms of PCR-RAPD bands made after reacted with the random primers. The polymorphic variations were observed within the species of the basidiocarps, but not between genomic DNA's of the mycelia obtained and the basidiocarps. Several different bands made from the primers (28 and 36) in PCR-RAPD reactions were identified within the genus of Pholiota and speculated to be specific for the individual basidiocarp of P. adiposa collected. The primers employed here were considered to be very useful for distinguishing the individual isolates or basidiocarps collected from the fields. Also, the basidiospores were obtained from the sporeprints of the above basidiocarps as a simple agar and confirmed with observations of clamp connection under microscopes. The matings of them indicated the 'tetrapolar' type, being different from the 'bipolar' type reported by Japanese basidiocarps of P. adiposa or P. nameko. Based on our work, the edible fungi collected were speculated to be a new breeding resource for those of Pholiota commercialized in Japan.

• 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.

• 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.20 no.3
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• pp.178-182
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• 2022

Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

• Journal of Mushroom
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• v.20 no.2
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• pp.50-54
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• 2022

This study was conducted to reduce the phenomenon of the biased cultivation of certain mushroom varieties and to develop a competitive variety of Grifola frondosa. We developed the first Korean white commercial mushroom strain, 'Bakyeon', by crossing monokaryons derived from brown strains. We have collected and tested the characteristics of mushrooms from domestic and international genetic resources since 2018. We bred the unique domestic variety, 'Bakyeon', which has the following characteristics. The optimal temperature for mycelial growth was 25~28℃ and the optimal temperature for fruit body growth was 16~18℃. The new variety was similar to the control variety (Daebak) in terms of the pileus, which formed a pine cone shape, and the number of days of cultivation. The yield was 94.1 g/bottle, which was 23% lower than the 108.5 g/bottle yield of the control variety. When incubating the parent and control varieties, the replacement line was clear. Moreover, polymerase chain reaction analysis of mycelial DNA resulted in different band patterns between the parent and control varieties, confirming the hybrid species.