11 종의 목이버섯 자실체로부터 얻은 50% EtOH추출물을 사용하여 지방전구세포인 3T3-L1의 지방세포 분화억제 효과를 확인한 모든 strain에서 억제효과가 존재하였다. 털목이에서는 21001 균주가 가장 우수하였고 흑목이에서는 21002 균주가 억제효과가 우수하였다. 전반적으로 털목이가 흑목이에 비해 지방세포 분화억제 효과가 우수하였다. 또한 Adipocyte RNA 발현량과 protein 발현량 측정 실험에서도 털목이 21001 균주가 지방세포 분화억제효과에 있어서 가장 우수하였다. 털목이 21001 버섯을 사용하여 동물실험을 진행한 결과 털목이 자실체 분말이나 50% EtOH 추출물을 급여한 실험에서 체중 감소효과가 나타났으며, 특히 EtOH 추출물 0.2%를 급여한 군에서 유의하게 감소함을 확인하였다(*P<0.05). 동물 희생 후 적출한 간 중량 및 부고환지방량과 혈청 내 중성지방 및 총 콜레스테롤, 혈당, ALT, AST 등도 털목이버섯 21001 EtOH 추출물 0.1% 이상 급여 시 고지방식이만을 급여한 군에 비해 유의차(*P<0.05 이상) 있게 감소함을 확인하였다.

The Pleurotus tuoliensis is a mushroom named Chinese ‘Bai-Ling-Gu’ from species of Pleurotus nebrodensis. we are mated for selection of cytoplasmic hybrid by mitochondria microsatellite marker and the method of Mon-Mon mating between monokaryotic strains derived from Pleurotus ferula ASI 0629 (Beesan No.1) and Pleurotus tuoliensis ASI 0663 (Baekryung 20). The cytoplasmic hybrids were identified 8 strains contained nuclear DNA bands of 'Baekryung 20' and mitochondrial DNA of monokaryon strains derived from Pleurotus ferula ASI 0629 (beesan No.1). The KiMB-Plft-15-81 was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Baekwhang'. The 'Baekwhang' cab be grown without an extra after-ripening period, can utilize bottle cultivation of Pleurotus eryngii. And also the ‘Seolwon’ is similar to the existing Pleurotus tuoliensis in shape and physiological characteristics, formed high stipe. We therefore expect that this new strains will substitutional goods of Pleurotus eryngii.

There are three kinds of Pleurotus eryngii : P. eryngii var. eryngii (P. eryngii), P. eryngii var. ferula (P. ferula : A-Wei-Mo) P. eryngii var. nebrodensis (P. nebrodensis : Bai-Ling-Gu). We bred P. ferula using Di-Mon mating and physiological assay and selected as follow (1) ‘Beesan No.1’ produced high yields, (2) ‘Beesan No.2’ was excellent morphological shape and anti-adipogenic Activit, (3) ‘Ergo’ included high ergothioneine such as anto-oxidant material, (4) ‘GW10-45’ included highest ergothioneine such as anto-oxidant material, (5) ‘Maeksong’ included high ergothioneine and effect of anti-inflammatory. We suggest that there are able to food-medicine materials.

The purpose of this study is to find out a food material having anti-obesity effect and suggest the effect of the material to people who are interested in anti-obesity. For this study, Auricularia (wood ear) has been used. It is world widely spread mushroom, especially in east north Asia countries. It is also called as wood ear in western countries because of its shape. The most considerable fact is that Auricularia is being used for popular food materials for cooking because of its good taste. Totally 11 strains of Auricularia were offered by Doctor Kim, Kil-ja in Jeollanam do Agricultural Research & Extension Services. There were 8 strains of Auricula-polytricha, and 3 strains of Auricula-judae. Each Auricularia strains were extracted with 50% Ethanol for 12 hours in 40°C and then 11 strains were prepared for in vitro study. In vitro study focused on the inhibition effect on adipocyte differentiation of Auricularia spp. 3T3-L1 (preadipocyte cell) was used for identifying the inhibition effect on adipocyte differentiation. Before that, MTT assay was used for measurement of cell viability. The result is essential for fixing the treat concentration of the extracts. After adipocyte differentiation, cells were stained with Oil Red O solution to dye differentiated adipocyte cells. Spectrophotometer was used to quantify the adipocyte cells. There are few adipocyte differentiation markers. From the markers, this study selected four markers (PPAR γ, FABP4(ap2), CD36, C/EBPα) and used them to measure mRNA expression using real-time PCR and Western blot experiment. As a result, this study found out that all the extract of strains has inhibition effects on adipocyte differentiation, especially the adipocyte differentiation control probability of Auricula-polytricha strain 21001 was 24.6%. Auricula-judae strain 21002 (19.3%), Auricula-polytricha strain 249 (14.0%) followed. The result concerning the inhibition effect was identified through Western blot experiment as well. As a further study, in vivo study is on progressing. 56 ICR mice (6w, male) are being used for the animal study. It consists of 8 groups. One is control, another is positive control, the other groups are Auricula-polytricha strain 21001 treatment group. Different concentration of powder or extract of Auricularia is treated in each treatment group. We are anticipating that the result of this further study will be correspond to that of in vitro study.

Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From SSR-enriched library, 490 white colonies were randomly selected and sequenced. In the 490 sequenced clones, 85 clones (17.35%) were redundant. Among the remaining 405 unique clones, 201 clones (49.6%) contained microsatellite sequences. As a result, 12 primer pairs produced reproducible polymorphic bands within diverse 4 strains and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, while for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among 32 Flammulina velutipes strains based on SSR data were generated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

‘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 25oC, respectively, for primordia formation where in fruiting body development occurred from 11±1 days at 14oC and 14±1 days at 7oC. The length of the pilei and stipes of the Baekseung harvested in optimal stag were 11.3±0.4 and 89.2±7.1 mm, respectively, whereas the values for Uri1ho were 10.7±1.0 and 91.3±20.8 mm, respectively. The yield of the Baekseung and Uri1ho strain grown on the sawdust substrate was 153.7±12.5 and 139.8±17.8 g, 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.

Worldwide production of mushrooms has been increasing by 10–20% every year. Recently, Pleurotus eryngii and P. nebrodensis have become popular mushroom species for cultivation. In particular, China exceeded 8.7 million tons in 2002, which accounted for 71.5% of total world output. A similar trend was also observed in Korea. Two kinds of mushrooms— Gumji (金芝; Ganoderma) and Seoji—are described in the ancient book 'Samguksagi' (History of the three kingdoms; B.C 57~A.D 668; written by Bu Sik Kim in 1145) during the Korea-dynasty. Many kinds of mushrooms are also described in more than 17 ancient books during the Chosun-dynasty (1392~1910) in Korea. Approximately 200 commercial strains of 38 species of mushrooms were developed and distributed to cultivators. The somatic hybrid variety of oyster mushroom, ‘Wonhyeongneutari,’ was developed by protoplast fusion, and distributed to growers in 1989. Further, the production of mushrooms as food was 199,829 metric tons, valued at 850 billion Korean Won (one trillion won if mushroom factory products are included) in 2015. In Korea, the major cultivated species are P. ostreatus, P. eryngii, Flammulina velutipes, Lentinula edodes, Agaricus bisporus, and Ganoderma lucidum, which account for 90% of the total production. Since mushroom export was initiated in 1960, the export and import of mushrooms have increased in Korea. Technology was developed for liquid spawn production, and automatic cultivation systems led to the reduction of production cost, resulting in the increase in mushroom export. However, some species were imported owing to high production costs for effective cultivation methods. In academia, RDA scientists have conducted mushroom genome projects since 1997. One of the main outcomes is the whole genome sequencing of Flammulina velutipes for molecular breeding. With regard to medicinal mushrooms, we have been conducting genome research on Cordyceps and its related species for developing functional foods. There are various kinds of beneficial substances in mushrooms; mushroom products, including pharmaceuticals, tonics, healthy beverages, functional biotransformants, and processed foods have also became available on the market. In addition, compost and feed can likewise be made from mushroom substrates after harvest.

The primary objective of the present study is the characterization of the hybrids of dikaryon-monokaryon(di-mono) and monokaryon-monokaryon(mono-mono) crosses in mushroom breeding. We employed this technique for developing develop superior species from Pleurotus spp. varieties with 56 Di-mono intraspecific hybrids of 14 combinations and 85 mono-mono intraspecific hybrids of 7 combinations between six Pleurotus ostreatus varieties and one Pleurotus florida variety. In this study, the results of analysis on hybridization rate, nuclear and mitochondrial DNA patterns, and colors and yields of fruit-bodies, are presented as follows. In di-mono crosses, hybrids between Pleurotus ostreatus and Pleurotus florida showed 100% of crossability as seen in those between Pleurotus ostreatus and Pleurotus ostreatus indicating that nuclei and mitochondria of a dikaryon migrated to a recipient of monokaryon. The mitochondrial DNA patterns of the hybrid strain were composed of 75% dikaryon donors and 25% monokaryon recipient. The crossability between mono-mono crossing ranges between 50 and 93.75%. 82.4% of the hybrid strain showed mitochondrial DNA patterns predominated by either parent, while the remaining 17.6% had recombinant or half-and-half combined patterns of both parents.

The primary objective of the present study is the characterization of the somatic hybrids of dikaryon-monokaryon (di-mono) crosses in mushroom breeding. We employed this technique for developing superior strain from Pleurotus ostreatus strains with 56 intraspecific hybrids of 14 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 morphology of fruit-bodies, are presented as follows. In di-mono crosses, somatic hybrids among Pleurotus strains showed 100% of crossability as seen in those among Pleurotus strains indicating that nuclei of a dikaryon migrated to a recipient. 89.3% of the somatic hybrids among Pleurotus strains were similar to the donor dikaryons, and 10.7% had combined DNA patterns of both parents. In the 14.3% di-mono cross between P. ostreatus and P. florida, the nuclear DNA patterns of the all hybrid strain showed the same or similar patterns compared to the donor dikaryons. 75.0% of the hybrid between P. ostreatus and P. ostreatus were similar to the donor dikaryons; 10.7% had combined DNA patterns of both parents. 82.2% of fruiting body morphology of the hybrids among Pleurotus strains were similar to the dikaryons, and 17.8% had combined DNA patterns of both parents. All hybrid strains between dikaryon P. florida and monokaryon P. ostreatus showed the fruiting body whose colors were similar to those of the dikaryon, while the hybrids between dikaryon P. ostreatus and monokaryon P. florida were all showed combined colors of both parents but are more similar to the dikaryon. Therefore, the fruiting body color of P. florida tends to be generally dominant. The present study was able to find out and suggest superior hybrid strains by identifying the nuclear DNA patterns of hybrids between Pleurotus strains as well as the characteristics of their fruiting bodies. This study expects that the advantages of the di-mono crossing are needs to be fully utilized in mushroom breeding and it is better to develop superior strains of Pleurotus strains.

Microsatellite loci are increasingly used as markers in the human, animal and plant genomes. Being highly mutable, microsatellite regions are able to differentiate between related taxa, even at the level of individual isolates in a single species. Studies on mushroom population structure, gene flow and dispersal between natural and cultivated species have become central in breeding programmes and the knowledge of new polymorphic, codominant markers will be a promising avenue to exploit wild genetic resources. The molecular phylogeny in 50 different commercial cultivated strains of Pleurotus eryngii using PCR amplification with URP primers and mitochondrial microsatellite primer was studed. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD analysis techniques were able to detect genetic variation among the tested strains. With these isolated PCR amplification with URP primers we intend to analyse the population structure of the P. eryngii species complex and investigate the structure of the basidiomycete genome which deserves. A few single-locus microsatellite markers have been isolated in Pleurotus eryngii and Pleurotus ferulae. This technique is useful in those species where microsatellite loci are rare in the mitochondria.

The mating-type genes control formation of the dikaryon from two haploid strains. These genes are now used in mating-type-assisted breeding programs for economically important mushrooms, especially the oyster mushroom, Pleurotus ostreatus, aiming at high-yield and high-quality standard mushroom production. However, it improves the breeding program when the breeder is able to quickly identify compatible strains in a given set of progeny. The two mating factors with their mating-type loci are used as markers for breeding and have been incorporated in a chromosome mapping investigation. The linkage maps include not only genetic markers such as the mating types that can be cored, but also molecular markers such as PCR-assisted approaches, e.g. RAPD analyses, or RFLP markers. Once mating-type genes within progeny may be more easily identified by the use of PCR-directed cloning of partial mating-type genes. We analyzed homeodomain (HD1 and HD2) and pheromone receptor(rcb1, 2 and 3) genes as molecular markers for breeding using mating type A and B of Pleurotus eryngii and Pleurotus ferulae by direct PCR.

This study was conducted to develop superior hybrids of Pleurotus ostreatus with di-mono and mono-monoka-ryon crosses. Random Amplified Polymorphic DNA-PCR (RAPD-PCR) was used to compare its mitochondrial DNA profiles of hybrids using specific primers designed from microsatellite markers of Pleurotus salmoneo-stramineus. A total of fifty-six dikaryon-monokaryon hybrids were sampled for RAPD-PCR experiments and the results show that twenty-four hybrids were dikaryon and thirty-one hybrids were monokaryon. Interestingly, one hybrid was an intermediate form with the DNA profiles that are different from those of its parents. The DNA profiles from eighty-eight monokaryon-monokaryon hybrids were also analyzed by RAPD-PCR. The results of mitochondria DNA profiles show that seventy-one hybrids are the same to one of their parents, but seventeen hybrids show DNA profiles of both parents.

World wide mushroom productions have been increased to 10-20% and more various mushrooms have been attempted to cultivate. Similar trends were also observed in Korea. More diverse mushroom varieties such as Pleurotus eryngii, Hericium erinaceus and Agrocybe aegerita have been attempted to cultivate in larger areas. In these days, 235 varieties of 33 different mushroom species have been cultivated. However only 50 varieties were protected by the law. Since 1960s, mushroom industry had been considered as one of the major export industries in Korea and mushroom production has been rapidly increased. In 2008, total mushroom production was about 198,209 ton, which were about 800 billion won (one trillion won if include mushroom factory products). Major cultivated species are Pleurotus ostreatus, P. eryngii, Flammulina velutipes, Lentinula edodes and Agaricus bisporus, which cover 90% of total production. We believed that the protection of mushroom vavarieties by the law is one of the main problems to be solved. Also, more studies to develop better mushroom production methods with low cost and improve the distribution structures are certainly required. In these days, the export of mushroom has been increased in Korea. We imported more mushrooms than exported for last 4 years, however, it has been changed that export is getting bigger in these days. Because we developed the liquid spawning and automatic cultivation systems, which lead the reduction of mushroom production cost and resulted in the increasement of export. If we develop better post-harvest system, it is certain that the mushroom export should be more important in the future. As an alternative way, some Korean companies are planning to build the plants in main export countries. Mushroom industry has promising future because mushroom is good for your health.