A new commercial strain “Mongdol” of oyster mushroom was developed by hyphal anastomosis. It was improvedwith hybridization between monokaryotic strain derived from Pleurotus ostreatus ASI 0627 and dikaryotic strain derived from P.ostreatus ASI 2929. The optimum temperature of mycelial growth and fruiting body development were 25~30oC and 12~18oC,respectively. When two different media including PDA (potato dextrose agar medium) and MCM (mushroom complete medium)were compared, the mycelial growth of this mushroom was faster in MCM than in PDA. Similar result was observed with thecontrol strain P. ostreatus ASI 2504. Analysis of the genetic characteristics of the new cultivar “Mongdol” showed a different DNAprofile as that of the control strain ASI 2504, when RAPD (Random Amplified Polymorphic DNA) primer URP3 and URP6 wereused. Fruiting body production per bottle was about 106 g using demonstration farms. The color of pileus was blackish gray andthe stipe was long. Therefore, we expect that this new strain “Mongdol” will satisfy the consumer’s demand for high qualitymushrooms.

‘Baekjung’ adaptable to high temperature was made by crossing between monokaryon derived from selfing of brownstrain and monokaryon derived from Korea white strain. In the condition that temperature is maintained at 10oC without lowtemperature of 4oC suppressing treatment and wrapping during cultivation period, it showed good productivity thanUri1ho(control). The optimum temperature of mycelial growth was 30oC and that of fruiting body initiation and developmentwere 14oC and 7oC, respectively. The days for the fruiting and yield were 7days and 277±11.2g per 1,100ml bottle,respectively. This variety needed high concentration of carbon dioxide up to 4,000 ppm for the good quality.

Twenty Inter simple sequence repeat (ISSR) primers were used to assess genetic diversity of 64 Agaricus strainsincluding 45 A. bisporus strains and other 19 Agaricus spp. ISSR primers, (GA)T, (AG)YC, (GA)C and (CTC) amplified PCRpolymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted forUPGMA cluster analysis. The varieties, Saea, Saedo, Saejeong and Saeyeon that have recently been developed in Korea wereinvolved in the same group with closely genetic relationship of coefficient similarity over 0.92, whereas, other Korean strains weregenetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese. Furthermore, ISSR-PCRpolymorphism could potentially be used to identify homokaryon isolates.

느타리버섯(Pleurotus ostreatus)의 품종 육성을 위해서는 많은 노력과 시간을 필요로 하고 있어, 최근에는 분자육종방법으로 마커를 이용하여 빠르고 정확하게 교잡주를 선발하고 있다. 본 실험에서는 육종효율을 증진하기 위해 교잡효율이 높은 Di-Mon 교배방법으로 부친의 핵과 모친의 미토콘드리아를 가진 세포질전환 교잡주를 선발하였다. 느타리버섯 수한3호와 흑변이체의 교잡종인 ‘다굴’의 이핵균주에 춘추2호의 단핵균주와 교잡하여 120여개의 교잡주를 얻고 여기에 1차로 미토콘드리아 microsatellite DNA 마커 (MtPO1)를 이용하여 단핵균주 춘추2호의 미토콘드리아 밴드를 형성하는 교잡주 80개를 1차 선발하였다. 선발된 80균주중에서 URP 프라이머를 이용하여 ‘다굴’의 2핵균주 핵 DNA 패턴을 가진 26교잡주를 선발하였다. 이 중에서 수량성이 우수한 3개 교잡주의 자실체 특성을 조사한 결과 초발이일수가 2일이나 빠르면서 수량성이 좋은 품종을 선발하여 ‘천화심’이라 명명하였다. 천화심은 수량이 높으면서 여름에도 재배가 가능해 춘추2호의 약점을 보완함으로써 춘추대체품종으로도 재배가 가능하리라 본다.

버섯 세균갈성색무늬병원균인 Pseudomonas tolaasii에 대한 길항미생물로 보고된 Pseudomonas azotoformans HC5 균주의 배양적 특성과 대량 배양을 위한 최적배양조건을 설정하였다. HC5 균주의 생육온도는 10~20oC, pH는 6.0~9.0의 범위에서 왕성한 생육을 보였다. 대량배양을 위한 효율적인 영양원 선발을 위하여 기본배지에 탄소원 fructose 등 18종, 무기질소원 NH4Cl 등 6종, 유기질소원 peptone 등 6종 그리고 아미노산 asparagine 등 11종을 각각 1%씩 첨가하였고, 무기염류 13종을 1 mM 농도로 첨가하여 각각에 대한 생육에 미치는 영향을 조사하였다. 또한 선발된 각각의 영양성분들에 대한 최적 농도를 조사하기 위하여 각각의 성분을 최소 0.1%에서 최대 4.0%까지 배지에 첨가하여 배양 후 생육정도를 조사하였다. 그 결과, 대량배양을 위한 생육최적조건은 온도 15oC, pH 6, 탄소원 0.6% adonitol, 유기질소원 1.5% yeast extract, 무기질소원 0.8% NH4H2PO4,아미노산 0.2% asparagine 그리고 무기염류는 5 mM MgSO4에서 왕성한 생육을 보였다.

팽이버섯과 느타리버섯의 색깔이 다른 계통과 부위별로 갓과 대로 나누어 아미노산 및 유리아미노산 분석을 실시 하여 성분 조성과 함량을 비교하였다. 그 결과 버섯 종류 와 계통, 그리고 부위에 따라 서로 다른 아미노산 종류가 검출되었다. 팽이, 느타리 모두에서 tryptophane은 검출되 지 않았으며, 느타리에서는 특이적으로 alanine이 검출되 지 않았다. 또한 팽이에서는 glutamic acid 함량이 가장 높았으며 methionine과 phenylalanine은 극히 적게 검출 되었다. 느타리버섯에서는 glutamic acid 함량이 가장 높 았고 phenylalanine과 cysteine이 낮았으며 특이하게 흑색 변이계통에서 leucine이 검출되지 않았다. 또한 부위별로 는 팽이와 느타리 모두 아미노산 종류별 비율은 동일한 경향을 보였으며, 일반적으로 대보다는 갓에서 아미노산 함량이 더 높은 경향을 보였다. 유리아미노산 분석 결과 에서는 버섯계통별로 검출되는 아미노산 함량이 서로 달 랐으며, 버섯의 갓과 대에서도 검출되는 아미노산 조성이 일치하지 않는 것으로 나타났다.

수집한 46개의 균주를 배양하여 rDNA의 ITS 염기서열 분석으로 유전적 계통분류 한 결과, Heiricum속이 아닌 균 주가 2균주가 있었음을 확인하였다. 본 결과를 바탕으로 효능이 있는 것으로 밝혀진 H.erinaceus와 가까운 유연관 계를 지니는 종들을 위주로 계통별로 균사 생장실험, 자실 체 발생조사 등을 연구할 예정이다. 또한 H.erinaceus은 아니지만, 생리활성효능이 뛰어난 종의 유무를 테스트하여 기존의 품종보다 효능 면에서 뛰어난 균주를 선발 교잡할 예정이다. 이와 같은 연구를 통해 우수품종을 선발하여 최 적 환경조건 테스트까지 거친다면 노루궁뎅이 버섯 소비 시장 구축에 도움이 되리라 생각된다.

수집된 주름버섯류의 233점의 phylogenic relationship 을 분류하고 자실체형태를 조사하였다. 리보좀 DNA의 ITS 부분의 염기서열을 분석을 통해 38점이 다르게 동정 되었다. 수집된 주름버섯의 계통수에서는 양송이와 여름 양송이가 group A 내에서 주름버섯은 group B에서 가까 운 유연관계를 보였다. 또한 수집된 국가별 및 양송이내 갓색에 따른 유연관계가 있는 지 알아보았으나, 그 차이 는 없었다. 수집된 주름버섯류의 자실체 특성을 조사하기 위해 1차, 2차 두 번 재배를 하였다. 이 결과로 주름버섯 류 수집균주의 재배의 유무를 알 수 있었고, 버섯 종류와 시기에 따른 수확량 및 자실체의 특성을 비교를 통해 주 름버섯이 2차 재배시 수확량 및 경도에서 우수한 특성을 보이며, 양송이 중에는 비백색양송이가 단단하다는 것을 알 수 있었다. 또한, 여름양송이의 초발이 소요일수와 갈 색양송이의 갓 색이 외부환경에 영향을 받는 다는 것을 보여주었다.

Mushroom is cultivated as one of the major economical crops in many areas in Korea. The total production has steadily increased from approximately 198,563 M/T in 2009 to 208,941 M/T in 2012. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Trichoderma harzianum is the causal agent of brown blotch disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of brown or cream lesions on pileus and stipe. These lesions are slightly concave spots and can be round or spreading. Antagonists against Trichoderma harzianum, CAM33 were selected and their control efficacy of green mould disease was investigated in this study. The CAM33 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus methyrotrophicus. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Trichoderma harzianum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Trichoderma harzianum. Control efficacy of browning disease of strain CAM33 treatment was 77% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 3.0% Saccharose, 1.5% Soytone, 1.0% (NH4)2HPO4, 10 mmol MgSO4, and 2.0% Glutamic acid at pH 6.0 at 25°C. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Trichoderma harzianum.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 10,996 M/T in 2012. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Cladobotryum mycophilum is the causal agent of cobweb disease of commercial mushrooms. Early symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps. Late symptoms progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing, and covered entirely fruiting bodies. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Cladobotryum mycophilum, the most destructive pathogen of cultivated mushrooms. The HC7 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus altitudinis. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Cladobotryum mycophilum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Cladobotryum mycophilum. Control efficacy of browning disease of strain HC7 treatment was 78% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 3% Soluble startch, 10% Soytone, 1% (NH4)2HPO4, 1 mmol KCl, and 0.5% L-asparagin at pH 6.0 at 30°C. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Cladobotryum mycophilum.

The white button mushroom, Agaricus biporus is commercially the fifth important edible mushroom, accounting for a Korean production of 10,996 tons in Korea, 2012. The button mushroom, A. bisporus, mostly cultivated at mushroom farm controlled 16-19°C temperature during fruiting period in all seasons. For this reason, the production costs including the cost of energy, is very expensive to keep optimal culturing temperature. In this study, mycelial growth of strains collected from various countries was investigated at different cultivation temperature conditions to use baseline data for developing thermo-tolerance varieties. Mycelial growth was recorded strains cultivated in CDA (Compost Dextrose Agar) after two weeks. 12 strains among total 264 strains were cultivated well in high temperature. These strains will be cultivated in high temperature to confirm relationships between mycelial growth and fruit-body formation in same conditions.

A total of 25 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains, we found 9376 SNPs, of which 8178 were non-polymorphic and 1198 were polymorphic. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4120 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr08: 950kb-2650kb, chr09: 500kb-1400kb, chr09: 2800kb-4350kb, and chr11: 2450kb-3500kb regions were identified to be associated with white fruitbody forming.

Twenty Inter simple sequence repeat (ISSR) and 30 SSR primers were used to assess genetic diversity of 64 Agaricus strains including 45 A. bisporus strains and other 19 Agaricus spp. Of them, four ISSR primers, (GA)₈T, (AG)₈YC, (GA)₈C and (CTC)₆and seven SSR markers produced PCR polymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted for UPGMA cluster analysis. Forty five strains of A. bisporus are genetically clustered into 6 groups, showing coefficient similarity from 0.75 to 0.9 among them. The varieties, Saea, saedo, Saejeong and Saeyeon that have recently been developed in Korea were involved in the same group with closely genetic relationship of coefficient similarity over 0.96, whereas, other strains were genetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 10,996 M/T in 2012. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas agarici is the causal agent of browning disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of browning lesions on pileus. These lesions are superficial brown spots and can be round or spreading. But P. agarici never caused sunken lesions and rotting of the mushroom tissues. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. The HC42 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus safensis. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA.. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for P. agarici cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Agaricus bisporus. Control efficacy of browning disease of strain HC42 treatment was 66% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 1.5% D-galactose, 1.5% yest extract, 1% NH4Cl, 1.5% KCl, and 1.0% L-asparagin at pH 6.0 at 25℃. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by P. agarici.

Ganoderma lucidum has a long history for traditional medicine in Asian countries. However, taxonomy of Ganoderma species remains controversial, since it was initially classified on the basis of morphological characteristics. Recently, Ganoderma sichuanense was proposed as a re-proposed name for Ganoderma lucidum in China. Likewise, all of the Korean G. lucidum were clustered into 1 group together with G. sichuanense from China, when a phylogenetic analysis was undertaken based on the ITS rDNA sequences of the Ganoderma species. Furthermore, G. lucidum from Europe and North America were clustered into different group. Based on these results, re-consideration of naming Korean cultivated G. lucidum is needed.

A galactose fermentation bacterium producing lactose from red seaweed, which was known well to com-promise the galactose as main reducing sugar, was isolated from button mushroom bed in Buyeo-Gun, Chungchugnam-do province. The lactic acid bacteria MONGB-2 was identified as Lactobacillus paracasei subsp. tolerans by analysisof 16S rRNA gene sequence. When the production of lactic acid and acetic acid by L. paracasei MONGB-2 was inves-tigated by HPLC analysis with various carbohydrates, the strain MONGB-2 efficiently convert the glucose and galactoseto lactic acid with the yield of 18.86 g/L and 18.23 g/L, respectively and the ratio of lactic acid to total organic acidswas 1.0 and 0.91g/g for both substrates. However, in the case of acetic acid fermentation, other carbohydrates besidesgalactose and red seaweed hydrolysate could not be totally utilized as carbon sources for acetic acid production by thestrain. The lactic acid production from glucose and galactose in the fermentation time courses was gradually enhancedupto 60 h fermentation and the maximal concentration reached to be 16-18 g/L from both substrates after 48 h of fer-mentation. The initial concentration of glucose and galactose were completely consumed within 36 h of fermentation, ofwhich the growth of cell also was maximum level. In addition, the bioconversion of lactic acid from the red seaweedhydrolysate by L. paracasei MONGB-2 appeared to be about 20% levels of the initial substrates concentration and thisresults were entirely lower than those of galactose and glucose showed about 60% of conversion. The apparent resultsshowed that L. paracasei MONGB-2 could produce the lactic acid with glucose as well as galactose by the homof-ermentation through EMP pathway

Y-Aminobutyric acid(GABA) is a four carbon non-protein amino acid that has several well-known phys-iological functions, such as a postsynaptic inhibitory neurotransmitter in the brain and induction of hypotensive and tran-quilizer effects. A lactic acid bacterium was isolated from button mushroom bed, which is showing high GABAproductivity by TLC or HPLC analysis. The strain was identified as Lactobacillus hilgardii by analysis of 16S rDNAgene sequence. When the maximum production of GABA by L. hilgardii was investigated with various concentration ofmonosodium glutamate, the yield of GABA reached to be 53.65mM at 1% mono sodium glutamate (MSG) in flask cul-tivation. A Glutamate decarboxylase (GAD) enzyme, which was known to convert MSG to GABA, was purified froma cell-free extract of L. hilgardii and the molecular weights of purified GAD was estimated to 60,000 by SDS-PAGE.The optimum pH and temperature of GAD were at pH4.6 and at 37°C, respectively. The GAD activity was increasedby the addition of sulfate ions such as ammonium sulfate, sodium sulfate and magnesium sulfate, indicating that theincrease of hydrophobic interaction causes the increase of GAD activity.

Flammulina velutipes is a popular edible basidiomycete mushroom found in East Asia and is commonly known as winter mushroom. Mushroom development showing dramatic morphological changes by different environmental factors is scientifically and commercially interesting. We have sequenced the 35.6-megabase pair genome of F. velutipes KACC42780 using a next generation sequencing (NGS) approach. Then, HiSeq2000 sequencing system was used for comparing gene expression of F. velutipes genome at three different developmental stages (mycelium, primordium, and fruit body). Ninety one percents (10,116 genes) of the 10,999 predicted genes are expressed in at least one developmental stage. In addition, analysis of expressed genes in three different developmental stages showed that only 0.25–1.5% (28–165) of the total expressed genes (10,013) were uniquely expressed in a single developmental stage, whereas 83.6% (9,198) was shared in all stages.

신품종 백아는 갈색계통간 교잡에 의하여 우리 고유의 백색계통을 최초로 개발하여 이 계통을 이용하여 육성하였기 때문에 기존의 일본 유래의 백색계통에서 벗어난 한국형 백색 품종이라 할 수 있다. ‘백아’의 주요특성으로 균사배양 최적온도는 25~30oC이며,버섯발생온도는 12~13oC, 자실체 생육온도는 67oC이다. 자실체의 갓 색깔은 백색으로 다발성이며 품질이 우수하고, 자실체 대가 단단하고 곧으며 굵고 길며 식미감이 좋은 특성을 가지고 있다. 균긁기후 발이까지 7일정도 소요 되며 병당(850ml) 수량은 111 34g로 전형적인 병재배용 백색 팽이 품종이라고할 수 있다. 재배상 유의할 점은 환기량이 많으면 대가 짧아지고 갓이 커져 품질이 저하되므로 CO2 농도를 4,000ppm까지 높여 주는 것이 좋다.

A total of 35 phosphate solubilizing bacterial strains were isolated from waste bed of Agaricus bisporus in Buyeo-Gun, Chungchugnam-do province and screened for the production of indole acetic acid(IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 μg ml-1 after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 μg ml-1) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of 30°C followed by 25°C. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains’ ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same Rf value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.