This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30oC, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.

고추는 전 세계적으로 중요한 경제작물이다. 하지만, 매년 Colletotrichum species에 의해 발생하는 고추탄저병에 의해 상품성이 떨어지고 수량이 현저히 감소하여 많은 농가에 중대한 피해를 주는 주요 병해이기 때문에 방제대책이 절실히 요구되고 있다. 본 연구에서는 국내에 최근 발생한 C. scovillei에 대해 Burkholderia cepacia를 이용하여 길항 능력을 확인하였다. 16S rRNA와 recA 유전자 분석을 통해 분리된 균주가 B. cepacia임을 확인하였고, 식물 생장 촉진 물질인 siderophore와 항진균 가수분해 효소인 protease, cellulase 생산을 기내 실험을 통해 확인하였다. 또한, C. scovillei에 대한 KF1의 균사 생장 억제율은 66.0%로 나타났으며, 1×105, 106, 107, 108 cfu/mL의 길항 미생물 처리 농도에 따른 포자 발아율은 각각 7.8%, 4.3%, 1.2%, 0%로 확인되어 분리된 길항 미생물은 효과적으로 병원균의 균사 생장 및 포자 발아를 억제하였다. 따라서 본 연구는 길항 미생물 B. cepacia KF1의 고추탄저병균에 대한 생물학적 방제제 자원으로서의 활용가능성을 제시하였다.

A gram-positive bacterium was isolated from the spent substrate of Agaricus bisporus that showed a marked antagonistic activity against Pseudomonas agarici. It was identified as Bacillus safensis HC42 based on its cultural, biochemical, and physiological characteristics, and 16S rRNA sequence. B. safensis HC42 was saprophytic, but not parasitic or pathogenic, in cultivated mushrooms and showed strong inhibition of P. agarici in vitro. Moreover, it showed a control efficacy of 66 % against browning disease caused by P. agarici in Agaricus bisporus. Therefore, B. safensis HC42 may be useful in the future for the development of a biocontrol system.

Cladobotryum mycophilum is the causal agent of cobweb disease of commercial mushrooms. Symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps and progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing. Two gram-positive bacterium were isolated from mushroom media that markedly showed the antagonistic activity against C. mycophilum.. These isolates were identified as Bacillus altitudinis and Bacillus subtilis by analysis of the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacteria is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacteria cell was sufficient for inhibition in vitro for C. mycophilum. Control efficacy of browning disease of Bacillus altitudinis treatment was 78% on Agaricus bisporus. The optimal culture medium 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. Control efficacy of browning disease of Bacillus subtilis treatment was 71% on Agaricus bisporus. The optimal culture medium was determined as follows: 1.5% Xylose, 2% Soytone, 1% NH4H2PO4, 7 mmol CaCl2, and 0.5% Histidine at pH 6.0 at 25°C. Accordingly, 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 C. mycophilum.

This study was conducted to investigate optimum conditions for mass production of ntagonistic microbes Alcaligenes sp. HC12. Alcaligenes sp. HC12 had a potent biological control agent to control browning disease caused by Pseudomonas agarici. Alcaligenes sp. HC12 markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. To define the optimum conditions for the mass production of the Alcaligenes sp. HC12, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 9.0 and 30o, respectively. The optimal concentration of medium elements for the growth of pathogen inhibitor bacterium(Alcaligenes sp. HC12) was determined as follows: 0.5% dextrine, 1.5% yest extract, 1.0% NaNO3, 0.5% KH2PO4, and 1.5% asparagine.

The purpose of this study was to identify the effects of continuous muscle strengthening applied to the antagonist of the sternocleidomatoid, upper trapezius, and pectoralis major, which are the shortened muscles of forward head posture(FHP) subjects, and Evjenth-Hamberg stretching(EHS) applied to the shortened muscles on changes in pressure pain threshold(PPT). Twenty subjects were divided into the continuous antagonist strengthening( CAS) group(n=10) and the EHS group(n=10), and each group performed its respective exercise three times a week for a six week period. The results were as follows: The comparison of changes in PPT within each group before and after the treatment showed a statistically significant difference( p<.05) according to the treatment period and a statistically significant difference according to the treatment period and method(p<.05). While the comparison of the tests of between subjects effects between the groups did not show a statistically significant difference, the CAS group exhibited better effects. The above results suggest that the combined application of CAS and EHS generates better effects on changes in PPT than the single application of EHS. Given that stretching and muscle strengthening exercises even for the short research period of six weeks could change the PPT, continuous exercises and a correct postural habit for a longer period of time are likely to help prevent chronic pain and correct FHP

This research was conducted to investigate the effect of continuous antagonistic muscle strengthening exercise and Evjenth-Hamberg stretching on the pulmonary function of university students with abnormal transformation of forward head posture(FHP). The results of study subject to the continuous antagonistic muscle strengthening(CAS) group(n=10) and Evjenth- Hamberg stretching(EHS) group(n=10) that was conducted 3 times a week for 6 weeks are as follow. FVC, IVC, and MVV all were shown to be significant in the pre·post comparison between the CAS group and EHS group(p<.05), and significant difference was shown for MW between the two groups(p<.05) in which the CAS group showed better effect. Based on the results above, it is considered that combining continuous antagonistic muscle strengthening exercise has better effect on pulmonary function compared to application of only Evjenth-Hamberg stretching.

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.

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.

버섯배지의 발효를 효율적으로 행하면서 버섯의 재배 시 빈번하게 발생하는 푸른곰팡이 병의 원인 균주인 Trichoderma sp. 곰팡이 성장을 억제하는 세균의 분리를 행하였다. 균원시료로부터 1차 분리한 약 200여 균주 중에서 성장속도가 빠르고, SM, AM 및 CM의 평판배지 상에서 clear zone이 뚜렷한 6균주를 2차 분리하였다. 분리한 6균주 중 cellulase, amylase 및 protease의 효소활성이 높고 T. virens와 T. harzianum에 대하여 강한 항균활성을 나타낸 C-1균주를 최종 분리균주로 선정하였다. 분리한 C-1균주는 Bergey's Manual of Systematic Bacteriology에 의한 동정과 16S rDNA 염기서열 분석을 행한 결과 Paenibacillus polymyxa 밝혀져 P. polymyxa CK-1으로 명명하였다. P. polymyxa CK-1균주의 생육조건을 검토한 결과 최적배양온도는 45˚C, 생육을 위한 배지의 최적 pH는 6.0~7.0 범위로 나타났다. T. virens와 T. harzianum 곰팡이의 생육억제를 위한 P. polymyxa CK-1의 배양시간은 22~36시간이 적당하였다. 그리고 P. polymyxa CK-1균주의 24시간째 배양용액을 처리한 petri dish에 두 곰팡이를 각각 접종한 후 10일 동안 방치하여도 곰팡이의 생육은 관찰되지 않았다. P. polymyxa CK-1 균주가 생산한 길항물질의 열안정성을 검토한 결과 60˚C 및 100˚C로 20분 동안 처리한 시험구에서는 두 곰팡이의 균사성장이 전혀 관찰되지 않았지만 121˚C에서 20분 동안 처리한 시험구에서는 약간의 균사성장이 관찰되었다. P. polymyxa CK-1배양액이 버섯균사 생육에 미치는 영향을 검토한 결과 팽이버섯, 표고버섯 등의 다양한 버섯균사 생육에 전혀 영향을 미치지 않는 것으로 나타났다.

Biogenic amine은 주로 단백질이 풍부한 전통장류 중 된 장과 간장에서 높게 발생하고 있다. 현재까지 국내외 식중독 사고를 보면 biogenic amine중 tyramine, histamine과 putrescine에 의한 보고가 있다. 전통장류의 biogenic amine 에 대한 위생적인 문제를 해결하기 위해 전통장류 제조에 적합한 발효미생물 중에서 biogenic amine 비생성 및 분해하는 미생물을 선발하여 종균으로 적용한다면 biogenic amine에 대한 문제를 해결할 수 있을 것이다. 이러한 문제의 해결을 위해 본 연구에서는 우수한 발효능력과 함께 유해균에 대해 강한 길항 능력을 지니며, biogenic amine 을 생산하지 않지만 높은 분해능력을 보이는 균주를 선발하여 장류제조 종균으로의 이용가능성을 실증하였다. 결론적으로 선발된 장류발효균주를 종균으로 이용하여 메주 제조 및 청국장 제조 등에 이용한다면 biogenic amine과 Bacillus cereus를 안전한 수준으로 낮출 수 있을 것으로 사료되었고, 전통장류의 HACCP적용을 위한 HAZARD를 관리하는 우수한 수단으로 이용될 수 있을 것이다. 또한 향후에는 단일 균종만 종균으로 사용할 경우 전통장류의 우수한 풍미를 재현하는데 한계가 있기 때문에 종균들을 적절하게 조합한 혼합발효를 통해 제조한다면 풍미가 좋을 뿐만 아니라, 위생문제를 해결한 우수한 장류를 생산 할 수 있을 것으로 판단되었다.

We have screened Bacillus strains suitable for the fermentation of soybean products with respect to the control of Bacillus cereus and the reduction of biogenic amines. Of 26 isolates, a strain named as the SCK A08 carried antimicrobial activity against B. cereus and Staphylococcus aureus, major food poisoning species in soybean products. PCR analysis revealed that the SCK A08 strain did not contain genes for Bacillus cereus toxins including nonhemolytic enterotoxin, hemolytic enterotoxin, cytotoxin K, cereulide and certrax. The SCK A08 strain could degrade histamine, tyramine, putrescine, and cadaverine by 67.41%, 76.59%, 57.32%, and 50.69%, respectively, during fermentation in cooked soybeans containing 0.5% (w/w) of each biogenic amine. The morphological and biochemical properties and phylogenetic relationships based on 16S rRNA gene sequences indicated that the isolate was most closely related to Bacillus licheniformis. Use of the strain SCK A08 would be a potential measure to overcome two hygienic problems that were frequently faced during manufacture of traditionally fermented soybean products.

Mushroom is cultivated as one of the major economical crops in many areas in Korea. The total production has steadily increased from approximately 186,400 M/T in 2007 to 190,111 M/T in 2011. The button mushroom, Agaricus bisporus, showed the 5th production to 13,052 M/T in 2011. 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. In this study, antagonistic bacteria to P. agarici were selected in vitro tests using confrontation bioassay and paper disk diffusion assay. The most active bacteria, HC12 were selected and their control efficacy of brown blotch disease was investigated in this study. After proceeding antagonistic test, HC12 was selected as a strong antagonist against P. agarici and the HC12 strain was identified as Alcaligenes sp. with the cultural, physiological and biochemical properties and analysis of the 16S rRNA. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 0.5% dextrin, 1.5% yest extract, 1% NaNO3, 0.5% KH2PO4, and 1.5% L-asparagin at pH 9.0 at 30℃. Control efficacy of browning disease by HC12 treatment was 63% on Agaricus bisporus..

We analyzed the viable population changes of Paenibacillus polymyxa E681, a plant growth-promoting rhizobacterium, on seeds and roots after bioformulation at varying time intervals during the storage. The viable population of E681 on tested crop seeds sustained log 4-5 cfu/seed after 300 days of seed treatment. The ability of root colonization and inhibition of fungal mycellial growth was not influenced even after 300 days of seed treatment. The seed-soaking treatment returned better results than powder formulation, in increasing the initial population of E681 on plant roots. Collectively, it was found that E681 is a durable and stable biological control agent for application to crop seeds.

본 연구는 친환경적으로 시금치 노균병을 방제하기 위하여 식물정유, 길항미생물 및 난황유의 병발생 억제효과를 검정하여 화학농약 대신에 이용할 수 있는 친환경제제로서의 가능성을 검토하였다. 5종의 식물정유 가운데 노균병 방제효과는 1회 처리구와 1주 간격 2회 처리구 모두 Eucalyptus approximans 오일의 병원균 억제효과가 60% 정도로 가장 높은 것으로 나타났다. 반면에 토양에서 선발된 길항균 세 종류의 시금치 노균병 방제효과는 모두 20% 이하로 식물정유 처리보다 낮게 나타났다. 난황유 0.3%와 0.5%농도에서 1주일간격 2회 처리구에서 병원균 억제율이 각각 84.5%와 91.5%로 억제되어 가장 효과가 우수한 것으로 나타났다.

This study was carried out investigate to availability of commercial microbial pesticide and antibacterial activity of isolates isolated from different mushroom media. Ten commercial microbial pesticide and EM liquid were collected from different company. EM of these one has been used for control of mushroom disease and growth promotion at the mushroom farms. The density of bacteria and yeast in EM cultural liquid was higher than those of the crude liquid. The pH values of EM showed the low acid levels(pH 3.5~3.9) by organic acid secreted from microorganism. The kinds of organic acid was acetic acid, lactic acid and succinic acid. The dominant bacteria isolated from EM liquid was Lactobactillus sp.(21 strains), Acetobacter sp.(9 strains), PaeniBacillus sp.(9 strains) and others(12 strains). The organic acid bacteria isolated from fermentation foods( was 92 strains and the dominant genus was Weissella sp.(41 strains), Leuconostoc sp.(21 strains), Enterococcus sp.(9 strains), Lactococcus sp.(9 strains) and others(12 strains). And we isolated 2,500 bacteria from oyster mushroom and button mushroom cultural media for selection of antagonistic bacteria. Thirty five strains of these isolates showed very strong antagonistic activity. These strains were identified Bacillus amyloliquefaciens, Bacillus subtilis, Brevibacterium halotolerans, Pseudomonas libanensis, Stenotrophomonas maltophilia and Alcaligenes faecalis by 16S rDNA analysis.