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Changes in microbial phase by period after hepa filter replacement in King oyster(Pleurotus eryngii) mushroom cultivation (큰느타리 재배사에서 헤파필터 교체 이후 기간에 따른 미생물상 변화)

  • Park, Hye-Sung;Min, Gyong-Jin;Lee, Eun-Ji;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.18 no.4
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    • pp.398-402
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    • 2020
  • This study was conducted to set up a proper replacement cycle of High Efficiency Particulate Air (HEPA) filters by observing the microbial populations in the air of the cultivation house of Pleurotus eryngii, before and after HEPA filter replacement at different periods. The density of bacteria and fungi in the air during each cultivation stage was measured using a sampler before the replacement of the HEPA filter. The results showed that airborne microorganisms had the highest density in the mushroom medium preparation room, with 169.7 CFU/㎥ of bacteria and 570 CFU/㎥ of fungi, and the removed old spaun had 126.3 CFU/㎥ of bacteria and 560 CFU/㎥ of fungi. The density of bacteria and fungi in the air at each cultivation stage before the replacement of the HEPA filter was 169.7 CFU/㎥ and 570 CFU/㎥, and 126.3 CFU/㎥ and 560 CFU/㎥, during the medium production and harvesting processes, respectively. After the replacement of the HEPA filter, the bacterial density was the lowest in the incubation room and the fungal density was the lowest in the cooling room. The microbial populations isolated at each period consisted of seven genera and seven species before the replacement, including Cladosporium sp., six genera and six species after 1 month of replacement, including Penicillium sp., 5 genera and 7 species after 3 months of replacement, including Mucor plumbeus, and 5 genera and 12 species, 5 genera and 10 species, and 5 genera and 10 species, 4, 5, and 6 months after the replacement, respectively, including Penicillium brevicompactum. During the period after replacement, the species were diversified and their number increased. The density of airborne microorganisms decreased drastically after the replacement of the HEPA filter. Its lowest value was recorded after 2 months of replacement, and it increased gradually afterwards, reaching a level similar to or higher than that of the pre-replacement period. Therefore, it was concluded that replacing the HEPA filter every 6 months is effective for reducing contamination.

Studies on Fungal Contamination and Mycotoxins of Rice Straw Round Bale Silage (사료용 볏짚 곤포사일리지의 곰팡이 및 Mycotoxin 오염 연구)

  • Sung, Ha-Guyn;Lee, Joung-Kyong;Seo, Sung
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.4
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    • pp.451-462
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    • 2011
  • The purpose of this study was to investigate fungi and mycotoxin contamination of the rice straw bale silage in Korean. It was tested the 33 samples of rice straw round bale silage with various condition which fed cattle in the farm. The level of fungal contamination was $2.1{\times}10^6\;cfu\;g^{-1}$ in the average and $9.2{\times}10^8\;cfu\;g^{-1}$ in the maximum. The fungal contamination was detected in the all of normal samples which good condition of rice straw bale silage. When the fungi was isolate and identify, it was found 28 species and mycotoxin producing fungi were 8 species as following as Aspergillus flavus, Aspergillus fumigatus, Fusarium culmorum, Fusarium verticillioides, Penicillium carneum, Penicillium paneum, Penicillium roqueforti, Penicillium viridicatum. Specially, Penicillium paneum was found 42% of samples and Aspergillus sp. (A. flavus, A. fumigatus) are 21% of samples. In case of mycotoxin contamination, the 42% of samples are detected more than one kind of mycotoxin. Some samples are contaminated three kinds of mycotoxin. This study was not found aflatoxin ($B_1$, $B_2$, $G_1$, $G_2$) and fumonisin ($B_1$, $B_2$), but were detected the contamination of ochratoxin A (1.0~5.8 ug/kg), deoxynivalenol (DON, 156.0~776.7 ug/kg) and zearalenone (ZON, 38.0~750.0 ug/kg). Therefore, the above results show that rice straw round bale silage expose on hazard factors as mycotoxigenic fungi and mycotoxin contamination, and than need more research about mycotoxin in animal feed to protect animal and human healthy.

Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi

  • Lim, Seong Mi;Yoon, Mi-Young;Choi, Gyung Ja;Choi, Yong Ho;Jang, Kyoung Soo;Shin, Teak Soo;Park, Hae Woong;Yu, Nan Hee;Kim, Young Ho;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.488-498
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    • 2017
  • The aim of this study was to identify volatile and agardiffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Diversity of Endophytic Fungi from Different Verticillium-Wilt-Resistant Gossypium hirsutum and Evaluation of Antifungal Activity Against Verticillium dahliae In Vitro

  • Li, Zhi-Fang;Wang, Ling-Fei;Feng, Zi-Li;Zhao, Li-Hong;Shi, Yong-Qiang;Zhu, He-Qin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1149-1161
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    • 2014
  • Cotton plants were sampled and ranked according to their resistance to Verticillium wilt. In total, 642 endophytic fungi isolates representing 27 genera were recovered from Gossypium hirsutum root, stem, and leaf tissues, but were not uniformly distributed. More endophytic fungi appeared in the leaf (391) compared with the root (140) and stem (111) sections. However, no significant difference in the abundance of isolated endophytes was found among resistant cotton varieties. Alternaria exhibited the highest colonization frequency (7.9%), followed by Acremonium (6.6%) and Penicillium (4.8%). Unlike tolerant varieties, resistant and susceptible ones had similar endophytic fungal population compositions. In three Verticillium-wilt-resistant cotton varieties, fungal endophytes from the genus Alternaria were most frequently isolated, followed by Gibberella and Penicillium. The maximum concentration of dominant endophytic fungi was observed in leaf tissues (0.1797). The evenness of stem tissue endophytic communities (0.702) was comparatively more uniform than the other two tissues. Eighty endophytic fungi selected from 27 genera were evaluated for their inhibition activity against highly virulent Verticillium dahliae isolate Vd080 in vitro. Thirty-nine isolates exhibited fungistasis against the pathogen at varying degrees. Seven species, having high growth inhibition rates (${\geq}75%$), exhibited strong antifungal activity against V. dahliae. The antifungal activity of both volatile and nonvolatile metabolites was also investigated. The nonvolatile substances produced by CEF-818 (Penicillium simplicissimum), CEF-325 (Fusarium solani), CEF-714 (Leptosphaeria sp.), and CEF-642 (Talaromyces flavus) completely inhibited V. dahliae growth. These findings deepen our understanding of cotton-endophyte interactions and provide a platform for screening G. hirsutum endophytes with biocontrol potential.

Study on Pandoraea sp. BCNU 315 Isolated from Soil (토양으로부터 분리한 Pandoraea sp. BCNU 315 에 관한 연구)

  • Kim, Seon-A;Choi, Hye-Jung;Woo, Seung-Hee;Hwang, Min-Jung;Park, Mi-Ran;Kim, Dong-Wan;Moon, Ja-Young;Joo, Woo-Hong
    • Journal of Life Science
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    • v.18 no.2
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    • pp.255-263
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    • 2008
  • Bacteria that antagonize plant pathogenic fungi were isolated from the sediment soil at the Ansan industrial estate. One isolate of them showed growth inhibition of Rhizoctonia solani, Botrytis cenerea, and Fusarium oxysporum. This strain was identified as Pandoraea sp. based on phenotypic and phylogenetic characteristics and termed Pandoraea sp. BCNU 315. Tryptone as nitrogen source and sucrose as carbon source were found to be most effective for the microbial growth. In addition, the optimum temperature and pH for microbial growth were $30^{\circ}C$ and pH 7.0, respectively. The substances generated from Pandoraea sp. BCNU 315 were purified and analyzed by column chromatography, HPLC, GC-MS and NMR. As a result, one compound was determined to be indole, another compound was predicted as cyclopentadecaheptene. Detailed structural clarification of the all of the rest six compounds from Pandoraea sp. BCNU 315 has to be accompanied in the further studies.

The effect of sterilization condition and UV-C irradiation on the reduction of contamination rate for oyster mushroom bottle culture (살균조건 및 UV-C 조사가 느타리버섯 병재배 오염율 감소에 미치는 영향)

  • Baek, Il-Sun;Chi, Jeong-Hyun;Jeoung, Yun-Kyeoung;Kim, Jeong-Han;Lim, Jae-Wook
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.256-261
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    • 2015
  • This study was conducted to reduce contamination ratio of oyster mushroom bottle cultivation. The optimal conditions of substrate sterilization for reducing of contamination ratio were at $121^{\circ}C$ for 90 min. In addition, UV-C irradiation is good for lower contamination ratio to continue over 6 hours at cooling and inoculation room after sterilization. The contamination ratio and density of microorganisms of substrate were showed 0% after sterilization at $121^{\circ}C$ for 90 min. Trichoderma sp., main pathogen of mushrooms, was detected from substrate after sterilized during 2 or 4 hours at $101^{\circ}C$ and $105^{\circ}C$, respectively. The amount of electricity used was the lowest at $121^{\circ}C$ for 90 min than that of other sterilization conditions. The UV-C irradiation treatment was used UV-C lamp(40 watts) in the inoculation room($56m^3$). The density of bacteria did not detected after UV-C irradiation for 6 hours. And the death ratio of bacteria and Trichoderma sp. was 99.9% after UV-C irradiation for 6 hours. However, in the same UV-C irradiation time, the death ration of Cladosporium sp. was 90.9%. Therefore, the death ratio of fungi was lower than that of bacteria at the same UV-C irradiation treatment.

Isolation and Optimal Culture Conditions of Brevibacillus sp. KMU-391 against Black Root Pathogens Caused by Didymella bryoniae (덩굴마름병원균인 Didymella bryoniae의 생물학적 방제를 위한 길항세균의 분리 및 특성)

  • Park Sung-Min;Jung Hyuck-Jun;Kim Hyun-Soo;Yu Tae-Shick
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.135-141
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    • 2006
  • We isolated a bacterium which produces antifungal substances from the Sanktpeterburg soils at Russia. The iso-lated strain was identified as Brevibacillus sp. and shown a strong antifungal activity on plant pathogenic fungi. Brevibacillus sp. KMU-391 produced maximum level of antifungal substances under incubation aerobically at $30^{\circ}C$ for 48 hours in trypticase soybean broth containing 1.0% sucrose and 1.0% polypeptone at 180 rpm and initiated pH adjusted to 7.0. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against Didymella bryoniae by dry cell weight. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Botrytis cinerea KACC 40573, Botrytis fabae KACC 40962, Colletotrichum gloeosporioides KACC 40804, Colletotrichum orbiculare KACC 40808, Didymella bryoniae KACC 40669, Fusarium graminearum KACC 41040, Fusarium oxysporum KACC 40037, Fusarium oxysporum KACC 40052, Fusarium oxysporum f, sp. radicis-Iycopersici KACC 40537, Fusarium oxysporum KACC 40902, Monosporascus cannonballus KACC 40940, Phytophthora camvibora KACC 40160, Rhizoctonia solani AG-1(IA) KACC 40101, Rhizoctonia solani AG-4 KACC 40142, and Scleotinia scleotiorum KACC 41065 by agar diffusion method.

Control Efficacy of Phloretin Isolated from Apple Fruits Against Several Plant Diseases

  • Shim, Sang-Hee;Jo, Su-Jung;Kim, Jin-Cheol;Choi, Gyung-Ja
    • The Plant Pathology Journal
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    • v.26 no.3
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    • pp.280-285
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    • 2010
  • In the course of a searching natural antifungal compounds from plant sources, we found that the methanol extract ($3,000\;{\mu}g/ml$) of Malus domestica fruits had potential of control against rice blast (Magnaporthe grisea) and tomato late blight (Phytophthora infestans). Under bioassay-guided purification, we isolated phloretin, a phenolic compound, with in vivo antifungal activity against M. grisea. By 1-day protective application of phloretin ($500\;{\mu}g/ml$), the compound strongly inhibited the disease development of M. grisea and P. infestans on rice and tomato seedlings, respectively. And red pepper anthracnose caused by Colletotrichum coccodes also was moderately suppressed. However, rice sheath blight (Rhizoctonia solani AG1), and barley powdery mildew (Blumeria graminis f. sp. hordei) were hardly controlled. In addition, the compound showed in vitro antifungal activity against some plant pathogenic fungi including Phytophthora capsici, Alternaria panax, Sclerotinia sclerotiorum, R. solani AG4, and M. grisea. This is the first report on the antifungal activity of phloretin against plant pathogenic fungi.

Antimicrobial Properties of Cold-Tolerant Eucalyptus Species against Phytopathogenic Fungi and Food-Borne Bacterial Pathogens

  • Hur, Jae-Seoun;Ahn, Sam-Young;Koh, Young-Jin;Lee, Choong-Il
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.286-289
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    • 2000
  • Mechanol extracts of three cold-tolerant eucalyptus trees-Eucalyptus darlympleana, E. gunnii and E. unigera were screened for antimicrobial activity against twenty two phyto-pathogenic fungi and six food-borne bacterial pathogens. E. unigera showed the antagonistic activity against all the tested pathogens. Among the tested fungal pathogens, Pythium species were highly sensitive to the leaf extracts. Especially, P. vanterpoolii, a causal agent of leaf blight in creeping bentgrass (Agrostis palustris), was completely inhibited by the extracts. The eucalyptus extracts were also effective in inhibiting the fungal growth of Botrytis cinerea and Phomopsis sp. isolated from the lesions of kiwifruit soft rot during post-harvest storage. Escherichia coli O-157 was less sensitive to the inhibition than the other bacterial pathogens tested. It was likely that Gram positive bacteria-Bacillus subtilis and Streptococcus mutans were more sensitive to the eucalyptus extracts than Gram negative bacteria-Escherichia coli, Salmonella enteritidis and Pseudomonas aeruginosa. Our findings suggest that the cold-tolerant eucalyptus species have antimicrobial properties that can serve the development of novel fungitoxic agents or food preservatives.

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Identification of Streptomyces sp. Producing Antibiotics Against Phytopathogenic Fungi, and Its Structure

  • Kim, Jung-Han;Jeong, Do-Hyeon;Park, Ki-Duk;Kim, Sung-Han;Kim, Kyung-Rae;Choi, Sung-Won;Kim, Ji-Tae;Choi, Ki-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.212-215
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    • 2004
  • In order to develop a biocontrol agent that can effectively control Fusarium wilt on Cymbidium genus, the effectiveness of antagonistic microbes against the cause pathogen was screened. The selected microbe showed a broad spectrum of antifungal activity, and the culture broth of this microbe had better preventive effect on Fusarium wilt than the commercial chemical agent in the pot assay. This isolated strain, GBA-12, was identified as Streptomyces kasugaensis, and the antifungal substance was purified from a broth culture of GBA-12. This purified substance was identified as a polyene macrolide (YS-822A) that was newly discovered from Streptomyces kasugaensis, and it exhibited antifungal activity against several phytopathogenic fungi.