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Isolation and Characterization of Various Strains of Bacillus sp. having Antagonistic Effect Against Phytopathogenic Fungi (식물 병원성 곰팡이에 길항작용을 갖는 다양한 Bacillus sp.의 균주 분리와 특성에 관한 연구)

  • Kim, Hee Sook;Kim, Ji-Youn;Lee, Song Min;Park, Hye-Jung;Lee, Sang-Hyeon;Jang, Jeong Su;Lee, Mun Hyon
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.603-613
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    • 2019
  • This study was carried out to examine the antagonistic effect against phytopathogenic fungi of isolated strains from soil samples collected from Busan, Changwon, and Jeju Island: Botrytis cinerea, Colletotrichum acutatum, Corynespora cassiicola, Fusarium sp., Rhizoctonia solani, Phytophthora capsici, and Sclerotinia sclerotiorum. According to results of our studies, isolated strains showed an antagonistic effect against phytopathogenic fungi. Such an antagonistic effect against phytopathogenic fungi is seen due to the production of siderophores, antibiotic substances, and extracellular amylase, cellulase, protease, and xylanase enzyme activities. Extracellular enzymes produced by isolated strains were significant, given that they inhibited the growth of phytopathogenic fungi by causing bacteriolysis of the cell wall of plant pathogenic fungi. This is essential to break down the cell wall of plant pathogenic fungi and thus help plant growth by converting macromolecules, which cannot be used by the plant for growth, into small molecules. In addition, they are putative candidates as biological agents to promote plant growth and inhibit growth of phytopathogenic fungi through nitrogen fixation, indole-3-acetic acid production, siderophore production, and extracellular enzyme activity. Therefore, this study suggests the possibility of using Bacillus subtilis ANGa5, Bacillus aerius ANGa25, and Bacillus methylotrophicus ANGa27 as new biological agents, and it is considered that further studies are necessary to prove their effect as novel biological agents by standardization of formulation and optimization of selected effective microorganisms, determination of their preservation period, and crop cultivation tests.

Comparison of mycorrhizal fungi associated with Pinus species in cultural characteristics and artificial mycorrhizal synthesis on Pinus thunbergii seedlings (소나무류 균근균의 배양적 특성비교 및 인공접종에 의한 해송묘목에의 균근협성)

  • Lee, Jong Kyu;Lee, Hoon Yong;Lee, Sang Yong
    • Journal of Forest and Environmental Science
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    • v.15 no.1
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    • pp.77-88
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    • 1999
  • This experiment was carried out to compare the cultural characteristics of mycorrizal fungi associated with Pinus species, and to form mycorrhizal association with Pinus thunbergii by artificial inoculation of these fungi. Mycorrhizal fungi tested showed great variations in cultural characteristics. Most fungal isolates was best grown on MP medium, except PDA for Lepista sp.(Ln73/92). Hagem for Rhizopogon rubescens(FRI91017), and FDA for Paxillus sp.(Pa60/92). Optimum temperature for these fungi was $25^{\circ}C$, except $30^{\circ}C$ for Pisolithus tinctorius(FRI91004 and Pt1). The range of pH conditions favorable for these fungal isolates were also variable from weak acidic(pH5) to weak alkalic(pH8). Utilization of the carbon sources for these mycorrhizal fungi was different. Fructose, glucose, and maltose were all utilized well, while xylose was not utilized generally. Mycelial growth on the media supplemented with potassium nitrate was better than those on other media with urea, asparagine, or peptone as a nitrogen source, and the poor growth was observed on the media with urea. Pisolithus tinctorius(Pt1) among 7 mycorrhizal fungi artificially inoculated for the mycorrhizal synthesis on pinus thunbergii seedlings in the test tube containing a mixture of peat moss-vermiculite(2:1, v/v) formed mycorrhizae successfully after 3 months. P. tinctorius formed branched and unbranched roots covered with thick fungal mantle and radiating extemal hyphae. Mycorrhizal root cross-sectioned by hand, stained, and observed by Nomarski interference microscope showed typical characteristics of ectomycorrhizae: fungal mantle on epidermal cells and thick Hartig net hyphae around cortex cells.

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Enzymological Characteristics and Identification of Useful Fungi Isolated from Traditional Korean Nuruk (한국전통누룩에서 분리한 유용곰팡이의 효소학적 특성 및 동정)

  • Kim, Hyun-Soo;Hyun, Ji-Sook;Kim, Jung;Ha, Hyun-Pal;Yoo, Dae-Sik
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.456-464
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    • 1998
  • For the standardization and quality improvement of traditional Korean Nuruk, 10 strains of fungi, which were isolated from Nuruks and showed good productivity of the saccharogenic and dextrinogenic enzymes, acid and flavor, were selected and their enzymological characteristics and identification were carried out. Aspergillus spp. and Rhizopus sp. showed a high liquefying activity without regard to cultivation time, whereas the majority of strains except for Rhizopus sp. had decreasing saccharifying activity in proportion to the increase in cultivation time. Aspergillus spp. No.17-2, No.17-6 and Rhizopus sp. No.18-1 showed high liquefying and saccharifying activity after 15 and 30 day cultivation. The optimum temperature of most of these saccharogenic and dextrinogenic enzymes was from 40$^{\circ}C$ to 60$^{\circ}C$, and their optimum pH was extensive between pH 3 and pH 11. But Penicillium spp.(2 strains) and Rhizopus sp. showed low activity under the alkalic and acidic conditions. Among these isolated strains, 5 strains which had shown the high productivity of materials were identified as Aspergillus oryzae NR3-6 and Aspergillus oryzae NR17-6, Aspergillus penicilloides NR12-1, Penicillium expansum NR7-7 and Rhizopus oryzee NRl8-1, respectively. Five kinds of mixed culture were carried out and all of them showed a better productivity of saccharogenic and dextrinogenic enzymes than single culture. These results indicate that it is possible to make traditional Korean liquors of good quality by using these fungi.

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Biocontrol Potential of Fungal Endophytes against Fusarium oxysporum f. sp. cucumerinum Causing Wilt in Cucumber

  • Abro, Manzoor Ali;Sun, Xiang;Li, Xingchun;Jatoi, Ghulam Hussain;Guo, Liang-Dong
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.598-608
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    • 2019
  • Endophytic fungi have received much attention as plant growth promoters as well as biological control agents against many plant pathogens. In this study, 30 endophytic fungal species, isolated from various plants in China, were evaluated using in vitro dual culture assay against Fusarium oxysporum f. sp. cucumerinum, causing wilt in cucumber. The results of the present study clearly showed that all the 30 endophytic fungal isolates were highly capable of inhibiting the mycelial colony growth of Fusarium oxysporum f. sp. cucumerinum with inhibition % over 66% as compared to control treatments. Among all of them, 5 isolates were highly effective such as, Penicillium sp., Guignardia mangiferae, Hypocrea sp., Neurospora sp., Eupenicillium javanicum, and Lasiodiplodia theobromae, respectively. The Penicillium sp. and Hypocrea sp. were highly effective as compared to other isolates. From in vitro results 10 best isolates were selected for greenhouse studies. The results of the greenhouse studies showed that among all of them 3 endophytic fungal isolates successfully suppressed wilt severity when co-inoculation with pathogen Fusarium. oxysporum f. sp. cucumerinum. The endophytic fungi also enhanced plant growth parameters of the host plants, the antagonistic fungal isolates increased over all plant height, aerial fresh, and dry weight as compared to control.

Antifungfal Activity Against Plant Pathogenic Fungi on Insect Enterobacteriaceae (식물병원성 곰팡이에 대한 곤충장내세균의 항균활성)

  • Oh, San Na;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.19 no.1
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    • pp.71-79
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    • 2015
  • In order to investigating the effects of antifungal activity of intestinal bacteria obtained from insect, it was identified these bacteria isolated from the gut. In this result, total 49 isolates of intestinal bacteria were identified from 10 kinds of insect species. It was that 4 isolates including Cedecea sp. from Nesidiocoris tenuis, 3 isolates including Enterobacter sp. from Odontotaenius disjunctus, 4 isolates including Acinetobacter sp. from Reticulitermes speratus, 4 isolates including Clavibacter sp. from Riptortus clavatus, 11 isolates including Bacillus sp. from Lema decempunctata, 3 isolates including Enterococcus sp. from Henosepilachna vigintioctopunctata 2 isolates including Staphylococccus sp. from Harmonia axyridis, 5 isolates including Enterobacter asburiae from Popillia mutans, 7 isolates including Aeromonas sp. from Hydrophilus acuminatus, and 7 isolates including Brucella sp. from Anomala octiescostata. In order to investigating antifungal activity against plant-pathogenic fungi, Altanaria solani, Colletotrichum gloeosporioides, Botrytis cinerea, Fusarium oxysporum, Phytophthora capsici, Rhizoctonia solani and Selerotinia sclerotiorum were dual cultured with each 49 gut enterobacteriaceae. As these results showed that many isolates have the antifungal activities including 26 isolates against A. solani, 6 isolates against B. cinerea, 13 isolates against C. gloeosporioides, 11 isolates against F. oxysporum, 17 isolates P. capsici, 2 isolates against R. solani and 2 isolates against S. sclerotiorum. Pseudomonas aeruginosa was showed strong antifungal activity against all of tested plant pathogens. It might be taken a potential for application against plant-pathogenic fungi with useful control agent.

Studies on the Toxigenic Fungi and Detoxification Methods Isolated from Korean Foodstuffs (한국(韓國) 식품중(食品中)의 유독성(有毒性) 진균(眞菌)의 검색(檢索) 및 해독(解毒)에 관(關)한 연구(硏究))

  • Koh, Choon-Myung;Lee, Bong-Kee;Kim, Se-Jong;Lew, Joon
    • The Journal of the Korean Society for Microbiology
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    • v.12 no.1
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    • pp.33-49
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    • 1977
  • These studies were carried out to detect the presence of mycotoxin fungi and detoxification of toxins in various kinds of grain and foodstuffs in Korea. The experiments were divided into three parts: mycologic, toxicologic and electron microscopic study. The resuIts were summarized as follows: 1. From the 210 various, samples(local grains, 150 samples; rice-cakes, 50 samples; meju, 10 samples), 1,205 colonies or fungi were isolated. In 1,127 of 1,205 colonies, it was possible to identify 21 genera. Among the identified strains, the predominant genera were Aspergillus sp.(28.5%), Penicillium sp.(27.1%) and Mucor sp.(7.8%). 2. In cytotoxicity test on HeLa cells, 25 strains showed severe toxic effects among the 240 strains of experiments. 3. In histopathologic test on mice, 21 strains showed severe toxic effect to mouse liver cells among the 240 tested strains. 4. In electron microscopic studies of HeLa cells and mouse liver cells from animal which had been treated with crude toxin, the liver cells showed the cytoplasmic change: dillatation or vacuolization of endoplasmic reticulum, swelling of mitochondria and disappearance of mitochondrial cristae, increased number of lipid and glycogen particles. Nucleus and nucelar envelope alterations, were also noted. 5. In the detoxification study with ultraviolet irradiation, 90% of the toxic substances were denatured by the ultraviolet light irradiation for 24 hours. 6. As a mass screening, the cytotoxicity test of HeLa cells and histopathologic study of mice liver cells treated with culture filtrates, employed and feasible to detect mycotoxin producing fungi.

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Detection of Fungi Associated with Ear Blighting of Rice Plants and Their Effect on Seed Discoloration and Yield (벼 이삭마름병해(病害)에 관여(關與)하는 균류(菌類)의 검출(檢出)과 검출(檢出)된 균류(菌類)가 종자(種子) 변색(變色) 및 수량(收量)에 미치는 영향(影響))

  • Kwon, Jin Sook;Park, Jong Seong;Yu, Seung Hun
    • Korean Journal of Agricultural Science
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    • v.12 no.2
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    • pp.199-205
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    • 1985
  • 1. Alternaria tenuis, Cladosporium cladosporioides, Curvularia lunata Fusarium semitectum, Gerlachia oryzae, Phoma sp. Trichoconiella padwickii were found associated with ear blighting of rice plants. The frequency of isolation of the fungi was found to be higher from the severely discolored rice grains than that from the brownish spotted grains, but H. oryzae was encounted highly from the latter. 2. Higher counts of Alternaria tenuis, Helminthosporium oryzae, Trichoconiella padwickii, Fusarium moniliforme, Fusarium semitectum were observed under alternating cycles of 12hrs of light and darkness than under complete darkness but G. oryzae was encountered highly under darkness in the blotter. While no difference in the occurrence of the other fungi was recorded under l2hr NUV light-12hr darkness cycles and darkness. 3. A. tenuis and C. cladosporioides were encountered highly at the lower temp. $(15^{\circ}C)$ but Phoma sp. was occurred highly at the higher temp. $(30^{\circ}C)$ when the seeds were incubated in the blotter. 4. Artificial inoculation of the fungi in the ears of rice plants caused severe discoloration of rice grains and reduced 1,000 grain weights and grain ripening percentage.

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Antifungal activity of Streptomyces costaricanus HR391 against some plant-pathogenic fungi (여러 식물병원성 진균을 억제하는 Streptomyces costaricanus HR391의 항진균능)

  • Kim, Hae-Ryoung;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.437-443
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    • 2016
  • In this study Streptomyces strains were isolated from soils and their antifungal activities and involved mechanisms were investigated. Among over 400 isolates of actinomycetes, Streptomyces costaricanus HR391 was selected as a potential antagonist to control several plant-pathogenic fungi. S. costaricanus HR391 inhibited mycelial growth of Fusarium oxysporum f. sp. raphani, F. oxysporum f. sp. niveum, F. oxysporum f. sp. lycopersici, and Rhizoctonia solani by 26.5, 26.2, 21.2, and 23.8%, respectively compared to those of uninoculated control after 7-day incubation on PDB medium. S. costaricanus HR391 produced $89{\mu}M$ of siderphore, and showed fungal cell wall-degrading activity including $0.46{\mu}mol/min/mg$ of chitinase and $0.83{\mu}mol/min/mg$ of ${\beta}$-1,3 glucanase. S. costaricanus HR391 secreted 87.49 mg/L of rhamnolipid, and produced 9.49 mg/L and 4.3 mM of lipopeptide, iturin A and surfactin, respectively, all they are membrane-disrupting biosurfactants. It also produced antimicrobial peptide and antibiotics phenazine. In addition to antifungal substances, S. costaricanus HR391 secreted plant growth-promoting phytohormones, zeatin, gibberellins and IAA. These results suggest that S. costaricanus HR391 may be utilized as an environment-friendly biocontrol agent against some important pathogenic fungi.

Screening of Ochratoxin A Producing Fungi from Greenhouse Horticulture (시설원예산물로부터 Ochratoxin A 생성 곰팡이의 검색)

  • Kang, Sung-Jo;Park, Bong-Jung;Lee, Jong-Ok;Kang, Jin-Soon;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1415-1419
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    • 1998
  • In order to evaluate the safety of greenhouse horticultures, a large number sample sources were collected, and the fungi of Aspergillus sp. and Penicillium sp. were isolated from them. Indirect competitive ELISA method and high performance liquid chromatography (HPLC) were applied to confirm the ochratoxin A producing abilities of isolated strains. One hundred ninety two sample sources including soil, pepper, strawberry and water mellon were collected for fungi isolation from western Gyeongnam, Andong and Gyeongbok. One hundred forty two strains of Aspergillus sp. and one hundred fifty three strains of Penicillium sp. were isolated respectively from them. The isolated fungi were tested for the production of ochratoxin A by ELISA. After culture of them on the modified sucrose low salt medium at $28^{\circ}C$ for 15 days, we found that five strains of Penicillium sp. produced ochratoxin A at the levels of $0.084{\sim}2.128\;{\mu}g/mL$. Among them, #129-2 strain isolated from water melon, showed the highest level of ochratoxin A as $2.128\;{\mu}g/mL$ broth. However, all of isolated Aspergillus sp. didn't produce ochratoxin A. When we compared the results of ELISA method with HPLC method, ochratoxin A production of each isolated strains showed very similar levels.

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Preparation of Cheeselike Product front Soybean (식물성(植物性) 치즈(Sufu)의 제조)

  • Park, Kwan-Hwa;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.23 no.2
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    • pp.115-122
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    • 1980
  • An attempt was made to isolate suitable kinds of fungi for the preparation of Sufu, one of the fermented soybean products, and also to investigate the chemical changes during the aging period of Sufu. 1. First, from the isolated fungi 14 strains of Mucor sp. were selected on the basis of their morphological texture and agreeable odor. Secondly, 7 strains of these fungi with relatively high proteolytic activity were selected. 2. All of the pehtzes that were fermented with the selected fungi have raised the water soluble protein content to about 10% on average. 3. The amount of hydrolyzed protein increased to the 18th. day and then did not show any visible increase during the aging period. Finally, the 3 strains of suitable fungi for preparing Sufu could be selected. It seemed that one of them was probably better than the other two which were almost same as the standard fungi, Actinomucor elegans, in protein hydrolyzation.

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