• 제목/요약/키워드: bacterial isolation

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Development of Molecular Biological Methods to Analyze Bacterial Species Diversity in Freshwater and Soil Ecosystems

  • Lee, Dong-Hun;Noh, Sung-Ae;Kim, Chi-Kyung
    • Journal of Microbiology
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    • 제38권1호
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    • pp.11-17
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    • 2000
  • A new method was developed for the rapid analysis of diverse bacterial species in the natural environment. Our method is based on PCR-single-strands-conformation polymorphism (PCR-SSCP) and selective isolation technique of single-stranded DNA. Variable V3 fragments of 16S rDNA were amplified by PCR with bacterial 16S rDNA primers, where one of the primers was biotinylated at the 5'-end. The biotinylated strands of the PCR products were selectively isolated by using streptavidin paramagnetic particles and a magnetic stand, to prevent SSCP analysis producing heteroduplexes from heterogeneous DNA samples. The selected strands were separated by electrophoresis on a polyacrylamide gel, and detected by silver staining. Analysis of PCR products from 8 bacterial strains demonstrated their characteristic DNA band patterns. In addition, changes in the structure of the bacterial community and species diversity in the microcosm treated with phenol could be monitored. After 3 weeks of incubation, phenol and its intermediate, 2-hydroxy-muconic-semialdehyde, were degraded by indigenous bacteria. These dominating bacterial populations were identified as strong bands on an SSCP gel. Therefore, this study provides useful tools for microbial community analysis of natural habitats.

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대전 대덕구 출토 복식 유물에 부착된 세균의 종류 및 섬유소분해효소 활성 (Bacterial Strains and Their Cellulase Activity from the Excavated Clothes at Daedeok-gu, Daejeon)

  • 이상준;차미선;조현혹;백영미;권영숙
    • 한국의류산업학회지
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    • 제7권1호
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    • pp.70-74
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    • 2005
  • This study was aimed to isolation and identification of attached bacteria on the clothes excavated from Deajeon area dating on the 16th century. From the observation with colony shape, 17 bacterial strains were isolated, and then 7 bacterial strains were identified with morphological and biochemical characteristics. Streptococcus sp., Alcaligenes faecalis, Gemella sp., Acinetobacter sp., Pseudomonas vesicularis, Aeromonas sain. salmonicida, Moraxella spp. In observation of the bacterial strains by the sort of textile, more bacterial strains were found in silk, cotton, and cotton batt than in ramie and hemp. It is suggest that hemp has antibacterial characteristics due to the presence of lignin. In the comparison washed samples with unwashed ones, there were more kinds of bacterial strains in washed samples. In the cellulase activity tests, all isolated bacteria had low level cellulase activity.

Isolation of Rhizobacteria in Jeju Island Showing Anti-Fungal Effect against Fungal Plant Pathogens

  • Lee, Chung-Sun;Kim, Ki-Deok;Hyun, Jae-Wook;Jeun, Yong-Chull
    • Mycobiology
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    • 제31권4호
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    • pp.251-254
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    • 2003
  • To select active bacterial strains to control plant diseases, 57 bacterial strains were isolated from the rhizosphere of the plants growing in various areas such as coast, middle and top of Halla Mountain in Jeju Island. Anti-fungal effect of isolated bactrial strains was tested in vitro by incubating in potato dextrose agar with isolates of four fungal plant pathogens Rhizoctonia solani, Fusarium oxysporum, Colletotrichum gloeosporioides and C. orbiculare, respectively. Thirty-four bacterial strains inhibited the hyphal growth of the plant pathogens, from which 17 strains inhibited one of the tested fungi, 10 strains two fungi, six strains three and a strain TRL2-3 inhibited all of the tested fungi. Some bacterial strains could inhibit weakly the hyphal growth of the plant pathogens, whereas some did very strongly with apparent inhibition zone between the plant pathogens and bacterial strains indicating the unfavorable condition for hyphal growth. Although there was no apparent inhibition zone, some bacterial strains showed a strong suppression of hyphal growth of plant pathogens. Especially, the inhibition by TRL2-3 was remarkably strong in all cases of the tested plant pathogens in this study that could be a possible candidate for biological control of various plant diseases.

A report of 31 unrecorded bacterial species belonging to the class Alphaproteobacteria in Korea

  • Kim, Kyung Hyun;Yoon, Jung-Hoon;Kim, Seung-Bum;Jahng, Kwang-Yeop;Cho, Jang-Cheon;Joh, Ki-seong;Cha, Chang-Jun;Seong, Chi-Nam;Bae, Jin-Woo;Im, Wan-Taek;Jeon, Che Ok
    • Journal of Species Research
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    • 제6권2호
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    • pp.129-140
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    • 2017
  • During a comprehensive investigation of indigenous prokaryotic species in Korea, a total 31 bacterial strains assigned to the class Alphaproteobacteria were isolated from diverse environmental habitats including freshwater, seawater, brackish water, ginseng soil, plant roots, natural caves, and tidal flats. Based on their high 16S rRNA gene sequence similarities(>99.1%) and formation of robust phylogenetic clades with the closest type species, each strain was assigned to an independent and predefined bacterial species. Because there were no published or official reports regarding the isolation of these 31 species in Korea, this study identified three species in two genera in the order Caulobacterales, 12 species in 10 genera in the order Rhodobacterales, three species in two genera in the order Rhizobiales, two species in two genera in the order Rhodospirillales and 11 species in seven genera, all in the order Sphingomonadaceae within the Alphaproteobacteria are reported as new alphaproteobacterial species in Korea. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source, and strain IDs are described in the species description section.

대황으로부터 세균성 질염 치료를 위한 항균성 물질의 분리 및 특성 (Isolation and Characterization of an Antibacterial Substance from Rheum palmatum for Treatment of Bacterial Vaginosis)

  • 장지은;강동희;윤재우;김현수
    • KSBB Journal
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    • 제32권2호
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    • pp.133-139
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    • 2017
  • Rheum palmatum has traditionally been used as a preventive agent and medication against fever and infection. The aim of this study was to isolate and characterize an antibacterial substance from R. palmatum that is effective against bacterial vaginosis. A methanol extract from R. palmatum showed antibacterial activity against Lactobacillus vaginalis KC TC 3515, Chryseobacterium gleum KCTC 2904, and Sphingomonas paucimobilis KCTC 2834, which cause bacterial vaginosis. After extraction and pH control of the methanol extract from R. palmatum, we found that acidic and alkaline extracts did not show antibacterial activity. A neutral extract (50 mg/mL) displayed an inhibitory zone of 18 mm on a nutrient agar plate with C. gleum KCTC 2904. Fractions No. 11 and 12 among 41 fractions obtained by silica gel column chromatography produced inhibitory zones of 10 mm on nutrient agar plates with C. gleum KCTC 2904. $R_f0.15$ and $R_f0.17$ spots produced by TLC of fraction No. 11 showed antibacterial activity against C. gleum KCTC 2904. Isolation and purification of the peak at a retention time (Rt) of 9.427 min was achieved by HPLC of $R_f0.29spots$. The peak at Rt 9.427 min showed antibacterial activity against C. gleum KCTC 2904.

Composition and Diversity of Gut Bacteria Associated with the Eri Silk Moth, Samia ricini, (Lepidoptera: Saturniidae) as Revealed by Culture-Dependent and Metagenomics Analysis

  • MsangoSoko, Kondwani;Gandotra, Sakshi;Chandel, Rahul Kumar;Sharma, Kirti;Ramakrishinan, Balasubramanian;Subramanian, Sabtharishi
    • Journal of Microbiology and Biotechnology
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    • 제30권9호
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    • pp.1367-1378
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    • 2020
  • The polyphagous eri silk moth, Samia ricini, is associated with various symbiotic gut bacteria believed to provide several benefits to the host. The larvae of S. ricini were subjected to isolation of gut bacteria using culture-dependent 16S rRNA generic characterization, metagenomics analysis and qualitative enzymatic assays. Sixty culturable aerobic gut bacterial isolates comprising Firmicutes (54%) and Proteobacteria (46%); and twelve culturable facultative anaerobic bacteria comprising Proteobacteria (92%) and Firmicutes (8%) were identified inhabiting the gut of S. ricini. The results of metagenomics analysis revealed the presence of a diverse community of both culturable and un-culturable gut bacteria belonging to Proteobacteria (60%) and Firmicutes (20%) associated with seven orders. An analysis of the results of culturable isolation indicates that these bacterial isolates inhabited all the three compartments of the gut. Investigation on persistence of bacteria coupled with metagenomics analysis of the fifth instar suggested that bacteria persist in the gut across the different instar stages. In addition, enzymatic assays indicated that 48 and 75% of culturable aerobic, and 75% of anaerobic gut bacterial isolates had cellulolytic, lipolytic and nitrate reductase activities, thus suggesting that they may be involved in food digestion and nutritional provision to the host. These bacterial isolates may be good sources for profiling novel genes and biomolecules for biotechnological application.

A report of 46 unrecorded bacterial species in Korea belonging to the classes Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria and Epsilonproteobacteria

  • Jung, Hye Su;Yoon, Jung-Hoon;Kim, Seung-Bum;Yi, Hana;Cho, Jang-Cheon;Joh, Kiseong;Cha, Chang-Jun;Seong, Chi-Nam;Bae, Jin-Woo;Im, Wan-Taek;Kim, Myung Kyum;Lee, Soon Dong;Jeon, Che Ok
    • Journal of Species Research
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    • 제8권2호
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    • pp.161-175
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    • 2019
  • During a comprehensive investigation of indigenous prokaryotic species in Korea, a total of 46 bacterial strains assigned to the classes Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, and Epsilonproteobacteria were isolated from a diversity of habitats including freshwater, seawater, brackish water, ginseng soil, plant roots, natural caves, and tidal flats. Based on their high 16S rRNA gene sequence similarities (>98.7%) and formation of strongly-supported phylogenetic clades with the closest type species, each strain was assigned to an independent, predefined bacterial species. Since there were no published or official reports regarding the isolation of these 46 species in Korea, here we report them as new species to Korea: 34 species in 14 families in the five orders of Alphaproteobacteria, 10 species in five families in the three orders of Betaproteobacteria, one species of Deltaproteobacteria and one species of Epsilonproteobacteria. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source, and strain IDs are described in the species description section.

Rapid identification of Burkholderia glumae from diseased seeds

  • Noh, Tae-Hwan;Song, Wan-Yeob;Kang, Mi-Hyung;Hyung Moo kim;Lee, Du-Ku;Park, Jong-Cheol;Shim, Hyeong-Kwon
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.136.1-136
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    • 2003
  • Bacterial grain rot by Burkholderia gluae cause severe damage in seedling and grain of rice after heading season. This seed-borne pathogen play a role as first infection agent that could be cause disease following cropping season. Until now the direct isolation of the bacteria has some trouble by interference of other bacteria existed inside seed. This study established convenient identification method as simple isolation with KB medium from seed showing symptom and using PCR identification. By this isolation method, B. glumae was isolated from 40 to 50% in brown rice and inner hull, however, there were saprophytic bacteria and fungi outer hull. In PCR identification with Ogf4 and Ogr3 primer to these 25 isolates, the amplified products were presented in all of the collections but not in 10 saprophytic germs. The isolation rate was constant to 3 months stored seeds. This result provide a rapid and convenient isolation and identification of B. glumae.

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Bacterial Carotenoids를 생산하는 광합성세균 Erythrobacter longus SY-46의 분리 및 특성 (Isolation and Characteristics of Photosynthetic Bacterium, Erythrobacter longus SY-46 which Produces Bacterial Carotenoids)

  • 김윤숙;이대성;정성윤;이원재
    • 한국환경과학회지
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    • 제17권4호
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    • pp.469-477
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    • 2008
  • The aerobic photosynthetic bacterium, which produces bacterial carotenoids was isolated and identified from coastal marine environments. This bacterium was identified by 16S rDNA sequencing and designated as Erythrobacter longus SY-46. E. longus SY-46 was Gram negative and rod shape, and the optimal culture conditions were $25^{\circ}C$, pH 7.0, and 3.0% NaCl concentration, respectively. The carbon and nitrogen sources required for the optimal growth were lactose and tryptone, respectively. Fatty acid compositions of E. longus SY-46 were $C_{18:1}$(78.32%), v-linolenic acid($C_{18:3n9.12.15c}:3.83%$), margaric acid($C_{17:0}$: 3.38%), palmitic acid($C_{16:0}$: 3.07%), and docosahexaenoic acid($C_{22:6n3}$: 2.21%). In addition, E. longus SY-46 showed the characteristic absorption peaks of bacterial carotenoids(in the region of 450 to 480 nm) and bacteriochlorophyll(770 to 772 nm). Major carotenoids of E. longus SY-46 were polyhydroxylated xanthophylls such as fucoxanthin and zeaxanthin.

Report of 20 unrecorded bacterial species in Korea belonging to the phylum Firmicutes during surveys in 2020

  • Park, Eun-Hee;Yoon, Jung-Hoon;Joh, Kiseong;Seong, Chi-Nam;Kim, Wonyong;Kim, Seung-Bum;Im, Wan-Taek;Cha, Chang-Jun
    • Journal of Species Research
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    • 제10권3호
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    • pp.217-226
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    • 2021
  • During a project aiming to comprehensively investigate indigenous prokaryotic species in Korea, a total of 20 bacterial strains phylogenetically belonging to the the class Bacilli of the phylum Firmicutes were isolated from various environmental sources such as soil, air, tidal flat, sea water, grain, wetland, breast milk and healthy human urine. Phylogenetic analysis based on 16S rRNA gene sequences revealed that 20 bacterial strains showed the high sequence similarities (≥98.7%) to the closest type strains and formed robust phylogenetic clades with closely related species of validly published names in the class Bacilli of the phylum Firmicutes. In the present study, we report 20 species of 13 genera of seven families of two orders of one class in the phylum Firmicutes, which have not been previously reported in Korea. Morphological, biochemical, and physiological characteristics, isolation sources, and NIBR deposit numbers of these unrecorded bacterial species are described in the species descriptions.