• 제목/요약/키워드: 16S rRNA analysis

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작물병원 진균에 대하여 항균 활성을 보이는 Paenibacillus polymyxa DY5의 동정 및 특성 (Identification and Characterization of Paenibacillus polymyxa DY5 with Antifungal Activity against Crop Pathogenic Fungi)

  • 김효윤;원항연;김완규;유관희
    • 한국균학회지
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    • 제37권2호
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    • pp.181-188
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    • 2009
  • A Gram-positive, rod-shaped bacteria named DY5 was isolated from a peat sample collected from Daeam mountain in Korea. The culture filtrate of the bacterial isolate DY5 showed a broad spectrum of antifungal activity on various crop pathogenic fungi such as Trichoderma koningii, Fusarium oxysporum, Colletotrichum gloeosporioides, Sclerotinia sclerotiorum, Rhizoctonia solani AG-1(IA) For the identification of the DY5, morphological, biochemical, API 50 CHB test, analysis of fatty acid and molecular phylogenetic approaches were performed. The DY5 was found to be a member of the genus Paenibacillus on the basis of morphological and biochemical analysis. The 16S rRNA of DY5 showed high similarity(98%) with Paenibacillus polymyxa. On the basis of these results, the DY5 was identified as Paenibacillus polymyxa. Antifungal substance of the DY5 would be mild alkaline proteine molecule. The DY5 seems to have a great potential to be a biocontrol agent against various crop pathogens.

Aureivirga callyspongiae sp. nov., Isolated from Marine Sponge Callyspongia elegans

  • Park, So Hyun;Kim, Ji Young;Heo, Moon Soo
    • 한국미생물·생명공학회지
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    • 제49권3호
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    • pp.384-390
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    • 2021
  • A Gram-negative, aerobic, motile by gliding, and rod-shaped marine bacterium, designated CE67T was isolated from the marine sponge Callyspongia elegans on Biyang-do in Jeju Island. The CE67T strain grew optimally at 25℃, pH 7.5, and in the presence of 2-3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain CE67T was related to the genus Aureivirga and had the highest 16S rRNA gene sequence similarity to the Aureivirga marina VIII.04T type strain (96.3%). The primary fatty acids (>10%) of strain CE67T were iso-C15:0 (35.3%) and iso-C17:0 3OH (21.8%). The polar lipid profile of strain CE67T contained phosphatidylethanolamine, unidentified aminolipids, and unidentified lipids. The predominant menaquinone was MK-6. The DNA G+C content was 29.1 mol%. Based on the polyphasic taxonomic analysis, strain CE67T was determined to be a representative novel species of the genus Aureivirga for which we propose the name Aureivirga callyspongiae sp. nov., whose strain type is CE67T (=KCTC 42847T=JCM 34566T).

In situ analysis of the bacterial community associated with the Korean salty fermented seafood jeotgal

  • Hyunjun Kim;Yoomin Ahn;Chulhee Park;Eungbin Kim
    • 환경생물
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    • 제39권4호
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    • pp.515-522
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    • 2021
  • Jeotgal is a salty and fermented traditional Korean fish sauce. Unlike most other previous studies that investigated samples purchased from retail markets, this study focused on samples of jeotgal with traceable history to Yeonggwang, a timehonored fishing village in Korea. Three jeotgal samples, which were made from small yellow croakers, largehead hairtail, and miscellaneous fish, were selected based on information obtained from interviews with local craftsmen and literature reviews. Bacterial community profiles of the three jeotgal samples were investigated to identify indicator (and potentially core) bacteria for jeotgal ripening. The 16S rRNA gene-based metagenomic analysis revealed that the dominant phyla and classes, (Gammaproteobacteria, Betaproteobacteria, Bacilli, and Clostridia) of the three different jeotgal were identical, albeit with different composition ratios. Diversification was evident beginning at the order level. Interestingly, each dominant order was mainly comprised of single members even at the genus level. The dominant genera included Halomonas, Tetragenococcus, Halanaerobium, Pseudomonas, Massilia, and Lentibacillus. This observed genus-level heterogeneity suggests that there are diverse bacterial signatures in jeotgal and that these can be used as indicators for jeotgal ripening and/or as starters to increase its sensory quality and functionality.

Strain Identification and Comparative Analysis of Toxigenic Cyanobacteria Determined by PCR

  • Jung Jong-Mun;Jung Eun-Young;LEE You-Jung;Park Hong-Ki;Jung Mi-Eun;Ji Ki-Won;Joo Gea-Jae
    • 한국환경과학회지
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    • 제14권7호
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    • pp.649-655
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    • 2005
  • Microcystis aeruginosa is common form of cyanobacteria (blue-green algae) capable of producing toxic heptapeptide (microcystin) that cause illness or death. The comparison of molecular genetic method with the morphological characteristics of cyanobacteria was conducted. We have designed PCR primers (JJM98F, JJM1141R) for cyanobacterial 16S rRNA and phycocyanin intergenic spacer (PC-IGS) gene domain. To confirm the production of microcystins, PCR primers for the N-methyltransferase (NMT) domain of microcystin synthetase gene mcyA were designed using 21 cyanobacteria strains Most of isolated strains from the Nakdong River was classified as Microcystis aeruginosa and the similarities were $99\%$ with M. aeruginosa AF 139292. $38.1\%$ of isolated strains contained microcystin synthesis gene. NMT (N-methyltransferase) were not detected in isolated strain in several strains, which means non-toxic. However, the NMTs of the strains were detected during the cultivation.

한강물로부터 분리된 방사선 내성 세균들의 계통학적 다양성 및 UV 내성 분석 (Phylogenetic diversity and UV resistance analysis of radiation-resistant bacteria isolated from the water in Han River)

  • 이재진;주은선;이도희;정희영;김명겸
    • 미생물학회지
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    • 제52권1호
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    • pp.65-73
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    • 2016
  • 이 논문은 서울 한강물에서 분리한 방사선 내성 세균군집과 분리된 신종 세균의 UV 내성 특성에 관한 내용이다. 세균은 R2A agar와 1/10 R2A agar를 사용하여 3 kGy가 조사된 한강물에서 분리되었다. 그 결과 방사선에 내성을 가지는 것으로 추측되는 균주를 60주 분리하였고, 본 연구 분석 자료로 사용하였다. 16S rRNA 유전자 염기서열 분석을 통해 분리균주 60개의 계통수를 파악한 결과, 3개의 문(4개의 속)이 확인되었고, Deinococcus-Thermus (Deinococcus)가 61.7%, Firmicutes(Exiguobacterium)는 15%, Bacteroidetes (Hymenobacter, Spirosoma)는 23.4%의 비중을 나타냈다. 분리균주 중 29개 균주가 Deinococcus, Hymenobacter, Spirosoma에 속하는 신종 또는 다른 신속으로 분류될 가능성을 보였으며, 앞으로 추가적인 신종 실험이 진행될 예정이다. 그리고 신종 예상균주를 9개 선정하여 UV 내성 실험을 진행한 결과, 9개 균주 모두 D. radiodurans $R1^T$ 균주 만큼 높은 UV 내성을 가지는 것으로 나타났다. 또한 분리된 Firmicutes (Exiguobacterium) 균주는 아직까지 방사선 내성 연구 보고가 되어 있지 않아서 추가적인 방사선 내성 연구가 필요하다.

Occurrence of Bacterial Stem Rot of Ranunculus asiaticus Caused by Pseudomonas marginalis in Korea

  • Li, Weilan;Ten, Leonid N.;Kim, Seung-Han;Lee, Seung-Yeol;Jung, Hee-Young
    • 식물병연구
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    • 제24권2호
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    • pp.138-144
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    • 2018
  • In December 2016, stem rot symptoms were observed on Persian buttercup (Ranunculus asiaticus) plants in Chilgok, Gyeongbuk, Korea. In the early stage of the disease, several black spots appeared on the stem of infected plants. As the disease progressed, the infected stem cleaved and wilted. The causal agent was isolated from a lesion and incubated on Reasoner's 2A (R2A) agar at $25^{\circ}C$. Total genomic DNA was extracted for phylogenetic analysis. Based on the 16S rRNA gene analysis, the isolated strain was found to belong to the genus Pseudomonas. To identify the isolated bacterial strain at the species level, the nucleotide sequences of the gyrase B (gyrB) and RNA polymerase D (rpoD) genes were obtained and compared with the sequences in the GenBank database. As the result, the causal agent of the stem rot disease was identified as Pseudomonas marginalis. To determine the pathogenicity of the isolated bacterial strain, it was inoculated into the stem of healthy R. asiaticus plant, the inoculated plant showed a lesion with the same characteristics as the naturally infected plant. Based on these results, this is the first report of bacterial stem rot on R. asiaticus caused by P. marginalis in Korea.

Expression of microRNA-218 and its Clinicopathological and Prognostic Significance in Human Glioma Cases

  • Cheng, Mao-Wei;Wang, Ling-Ling;Hu, Gu-Yu
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권5호
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    • pp.1839-1843
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    • 2015
  • Background: MicroRNAs are a class of noncoding RNAs which regulate multiple cellular processes during tumor development. The purpose of this report is to investigate the clinicopathological and prognostic significance of miR-218 in human gliomas. Materials and Methods: Quantitative RT-PCR (qRT-PCR) was conducted to detect the expression of miR-218 in primary normal human astrocytes, three glioma cell lines and 98 paired glioma and adjacent normal brain tissues.Associations of miR-218 with clinicopathological variables of glioma patients were statistically analyzed. Finally, a survival analysis was performed using the Kaplan-Meier method and Cox's proportional hazards model. Results: The expression level of miR-218 in primary normal human astrocytes was significantly higher than that in glioma cell lines (p<0.01). Also, the expression level of miR-218 in glioma tissues was significantly downregulated in comparison with that in the adjacent normal brain tissues (p<0.001). Statistical analyses demonstrated that low miR-218 expression was closely associated with advanced WHO grade (p=0.002) and low Karnofsky performance score (p=0.010) of glioma patients. Kaplan-Meier analysis with the log-rank test showed that patients with low-miR-218 expression had poorer disease-free survival and overall survival (p=0.0045 and 0.0124, respectively). Multivariate analysis revealed that miR-218 expression was independently associated with the disease-free survival (p=0.009) and overall survival (p=0.004) of glioma patients. Conclusions: Our results indicate that miR-218 is downregulated in gliomas and that its status might be a potential valuable biomarker for glioma patients.

Molecular Identification of Vaginal Lactobacillus spp. Isolated from Korean Women

  • CHANG, CHUNG EUN;SYLVIA I. PAVLOVA;LIN TAO;EUN-KI KIM;SEUNG CHUL KIM;HYUN SHIK YUN;JAE-SEONG SO
    • Journal of Microbiology and Biotechnology
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    • 제12권2호
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    • pp.312-317
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    • 2002
  • Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • 제42권1호
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

A report of 27 unrecorded bacterial species within the class Alphaproteobacteria isolated from various sources of Korea in 2021

  • Haneul Kim;Heeyoung Kang;Wonyong Kim;Myung Kyum Kim;Jung-Hoon Yoon;Seung Bum Kim;Taegun Seo;Che Ok Jeon;Wan-Taek Im;Kiseong Joh
    • Journal of Species Research
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    • 제12권spc2호
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    • pp.33-44
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    • 2023
  • In 2021, a total of 27 bacterial strains were isolated from soil, tree bark, moss, wetland, sea sediment, tidal flat, seawater and seaweed within Republic of Korea. Based on the analysis of 16S rRNA gene sequence (>98.7% sequence similarity), these isolates were assigned to the class Alphaproteobacteria as unrecorded species in Korea. The 27 strains were classified into the 10 families: Maricaulaceae of the order Caulobacterales; Brucellaceae, Methylobacteriaceae, Nitrobacteraceae and Rhizobiaceae of the order Hyphomicrobiales; Micropepsaceae of the order Micropepsales; Rhodobacteraceae of the order Rhodobacterales; Azospirillaceae of the order Rhodospirillales; and Erythrobacteraceae and Sphingomonadales of the order Sphingomonadaceae. There is no official report of these 27 species in Korea. Therefore, we report 27 isolates as unrecorded species, and described isolation sources, Gram-stain reactions, physiological and biochemical properties and morphologies of these strains.