• Title/Summary/Keyword: rRNA sequence

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A report of 36 unrecorded bacterial species belonging to the phyla Actinomycetota, Bacillota, Bacteroidota, Deinococcota, and Pseudomonadota isolated in Republic of Korea

  • Che-Ok Jeon;Wonyong Kim;Jin-Woo Bae;Chi-Nam Seong;Wan-Taek Im;Seung-Bum Kim;Jang-Cheon Cho;Myung Kyum Kim;Chang-Jun Cha;Taegun Seo;Jung-Hoon Yoon
    • Journal of Species Research
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    • v.12 no.4
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    • pp.415-429
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    • 2023
  • As part of a comprehensive investigation of indigenous prokaryotic species in the Republic of Korea, 37 bacterial strains belonging to 36 species were isolated from diverse environmental habitats. These strains were assigned to five phyla, namely Deinococcota, Actinomycetota, Bacillota, Bacteroidota, and Pseudomonadota. Each strain was identified based on 16S rRNA gene sequence similarity (>98.7%) and the formation of definite phylogenetic clades with their closest reported species. Among isolates, there is one species belonging to the phylum Deinococcota, five species belonging to the phylum Actinomycetota, four species belonging to the phylum Bacillota, nine species belonging to the phylum Bacteroidota, and 17 species belonging to the phylum Pseudomonadota (comprising eight species of the class Alphaproteobacteria, one species of the class Betaproteobacteria, and eight species of the class Gammaproteobacteria). Based on 16S rRNA gene sequence analysis, each strain was assigned to independent and predefined bacterial species. Since there were no published or official reports regarding these 36 species in the Republic of Korea, they have been reported as unrecorded species in the Republic of Korea. Their Gram stain, cell morphology, colony, basic biochemical characteristics, strain ID, and isolation source of each species are described in the species descriptions.

A streamlined pipeline based on HmmUFOtu for microbial community profiling using 16S rRNA amplicon sequencing

  • Hyeonwoo Kim;Jiwon Kim;Ji Won Cho;Kwang-Sung Ahn;Dong-Il Park;Sangsoo Kim
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.40.1-40.11
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    • 2023
  • Microbial community profiling using 16S rRNA amplicon sequencing allows for taxonomic characterization of diverse microorganisms. While amplicon sequence variant (ASV) methods are increasingly favored for their fine-grained resolution of sequence variants, they often discard substantial portions of sequencing reads during quality control, particularly in datasets with large number samples. We present a streamlined pipeline that integrates FastP for read trimming, HmmUFOtu for operational taxonomic units (OTU) clustering, Vsearch for chimera checking, and Kraken2 for taxonomic assignment. To assess the pipeline's performance, we reprocessed two published stool datasets of normal Korean populations: one with 890 and the other with 1,462 independent samples. In the first dataset, HmmUFOtu retained 93.2% of over 104 million read pairs after quality trimming, discarding chimeric or unclassifiable reads, while DADA2, a commonly used ASV method, retained only 44.6% of the reads. Nonetheless, both methods yielded qualitatively similar β-diversity plots. For the second dataset, HmmUFOtu retained 89.2% of read pairs, while DADA2 retained a mere 18.4% of the reads. HmmUFOtu, being a closed-reference clustering method, facilitates merging separately processed datasets, with shared OTUs between the two datasets exhibiting a correlation coefficient of 0.92 in total abundance (log scale). While the first two dimensions of the β-diversity plot exhibited a cohesive mixture of the two datasets, the third dimension revealed the presence of a batch effect. Our comparative evaluation of ASV and OTU methods within this streamlined pipeline provides valuable insights into their performance when processing large-scale microbial 16S rRNA amplicon sequencing data. The strengths of HmmUFOtu and its potential for dataset merging are highlighted.

Genetic Relationship of the Ampelopsis brevipedunculata var. heterophylla and Vitis thunbergii var. sinuata with the Other Vitis Plants (개머루와 까마귀머루의 유전적 유연관계 분석)

  • Bae, Young-Min
    • Journal of Life Science
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    • v.27 no.1
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    • pp.89-94
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    • 2017
  • DNA sequences of the intergenic spacer 1 and intergenic spacer 2 of the nineteen plants belonging Vitis genus were collected from the Genbank. DNA sequences of the same regions of Vitis thunbergii var. sinuata and Ampelopsis brevipedunculata var. heterophylla, both common plants in Korea, were not available in Genbank. Those two plants were collected, their genomic DNA encoding 18S rRNA, intergenic spacer 1, 5.8S rRNA, intergenic spacer 2 and part of 28S rRNA amplified and DNA sequence determined. DNA sequences of twenty-one plants including two Korean plants were aligned by the Multiple sequence comparison by log-expectation(MUSCLE) algorithm and the alignment was used to calculate neighbor-joining tree and pairwise distance. The results indicate DNA sequences of the two Korean plants are highly homologous with each other, but they are quite distantly related to the other Vitis plants. Distant relationship of the two Korean plants with the other Vitis plants might be due to independent evolution of those two plants in geographically isolated environment. Those two Korean plants are classified in different genera based on the morphology, one in Vitis genus and the other in Ampelopsis genus, providing another example of discrepancy between morphological and genetic classification.

Draft genome sequence of a bacterial plant pathogen Erwinia pyrifoliae strain EpK1/15 isolated from an apple twig showing black shoot blight (가지검은마름병 병징을 보이는 사과나무 가지에서 분리한 식물병원세균인 Erwinia pyrifoliae EpK1/15 균주의 유전체 해독)

  • Lee, Gyu Min;Oh, Eom-Ji;Ko, Seyoung;Park, Jungkum;Park, Duck Hwan;Kim, Donghyuk;Oh, Chang-Sik
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.69-70
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    • 2018
  • Erwinia pyrifoliae is a Gram-negative bacterium causing black shoot blight in apple and Asian pear trees. E. pyrifoliae strain EpK1/15 was isolated in 2014 from an apple twig from the Pocheon, Gyeonggi-do, South Korea. In this study, we report the draft genome sequence of E. pyrifoliae EpK1/15 using PacBio RS II platform. The draft genome is comprised of a circular chromosome with 4,027,225 bp and 53.4% G + C content and a plasmid with 48,456 bp and 50.3% G + C content. The draft genome includes 3,798 protein-coding genes, 22 rRNA genes, 77 tRNA genes, 13 non-coding RNA genes, and 231 pseudo genes.

The complete genome sequence of a marine sponge-associated bacteria, Bacillus safensis KCTC 12796BP, which produces the anti-allergic compounds (해양 해면체로부터 분리한 세균으로 항알러지성물질을 생산하는 Bacillus safensis KCTC 12796BP의 유전체 해독)

  • Hanh, Nguyen Phan Kieu;Kim, Soo Hee;Kim, Geum Jin;Choi, Hyukjae;Nam, Doo Hyun
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.448-452
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    • 2018
  • The full genome sequence of Bacillus safensis KCTC 12796BP which had been isolated from the marine sponge in the seawater of Jeju Island, was determined by Pac-Bio next-generation sequencing system. A circular chromosome in the length of 3,935,874 bp was obtained in addition to a circular form of plasmid having 36,690 bp. The G + C content of chromosome was 41.4%, and that of plasmid was 37.3%. The number of deduced CDSs in the chromosome was 3,980, whereas 36 CDS regions were determined in a plasmid. Among the deduced CDSs in chromosome, 81 tRNA genes and 24 rRNA genes in addition to one tmRNA were allocated. More than 30 CDSs for sporulation, 16 CDSs for spore coat, and 20 CDSs for germination were also assigned in the chromosome. Several genes for capsular polysaccharide biosynthesis and for flagella biosynthesis and chemotaxis in addition to genes for osmotic tolerance through glycine-choline betaine pathway were also identified. Above all, the biosynthetic gene cluster for anti-allergic compounds seongsanamides were found among two non-ribosomal peptide synthetase (NRPS) gene clusters for secondary metabolites.

Molecular Authentication and Phylogenetic Relationship of Bupleurum Species by the rDNA-ITS Sequences (rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석)

  • Moon, Byeong-Cheol;Choo, Byeong-Kil;Ji, Yun-I;Yoon, Tae-Sook;Lee, A-Young;Cheon, Myeong-Sook;Kim, Bo-Bae;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.3
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    • pp.59-68
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    • 2009
  • Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.

Isolation and Identification of Microorganism with Potent Fibrinolytic Activity from Korean Traditional Deonjang (재래식 된장으로부터 혈전용해활성을 나타내는 세균의 분리 및 동정)

  • Hyun Kwang-Wook;Lee Jong-Soo;Ham Jung-Hee;Choi Shin-Yang
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.24-28
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    • 2005
  • A bacterial strain D­1 found to have potent fibrinolytic activity was isolated from Korean traditional Doenjang. It was identified as Bacillus sp. based on its 16S rRNA sequence analysis, morphological and physiological characteristics.

Cohnella panacarvi sp. nov., a Xylanolytic Bacterium Isolated from Ginseng Cultivating Soil

  • Yoon, Min-Ho;Ten, Leonid N.;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.913-918
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    • 2007
  • A Gram-positive, aerobic, rod-shaped, nonmotile, endospore-forming bacterium, designated Gsoil $349^T$, was isolated from soil of a ginseng field and characterized using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences revealed that the strain Gsoil $349^T$ belongs to the family Paenibacillaceae, and the sequence showed closest similarity with Cohnella thermotolerans DSM $17683^T$ (94.1%) and Cohnella hongkongensis DSM $17642^T$ (93.6%). The strain showed less than 91.3% 16S rRNA gene sequence similarity with Paenibacillus species. In addition, the presence of MK-7 as the major menaquinone and $anteiso-C_{15:0},\;iso-C_{16:0},\;and\;C_{16:0}$ as major fatty acids suggested its affiliation to the genus Cohnella. The G+C content of the genomic DNA was 53.4 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil $349^T$ should be treated as a novel species within the genus Cohnella for which the name Cohnella panacarvi sp. nov. is proposed. The type strain is Gsoil $349^T\;(=KCTC\;13060^T=\;DSM\;18696^T)$.

Cohnella damensis sp. nov., a Motile Xylanolytic Bacteria Isolated from a Low Altitude Area in Tibet

  • Luo, Xuesong;Wang, Zhang;Dai, Jun;Zhang, Lei;Fang, Chengxiang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.410-414
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    • 2010
  • A bacterial strain, 13-$25^T$ with xylanolytic activity isolated from a single soil sample, was characterized with respect to its phenetic and phylogenetic characteristics. The cells of the isolate are Gram-staining variable rods, but spore formation was not observed. This strain is catalase- and oxidase-positive, and able to degrade starch and xylan. The predominant fatty acids are anteiso-$C_{15:0}$, $C_{16:0}$, and iso-$C_{16:0}$. The major respiratory quinone is menaquinone 7(MK-7), with a polar lipid profile consistent with the genus Cohnella. The DNA G+C content is 54.3 mol%. The 168 rRNA gene sequence analysis indicates that this organism belongs to the genus Cohnella, with Cohnella panacarvi as the closest phylogenetic neighbor. Low levels of 168 rRNA gene sequence similarity (<97.0%) with respect to other taxa with published names and the identification of distinctive phenetic features in the isolate indicate that the strain 13-$25^T$ represents a novel species of the genus Cohnella, for which the name Cohnella damensis sp. novo is proposed. The type strain is 13-$25^T$ (=CCTCC AB $208103^T$=KCTC $13422^T$).

Unrecorded bacterial species belonging to the phylum Actinobacteria originated from Republic of Korea

  • Kim, Mi-Sun;Lee, Ji-Hee;Kim, Seung-Bum;Cho, Jang-Cheon;Lee, Soon Dong;Joh, Ki-seong;Cha, Chang-Jun;Im, Wan-Taek;Bae, Jin-Woo;Jahng, Kwangyeop;Yi, Hana;Seong, Chi-Nam
    • Journal of Species Research
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    • v.6 no.1
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    • pp.25-41
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    • 2017
  • As a subset study for the collection of Korean indigenous prokaryotic species, 62 bacterial strains belonging to the phylum Actinobacteria were isolated from various sources. Each strain showed higher 16S rRNA gene sequence similarity (>98.75%) and formed a robust phylogenetic clade with closest species of the phylum Actinobacteria which were defined with valid names, already. There is no official description on these 62 actinobacterial species in Korea. Consequently, unrecorded 62 species of 25 genera in the 14 families belonging to the order Actinomycetales of the phylum Actinobacteria were found in Korea. Morphological properties, basic biochemical characteristics, isolation source and strain IDs are described in the species descriptions.