• Title/Summary/Keyword: 16S rDNA analysis

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Multilocus sequence analysis of the genus Aliivibrio: Identification and phylogeny of Aliivibrio species isolated from cultured walleye pollock (Gadus chalcogrammus) in Korea

  • Nam, U-Hwa;Seo, Hyun-Joon;Jang, Su-Rim;Kim, Mi-Ri;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.32 no.2
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    • pp.69-80
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    • 2019
  • We performed MLSA (multilocus sequence analysis) and phenotypic characterization of Aliivibrio species isolated from walleye pollock (Gadus chalcogrammus) maintained in 3 different facilities of Gangwon Province, the east coast of Korea. Of 38 Aliivibrio species identified by 16S rDNA sequences, 12 strains were randomly selected and MLSA was conducted with 5 house-keeping genes (gapA, gyrB, pyrH, recA and rpoA) and 16S rDNA gene. Phylogenetic analysis and homology of the concatenated sequences (4,580 bp) with other Vibrionaceae genera revealed that 4 strains (GNGc16.1, YYGc16.1, YYGc16.2, GSGc18.1) were identified as Aliivibrio logei and one strain (GSGc16.1) as A. wodanis. One strain (GSGc17.1) was tentatively identified as A. logei, but needs further analysis because it did not belong to the same clade with A. logei type strain. 6 strains (GSGc17.2, GNGc16.2, GSGc16.2, GSGc17.3, GSGc18.2, GSGc17.4) need further investigation as potential novel species. Either phenotypic characterization or 16S rDNA sequence alone did not provide enough information for identification of Aliivibrio strains at the species level. A. logei and A. wodanis are generally known as non-pathogenic bacteria, but also known as opportunistic or secondary pathogens of cold water fishes. Cares should be taken to prevent potential outbreaks due to these bacteria, although there was no outbreaks during the sampling period.

Identification of Fusobacterium nucleatum isolated from Korean by F. nucleatum subspecies-specific DNA probes (Dot blot hybridization법을 이용한 Fusobacterium nucleatum 아종-특이 DNA 프로브의 특이성 평가)

  • Kim, Hwa-Sook;Kook, Joong-Ki
    • Journal of Korean society of Dental Hygiene
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    • v.6 no.4
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    • pp.311-324
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    • 2006
  • The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

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Bacterial Diversity and its Phylogenetic Analysis in Lake Sapgyo (삽교호의 세균 다양성과 계통분류학적 분석)

  • Kim, Myeong;Jeon, Eun-Hyeong;An, Tae-Yeong
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.272-276
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    • 2003
  • Sapgyo Lake is an artificial freshwater reservoir which is located to the midwest of Korea and is the main water reservoir for industry and agriculture of the region. In this study we investigated environmental factors and the change of bacterial community with the influence of surrounding inflow water and the seasonal variation using the molecular ecological approach. Water samples were collected at front of the dike in May and August, 2001. Bacterial genomic DNAs were extracted directly and purified for the amplification of bacterial 16S rDNA. Clone libraries of the 16S rDNA were constructed using pGEM-T easy vector and RFLP analysis was performed to make a group as OTUs with 4 base recognizing enzymes (MspI and HaeIII). The estimated values of richness in August sample was higher than in May. Thirty-three of 153 clones in May and thirty-eight of 131 clones in August were sequenced from forward region of bacterial 16S rDNA for about 600~800 bp. Proteobacteria, Cytophaga, gram positive bacteria and Verrucomicrobia were observed both months. Especially, Planctomyces, cyanobacteria and chloroplast appeared in August when algal bloom occurred. On the whole investigation, Sapgyo lake showed a typical community structure of estuarine and was influenced by heterochthonous organic matters from the surrounding stream.

16S rDNA Analysis 9f Bacterial Diversity in Three Fractions of Cow Rumen

  • Cho, Soo-Jeong;Cho, Kye-Man;Shin, Eun-Chule;Lim, Woo-Jin;Hong, Su-Young;Choi, Byoung-Rock;Kang, Jung-Mi;Lee, Sun-Mi;Kim, Yong-Hee;Kim, Hoon;Yun, Han-Dae
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.92-101
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    • 2006
  • The bacterial diversity of the bovine rumen was examined using a PCR-based approach. 16S rDNA sequences were amplified and cloned from three fractions of rumen (solid, fluid, and epithelium) that are likely to represent different bacterial niches. A total of 113 clones were sequenced, and similarities to known l6S rDNA sequences were examined. About $47.8\%$ of the sequences had $90-97\%$ similarity to 16S rDNA database sequences. Furthermore, about $62.2\%$ of the sequences were $98-100\%$ similar to 16S rDNA database sequences. For the remaining $6.1\%$, the similarity was less than $90\%$. Phylogenetic analysis was also used to infer the makeup of the bacterial communities in the different rumen fractions. The Cytophaga-Flexibacter-Bacteroides group (CFB, $67.5\%$), low G+C Gram-positive bacteria (LGCGPB, $30\%$), and Proteobacteria $(2.5\%)$ were represented in the rumen fluid clone set; LGCGPB $(75.7\%)$, CFB$(10.8\%)$, Proteobacteria $(5.4\%)$, high G+C Gram-positive bacteria (HGCGPB, $5.4\%$), and Spirochaetes $(2.7\%)$ were represented in the rumen solid clone set; and the CFB group $(94.4\%)$ and LGCGPB $(5.6\%)$ were represented in the rumen epithelium clone set. These findings suggest that the rumen fluid, solid, and epithelium support different microbial populations that may play specific roles in rumen function.

Genetic Similarity Between Jujube Witches¡?Broom and Mulberry Dwarf Phytoplasmas Transmitted by Hishimonus sellatus Uhler

  • Cha, Byeongjin;Han, Sangsub
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.98-101
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    • 2002
  • Using phytoplasma universal primer pair Pl and P7, a fragment of about 1.8 kb nucleotide sequences of 16S rRNA gene and 16S-23S rRNA intergenic spacer region, and a portion of 23S rRNA gene of jujube witches'broom (JWB) and mulberry dwarf(MD) phytoplasmas were determined. The nucleotide sequences of JWB and MD were 1,850 bp and 1,831 bp long, respectively. The JWB phytoplasma sequence was aligned with the homologous sequence of MD phytoplasma. Twenty-eight base insertions and nine base deletions were found in the JWB phytoplasma sequence compared with that of MD phytoplasma. The similarity of the aligned sequences of JWB and MD was 84.8%. The near-complete 16S rRNA gene DNA sequences of JWB and MD were 1,529 bp and 1,530 bp in length, respectively, and revealed 89.0% homology. The 16S-23S rRNA intergenic spacer region DNA sequences were 263 bp and 243 bp in lengths respectively, while homology was only 70% and the conserved tRNA-lle gene of JWB and MD was located into the intergenic space region between 16S-23S rRNA gene. The nucleotide sequences were 77 bp long in both JWB and MD, and showed 97.4% sequence homology. Based on the phylogenetic analysis of the two phytoplasmas, the JWB phytoplasma belongs to the Elm yellow phytoplasma group (16S rV), whereas, the MD phytoplasma belongs to the Aster yellow group (16S rI).

A New Report of Prionospio kirrae (Annelida: Spionidae) from Korea

  • Lee, Geon Hyeok;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.38 no.2
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    • pp.91-97
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    • 2022
  • Spionid polychaete Prionospio kirrae Wilson, 1990 is newly reported from the Yellow Sea in Korea. This species is characterized by four pairs of branchiae, which are apinnate on chaetigers 2-4 and pinnate on chaetiger 5, a caruncle extending to the posterior end of chaetiger 1, the presence of a distinctly high dorsal crest on chaetiger 11, and the presence of tridentate hooded hooks with rounded apical teeth. Sequences of partial mitochondrial cytochrome c oxidase subunit I (COI), 16S ribosomal DNA(16S rDNA), and the nuclear 18S ribosomal DNA(18S rDNA) of the species are determined from Korean specimens.

Bacterial Community Structure and Diversity of the Zoysia japonica Soil Treated with Liquid Fertilizer Containing Amino Acids (아미노산 액비를 처리한 들잔디 토양 미생물 군집구조 및 다양성)

  • Kim Dong-Il;Kim Dong-Hun
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.103-110
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    • 2006
  • T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in Zoysia japonica soil treated with liquid fertilizer containing amino acids(LFcAA) after spray with herbicide. The results of T-RFLP (terminal restriction fragment length poly-morphism) analysis using restriction enzyme Hae III showed that the T-RFs of various size appeared evenly in the 32 clones of KD3 and 38 clones of KD4 respectively that had been treated with liquid fertilizer containing amino acid(LFcAA) compared to 23 clones of KD2 hat had not been treated with LFcAA. The microbial com- munity structure in KD2 appeared less diverse than those in KD3 and KD4. Analysis of partial sequences for 110 clones from KDI (control), KD2 (non-treated), KD3 (LFcAA 1X), KD4 (LFcAA 2X), respectively, revealed that most bacteria were related with uncultured bacteria in a 16S rDNA sequence similarity range of 91-99% through blast search. Otherwise, the other clones were members of proteobacteria, Acidobacteria, Act-inobacteria, Sphingobacteria and Planctomyces groups. Especially in KD4, members of Alpha Proteobacteria, Rhizobiales, Sphigomonadales, Caulobacterales, Gamma Proteobacteria, the genus Pseudomonas, Betapro-teobacteria, Nitrosomonadales and genus Nitrosospira appeared to be dominant. In addition, Acidobacteria group, Actinobacteria group, Planctomycetacia and Sphingobacteria were also shown. The microbial com-munity structure in Z. japonica soil sprayed with herbicide was affected by LFcAA.

Bacterial Community Diversity Associated with Two Marine Sponges from the South Pacific Ocean based on 16S rDNA-DGGE analysis (남태평양에 서식하는 두 종의 해면 Hyrtios sp.와 Callyspongia sp.의 공생세균 군집의 다양성)

  • Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.255-261
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    • 2010
  • The bacterial community structure associated with two marine sponges, Hyrtios sp. 604 and Callyspongia sp. 612 collected from the South Pacific Ocean were analyzed by 16S rDNA-denaturing gradient gel electrophoresis (DGGE). The phylogenetic analysis showed that the bacterial community associated with Hyrtios sp. 604 contained diverse bacterial groups such as Chloroflexi, Firmicutes, Cyanobacteria, Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, and Acidobacteria. Callyspongia sp. 612 harbored Chloroflexi, Cyanobacteria, Alphaproteobacteria, and Gammaproteobacteria. Hyrtios sp. 604 belonging to genus Hyrtios known to produce natural products showed greater bacterial diversity than Callyspongia sp. 612. Phylum Actinobacteria was shown to be one of dominant bacterial groups in Hyrtios sp. 604. Although the same phyla of bacteria were found in both sponge species, the spongeassociated predominant bacterial groups differed between the two sponges with different chemical characteristics from the same geographical location. Uncultured bacteria represented over 90% of the bacteria diversity present in all bacterial communities of the sponges.

Isolation and Identification of Bacteria Lysing Anabaena cylindrica (Anabaena cylindrica 분해세균의 분리 및 동정)

  • Choi, Yong-Keel;Hong, Yup;Shin, Kyu-Chul;Kim, Min-Seong;Han, Myung-Soo
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.107-112
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    • 2002
  • To isolate the bacteria lysing cyanobacteria, the sediment samples were collected from Dochang and Pal'tang Reservoir and Seokchon Lake. Each sample was smeared on the Anabaena cylindrica lawn and incubated in light chamber for 11 days. Bacteria having cyanobacteria-Iysing activity were isolated from the samples of Seokchon reservoir. Confirmation of cyanobacteria-Iysing activity was carried out to measure chlorophyll a and bacterial cell counting in mixed culture of Anabaena cylindrica and bacteria. Lysis was detected when extracellular meterials was added to the Anabaena cylindrica culture. The isolate was identified by analysis based on 16S rDNA sequence and morphological and physiological properties. The bacterial strain was taxonomically studied by the phylogenetic analysis based on 165 rDNA sequence. This strain was identified as a member of the genus Bacillus and designated as Bacillus sp. CHS1.