• Title/Summary/Keyword: 16S rRNA gene sequences

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Bacterial Diversity and Distribution of Cultivable Bacteria Isolated from Dokdo Island (독도 주변의 해수에서 분리한 세균의 다양성과 군집구조 분석)

  • Sung, Hye-Ri;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.263-272
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    • 2010
  • One hundred sixty three strains showing different colony morphological characteristics on different concentration of marine agar (MA) plates were isolated from ambient seawater near Dokdo island. Bacterial diversity and distributions were studied by phylogenetic analysis of the partial 16S rRNA gene sequences. One hundred sixty three strains were partially sequenced and analyzed phylogenetically. They were composed of 5 phyla, of which gamma-proteobacteria (58%), alpha-proteobacteria (20%), bacteriodetes (16%) were predominant. They were affiliated with 90 species. The 16S rRNA sequence similarity of the isolates was in 93.3 to 100 % range to reported sequence data. Thirty six isolates of among them were assumed to be novel species candidates based on similarity analysis of the 16S rRNA gene sequences. Overall, Proteobacteria and Bacteriodetes of the Dokdo coastal sea water showed a high diversity.

Discrepancies in genetic identification of fish-derived Aeromonas strains

  • Han, Hyun-Ja;Kim, Do-Hyung
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.391-400
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    • 2009
  • Genetic identification of 17 fish-derived Aeromonas strains was attempted using 5 housekeeping genes. 16S rRNA, gyrB, rpoD, dnaJ and recA genes from the 17 strains were amplified, and total of 85 amplicons were sequenced. DNA sequences of the strains and type strains of the 17 Aeromonas homology groups were used for genetic identification and phylogenetic analyses. None of the strains was identified as a single species using the 16S rRNA gene, showing the same identities (average = 99.7%) with several Aeromonas species. According to gyrB, rpoD, dnaJ, and recA, 9 strains and RFAS-1 used in this study were identified as A. hydrophila and A. salmonicida, respectively. However, the other strains were closely related to 2 or more Aeromonas species (i.e., A. salmonicida, A. veronii, A. jandaei, A. media and A. troda) depending on the genetic marker used. In this study, gyrB, rpoD, dnaJ and recA gene sequences proved to be advantageous over 16S rRNA for the identification of field Aeromonas isolates obtained from fish. However, there are discrepancies between analyses of different phylogenetic markers, indicating there are still difficulties in genetic identification of the genus Aeromonas using the housekeeping genes used in this study. Advantages and disadvantages of each housekeeping gene should be taken into account when the gene is used for identification of Aeromonas species.

Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio ichthyoenteri Species-specific Primer 개발)

  • Moon Young-Gun;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.117-124
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    • 2005
  • Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

Marine Bacteria Associated with the Korean Brown Alga, Undaria pinnatifida

  • Lee, Yoo-Kyung;Jung, Hyun-Jung;Lee, Hong-Kum
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.694-698
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    • 2006
  • Several marine bacterial strains were isolated from Undaria pinnatifida (Miyok in Korean). Sixty-six strains were isolated on R2A agar media at $10^{\circ}C$ and identified by a phylogenetic analysis of the 16S rRNA gene sequences. They were grouped into 10 different sequence types based on the initial sequence analysis of the 5' domain of the gene (approximately 500 bp). Full sequences of 16S rRNA gene, were obtained from one strain in each sequence type and the species-affiliation was determined using phylogenetic and sequence similarity analyses. The results of the analyses indicated that they were closely related to Psychrobacter aquimaris, P. celer, P. nivimaris, P. pulmonis, Psychromonas arctica or Bacillus psychrodurans. These bacteria are marine or psychrotrophic bacteria. Because the sporophytes of U. pinnatifida are cultured on the costal area during winter, the U. pinnatifida-associated bacteria appeared to grow at low temperatures. U. pinnatifida sporophytes can be a good source for the isolation of psychrotrophic bacteria.

First Report on the Witches' Broom in Annual Statice (Limonium sinuatum) in Korea

  • Chung, Bong-Nam;Huh, Kun-Yang;Jeong, Myeong-Il
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.383-386
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    • 2005
  • In 2003 typical phytoplasma symptoms of witches' broom and flower malformation were observed on statice (Limonium sinuatum) plants grown at commercial greenhouses in Busan, South Korea. The DNA extracted from the infected leaves was amplified using universal primer pair of Pl/P6 derived from conserved 16S rRNA gene of Mollicutes giving the expected Polymerase chain reaction (PCR) product of 1.5 kb. In the nested PCR assays, the expected DNA fragment of 1.1 kb was amplified with the specific primer pair 16Fl/Rl that was designed on the basis of aster yellows (AY) phytoplasma 16S rDNA sequences. The 1.1 kb PCR products were cloned and nucleotide sequences were determined. The sequences were identical to that of Onion yellows OY phytoplasma (GenBank accession no. D12569) isolated from Onion in Japan. Electron microscopy of thin sections of leaf veins showed phytoplasma bodies in the phloem. Statice witches' broom symptom occurred on statice in commercial greenhouses in Korea was confirmed as infection of AY phytoplasma by transmission electron microscopy observation, and by determination of 16S rRNA gene sequences of phytoplasma.

Phylogenetic Differentiation of Streptomyces spp. Isolated from Potato Scab Lesions in Jeju Island of Korea on the Basis of 16S rRNA Gene Sequences (제주도 감자 더뎅이병징에서 분리된 Streptomyces spp.의 16S rRNA 유전자 염기서열 분석)

  • Lee, Soo-Hyun;Ko, Young-Hwan;Kim, Chang-Jin;Kim, Bum-Joon;Lee, Keun-Hwa
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.347-351
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    • 2007
  • Potato scab is prevalent in all potato-growing areas of Jeju Island and causes economically significant losses. Streptomyces species are known as pathogens of potato scab. In this study, we analyzed the 16S rRNA sequences of Streptomyces spp, which are isolated from potato scab lesions in Jeju Island, and constructed 16S rRNA phylogenetic tree. All isolates were clearly differentiated into the genus Streptomyces, and the tree also showed that new scab-causing Streptomyces spp or not yet named species of Streptomyces are existed in Jeju Island, Korea.

Bacterial Diversity in the Human Saliva from Different Ages

  • Kang, Jung-Gyu;Kim, Seong-Hwan;Ahn, Tae-Young
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.572-576
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    • 2006
  • To obtain primary idea on oral bacterium species that are generally present in periodotally healthy Koreans, the oral bacterial flora in the saliva of four periodontally healthy Koreans at different ages (5, 32, 35, 65) was investigated in this study. For this investigation, 16S rRNA gene clone libraries were generated from the saliva of the four healthy Koreans, and 50 clones were randomly selected from each saliva clone library and sequenced. Totally, 37 different kinds of bacterial 16S rRNA gene sequences were identified based on sequence homology search through GenBank database. The 37 kinds of saliva clone sequences were classified to 14 genera and 2 uncultured and 1 unidentified bacteria. Among the 14 identified genera, Streptococcus, Prevotella, and Veillollella were common genera, and Streptococcus was dominant genus that accounted for 7 different species. Among the seven Streptococcus species, S. salivarius appeared as the most common species. More numbers of species belonging to the genera Streptococcus and Prevotella was present in saliva from ages 32 and 35. While saliva from ages 5 and 65 showed more numbers of species belonging to the genera Rothia, including potential pathogenic species. Overall, saliva of a young child and a senior showed higher bacterial diversity than that of young adults.

Inferring the Molecular Phylogeny of Chroococcalian Strains (Blue-green algae/Cyanophyta) from the Geumgang River, Based on Partial Sequences of 16S rRNA Gene

  • Lee, Wook-Jae;Bae, Kyung-Sook
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.335-339
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    • 2002
  • Partial sequences of 16S rRNA gene of five chroococcalian blue-green algal strains, Aphanothece nidulans KCTC AG10041, Aphanothece naegelii KCTC AG10042, Microcystis aeruginosa KCTC AG10159, Microcystis ichthyoblabe KCTC AG10160, and Microcystis viridis KCTC AG10198, which were isolated from water from the Geumgang River, were determined and were inferred their phylogenetic and taxonomic positions among taxa of order Chroococcales. Most taxa of Chroococcales whose partial 16S rRNA gene sequences were aligned in this study, are clustered with other related taxa. Aphanothece nidulans KCTC AG10041 and Aphanothece naegelii KCTC AG10042 made a cluster with other European species of these genera, which supported 100% of the bootstrap trees with a very high sequence similarity (97.4-99.4%) in this study. Three strains, Microcystis aeruginosa KCTC AG10159, M. ichthyoblabe KCTC AG10160, and M. viridis KCTC AG10198, formed a cluster with other Microcystis spp. supported 100 % of the bootstrap trees with a similarity of 97.0-99.9% except for two strains. However, this phylogentic tree made no resolution among the species of Microcystis spp. The topology of the tree reconfirmed the taxonomic status of three species of Microcystis, identified in this study based on the morphology, as three colonial types of Microcystis aeruginosa com. nov. Otsuka et al. (1999c). The genera of chroococcalian cyanophytes are heterogeneously clustered in these sequence analyses. We suggest that more molecular studies on the genera of Chroococcales with reference strains, widely collected from restricted geographic or environmental ranges, get accurate taxonomic or phylogenetic determinations.

Bacterial Diversity at Different Depths in Lead-Zinc Mine Tailings as Revealed by 16S rRNA Gene Libraries

  • Zhang, Han-Bo;Shi, Wen;Yang, Ming-Xia;Sha, Tao;Zhao, Zhi-Wei
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.479-484
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    • 2007
  • Bacterial communities at 10 cm, 100 cm, and 200 cm depths in a 100-year-old lead-zinc tailing heap were evaluated by constructing 16S rRNA gene libraries. In total, 98 operational taxonomic units (OTUs) were identified from 193 clones at a 3% sequence difference level. The OTU number and species richness decreased with the depth. Species composition was significantly different between the three libraries. Fifty-seven percent of the examined clones were Acidobacteria and 27% belonged to Proteobacteria. Other sequences included Chloroflexi, Firmicutes, Chlamydiae, Actinobacteria, Gemmatimonadetes, Nitrospira, and three unclassified OTUs. Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were mainly distributed in the rhizosphere of naturally colonizing plants; however, Deltaproteobacteria, Acidobacteria, and Chloroflexi tended to inhabit the deeper tailings (below the 100 cm-depth).

Molecular Identification of the Bloom-forming Cyanobacterium Anabaena from North Han River System in Summer 2012 (북한강 수계 조류대발생 원인종 남조 Anabaena의 분자계통학적 검토)

  • Li, Zhun;Han, Myung-Soo;Hwang, Su-Ok;Byeon, Myeong-Seop;Hwang, Soon-Jin;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.301-309
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    • 2013
  • Between May and August 2012, a massive cyanobacterial bloom with Anabaena has been occurred throughout the North Han River. Sampling was conducted at one station on each lake, L. Uham, L. Cheongpyung, and L. Paldang, where occurred a dense bloom, in 13 July. According to the microscopic examination, the blooms was dominated by one specific filamentous cyanobacterium Anabaena and other phytoplankton. Morphologically, previous literature proven that this Anabaena species is A. crassa (Lemmermann) Komark.-Legn. & Cronberg. However, identification of species in a mixed population is complicated due to limited morphological differences. Therefore, with live sample including trichome, akinete and heterocyst, the sequences of 16S rRNA gene of Anabaena isolates were cloned and analyzed, and three 16S rRNA gene sequences of 1188~1520 bp in length were obtained. It was shown from the homologous analysis results that the obtained 16S rRNA sequences were highly homologous to the relevant sequences of A. crassa in GenBank. The 16S rRNA sequences of 63 species were retrieved from GenBank, and the phylogenetic tree was constructed by using these sequences.