• Title/Summary/Keyword: 16s rDNA

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PCR-based identification of uncultural bacterium from malformed Agaricus blazei (신령버섯의 기형으로부터 배양불능세균의 PCR 검정)

  • Shin, Pyung-Gyun;Park, Yun-Jung;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Se-Jong;Lee, Keum-Hee
    • Journal of Mushroom
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    • v.8 no.4
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    • pp.157-160
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    • 2010
  • Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.

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Identification and Comparison of the Nucleotide Sequence of 16S-23S rRNA Gene Intergenic Small SR(Spacer Region) of Lactobacillus rhamnosus ATCC 53103 with Those of L. casei, L. acidophilus and L. helveticus

  • Byun, J.R.;Yoon, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1816-1821
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    • 2003
  • Reliable PCR based identification of lactobacilli has been described utilizing the sequence of 16S-23S rRNA intergenic spacer region. Those sequence comparisons showed a high degree of difference in homology among the strains of L. rhamnosus, L. casei, L. acidophilus and L. helveticus whose 16S-23S rRNA intergenic small SR's sizes were 222 bp, 222 bp, 206 bp and 216 bp respectively. The sequence of 16S-23S rRNA intergenic spacer region of L. rhamnosus ATCC 53103 revealed the close relatedness to those of L. casei strains by the homology ranges from 95.4% to 97.2%. 16S-23S rRNA intergenic spacer region nucleotide sequence of L. acidophilus showed some distant relatedness with L. rhamnosus ATCC 53103 with the homology ranges from 40.3% to 41.8% and that with L. helveticus was shown to be 30% of homology, which exists at the most distant phylogenetic relatedness. The identification of species and strain of lactobacilli was possible on the basis of these results. The common sequences among the 17 strains were CTAAGGAA located in the initiating position of the DNA and some discrepancies were found between the same strains based on these results.

Detection of Nocardia sp. Hl7-1 by PCR during Bioremediation of Crude Oil-Contaminated Soil (원유 오염토양의 Bioremediation과정 동안 PCR을 이용한 Nocardia sp. Hl7-1의 검출)

  • Baek, Kyung-Hwa;Lee, Young-Ki;Lee, In-Sook;Oh, Hee-Mock;Yoon, Byung-Dae;Kim, Hee-Sik
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.91-95
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    • 2004
  • For the detection of the oil-degrading bacterium, Nocardia sp. Hl7-1, inoculated during the bioremediation of oil-contaminated soil, a species-specific primer was constructed based on the 16S rDNA sequence of this strain. Two forward primers and two reverse primers were designed and tested against both closely and distantly related bacterial strains. All the primers designed were specific to the Nocardia sp. H17-1. Particularly, primer sets NH169F-NH972R and NH575F-NH972R could be used to detect 50 fg of template DNA and TEX>$1.2${\times}$10^4$ CFU/g of sandy soil. These two PCR primer sets successfully detected the H 17-1 strain in the oil-con-laminated soil samples containing heterogeneous DNA. We also conformed the primer specificity by restriction-enzyme cleavage of the PCR products and denaturing gradient gel electrophoresis.

Monitoring the Bacterial Community Dynamics in a Petroleum Refinery Wastewater Membrane Bioreactor Fed with a High Phenolic Load

  • Silva, Cynthia C.;Viero, Aline F.;Dias, Ana Carolina F.;Andreote, Fernando D.;Jesus, Ederson C.;De Paula, Sergio O.;Torres, Ana Paula R.;Santiago, Vania M.J.;Oliveira, Valeria M.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.21-29
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    • 2010
  • The phenolic compounds are a major contaminant class often found in industrial wastewaters and the biological treatment is an alternative tool commonly employed for their removal. In this sense, monitoring microbial community dynamics is crucial for a successful wastewater treatment. This work aimed to monitor the structure and activity of the bacterial community during the operation of a laboratory-scale continuous submerged membrane bioreactor (SMBR), using PCR and RT-PCR followed by denaturing gradient gel electrophoresis (DGGE) and 16S rRNA libraries. Multivariate analyses carried out using DGGE profiles showed significant changes in the total and metabolically active dominant community members during the 4-week treatment period, explained mainly by phenol and ammonium input. Gene libraries were assembled using 16S rDNA and 16S rRNA PCR products from the fourth week of treatment. Sequencing and phylogenetic analyses of clones from the 16S rDNA library revealed a high diversity of taxa for the total bacterial community, with predominance of Thauera genus (ca. 50%). On the other hand, a lower diversity was found for metabolically active bacteria, which were mostly represented by members of Betaproteobacteria (Thauera and Comamonas), suggesting that these groups have a relevant role in the phenol degradation during the final phase of the SMBR operation.

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.

Molecular characterization of bacterial endosymbionts of Acanthamoeba isolates from infected corneas of Korean patients

  • Xuan, Ying-Hua;Yu, Hak-Sun;Jeong, Hae-Jin;Seol, Sung-Yong;Chung, Dong-Il;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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    • pp.1-9
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    • 2007
  • The endosymbionts of 4 strains of Acanthamoeba(KA/E9, KA/E21, KA/E22, and KA/E23) isolated from the infected corneas of Korean patients were characterized via orcein stain, transmission electron microscopic examination, and 16S rDNA sequence analysis. Double membrane-bound, rod-shaped endosymbionts were distributed randomly throughout both the trophozoites and cysts of each of Acanthamoeba isolates. The endosymbionts of KA/E9, KA/E22, and KA/E23 were surrounded by electron-translucent areas. No lacunae-like structures were observed in the endosymbionts of KA/E21, the bacterial cell walls of which were studded with host ribosomes. Comparative analyses of the 16S rDNA sequences showed that the endosymbionts of KA/E9, KA/E22 and KA/E23 were closely related to Caedibacter caryophilus, whereas the KA/E21 endosymbiont was assigned to the Cytophaga-Flavobacterium-Bacteroides(CFB) phylum. In the 4 strains of Acanthamoeba, the hosts of the endosymbionts were identified as belonging to the Acanthamoeba castellanii complex, which corresponds to the T4 genotype. Acanthamoeba KA/E21 evidenced characteristics almost identical to those of KA/E6, with the exception of the existence of endosymbionts. The discovery of these endosymbionts from Acanthamoeba may prove essential to future studies focusing on interactions between the endosymbionts and the amoebic hosts.

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.

Deinococcus rubrus sp. nov., a Bacterium Isolated from Antarctic Coastal Sea Water

  • Srinivasan, Sathiyaraj;Lim, Sangyong;Lim, Jae-Hyun;Jung, Hee-Young;Kim, Myung Kyum
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.535-541
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    • 2017
  • Two Gram-staining-negative, red-pinkish, coccus-shaped, non-motile, and aerobic bacterial strains, designated $Ant21^T$ and Ant22, were isolated from the Antarctic coastal sea water. Strains $Ant21^T$ and Ant22 showed UVC and gamma radiation resistance. Phylogenetic analyses based on 16S rRNA gene sequences determined that these strains belong to the genus Deinococcus. Through the analyses of the 16S rRNA gene sequences, strains $Ant21^T$ and Ant22 were found to have 97.7% and 97.8% similarity to Deinococcus marmoris DSM $12784^T$ and 97.0% and 97.2% similarity to Deinococcus saxicola AA-$1444^T$, respectively. The sequence similarity with the type strains of other Deinococcus species was less than 96.9% for both strains. Strains $Ant21^T$ and Ant22 shared relatively high 16S rRNA gene sequence similarity (99.3%) and had a closely related DNA reassociation value of $84{\pm}0.5%$. Meanwhile, they showed a low level of DNA-DNA hybridization (<30%) with other closely related species of the genus Deinococcus. The two strains also showed typical chemotaxonomic features for the genus Deinococcus, in terms of the major polar lipid (phosphoglycolipid) and the major fatty acids ($C_{16:0}$, $C_{16:1}$ ${\omega}6c/{\omega}7c$, $iso-C_{17:0}$, and $iso-C_{15:0}$). They grew at temperatures between $4^{\circ}C$ and $30^{\circ}C$ and at pH values of 6.0-8.0. Based on the physiological characteristics, the 16S rRNA gene sequence analysis results, and the low DNA-DNA reassociation level with Deionococcus marmoris, strains $Ant21^T$ ($=KEMB\;9004-167^T$ $=JCM\;31436^T$) and Ant22 (KEMB 9004-168 =JCM 31437) represent novel species belonging to the genus Deinococcus, for which the name Deinococcus rubrus is proposed.

Phylogenetic Inter- and Intrarelationships of the Genus Microbispora of the Family Streptosporangiaceae Based on 16S Ribosomal DNA Sequences (16S Ribosomal DNA 염기서열 분석에 근거한 Streptosporangiaceae과 Microbispora 속의 계통 관계)

  • Lee, Soon-Dong
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.429-434
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    • 2003
  • The 16S rDNA sequences of nine strains, two type strains of validated Microbispora species and a strain of invalidated Microbispora species, and six soil isolates, were determined and compared with those of representatives of the family Streptosporangiaceae. The phylogenetic analysis indicated that all of the validated species of the genus Microbispora consistently formed a monophyletic unit and were well separated from the other genera of the family Streptosporangiaceae. All the isolates were placed to the genus Microbispora, whereas an invalidated Microbispora species, Microbispora griseoalba IMSNU $22049^{T}$ (= KCTC $9314^{T}$), was closely related to members of the genus Nocardia.

Occurrence of the B- and Q-biotypes of Bemisia tabaci in Korea (담배가루이 B와 Q 계통의 국내 발생 상황)

  • Lee, Min-Ho;Kang, Seok-Young;Lee, Sun-Young;Lee, Heung-Su;Choi, June-Yeol;Lee, Gwan-Seok;Kim, Whang-Yong;Lee, Si-Woo;Kim, Seon-Gon;Uhm, Ki-Baik
    • Korean journal of applied entomology
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    • v.44 no.3 s.140
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    • pp.169-175
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    • 2005
  • The occurrence of tobacco whiteflies, Bemisia tabaci, in greenhouses was monitored in Korea in 2005. Bemisia tabaci occurred in the rose, sweet pepper, tomato, and cucumber greenhouses of Chungbuk, Chungnam, Gyongnam, and Jeonnam Provinces, but not in Jeonbuk and Gyongbuk Provinces. The biotypes and genetic differentiation of the whiteflies collected in each regions were analyzed by mitochondrial 16S DNA sequences. The 16S DNA sequences of Jincheon (Chungbuk Province) samples were similar to DNA data reported from Japan and Israel which were known as the B biotype. However, the DNA sequences of the Buyeo (Chungnam), Geoje (Gyongnam) and Boseong (Jeonnam) collections, which were 100% homologous showed over 99% similarity to the DNA of Q biotype from Spain and Egyrt. Here we report the first founding of the Q biotype in Korea. It is assumed that, unlike the B biotype reported from Jincheon since 1998, the Q biotype might have been introduced recently from the certain foreign region/country to the greenhouses in those provinces.