• Title/Summary/Keyword: 16S rDNA(16S rRNA)

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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.

Morphological Characteristics of Bemisia tabaci(Gennadius) (Homoptera: Aleyrodidae) and Discrimination of Their Biotypes in Korea by DNA Makers (담배가루이 Bemisia tabaci(Gennadius)(Homoptera: Aleyrodidae)의 형태적 특징과 DNA 표식자에 의한 biotype 판별)

  • 이명렬;안성복;조왕수
    • Korean journal of applied entomology
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    • v.39 no.1
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    • pp.5-12
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    • 2000
  • The sweetpotato whiteflies, Bemisia tabaci(Gennadius), were found recently in Korea on Glycine max, Euphorbia pulcherrima, and Rosa hybrida. The biotype identity of Bemisia tabaci in Korea was determined by several DNA markers including the random amplified polymorphic DNAs, and restriction fragments length polymorphism of mitochondrial 12S and 16S rRNA genes. The electromorph profiles of DNA fragments from the rose(Jincheon) and poinsettia(Seoul) populations in Korea are both identical to those of B biotypes distributed in Australia, Israel, and Japan. The populations of B. tabaci collected on Glycine max, Ipomea batatas, and Perilla frutescens in different localities retained the same DNA markes with the population from Lonicera japonica and shikoku of Japan. These populations are non-B biotype and considered as an indigenous type in the Far Eastern Asia Region including Korea and Japan, Morphological Characteristics of B. Tabaci were also observed by the scanning electron microscope and described with the comparison to the other important whitefly pest, Trialeurodes vaporariorum (Westwood).

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Community Structure of Bacteria Associated with Two Marine Sponges from Jeju Island Based on 16S rDNA-DGGE Profiles (16S rDNA-DGGE를 이용한 2종의 제주도 해양 해면의 공생세균의 군집 구조)

  • Park, Jin-Sook;Sim, Chung-Ja;An, Kwang-Deuk
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.170-176
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    • 2009
  • Culture-independent 16S rDNA-DGGE profiling and phylogenetic analysis were used to examine the predominant bacterial communities associated with the two sponges, Dictyonella sp. and Spirastrella abata from Jeju island. The culture-independent approach involved extraction of total bacterial DNA, PCR amplification of the 16S ribosomal DNA using primer pair 341f-GC and 518r, and separation of the amplicons on a denaturing gradient gel. Denaturing gradient gel electrophoresis banding patterns indicated 8 and 7 bands from the two sponge species, Dictyonella sp. and Spirastrella abata, respectively. There were not common major bands in two different sponges. Comparative sequence analysis of variable DGGE bands revealed from 93% to 98% similarity to the known published sequences. The dominant bacterial group of Dictyonella sp. belonged to uncultured Gammaproteobacteria, while, that of Spirastrella abata belonged to uncultured Alphaproeobacteria and Firmicutes. DGGE analysis indicated predominant communities of the sponge-associated bacteria differ in the two sponges from the same geographical location. This result revealed that bacterial community profiles of the sponges were host species-specific.

Archaeal Communities in Mangrove Soil Characterized by 16S rRNA Gene Clones

  • Yan, Bing;Hong, Kui;Yu, Zi-Niu
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.566-571
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    • 2006
  • An archaeal 16S rRNA gene library was constructed from mangrove soil. Phylogenetic analysis revealed archaea in mangrove soil including the Crenarchaeota (80.4%) and Euryarchaeota (19.6%) phyla. The archaeal community in mangrove soil appears to be a mixture of organisms found in a variety of environments with the majority being of marine origin.

Phylogenetic Analysis of Pectobacterium Species Using the 16S-23S rRNA Intergenic Spacer Regions

  • Kwon, Soon-Wo;Cheun, Meung-Sook;Kim, Sang-Hee;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • v.16 no.2
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    • pp.98-104
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    • 2000
  • For the taxonomic evaluaition, 15 strains of the genus Pectobacterium and Erwinia were analyzed for 16S-23S rDNA intergenic spacer regions (ISRs). These species contained two types of ISRs, large and small ISRs. Large ISRs were on the range of 474-569 bp size, and coding transfer $\textrm{RNA}^{11e}$($\textrm{tRNA}^{11e}$) and $\textrm{tRNA}^{Ala}$. Small ISRs were 354-459 bp in length and coding $\textrm{tRNA}^{Glu}$. The sequence variations of two ISRs among species and strains were very high as compared with 16S rRNA gene sequences. By phylogenetic trees on the basis of two ISRs, Pectobacterium ere differentiated into P. carotovorum-P. cactiaidum group and P. chrysanthemi group. However, the taxonomic position of E. cypripedii and E. rhapontici, which were not clear on taxonomic delineation between Pectobacterium and Erwinia, were not clearly resolved on the basis of ISRs.

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Identification of Bacteria from Periapical Abscess Using 16S rDNA Clone Libraries. (16S rDNA 클론 Libraries를 이용한 치근단 농양 병소의 세균 동정)

  • 유소영;김미광;김화숙;황호길;김평식;임성훈;오상호;민정범;국중기
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.195-198
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    • 2004
  • Molec-ular analysis was performed on the microflora found In the necrotic pulpal tissue collected from 5 infected root canals that were diagnosed as a periapical abscess. 16S rRNA coding gene (rDNA) library construction and sequencing were performed in order to identify the microflora, The 16S rDNA sequences from 278 clones were identified by a comparison with the database sequence in GenBank. Three phylum and 31 species, which were related to the oral microflora, were identified from the 3 samples (No. 87, 105, and 115). Dialister invisus (5.6%), Peptostreptococcus micron (18.3%), and Veillonella sp. (3.3%) were the organism present in all tee samples. Lac-tobacillusfementum (2.8%),Eubacterumsp./E. infirmum (6.7%), Shuttleworthiasatelles (3.9%), Psudorarnihacfer alactoiyticus (13.3%), Bulleidia moorei (2.8%), and Prevotella denticola (1.1%) were found in two samples. Two phylum and low species of environmental microflora were identified from 2 samples (No.95 and 101). The reason for this might be contamination of the samples with dental water. These results showed that molecular analysis could reveal more diverse microflora that are associated with endodontic infections than that revealed by conventional cultural methods. In addition, these results may of for the basic data to epidemiological studies related with endodontic infection.

Analysis of 16S-23S rRNA Intergenic Spacer Region of Vibrio vulnificus (Vibrio vulnificus의 16S-23S rRNA Intergenic Spacer Region 분석)

  • PARK Young Mi;LEE Jehee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.3
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    • pp.239-246
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer region (ISR) of Vibrio vulnificus KCTC 2959. ISRs were amplified by primers complementary to conserved regions of 16S and 23S rRNA genes. ISR amplicons were cloned and sequenced. Analysis of the ISR sequences showed that V. vulnificus KCTC 2959 contains five types of polymorphic ISRs. Size of ISRs ranged from 424 to 741 bp in length and the number of tRNA genes ranged from one to four. The ISRs were designated as ISR-E $(tRNA^{Glu}),\;ISR-IA\;(tRNA^{Ile}-tRNA^{Ala})$, ISR-EKV $(tRNA^{Glu}-tRNA^{Lys}-tRNA^{Val})$, ISR-IAV $(tRNA^{Ile}-tRNA^{Ala}-tRNA^{val})$ and ISR-EKAV $(tRNA^{Glu}-tRNA^{Lys}-tRNA^{Ala}-tRNA^{Val})$ based on their tRNA genes. Multiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability. We used the sequences of variable domains to design species-specific primer for detection PCR. Specificity of the primers was examined using genomic DNA prepared from 18 different Vibrio species. The results showed that the PCR using primers designed in this study can be used to detect V. vulnificus from other Vibrio species.

Detection of Pectobacterium chrysanthemi Using Specific PCR Primers Designed from the 16S-23S rRNA Intergenic Spacer Region

  • Kwon, Soon-Wo;Myung, In-Sik;Go, Seung-Joo
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.252-256
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    • 2000
  • The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

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Analysis of RNA Polymerase Beta Subunit (rpoB) Gene Sequences for the Discrimination of Cyanobacteria Anabaena Species (남조세균 Anabaena 종 구분을 위한 RNA Polymerase Beta Subunit (rpoB) 유전자 염기서열 분석)

  • Cheon, Ju-Yong;Lee, Min-Ah;Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.268-274
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    • 2011
  • Anabaena (Cyanobacteria, Nostocales) are important for water quality controls, because they are often responsible for freshwater green tides; moreover, some species are reported to produce hepatotoxin. In this study, we sequenced RNA polymerase beta subunit (rpoB) gene of Anabaena, and evaluated their sequences for the potential use of a molecular taxonomic marker in this taxon. Anabaena rpoB showed low DNA similarity and high genetic divergences when compared those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.01). Parsimony analyses showed the rpoB gene evolves 4.8-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each Anabaena strain more clearly compared with a 16S rRNA tree. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the species discrimination of Anabaena.

16S rDNA-PCR and RFLP Analysis for rapid identification of Spoilage Bacteria from low Salt Cucumber Brine (저염 발효오이로부터 16S rDNA-PCR과 RFLP분석을 통한 부패균의 신속한 확인)

  • 김재호;장혜영
    • KSBB Journal
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    • v.19 no.1
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    • pp.72-77
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    • 2004
  • The aim of this study was to isolate and identify the spoilage bacteria in the low salt cucumber brine. The PCR amplicons comprising a portion of the 16S rRNA gene of the isolated colonies were directly sequenced and the untrimmed whole sequencing results of the unknown strains were aligned with the type strains using BLAST of NCBI. Then Sequence Aligner and Sequence Match of RDP confirmed the outcome. The identified isolates were eight species and belong to three genuses: Clostridium, Lactobacillus, and Bacillus. The RFLP pattern of the 16S rRNA gene of isolates verified the identified species. From now on the complex spoiling process of law salt fermented cucumber could be analyzed using the isolated species individually or with certain combinations.