• Title/Summary/Keyword: 16S rDNA sequences

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Antifungal Activity of Streptomyces griseofuscus 200401 against Pathogens causing Late Blight and Anthracnose on Pepper (역병균과 고추 탄저병에 대한 Streptomyces griseofuscus 200401의 항균활성)

  • Lim, Tae-Heon
    • The Korean Journal of Pesticide Science
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    • v.9 no.1
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    • pp.102-107
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    • 2005
  • To select microorganisms that exhibit antifungal activity against the fungal pathogens of pepper, Phytophthora capsici and Colletotrichum acutatum, soil samples from a forest and natural fields of Gajang-Dong, Sangju-city were tested in vitro and in vivo. Streptomyces griseofuscus 200401 was finally selected throughout antifungal activity test with dual culture, culture broth, and fruits. For the identification of the strain, nucleotide sequences of 16S rDNA and whole cell fatty acids were analyzed. It is like that the strain 200401 may be a novel biological control agent that can reduce application of chemical fungicides to control late blight and anthracnose on pepper.

Genetic Phylogeny among Three Species Red Seabream, Black Seabream and Rock Bream Based on Mitochondrial DNA Sequences

  • Kim, Mi-Jung;An, Hye-Suck;Kim, Kyung-Kil;Park, Jung-Youn
    • Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.171-178
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    • 2009
  • The Perciformes include approximately 40% of all bony fishes and are the largest order of vertebrates. This order includes some of the most economically relevant marine fishes, particularly the red seabream, black seabream and rock bream. A 409 bp fragment of the cytochrome b (cyt b) gene and 403 bp and 518 bp fragments of ribosomal RNA (12S and 16S rRNA, respectively) were sequenced from five populations of natural and cultured red seabreams, natural black seabream, and natural and cultured rock breams. The mitochondrial DNA sequences were utilized for the genetic identification and population structural analyses of these three species. Phylogenetic relationships of intra- and inter-species were elucidated using three types of molecular genetic markers from three species of the order Perciformes in Korea. We noted no significant differences in the intra-specific variation of the cyt b and rRNA genes in each population however, inter-specific divergences were greater than intra-specific variation. Inter-specific variation was induced more by transition than transversion type in the cyt b and rRNA genes. The cyt b gene and rRNA genes make it possible to determine the inter-species divergence. The rRNA genes have more conserved sequences than the cyt b gene. Therefore, these genes are expected to prove useful among species belonging to the different genera or families.

A Phylogenetic Study in Some Long-Horned Beetles (Coleoptera: Cerambycidae) Using Mitochondrial COI Gene and 16S rRNA Sequences

  • Yoon, Hyung-Joo;Bae, Jin-Sik;Kim, Iksoo;Jin, Byung-Rae;Mah, Young-Il;Moon, Jae-Yu;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.37-53
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    • 2001
  • Two regions of mtDNA genome, cytochrome oxidase subunit I (COI) and 165 ribosomal RNA (165 rRNA) genes, were sequenced for 15 species of the long-horned beetle belonging to four subfamilies and geographic samples of mulberry longicorn beetle, Apriona germari, from two localities in Korea. Ten samples of A. germari collected from Suwon and Busan revealed three COI haplotypes ranging in nucleotide divergence of 0.3% to 0.5%, and the two populations shared one common COI haplotype (80%). The sequence divergence among 15 species of the long-horned beetle was much higher in COI gene (12.3%∼39.4%) than 16S rRNA gene (7.2% to 23.1), and the maximum value in the COI gene is exceptional compared with other relevant studies, including that of Coleoptera. The greatly increased divergence in the COI gene, in facto was stemmed from a peculiar sequence of Prionus insularis belonging to Prioninne, divergence of which ranges from 31.2% to 39.3% from other species. We discussed possible reason of the divergence in this species. Due to the abnormality of COI gene divergence, decrease in phylogenetic signal was severe in COI nucleotide and, subsequently, the converted amino acid sequences, rendering us to put more confidence on the 16S5 rRNA gene data. Although the molecular phylogeny confidently supports the monophyletic origin of Lepturinae, the presence of discrepancy between molecular data and traditional taxonomic views also is a testable hyothesis. One such discrepancy includes taxonomic position of Sophronica obrioides and Theophilea cylindricollis belonging to Lamiinae.

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Analysis of Bacterial Community Structure in the Soil and Root System by 168 rRNA Genes (16S rDNA를 이용한 토양, 작물근계의 세균군집 구조해석)

  • Kim, Jong-Shik;Kwon, Soon-Wo;Ryu, Jin-Chang;Yahng, Chang-Sool
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.4
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    • pp.266-274
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    • 2000
  • Understanding of microbial community structure in soil-root system is necessary to use beneficial soil and rhizosphere microbes for improvement of crop production and biocontrol. The knowledge of behavior and function of microbes in soil-root system plays a key role for the application of beneficial inocula. Because the majority of the intact bacteria in soil are unable to grow on nutrient media, both culturable and nonculturable bacteria have to be studied together. In our study, culture-independent survey of bacterial community in the soil-root system of red pepper fields was conducted by the sequence analysis of three universal clone libraries of genes which code for small-subunit rRNA (rDNA). Universal small subunit rRNA primers were used to amplify DNA extracted from each sample and PCR products were cloned into pGEM-T. Out of 27 clones sequenced, 25 clones were from domain bacteria. Two of the rDNA sequences were derived from eukaryotic organelles. Within the domain bacteria, several kingdoms were represented : the Proteobacteria (16 clones). Cytophyga-Flexibacter-Bacteroides group (2 clones). the high G+C content gram-positive group(1 clone) and 4 unknown clones.

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Isolation, Identification, and Characterization of Bacillus strains from the Traditional Korean Soybean-fermented Food, Chungkookjang

  • Joo, Myeong-Hoon;Hur, Sung-Ho;Han, Yong-Soo;Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.202-210
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    • 2007
  • A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

Anaplasma marginale and A. platys Characterized from Dairy and Indigenous Cattle and Dogs in Northern Vietnam

  • Chien, Nguyen Thi Hong;Nguyen, Thi Lan;Bui, Khanh Linh;Van Nguyen, Tho;Le, Thanh Hoa
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.43-47
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    • 2019
  • Anaplasma marginale and A. platys were detected and characterized (16S rDNA sequence analysis) from dairy and indigenous cattle, and the latter in domestic dogs in Vietnam. A phylogenetic tree was inferred from 26 representative strains/species of Anaplasma spp. including 10 new sequences from Vietnam. Seven of our Vietnamese sequences fell into the clade of A. marginale and 3 into A. platys, with strong nodal support of 99 and 90%, respectively. Low genetic distances (0.2-0.4%) within each species supported the identification. Anaplasma platys is able to infect humans. Our discovery of this species in cattle and domestic dogs raises considerable concern about zoonotic transmission in Vietnam. Further systematic investigations are needed to gain data for Anaplasma spp. and members of Anaplasmataceae in animal hosts, vectors and humans across Vietnam.

Phylogenetic Analysis of Culturable Arctic Bacteria

  • Lee, Yoo-Kyung;Kim, Hyo-Won;Cho, Kyeung-Hee;Kang, Sung-Ho;Lee, Hong-Kum;Kim, Yea-Dong
    • Ocean and Polar Research
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    • v.26 no.1
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    • pp.51-58
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    • 2004
  • We isolated and identified culturable Arctic bacteria that had inhabited soils around the Korean Arctic Research Station Dasan located at Ny-Alsund, Svalbard, Norway $(79^{\circ}N,\;12^{\circ}E)$. The collected soils were diluted in distilled water; the diluted soil-water was spread on 3M petri-films at Dasan Station. The petri-films were transported to the laboratory at KORDI, and cultured at $4^{\circ}C$. Colonies grown on the petri-films were subsequently cultured on nutrient agar plates at $4^{\circ}C$ every 7 days. The pure colonies were inoculated into nutrient liquid media, genomic DNA was extracted, and phylogenetic analysis was performed on the basis of 165 rDNA sequences. A total of 227 strains of bacteria were isolated. Among them, 16S rDNA sequences of 185 strains were identical with those of known strains isolated in this study, and 42 strains were finally identified. Phylogenetic analysis using 16S rDNA indicated that the 30 strains belonged to Pseudomonas, 7 strains to Arthrobacter, two strains to Flavobacterium, and the remaining to Achromobacter, Pedobacter, and Psychrobacter. Among the 42 strains, 14 bacteria produced protease: they were 6 strains of Pseudomonax, 4 strains of Arthrobater, an Achromobacter strain, 2 strains of Flavobacterium, and a Pedohacter strain. We expect these Arctic bacteria can be used for screening to develop new industrial enzymes that are active at low temperatures.

Isolation and Genetic Characterization of Protease-Producing Halophilic Bacteria from Fermenting Anchovy (발효중인 멸치액젓에서 분리한 단백질분해효소 생산 호염성 세균의 유전적 특성)

  • Lee, Jin-Ho
    • Journal of Life Science
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    • v.22 no.2
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    • pp.167-176
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    • 2012
  • Three protease-producing halophilic bacteria were isolated from fermenting anchovy. Isolated FAM 10, FAM 114, and FAM 115 were found to grow optimally at salt concentrations of 2-4%, 10%, and 6%, respectively, and could grow in salinity of up to 18-22%. The salinity conditions for optimum protease production were 6% in FAM 10 and 10% in FAM 114 and FAM 115. The protease activity of FAM 10 was gradually inhibited by the addition of NaCl up to 10%, and was not evident at 14%, whereas FAM 114 and FAM 115 displayed protease activity at 14% NaCl and could not be measured at 18%. These results demonstrated that the three isolated strains belong to protease-producing, moderately halophilic bacteria. Strain FAM 10, FAM 114, and FAM 115 were identified as Salinivibrio sp., Halobacillus sp., and Halobacillus sp. respectively, based on comparative analyses of the 16S rRNA gene and the 16S-23S intergenic space sequence (IGS), biochemical testing, and Gram staining. Salinivibrio sp. FAM 10 had two 16S rDNAs containing different sequences at position 191 and four IGSs that harbored no tRNA gene and tRNA genes for isoleucine, alanine, glutamate, lysine, and/or valine. Halobacillus sp. FAM 114 and FAM 115 had completely identical 16S rRNA gene sequences and showed 99% identity to the sequences of various Halobacillus strains. The three IGSs found in the genome of both strains displayed 99% sequence identity with Halobacillus aidingensis and Halobacillus sp. JM-Hb, and had $IGS^0$ with no tRNA gene and $IGS^{IA}$ with tRNA genes for isoleucine and alanine.

An Antifungal Activity of Streptomyces sp. against Cryphonectria parasitica

  • Lee, Sang-Hyun;Kim, Kyung-Hee;Lee, Jae-Pil;Yi, Yong-Sub;Kim, Hye-Sook;Lim, Yoong-Ho
    • Journal of Applied Biological Chemistry
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    • v.48 no.4
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    • pp.222-225
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    • 2005
  • Strains showing antifungal activity against Cryphonectria parasitica were isolated from coast soil of Taean , Korea. Of 152 strains isolated, 6 strains showed antifungal activity in vitro against C. parasitica. Ta24 strain showed highest activity with 1.6 cm clean inhibition zone. For strain identification, the morphological characteristic and 16S rDNA sequences were determined. Ta24 strain showed 99% homology with Streptomyces sampsonii and was identified as Streptomyces sp.

Detection of Gram-negative Bacteria in Broad-range PCR Amplifying 16S rRNA Gene with Semi-nested Primers and Its Application in Market Milk (16S rRNA 유전자의 Semi-nested Primer를 이용한 Broad-range PCR에 의한 그람음성세균의 검출과 시유에서의 응용)

  • Choi, Suk-Ho;Choi, J.J.;Lee, S.B.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.465-474
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    • 2005
  • A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.