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

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Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.

Morphogenetic Identification of Eel's Larva (Leptocephalus) Collected by Set net in Namhae, Korea (남해 정치망에서 채집한 엽상자어(Leptocephalus)의 형태 및 유전학적 특성)

  • Chang-Gi Hong;Kyeong-Ho Han
    • Journal of Marine Life Science
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    • v.8 no.2
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    • pp.128-135
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    • 2023
  • The present study was tried to identify whether the eel's larva was close to a conger (Conger myriaster), a pipe conger (Muraenesox cinereus) or four species of Anguilla. Experimental fishes were collected by set net in the gulf of enggang, Namhae, Korea from May to June. Their morphological characteristics were compared with adult fishes of a conger, a pipe conger and four species of Anguilla. For genetic classification, DNA was isolated and amplified by using 12S rRNA and 16S rRNA primer set. The PCR products were direct sequencing in both directions. The nucleotide sequences were analyzed using softwares. As results of morphological measurement on eel's larva, the percentages of head length and preanal length against total length were similar with a conger. Based on the nucleotide sequences, the phylogenetic tree also revealed a close relationship to a conger. Therefore, eel's larva, caught in Namhae from May to June, was identified into a conger's larva.

Microbial Community Analysis using RDP II (Ribosomal Database Project II):Methods, Tools and New Advances

  • Cardenas, Erick;Cole, James R.;Tiedje, James M.;Park, Joon-Hong
    • Environmental Engineering Research
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    • v.14 no.1
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    • pp.3-9
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    • 2009
  • Microorganisms play an important role in the geochemical cycles, industry, environmental cleanup, and biotechnology among other fields. Given the high microbial diversity, identification of the microorganism is essential in understanding and managing the processes. One of the most popular and powerful method for microbial identification is comparative 16S rRNA gene analysis. Due to the highly conserved nature of this essential gene, sequencing and later comparison of it against known rRNA databases can provide assignment of the bacteria into the taxonomy, and the identity of its closest relatives. Isolation and sequencing of 16S rRNA genes directly from natural environments (either from DNA or RNA) can also be used to study the structure of the whole microbial community. Nowadays, novel sequencing technologies with massive outputs are giving researchers worldwide the chance to study the microbial world with a depth that was previously too expensive to achieve. In this article we describe commonly used research approaches for the study of individual microorganisms and microbial communities using the tools provided by Ribosomal Database Project website.

Gluconacetobacter persimmonis sp. nov., Isolated from Korean Traditional Persimmon Vinegar

  • Yeo, Soo-Hwan;Lee, Oh-Seuk;Lee, In-Seon;Kim, Hyun-Soo;Yu, Tae-Shick;Jeong, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.276-283
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    • 2004
  • Screening was performed to isolate cellulose-producing microorganisms from the Korean traditional fermented persimmon vinegar. The resulting strain, KJ $145^{T}$, was then taxonomically investigated by phenotypic characterization, particularly chemotaxonomic, and by phylogenetic inference based on a 16S rDNA sequence analysis including other related taxa. Strain KJ $145^{T}$ was found to grow rapidly and form pale white colonies with smooth to rough surfaces on a GYC agar. Strain KJ $145^T$ also produced acetate from ethanol, and was tolerable to 10% ethanol in SM medium. In a static culture, a thick cellulose pellicle was produced, and in GYC broth, the strain grew at temperatures ranging from 28 to $40^\circ{C}$ with an optimum pH of 4.0. The genomic DNA G+C content of strain KJ $145^T$ was 61.9 mol%, and the predominant ubiquinone was Q 10 as the major quinone and Q9 as the minor quinone. The major cellular fatty acids were $C_{16:0}$ and the sum in feature 7 ($C_{18:1}$ w9c, w12t and/or w7c). A 16S rRNA-targeted oligonucleotide probe specific for strain KJ $145^T$was constructed, and the phylogenetic position of the new species was derived from a 16S rDNA-based tree. When comparing the 16S rDNA nucleotide sequences, strain KJ $145^T$ was found to be most closely related to G. hansenii LMG $1527^T$ (99.2%), although KJ $145^T$ was still distinct from G. hansenii LMG $l527^T$ and G. xylinus LMG $1515^T$ in certain phenotypic characteristics. Therefore, on the basis of 16S rDNA sequences and taxonomic characteristics, it is proposed that strain KJ $145^T$ should be placed in the genus Gluconacetobacter as a new species, Gluconacetobacter persimmonis sp. nov., under the type-strain KJ $145^T$ (=KCTC =$10175BP^T$=KCCM=$10354^T$).

Rapid and Efficient Detection of 16SrI Group Areca Palm Yellow Leaf Phytoplasma in China by Loop-Mediated Isothermal Amplification

  • Yu, Shao-shuai;Che, Hai-yan;Wang, Sheng-jie;Lin, Cai-li;Lin, Ming-xing;Song, Wei-wei;Tang, Qing-hua;Yan, Wei;Qin, Wei-quan
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.459-467
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    • 2020
  • Areca palm yellow leaf (AYL) disease caused by the 16SrI group phytoplasma is a serious threat to the development of the Areca palm industry in China. The 16S rRNA gene sequence was utilized to establish a rapid and efficient detection system efficient for the 16SrI-B subgroup AYL phytoplasma in China by loop-mediated isothermal amplification (LAMP). The results showed that two sets of LAMP detection primers, 16SrDNA-2 and 16SrDNA-3, were efficient for 16SrI-B subgroup AYL phytoplasma in China, with positive results appearing under reaction conditions of 64℃ for 40 min. The lowest detection limit for the two LAMP detection assays was the same at 200 ag/μl, namely approximately 53 copies/μl of the target fragments. Phytoplasma was detected in all AYL disease samples from Baoting, Tunchang, and Wanning counties in Hainan province using the two sets of LAMP primers 16SrDNA-2 and 16SrDNA-3, whereas no phytoplasma was detected in the negative control. The LAMP method established in this study with comparatively high sensitivity and stability, provides reliable results that could be visually detected, making it suitable for application and research in rapid diagnosis of AYL disease, detection of seedlings with the pathogen and breeding of disease-resistant Areca palm varieties.

Molecular Characterization of the Bacterial Community in Activated Sludges by PCR­RFLP (PCR-RFLP 방법을 이용한 활성 슬러지의 세균군집 분석)

  • Lee Hyun-Kyung;Kim Jun-Ho;Kim Chi-Kyung;Lee Dong-Hun
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.307-312
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    • 2004
  • Diversity of the bacterial communities and the relation between community structure and components of waste­water were analyzed by 16S rRNA-based molecular techniques. Clone libraries of the 16S rDNAs from the sludges were constructed by PCR and cloning. The 1,151 clones from a sludge sample of sewage treatment plant were clustered into 699 RFLP phylotypes and the 1,228 clones from the wastewater disposal plant of chemical industry were clustered into 300 RFLP phylotypes. Shannon-Weiner diversity indices of two sampling sites were 8.7 and 6.1, indicating that the bacterial community structure of sewage treatment plant was more diverse than that of wastewater disposal plant of chemical industry. Forty clones belonging to predominant RFLP types were selected and sequenced. Seventy percent (28 clones) of the sequenced clones were related to the uncultured bacteria in public databases. The ${\beta}-Proteobacteria$ dominated in the bacterial communities of investigated two sludge samples. 16S rDNA sequences of the sewage treatment plant were similar to those of other activated sludges, while the bacterial community in wastewater disposal plant of chemical industry rep­resented the strains identified from high-temperature, anaerobic, hydrocarbon-rich, and sulfur-rich environ­ments. This result suggested that bacterial communities depended upon the components of wastewater.

Occurrence of Natural Hybrid between Oplegnathus fasciatus and Oplegnathus punctatus from the South Sea of Korea (한국 남해에서 출현한 돌돔 (Oplegnathus fasciatus)과 강담돔 (Oplegnathus punctatus) 사이의 자연교잡종)

  • Kwun, Hyuck-Joon;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.22 no.3
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    • pp.201-205
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    • 2010
  • One specimen of a natural hybrid of an Oplegnathus (Oplegnathus fasciatus $\times$ Oplegnathus punctatus) was found in Tongyeong, Korea in August 2008. We, herein, describe its morphological and genetic characteristics and compare them with those of O. fasciatus and O. punctatus. In morphology, the hybrid showed many distinctive black rounded blotches on body sides and four faint vertical bars, being in those features similar to O. punctatus. Although the counts and measurements of the hybrid mostly overlapped between O. fasciatus and O. punctatus, the Oplegnathus hybrid resembled O. punctatus in the ratio of pelvic-fin length in standard length: Oplegnathus hybrid (26.7%) was closer to O. punctatus (26.4%) than to O. fasciatus (17.2~23.6%). In genetics, as a result of analysis of 510 base pair sequences of mitochondrial DNA 16S rRNA, the hybrid was closer to O. fasciatus (d=0.000~0.010) than to O. punctatus (d=0.020). Our results suggest that the natural hybridization represented by the subject specimen occurred between an O. fasciatus female and an O. punctatus male.

Antibiotic and Insecticidal Activities of Metabolites Derived From an Entomopathogenic Bacterium, Xenorhabdus nematophila, Against The Bean Bug, Riptortus clavatus (곤충병원세균(Xenorhabdus nematophila) 유래물질의 톱다리개미허리노린재(Riptortus clavatus) 장내세균 증식억제 및 살충효과)

  • Seo, Sam-Yeol;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.49 no.3
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    • pp.251-259
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    • 2010
  • A bacterial colony was isolated from the gut of the bean bug, Riptortus clavatus. From morphological and biochemical tests, the bacterial isolate showed the highest similarity to Staphylococcus succinus. DNA sequence of 16S rRNA gene of the bacterium supported the identification. Oral administration of penicillin G to adults of R. clavatus gave a dose-dependent mortality of adults of R. clavatus to adults along with significant decrease of the bacterial population in the gut. Similarly, three metabolites (benzylideneacetone, proline-tyrosine, and acetylated phenylalanine-glycine-valine) derived from an entomopathogenic bacterium, Xenorhabdus nematophila, also inhibited growth of the gut bacterial population and gave significant mortalities to R. clavatus. These results suggest that a gut bacterial population classified as Staphylococcus sp. is required for survival of R. clavatus and that the three bacterial metabolites had toxic effects on the bugs due to their antibacterial properties.

Identification and Biochemical Characterization of Xylanase-producing Streptomyces glaucescens subsp. WJ-1 Isolated from Soil in Jeju Island, Korea (제주도 토양에서 분리한 xylanase 생산균주 Streptomyces glaucescens subsp. WJ-1의 동정 및 효소의 생화학적 특성 연구)

  • Kim, Da Som;Jung, Sung Cheol;Bae, Chang Hwan;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.43-50
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    • 2017
  • A xylan-degrading bacterium (strain WJ-1) was isolated from soil collected from Jeju Island, Republic of Korea. Strain WJ-1 was characterized as a gram-positive, aerobic, and spore-forming bacterium. The predominant fatty acid in this bacterium was anteiso-$C_{15:0}$ (42.99%). A similarity search based on 16S rRNA gene sequences suggested that the strain belonged to the genus Streptomyces. Further, strain WJ-1 shared the highest sequence similarity with the type strains Streptomyces spinoveruucosus NBRC 14228, S. minutiscleroticus NBRC 13000, and S. glaucescens NBRC 12774. Together, they formed a coherent cluster in a phylogenetic tree based on the neighbor-joining algorithm. The DNA G+C content of strain WJ-1 was 74.7 mol%. The level of DNA-DNA relatedness between strain WJ-1 and the closest related species S. glaucescens NBRC 12774 was 85.7%. DNA-DNA hybridization, 16S rRNA gene sequence similarity, and the phenotypic and chemotaxonomic characteristics suggest that strain WJ-1 constitutes a novel subspecies of S. glaucescens. Thus, the strain was designated as S. glaucescens subsp. WJ-1 (Korean Agricultural Culture Collection [KACC] accession number 92086). Additionally, strain WJ-1 secreted thermostable endo-type xylanases that converted xylan to xylooligosaccharides such as xylotriose and xylotetraose. The enzymes exhibited optimal activity at pH 7.0 and $55^{\circ}C$.