• Title/Summary/Keyword: 16S ribosomal DNA

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Molecular Identification and Morphological Description of Juvenile of the Previously Unrecorded Species Long-snout Tonguefish, Symphurus longirostris (Cynoglossidae) from the Southern Sea of Jeju Island, Korea (한국 제주도 남부 해역에서 출현한 참서대과(Cynoglossidae) 어류 1미기록종, Symphurus longirostris 치어의 분자동정 및 형태기재)

  • Yeon-Ju Seo;Jin-Koo Kim;Hyo-Jae Yu;Se Hun Myoung
    • Korean Journal of Ichthyology
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    • v.36 no.3
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    • pp.288-294
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    • 2024
  • A juvenile of long-snout tonguefish, Symphurus longirostris belonging to the family Cynoglossidae, was collected from the waters around Jeju Island, Korea in August 2023. Molecular analysis revealed that our juvenile specimen was identical to S. longirostris adult from Japan (Kimura-2-parameter distance, d=0.002) based on mitochondrial DNA 16S ribosomal RNA region sequences 475 base pairs, and differed from its congeneric S. orientalis (d=0.199). The juvenile of S. longirostris has the following morphological traits: obvious dermal spots on both sides of the bases of the dorsal and anal fins; and peritoneum bluish black on its dorsalmost aspect. The juvenile of S. longirostris is also distinguished from S. orientalis in that they do not have a patch of melanophores above the abdominal cavity on the ocular-side. This is the first record of S. longirostris in Korea, and we propose a new Korean name "Gin-ko-bo-seop-seo-dae" for this species.

Chitinase Production and Isolation of Serratia plymuthica AL-1 Antagonistic to White Rot Fungi from Allium fistulosum Roots. (대파 뿌리로부터 흑색썩음균핵병균에 길항하는 Serratia plymuthica AL-1의 분리 및 Chitinase의 생산)

  • 주길재;이익희;김진호
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.135-141
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    • 2002
  • This study was carried out to isolate antagonistic bacterium against Sclerotium cepivorum causing Allium fistulosum white rot. Total of 146 strains were isolated from A. fistulosum roots. The isolates were screened for antagonism to S. cepivorum and the isolated strain No. AL-1 was selected among these bacteria. It was identified as Serratia plymuthica based on morphological and physiological characteristics according to the Bergey's mannual of systematic bacteriology and 16S rDNA sequences methods. Serratia plymuthica AL-1 showed broad spectrum of antifungal activities against plant pathogenic fungi Alternaria altrata, Colletotrichum gleosporioids, Phoma sp., Rhizoctonia solani, Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium niveum but not inhibited Didymella bryoniae. When S. plymuthica AL-1 cultivated in the TSB medium containing 1% colloidal chitin, the high molecular fraction (>10 kDa) have chitinase activity (3.2 units/ml) and the low molecular fraction (<10 kDa) have not chitinase activity. Oppositely, after heat treatment (80℃ for 30 min) of the cultivation supernatant, the high molecular fractions have not antifungal activity but the low molecular fractions have antifungal activity.

Occurrence and Molecular Phylogenetic Characteristics of Benthic Sand-dwelling Dinoflagellates in the Intertidal Flat of Dongho, West Coast of Korea (서해안 동호 사질 조간대에 서식하는 저서성 와편모류의 출현양상 및 분자계통학적 특성)

  • KIM, SUNJU;YOON, JIHAE;PARK, MYUNG GIL
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.3
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    • pp.141-150
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    • 2015
  • Dinoflagellates are ubiquitous and important primary producers in the oceans. They have diverse trophic modes, i.e., phototrophic, heterotrophic, and mixotrophic modes and thereby, play important ecological role in marine microbial food-web. While many studies have been focused on planktonic dinoflagellates in pelagic ecosystems, benthic, sand-dwelling dinoflagellates that inhabit in intertidal zone have been very poorly documented worldwide. We investigated biodiversity, occurrence, and molecular phylogeny of benthic, sand-dwelling dinoflagellates from the intertidal flat of Dongho, west coast of Korea during low-tide, monthly from November 2012 to February 2014. About 27 species of 13 genera in orders Gonyaulacales, Gymnodiniales, Peridiniales, Prorocentrales have been identified, of which members in the genus Amphidinium constituted a major part of the sand-dwelling dinoflagellates in this area. A total of 34 isolates from 16 species of the sand-dwelling dinoflagellates were isolated from Dongho, Mohang, Gamami, and Songho in the west coast and Hyupjae in Jeju of Korea, their 28S rDNA sequences were successfully amplified, and applied for molecular phylogenetic analyses. In the 28S rDNA phylogeny, Amphidinium species diverged across three major clusters within the order Gymnodiniales and formed polyphyletic group. Based on the unambiguously aligned partial 28S rDNA sequences including variable D2 region, the genotypes of Amphidinium mootonorum Korean strains greatly differed from that of Canadian strain with 19.2% of pairwise nucleotide difference, suggesting that further ultrastructural studies may provide additional characters to clearly separate these genotypes. Two potential toxic species, Amphidinium carterae and A. operculatum appeared occasionally during this study. Quantitative assessment and toxicity of those species should be addressed in the future.

IDENTIFICATION OF FUSOBACTERIUM NUCLEATUM AND FUSOBACTERIUM NECROPHORUM USING POLYMERASE CHAIN REACTION(PCR) (중합효소연쇄반응(Polymerase Chain Reaction)을 이용한 Fusobacterium nucleatum 및 Fusobacterium necrophorum의 동점에 관한 연구)

  • Kang, Chang-Woo;Park, Dong-Sung;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.40-48
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    • 1999
  • This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.

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Characterization of Bacterial Community in the Ecosystem Amended with Phenol (페놀이 첨가된 생태계에서 세균 군집구조 변화의 분석)

  • 김진복;김치경;안태석;송홍규;이동훈
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.72-79
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    • 2001
  • The effect of phenol on the change of bacterial community in the effluent water from a wastewater treatment plant was analyzed by PCR and terminal restriction fragment length polymorphism (T-RFLP). The fragments of 16S rDNA were amplified by PCR with bacterial primers, where one of the primers was biotinylated at the 5'-end. After digestion with restriction enzymes, HaeIII and AluI, the biotinylated terminal restriction tragments (T-RFs) of the digested products were selectively isolated by using streptavidin paramagnetic particles. The single-stranded DNA of T-RFs was separated by electrophoresis on a polyacrylamide gel and detected by silver staining technique. When 10 standard strains were analyzed by our method, each strain had a unique T-RF which corresponded to the calculated size from the known sequences of RDP database. The T-RFLP fingerprint generated from the effluent water was very complex, and the predominant T-RFs corresponded to members of the genus Acinetobacter, Bacillus and Pseudomonas. In addition, the perturbation of bacterial community was observed when phenol was added to the sample at the final concentration of 250 $l^{-1}$. The number of T-RFs increased and the major bacterial population could be assigned to the genus Acinetobacter, Comamonas, Cytophaga and Pseudomonas. A intense band assigned to the putative genera of Acinetobacter and Cytophaga was eluted, amplified, and sequenced. The nucleotide sequence of the T-RF showed close relationship with the sequence of Acinetobacter junii.

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Molecular Identification and First Morphological Description of Larvae and Juveniles of Neosalanx anderssoni (Salangidae) Collected from the Southwestern Sea of Korea (한국 서해 남부해역에서 채집된 도화뱅어, Neosalanx anderssoni (뱅어과) 자치어의 분자 동정 및 첫 형태기재)

  • Seo-Yeon Koo;Se-Hun Myoung;Jin-Koo Kim
    • Korean Journal of Ichthyology
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    • v.36 no.1
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    • pp.94-100
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    • 2024
  • During ichthyoplankton survey in the southwestern sea of Korea, we collected six individuals of noodlefish larvae and juveniles between April and May 2023. They were identified as Neosalanx anderssoni by mitochondrial DNA cytochrome c oxidase subunit I or 16S ribosomal RNA sequences, and their external morphological traits were described for the first time. All six individuals have a slender and elongated body. When preflexion and flexion larval stages (10.24 mm notochord length, NL and 15.47 mm total length, TL, respectively), oval-shaped black melanophores were distributed in a row along the ventral side of the gut. However, when postflexion larval and juvenile stages (23.58~25.90 mm TL, and 29.20~31.26 mm TL, respectively), melanophores on the ventral side of the gut were disappeared, and dark spot-shaped melanophores appeared along the dorsal side of the gut in a single row. Also, from the postflexion larval stage (23.58 mm TL), two large black spots began to appear symmetrically on the caudal fin. Our results suggest that N. anderssoni may use coastal area as spawning and/or nursery ground unlike previous study (Kim and Park, 2002).

Identification and Antimicrobial Activity Detection of Lactic Acid Bacteria Isolated from Corn Stover Silage

  • Li, Dongxia;Ni, Kuikui;Pang, Huili;Wang, Yanping;Cai, Yimin;Jin, Qingsheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.620-631
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    • 2015
  • A total of 59 lactic acid bacteria (LAB) strains were isolated from corn stover silage. According to phenotypic and chemotaxonomic characteristics, 16S ribosomal DNA (rDNA) sequences and recA gene polymerase chain reaction amplification, these LAB isolates were identified as five species: Lactobacillus (L.) plantarum subsp. plantarum, Pediococcus pentosaceus, Enterococcus mundtii, Weissella cibaria and Leuconostoc pseudomesenteroides, respectively. Those strains were also screened for antimicrobial activity using a dual-culture agar plate assay. Based on excluding the effects of organic acids and hydrogen peroxide, two L. plantarum subsp. plantarum strains ZZU 203 and 204, which strongly inhibited Salmonella enterica ATCC $43971^T$, Micrococcus luteus ATCC $4698^T$ and Escherichia coli ATCC $11775^T$ were selected for further research on sensitivity of the antimicrobial substance to heat, pH and protease. Cell-free culture supernatants of the two strains exhibited strong heat stability (60 min at $100^{\circ}C$), but the antimicrobial activity was eliminated after treatment at $121^{\circ}C$ for 15 min. The antimicrobial substance remained active under acidic condition (pH 2.0 to 6.0), but became inactive under neutral and alkaline condition (pH 7.0 to 9.0). In addition, the antimicrobial activities of these two strains decreased remarkably after digestion by protease K. These results preliminarily suggest that the desirable antimicrobial activity of strains ZZU 203 and 204 is the result of the production of a bacteriocin-like substance, and these two strains with antimicrobial activity could be used as silage additives to inhibit proliferation of unwanted microorganism during ensiling and preserve nutrients of silage. The nature of the antimicrobial substances is being investigated in our laboratory.

Bioremediation Potential of a Tropical Soil Contaminated with a Mixture of Crude Oil and Production Water

  • Alvarez, Vanessa Marques;Santos, Silvia Cristina Cunha dos;Casella, Renata da Costa;Vitae, RonaIt Leite;Sebastin, Gina Vazquez;Seldin, Lucy
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1966-1974
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    • 2008
  • A typical tropical soil from the northeast of Brazil, where an important terrestrial oil field is located, was accidentally contaminated with a mixture of oil and saline production water. To study the bioremediation potential in this area, molecular methods based on PCR-DGGE were used to determine the diversity of the bacterial communities in bulk and in contaminated soils. Bacterial fingerprints revealed that the bacterial communities were affected by the presence of the mixture of oil and production water, and different profiles were observed when the contaminated soils were compared with the control. Halotolerant strains capable of degrading crude oil were also isolated from enrichment cultures obtained from the contaminated soil samples. Twenty-two strains showing these features were characterized genetically by amplified ribosomal DNA restriction analysis (ARDRA) and phenotypically by their colonial morphology and tolerance to high NaCl concentrations. Fifteen ARDRA groups were formed. Selected strains were analyzed by 16S rDNA sequencing, and Actinobacteria was identified as the main group found. Strains were also tested for their growth capability in the presence of different oil derivatives (hexane, dodecane, hexadecane, diesel, gasoline, toluene, naphthalene, o-xylene, and p-xylene) and different degradation profiles were observed. PCR products were obtained from 12 of the 15 ARDRA representatives when they were screened for the presence of the alkane hydroxylase gene (alkB). Members of the genera Rhodococcus and Gordonia were identified as predominant in the soil studied. These genera are usually implicated in oil degradation processes and, as such, the potential for bioremediation in this area can be considered as feasible.

Isolation and Characterization of Endophytic Actinomycetes from Chinese Cabbage Roots as Antagonists to Plasmodiophora brassicae

  • Lee, Sun-Og;Choi, Gyung-Ja;Choi, Yong-Ho;Jang, Kyoung-Soo;Park, Dong-Jin;Kim, Chang-Jin;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1741-1746
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    • 2008
  • This study was conducted to select endophytic actinomycetes as biocontrol agents against Chinese cabbage clubroot caused by Plasmodiophora brassicae. A total of 81 endophytic actinomycetes were isolated from surface-sterilized roots of Chinese cabbage that was grown on paddy field and upland soils collected from various locations in Korea. By using 16S ribosomal DNA (rDNA) gene sequencing, they were classified to 8 actinobacterial genera. The genus Microbispora (67%) was most frequently isolated, followed by Streptomyces (12%) and Micromonospora (11%). Three of the 81 isolates, when inoculated in germinated Chinese cabbage seeds and then transplanted to pots, effectively suppressed the occurrence of a post-inoculated strain of P. brassicae in the pots. They showed control values of 58% for strain A004, 33% for strain A011, and 42% for strain A018. Based on cell wall components, morphological characteristics, and phylogenetic analyses, the three antagonistic isolates were identified as Microbispora rosea subsp. rosea (A004 and A011) and Streptomyces olivochromogenes (A018). Further researches on the field efficacy and action modes of the three actinomycetes are in progress.

Occurrence of Leptosphaerulina Leaf Blight on Kentucky Bluegrass Caused by Leptosphaerulina trifolii (Leptosphaerulina trifolii에 의한 Kentucky Bluegrass의 Leptosphaerulina 잎마름병 발생)

  • Kim, Jeong-Ho;Shim, Gyu-Yul;Kim, Young-Ho
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.94-96
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    • 2010
  • In May of 2004 through 2007, Leptosphaerulina leaf blight caused by Leptosphaerulina trifolii occurred on Kentucky bluegrass (Poa pratensis) at golf courses in Gangwon Province, Korea. Symptoms on the turfgrass caused by L. trifolii were leaf blights, dying from the leaf tip downwards to the crown, which appeared patches in the field because of local pockets of severely infected (blighted) grass. Perithecia were produced on old or weak leaves, including club-shaped asci, each of which contained 8 pale brown muriform ascospores with cross and longitudinal septa. Ascospores of the fungus isolated from the diseased leaf tissue and cultured on potato-dextrose agar (PDA) were muriform multicellular (composed of 3-6 cells) and $23.4-40.5{\times}7.8-15.6{\mu}m$ in size with 3-4 transverse and 0-3 longitudinal septa, which were morphologically identical to L. trifolii reported previously. DNA sequences of ribosomal RNA gene (internal transcribed spacer) of the fungus were homologous with similarity of 99% to those of L. trifolii isolates in GenBank database, confirming the identity of the causal agent of the disease. Pathogenicity of the fungus was also confirmed on the creeping bentgrass by Koch's postulates. This is first report of Leptosphaerulina leaf blight on turfgrass caused by L. trifolii in Korea.