• Title/Summary/Keyword: 18S rDNA sequence analysis

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Isolation and Identification of Yeast Strain from Fermented Tea (발효차로부터 효모의 분리 및 동정)

  • Kang, Ok-Ju
    • Korean journal of food and cookery science
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    • v.24 no.1
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    • pp.11-15
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    • 2008
  • In searching for yeast to be utilized as biocontrol agents, a single yeast strain was isolated from Camellia sinensis based on its morphological, cultural, physiological, and biochemical properties, as well as by molecular techniques. This single strain was pink to red in color and designated as strain JY-1. The effects of temperature, pH, NaCl concentration, and ethanol concentration on the growth of the JY-l strain were examined for the JY-1. Growth occurred at temperatures ranging from 20 to $35^{\circ}C$, and between pH 3.0 and 12.0, with optimal growth at $25-30^{\circ}C$ and pH 5.0. The yeast also grew in the presence of 0-2% (w/v) NaCl and 0-4% (v/v) EtOH. The isolate was further classified based on biochemical characteristics using the VITEK system. The biochemical data obtained using this system were similar to those of Rhodotorula glutinis/Rhodotorula mucilaginosa (exhibiting a 93% matching level). Molecular phylogenetic analysis based on l8S rDNA sequences indicated that the yeast represented a basidiomycetous species, and its highest degree of sequence similarity was with Rhodosporidium azoricum, strain JCM11251 (99%).

Isolation and Phylogenetic Characterization of Chitinase Producing Oligotrophic Bacteria (Chitinase생산 저영양세균의 분리 및 계통분류학적 특성)

  • Kim Soo-Jin;Kim Min-Young;Koo Bon-Sung;Yoon San-Hong;Yeo Yun-Soo;Park In-Cheol;Kim Yoon-Ji;Lee Jong-Wha;Whang Kyung-Sook
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.293-299
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    • 2005
  • Many isolates from soil of Korean ginseng rhizosphere did not show remarkable growth on full strength of the conventional nutrient broth (NB medium) but grew on its 100-fold dilution (DNB medium). Six hundred-forty strains were isolated as oligotrophic bacteria. In the course of screening for new bioactive compounds from oligotrophic bacteria from soil, 8 strains which had appeared to form of clear zone on a medium containing colloidal chitin as a sole carbon source were selected for further studies. Strain CR42 hydrolyzed a fluorogenic analogue of chitin, 4-methylumbelliferyl-D-glucosaminide (MUF-NAG) . Mo st of the culture supernatant of these isolates hydrolyzed 4-methylumbelliferyl-D-N,N'-diacetylchitobioside (MUF-diNAG). The isolates were heterogeneous and categorized to gamma- and beta-proteobacteria, Bacillaceae, Actinobactepia, and Bacteroides by 16S rRNA analysis. Two strains, WR164 and CR18, had a 16S rRNA sequence of $95-96\%$ identical to uncultured bacteria. It was observed that CR2 and CR75 could inhibit the growth of Colletotrichum gloeosporioides with hyphal extention-inhibition assay on PDA plate supplemented with $1\%$ colloidal chitin.

Characterization of Newly Recorded Talaromyces veerkampii Isolated from Field Soil in Korea based on Morphology and Multigene Sequence Analysis

  • Mahesh Adhikari;Hyun Seung Kim;Hyo Bin Park;Ki Young Kim;In Kyu Lee;Eun Jeong Byeon;Ji Min Woo;Hyang Burm Lee;Youn Su Lee
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.347-355
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    • 2022
  • A fungal isolate belonging to the phylum Ascomycota was isolated and identified as Talaromyces veerkampii in 2017 during a survey of fungal diversity in field soils in Korea. This fungal isolate was identified and described based on macro- and micromorphological and molecular characterization. The identification was also based on partial 18S-ITS1-5.8S-ITS2-28S rDNA and calmodulin (CaM)-encoding gene sequencing data. Talaromyces veerkampii has not been previously reported in Korea. Thus, we report here a newly discovered species from soil in Korea along with its morphological and molecular characteristics.

Paenibacillus donghaensis sp. nov., a Xylan-degrading and Nitrogen-fixing Bacterium Isolated from East Sea Sediment

  • Choi, Jeong-Hwa;Im, Wan-Taek;Yoo, Jae-Soo;Lee, Sang-Mahn;Moon, Deok-Soo;Kim, Hyeon-Ju;Rhee, Sung-Keun;Roh, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.189-193
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    • 2008
  • A Gram-positive and endospore-forming strain, $JH8^T$, was isolated from deep-sea sediment and identified as a member of the genus Paenibacillus on the basis of 16S rRNA gene sequence and phenotypic analyses. According to a phylogenetic analysis, the most closely related species was Paenibacillus wynnii LMG $22176^T$ (96.9%). Strain $JH8^T$ was also facultatively anaerobic and grew optimally at $20-25^{\circ}C$. The major cellular fatty acid was anteiso-$C_{15:0}$, and the DNA G+C content was 53.1mol%. The DNA-DNA relatedness between the isolate and Paenibacillus wynnii LMG $22176^T$ was 7.6%, indicating that strain $JH8^T$ and P. wynnii belong to different species. Based on the phylogenetic, phenotypic, and chemotaxonomic characteristics, strain $JH8^T$ would appear to belong to a novel species, for which the name Paenibacillus donghaensis sp. novo is proposed (type strain=KCTC $13049^T=LMG\;237S0^T$).

Structural Characterization of Non-reducing Oligosaccharide Produced by Arthrobacter crystallopoietes N-08

  • Bae, Bum-Sun;Shin, Kwang-Soon;Lee, Ho
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.519-525
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    • 2009
  • A bacterial strain (Strain N-08) capable of extracellularly producing high level of non-reducing oligosaccharide (NR-OS) isolated from soil. The strain was identified phylogenetically by 16S rDNA sequence analysis and found to be very close to Arthrobacter crystallopoietes. The high production of NR-OS was observed in the basal culture medium containing maltose as a sole carbon source. The NR-OS in culture supernatant was purified by glucoamylase treatment and Dowex-1 (OH.) ion exchange chromatography and its structure was characterized. This oligosaccharide consisted of only glucose. Methylation analysis indicated that this fraction was composed mainly of non-reducing terminal glucopyranoside. Matrixassisted laser-induced/ionization time-of-flight (MALDI-TOF) and electrospray ionization-mass spectrometry (ESI-MS)/MS analyses suggested that this oligosaccharide comprised non-reducing disaccharide unit with 1,1-glucosidic linkage. When this disaccharide was analyzed by $^1H$-NMR and $^{13}C$-NMR, it gave the same signals with $\alpha$-D-glucopyranosyl-(1,1)-$\alpha$-Dglucopyranoside. These results indicated that the NR-OS produced by A. crystallopoietes N-08 was ${\alpha}1$,${\alpha}1$-trehalose. This is the first report of the trehalose which can be produced directly from maltose by A. crystallopoietes N-08.

Gamma Radiation Induced Mutagenesis of Lysobacter enzymogenes for Enhanced Chitinolytic Activity

  • Lee, Young-Keun;Kim, Kyoung Youl;Senthilkumar, M.
    • Journal of Radiation Industry
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    • v.4 no.1
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    • pp.65-71
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    • 2010
  • Two chitinase producing strains CHI2 and CHI4 were isolated from soybean rhizosphere soil. Both the strains belonged to Lysobacter enzymogenes as indicated by 16S rDNA sequence analysis. Though strain CHI2 and CHI4 produced extracellular chitinase, they differ in their chitinolytic activity. CHI4 produced approximately three times the higher amounts of enzyme than that of CHI2 under specified conditions. CHI2 produced $535.67U\;l^{-1}$ of chitinase after 48 h incubation with a specific activity of $3.91U\;mg^{-1}$ of protein while strain CHI4 produced $1584.13U\;l^{-1}$ of chitinase with a specific activity of $10.88U\;mg^{-1}$ protein. SDS-PAGE analysis indicated that the molecular weight of chitinase enzyme was approximately 45 kDa. A faint band with a molecular weight of 55 kDa reveals the possibility for the presence of another kind of chitin binding protein. Mutant library was developed by exposing the isolates to gamma rays at their $LD_{99}$ value (0.23 kGy). Totally, 11 mutants of CHI2 and CHI4 are reported to have enhanced chitinase activity. Several leaky mutant clones with decreased enzyme activity and a defective mutant (CHI2-M16) with complete loss of chitinase activity were also identified. CHI4-M18, CHI4-M8 and CHI4-M29 showed 78.8, 41.5, and 31.9% increased chitinase activity over wild type CHI4.

Isolation and Identification of Oceanisphaera sp. JJM57 from Marine Red Algae Laurencia sp. (Ceramiales: Rhodomelaceae) (해양 홍조류 Laurencia sp. (Ceramiales: Rhodomelaceae)에서 분리한 Oceanisphaera sp. JJM57의 분리 및 동정)

  • Kim, Man-Chul;Dharaneedharan, S.;Moon, Young-Gun;Kim, Dong-Hwi;Son, Hong-Joo;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.58-63
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    • 2013
  • A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM $15406^T$ and Oceanisphera donghaensis KCTC $12522^T$, respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to $37^{\circ}C$. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2-OH, $C_{16:0}$, and $C_{18:1}$ ${\omega}7c$ fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

Variovorax terrae sp. nov. Isolated from Soil with Potential Antioxidant Activity

  • Woo, Chae Yung;Kim, Jaisoo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.855-861
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    • 2022
  • A white-pigmented, non-motile, gram-negative, and rod-shaped bacterium, designated CYS-02T, was isolated from soil sampled at Suwon, Gyeonggi-do, Republic of Korea. Cells were strictly aerobic, grew optimally at 20-28℃ and hydrolyzed Tween 40. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain CYS-02T formed a lineage within the family Comamonadaceae and clustered as members of the genus Variovorax. The closest members were Variovorax guangxiensis DSM 27352T (98.6% sequence similarity), Variovorax paradoxus NBRC 15149T (98.5%), and Variovorax gossypii JM-310T (98.3%). The principal respiratory quinone was Q-8 and the major polar lipids contain phosphatidylethanolamine (PE), phosphatidylethanolamine (PG), and diphosphatidylglycerol (DPG). The predominant cellular fatty acids were C16:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω6c). The DNA GC content was 67.7 mol%. The ANI and dDDH values between strain CYS-02T and the closest members in the genus Variovorax were ≤ 79.0 and 22.4%, respectively, and the AAI and POCP values between CYS-02T and the other related species in the family Comamonadaceae were > 70% and > 50%, respectively. The genome of strain CYS-02T showed a putative terpene biosynthetic cluster responsible for antioxidant activity which was supported by DPPH radical scavenging activity test. Based on genomic, phenotypic and chemotaxonomic analyses, strain CYS-02T was classified into a novel species in the genus Variovorax, for which the name Variovorax terrae sp. nov., has been proposed. The type strain is CYS-02T (= KACC 22656T = NBRC 00115645T).

The polymorphism of Theileria buffeli major surface protein associate with their clinical signs in holstein in Korea (국내 젖소에서 Theileria buffeli 주요 표면 단백질 유전자의 다양성 분석)

  • Yu, Do-Hyeon;Li, Ying-Hua;Chae, Joon-Seok;Park, Jin-Ho
    • Korean Journal of Veterinary Research
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    • v.51 no.2
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    • pp.107-115
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    • 2011
  • Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected Holstein in Jeju Island.

Occurrence of Powdery Mildew Caused by Erysiphe abeliicola on Glossy Abelia in Korea (Erysiphe abeliicola에 의한 꽃댕강나무 흰가루병 발생)

  • Cho, Sung-Eun;Park, Ji-Hyun;Lee, Seung-Kyu;Lee, Sang-Hyun;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.18 no.2
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    • pp.133-138
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    • 2012
  • In November 2009, a powdery mildew on glossy abelia (Abelia ${\times}$ grandiflora) was found in Seogwipo, Jeju Island, Korea. Further survey in the southern part of Korea, e.g., Jeju, Busan, and Tongyeong confirmed occurrence of the disease. White colonies were present on leaves, young stems, and flowers, detracting from their beauty in landscape plantings. Severely infected lesions were discolored to red-purplish. Based on the morphological characteristics and analysis of rDNA, the fungus associated with the symptoms was identified as Erysiphe abeliicola U. Braun & S. Takam. This work provides the morphological feature of its anamorph for the first time, which is characterized by having multi-lobed hyphal appressoria and short foot-cells of conidiophores. Morphological characteristics of mature chasmothecia were consistent with the previous Japanese record of this species. The sequence of internal transcribed spacer region of ribosomal DNA obtained from a Korean sample showed that this species places in the section Microsphaera of the genus Erysiphe in phylogenetic position, corresponding with the classical taxonomy. This is the first report of E. abeliicola and its host plant in Korea. The host plant A. ${\times}$ grandiflora is newly listed in the host range of E. abeliicola.