• Title/Summary/Keyword: Genetic relatedness

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Analysis of Genetic and Pathogenic Diversity of Ralstonia solanacearum Causing Potato Bacterial Wilt in Korea

  • Cho, Heejung;Song, Eun-Sung;Lee, Young Kee;Lee, Seungdon;Lee, Seon-Woo;Jo, Ara;Lee, Byoung-Moo;Kim, Jeong-Gu;Hwang, Ingyu
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.23-34
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    • 2018
  • The Ralstonia solanacearum species complex (RSSC) can be divided into four phylotypes, and includes phenotypically diverse bacterial strains that cause bacterial wilt on various host plants. This study used 93 RSSC isolates responsible for potato bacterial wilt in Korea, and investigated their phylogenetic relatedness based on the analysis of phylotype, biovar, and host range. Of the 93 isolates, twenty-two were identified as biovar 2, eight as biovar 3, and sixty-three as biovar 4. Applied to the phylotype scheme, biovar 3 and 4 isolates belonged to phylotype I, and biovar 2 isolates belonged to phylotype IV. This classification was consistent with phylogenetic trees based on 16S rRNA and egl gene sequences, in which biovar 3 and 4 isolates clustered to phylotype I, and biovar 2 isolates clustered to phylotype IV. Korean biovar 2 isolates were distinct from biovar 3 and 4 isolates pathologically as well as genetically - all biovar 2 isolates were nonpathogenic to peppers. Additionally, in host-determining assays, we found uncommon strains among biovar 2 of phylotype IV, which were the tomato-nonpathogenic strains. Since tomatoes are known to be highly susceptible to RSSC, to the best of our knowledge this is the first report of tomato-nonpathogenic potato strains. These results imply the potential prevalence of greater RSSC diversity in terms of host range than would be predicted based on phylogenetic analysis.

Genetic relatedness of Megalocytivirus from diseased fishes in Korea (국내 어류에서 분리된 Megalocytivirus의 유전형 분류 및 상관관계 분석)

  • Lee, Eun Sun;Cho, Miyoung;Min, Eun Young;Jung, Sung Hee;Kim, Kwang Il
    • Journal of fish pathology
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    • v.32 no.2
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    • pp.49-57
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    • 2019
  • In this study, we collected 39 megalocytiviruses isolated from diseased fish in Korea from 2012 to 2018. Major capsid protein (MCP) gene, a part of vascular endothelial growth factor (VEGF) gene and histidine triad motif-like protein (HIT) genes of Megalocytivirus were targeted for PCR amplification and analysis of those DNA nucleotide sequences. Korean strains revealed two genotypes (red sea bream iridovirus and turbot reddish body iridovirus types) based on the phylogeny of MCP gene. The red sea bream iridovirus type (RSIV-type) megalocytiviruses were divided into RSIV-subgroup 1 and 2. From the phylogenetic analysis of the VEGF genes, a genotypic variant of RSIV-type Megalocytivirus was identified. The HIT-like protein gene was detected in RSIVs, but not in TBRIV and ISKNV, suggesting that HIT-like protein gene may be specific in RSIV.

Genetic analysis of polyketide biosynthetic genes isolated from Streptomyces albus, a salinomycin producer.

  • JOO-WON SUH;KWON, HYOUNG-JIN;C.R. HUTCHINSON;HYUNG-JONG JIN;SOO-UN KIM;KYE-JOON LEE
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.74-79
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    • 1995
  • Sequence analysis of a DNA region encompassing the site of hybridization to actl, the gene for type II minimal polyketide synthase (PKS) for actinorhodin biosynthesis, from Streptomyces ablus revealed three more complete open reading frames additional to the already found two genes, plausibly encoding ${\beta}-ketoacyl$ synthase/acyl transferase (KS/AT) and chain length determining factor (ClF). The open reading frames (ORFs) were named salA, salD, and salE, from the upstream. In the homology analysis of the deduced amino acid sequences, SalA resembles the Streptomyces glaucescens Tcml, decaketide cyclase, SalD resembles acyl carrier protein in type II PKS, and SalE resembles the Actlll ketoreductase, The whole 4.4 kb of DNA sequence obeys the same conservation pattern as other type II PKSs. Therefore, we suggest that the 4.4 kb DNA from Streptomyces albus encompasses genes encoding enzymes for polyketide biogenesis in the organism and its organization is type II. The exsitence of SaIA, an analogue of the aromatic cyclase, revealed a relatedness of the 4.4 kb DNA with the aromatic PKS.

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Structure and Diversity of Arsenic-Resistant Bacteria in an Old Tin Mine Area of Thailand

  • Jareonmit, Pechrada;Sajjaphan, Kannika;Sadowsky, Michael J.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.169-178
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    • 2010
  • The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis. Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthrobacter koreensis and $\beta$-Proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic-contaminated soils. The majority of the As-resistant isolates were Gramnegative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. The PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using the Pearson product-moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. Although many isolates were genetically diverse, others were clonal in nature. Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.

Identification and Characterization of Agar-degrading Vibrio sp. GNUM08123 Isolated from Marine Red Macroalgae (한천분해 미생물 Vibrio sp. GNUM08123의 동정 및 agarase 생산의 발효적 특성)

  • Chi, Won-Jae;Kim, Yoon Hee;Kim, Jong-Hee;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.45 no.3
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    • pp.243-249
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    • 2017
  • An agar-degrading bacterium, designated as the GNUM08123 strain, was isolated from samples of red algae collected from the Yongil Bay near East Sea, Korea. The isolated GNUM08123 strain was gram-negative, aerobic, motile, and beige-pigmented, with $C_{16:0}$ (25.9%) and summed feature 3 (comprising $C_{16:1}{\omega}7c/iso-C_{15:0}2-OH$, 34.4%) as its major cellular fatty acids. A similarity search based on the 16S rRNA gene sequence revealed that it belonged to class Gammaproteobacteria and shared 97.7% similarity with the type strain Vibrio chagasii $R-3712^T$. The DNA G+C content of strain $GNUM08123^T$ was 46.9 mol%. The major isoprenoid quinone was ubiquinone-8. The results of DNA-DNA relatedness and 16S rRNA sequence similarity analyses, in addition to its phenotypic and chemotaxonomic characteristics, suggest that strain GNUM08123 is a novel species within genus Vibrio, designated as Vibrio sp. GNUM08123. Agarase production by strain GNUM08123 was induced by agar and sucrose, but was repressed probably owing to carbon catabolite repression by glucose and maltose.

Analysis of antimicrobial resistance and PFGE patterns of Salmonella spp. isolated from chickens at slaughterhouse in Incheon area (인천지역 닭 도축장에서 분리된 Salmonella spp.의 항생제 내성 및 PFGE 패턴분석)

  • Yang, Ha-Young;Lee, Sung-Mo;Park, Eun-Jeong;Kim, Jung-Hee;Lee, Jung-Goo
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.325-334
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    • 2009
  • Salmonella spp. are the important pathogens both economically and clinically in animals as well as human. Some of them have highly zoonotic potentials even though they are asymptomatic in animals. Therefore, the prevalence of Salmonella spp. in animals is highly concerned for human health. The present study was carried out to investigate the prevalence, antimicrobial resistance and PFGE patterns of Salmonella spp. isolated from chickens at slaughterhouse in Incheon area. The overall isolation rate of Salmonella spp. from cloaca and cecum specimens was 7.3 % (37/510). Thirty seven isolates of Salmonella spp. were identified to 5 serotypes; S. Enteritidis, S. Newport, S. Typhimurium, S. Gallinarum, and S. Derby with prevalence of 46.0%, 40.5%, 8.1%, 2.7%, and 2.7%, respectively. Resistance to nalidixic acid was found in 97.3% of Salmonella spp. isolated, followed by streptomycin (16.2%), tetracycline (16.2%), ampicillin (5.4%). Only 6 isolates (16.2%) showed resistance to more than two antimicrobials. In PFGE analysis of chicken and human isolates with Xba I, S. Enteritidis isolates from chicken showed very high similarity over 82.8% and also the similarity was very high in the comparison with human isolates. However, the higher similarity (100%) was observed among chicken isolates of S. Typhimurium. These results suggest the close genetic relatedness of Salmonella spp. isolated from chickens with human.

Comparative Analysis of Nucleotide Sequence and Codon Usage of Arylphorin Gene Cloned from Four Silk-Producing Insects and Their Molicular Phylogenetics

  • Lee, Sang-Mong;Hwang, Jae-Sam;Lee, Jin-Sung;Goo, Tae-Won;Kwon, O-Yu;Kim, Ho-Rak
    • Journal of Life Science
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    • v.9 no.1
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    • pp.84-89
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    • 1999
  • To determine phylogenetic relatedness of four silk-producing silkmoths (B. mori, B. mandarina, A. yamamai and A. pernyi), internal coding region of arylphorin which is a storage protein in hemolymph protein of insects were amplified by polymerase chain reaction and then sequenced and compared each other. The nucleotide composition was biased toward adenine and thymine(59% A+T) and a strong bias for use of C in the third position of codons was found for Phe and Tyr. Together TTC(Phe) and TAC(Tyr) account for about 16.8% (10 for TTC and 8 for TAC) of all codon usage. The nucleotide similarity of arylphorin gene from B. mori showed 99%, 98% and 97% homology with those of B. mandarina, A. yamamai and A. pernyi, respectively. Also, the nucleotide sequence of arylphorin gene from B. mandarina showed 98% and 97% homology with those of A. yamamai and A.pernyi, respectively. Between A. yamamai and A. pernyi, the sequence homology was 97%. The deduced amino acid sequences in B. mori, B. mandarina and A. yamamai showed almost 99% homology. Although the aryphorin gene provided insufficient variability among the four insect species, A UPGMA tree is generated that supported the monophyly of silk-producing insects, with M. sexta placed basal to it. It is suggest that silk-producing insects have a close relationship and a homogeneous genetic background from comparison with those of other insects.

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First Report on Isolation of Salmonella Enteritidis from Eggs at Grocery Stores in Korea

  • Kim, Young Jo;Song, Bo Ra;Lim, Jong Su;Heo, Eun Jeong;Park, Hyun Jung;Wee, Sung Hwan;Oh, Soon Min;Moon, Jin San
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.239-243
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    • 2013
  • Salmonella Enteritidis is responsible for causing foodborne diseases upon consumption of egg products. While cases of S. Enteritidis isolation from eggs have been reported in other countries, no such cases have previously been reported in Korea. In this study, we report the first isolation and identification of S. Enteritidis from domestically distributed eggs in Korea. Eggs were collected from eight countrywide grocery stores during different seasons between 2011 and 2012. Egg contents and washing solution of egg shells were incubated in buffered peptone water, and the enriched broth was further enriched in tetrathionate broth and Rappaport-Vassiliadis. The secondary enriched broth was streaked on xylose lysine desoxycholate agar. The suspected colonies were confirmed to S. Enteritidis by a biochemical test, serotyping, and PCR test. Genetic relatedness among the isolates was analyzed using Diversilab Salmonella kit. Three strains of S. Enteritidis were isolated from egg contents and egg shells collected from grocery stores of the Eumseong-city in the fall of 2011. All three stains showed resistance to chloramphenicol, streptomycin, nalidixic acid, and ampicillin by the disk diffusion method. In addition, the isolates showed more than 99% DNA homology, indicating that they were presumably identical strains. Therefore, there is a requirement to monitor and control against S. Enteritidis from eggs in Korea.

Characterization of Veterinary Hospital-Associated Isolates of Enterococcus Species in Korea

  • Chung, Yeon Soo;Kwon, Ka Hee;Shin, Sook;Kim, Jae Hong;Park, Yong Ho;Yoon, Jang Won
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.386-393
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    • 2014
  • Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n=465) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.

Pathgenicity on Ginseng and Sequence Assays of Ilyonectria radicicola Isolated from Chestnut Rhizosphere Soils (밤나무 근권토양에서 분리한 Ilyonectria radicicola 균주의 인삼에 대한 병원성 및 유전적 분석)

  • Seo, Mun Won;Song, Jeong Young;Kim, Sun Ick;Oh, Sang Keun;Kim, Hong Gi
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.4
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    • pp.302-307
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    • 2018
  • Background: A soil-borne pathogenic fungus, Ilyonectria radicicola (Cylindrocarpon destructans) causes root rot on ginseng (Panax ginseng C. A. Meyer) and is known to attack many other plants. The Nectria/Neonectria radicicola complex has been renamed as the I. radicicola complex after analysis of its multi-gene relatedness and morphological characteristics. The fungi in this complex have been reclassified into 16 species under the genus Ilyonectria based on characteristics analysis Methods and Results: To obtain useful data from the Korean ginseng root rot, I. radicicola was isolated from the rhizosphere soils of the chestnut tree. They were identified through a pathogenicity test and a survey of the morphological features. The existence of I. radicicola in soil samples was confirmed by PCR detections using nested PCR with species-specific primer sets. These were subsequenctly isolated on semi-selective media from PCR-positive soils. Genetic analysis of the I. radicicola complex containing these pathogens was done by comparing the DNA sequences of the histone h3 region. These isolates originating from the rhizosphere soils of chestnut constituted a clade with other closely related species or I. radicicola isolates originating from ginseng or other host plants, respectively. Additionally, the pathogenicity tests to analyze the characteristics of these I. radicicola isolates revealed that they caused weakly virulent root rot on ginseng. Conclusions: This is the first study reporting that I. radicicola isolates from chestnut rhizosphere soils can attack ginseng plant in Korea. Thus, these results are expected to provide informations in the selection of suitable fields for ginseng cultivation.