• 제목/요약/키워드: Multilocus sequence analysis

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Molecular Analysis of Carbapenem-Resistant Enterobacteriaceae at a South Korean Hospital

  • Lee, Miyoung;Choi, Tae-Jin
    • 한국미생물·생명공학회지
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    • 제48권3호
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    • pp.389-398
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    • 2020
  • The prevalence of carbapenem-resistant Enterobacteriaceae (CRE) is increasing globally, resulting in high mortality rates. Although CRE is a relatively recent problem in Korea (the first case was not diagnosed until 2010), it is responsible for serious morbidities at an alarming rate. In this study, we carried out a molecular genetic analysis to determine the incidence of CRE and carbapenemase-producing Enterobacteriaceae (CPE) at a general hospital in Korea between August 2017 and August 2019. Forty strains of CPE were isolated from various clinical specimens and analyzed via antimicrobial susceptibility testing, polymerase chain reaction to detect β-lactamase genes, deoxyribonucleic acid sequencing, multilocus sequence typing, curing testing, and conjugal transfer of plasmids. The results demonstrated that all 40 isolates were multidrug-resistant. The fluoroquinolone susceptibility test showed that 75% of the Enterobacteriaceae isolates were resistant to ciprofloxacin, whereas 72.5% were resistant to trimethoprim-sulfamethoxazole. Further, conjugation accounted for 57.5% of all resistant plasmid transfer events, which is 4.3-fold higher than that observed in 2010 by Frost et al. Finally, the high detection rate of transposon Tn4401 was associated with the rapid diffusion and evolution of CPE. Our results highlight the rapid emergence of extensively drugresistant strains in Korea and emphasize the need for employing urgent control measures and protocols at the national level.

Pectobacterium brasiliense as a Causative Agent for Soft Rot of Radish in Korea

  • Kyoung-Taek Park;Soo-Min Hong;Chang-Gi Back;In-Kyu Kang;Seung-Yeol Lee;Leonid N. Ten;Hee-Young Jung
    • 식물병연구
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    • 제29권1호
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    • pp.64-71
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    • 2023
  • In October 2021, soft rot disease seriously affected radish crop in Dangjin, Chungcheongnam-do, Korea. The infected radishes were stunted and turned dark green, with yellowish leaf foliage. A slimy, wet, and decayed pith region was observed in the infected roots. The bacterial strain KNUB-03-21 was isolated from infected roots. The biochemical and morphological characteristics of the isolate were similar to those of Pectobacterium brasiliense. Phylogenetic analysis based on the sequences of the 16S rRNA region and the concatenated DNA polymerase III subunit tau (dnaX), leucine-tRNA ligase (leuS), and recombinase subunit A (recA) genes confirmed that the isolate is a novel strain of P. brasiliense. Artificial inoculation of radish with P. brasiliense KNUB-03-21 resulted in soft rot symptoms similar to those observed in infected radish in the field; subsequently, P. brasiliense KNUB-03-21 was reisolated and reidentified. To our knowledge, this is the first report of P. brasiliense as a causal pathogen of radish soft rot in Korea.

Genetic Diversity and Population Structure of the Xanthomonas campestris pv. campestris Strains Affecting Cabbages in China Revealed by MLST and Rep-PCR Based Genotyping

  • Chen, Guo;Kong, Congcong;Yang, Limei;Zhuang, Mu;Zhang, Yangyong;Wang, Yong;Ji, Jialei;Fang, Zhiyuan;Lv, Honghao
    • The Plant Pathology Journal
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    • 제37권5호
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    • pp.476-488
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    • 2021
  • Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot for cruciferous vegetables worldwide, especially for the cole crops such as cabbage and cauliflower. Due to the lack of resistant cabbage cultivars, black rot has brought about considerable yield losses in recent years in China. Understanding of the pathogen features is a key step for disease prevention, however, the pathogen diversity, population structure, and virulence are largely unknown. In this study, we studied 50 Xcc strains including 39 Xcc isolates collected from cabbage in 20 regions across China, using multilocus sequence genotyping (MLST), repetitive DNA sequence-based PCR (rep-PCR), and pathogenicity tests. For MLST analysis, a total of 12 allelic profiles (AP) were generated, among which the largest AP was AP1 containing 32 strains. Further cluster analysis of rep-PCR divided all strains into 14 DNA groups, with the largest group DNA I comprising of 34 strains, most of which also belonged to AP1. Inoculation tests showed that the representative Xcc strains collected from diverse regions performed differential virulence against three brassica hosts compared with races 1 and 4. Interestingly, these results indicated that AP1/DNA I was not only the main pathotype in China, but also a novel group that differed from the previously reported type races in both genotype and virulence. To our knowledge, this is the first extensive genetic diversity survey for Xcc strains in China, which provides evidence for cabbage resistance breeding and opens the gate for further cabbage-Xcc interaction studies.

Occurrence of Leaf Spot Disease Caused by Alternaria crassa (Sacc.) Rands on Jimson Weed and Potential Additional Host Plants in Algeria

  • Bessadat, Nabahat;Hamon, Bruno;Bataille-Simoneau, Nelly;Chateau, Corentin;Mabrouk, Kihal;Simoneau, Philippe
    • The Plant Pathology Journal
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    • 제36권2호
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    • pp.179-184
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    • 2020
  • A leaf spot pathogen Alternaria sp. was recovered from jimson weed, tomato, parsley, and coriander collected during surveys of blight diseases on Solanaceae and Apiaceae in Algeria. This species produced large conidial body generating long apical beaks that tapered gradually from a wide base to a narrow tip and short conidiophores originating directly from the agar surface. This species exhibited morphological traits similar to that reported for Alternaria crassa. The identification of seven strains from different hosts was confirmed by sequence analyses at the glyceraldehyde-3-phosphate dehydrogenase, RNA polymerase second largest subunit, and translation elongation factor 1-alpha loci. Further the pathogen was evaluated on jimson weed, coriander, parsley, and tomato plants, and this fungus was able to cause necrotic lesions on all inoculated plants. A. crassa is reported for the first time as a new species of the Algerian mycoflora and as a new potential pathogen for cultivated hosts.

Occurrence and Epidemics of Bacterial Canker of Kiwifruit in Korea

  • Kim, Gyoung Hee;Jung, Jae Sung;Koh, Young Jin
    • The Plant Pathology Journal
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    • 제33권4호
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    • pp.351-361
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    • 2017
  • Bacterial canker is the largest limiting factor in the cultivation and production of kiwifruit worldwide. Typical symptoms comprise necrotic spots on leaves, canker and dieback on canes and trunks, twig wilting, and blossom necrosis. Pseudomonas syringae pv. actinidiae (Psa), which is the causal agent of kiwifruit bacterial canker, is divided into four biovars based on multilocus sequence analysis of different genes, additional PCR testing of pathogenic genes (argKtox cluster, cfl, and various effector genes), and biochemical and physiological characterization. Bacterial canker caused by Psa biovar 2 designated Psa2 was detected for the first time on the green-fleshed kiwifruit cultivar Hayward in 1988 and the yellow-fleshed kiwifruit cultivar Hort16A in 2006 in Korea. Psa biovar 3 designated Psa3, responsible for the current global pandemics of kiwifruit bacterial canker, began to appear in Korea in 2011 and caused tremendous economic losses by destroying many vines or orchards of yellow-fleshed kiwifruit cultivars in one or several growing seasons. Bacterial canker epidemics caused by both Psa2 and Psa3 are prevalent in Korea in recent years. In this review, we summarize the symptomatology, etiology, disease cycle, diagnosis, and epidemiology of kiwifruit bacterial canker in Korea.

Isolation and Identification of Ice Nucleation Active Fusarium Strains from Rapid Apple Declined Trees in Korea

  • Avalos-Ruiz, Diane;Ten, Leonid N.;Kim, Chang-Kil;Lee, Seung-Yeol;Jung, Hee-Young
    • The Plant Pathology Journal
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    • 제38권4호
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    • pp.403-409
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    • 2022
  • In biological particles such as Fusarium species, ice nucleation activity (INA) has been observed. Fusarium strains isolated from apple declined trees in Korea were identified with a multilocus sequence analysis using the tef1 and rpb1 genes. Droplet-freezing and tube-freezing assays were used to determine the INA of the strains, using Pseudomonas syringae pv. syringae KACC 21200 as a positive control and resulting in seven INA+ fungal strains that were identified as F. tricinctum (KNUF-21-F17, KNUF-21-F18, KNUF-21-F29, KNUF-21-F32, KNUF-21-F38, KNUF-21-F43, and KNUF-21-F44). The effect of Fusarium INA+ KNUF-21-F29 was compared to that of INA- strains on Chrysanthemum morifolium cv. Shinma explants. A higher callus formation and no-shoot formation were observed, suggesting that fungal INA could play a role in cold injuries and be a factor to consider in rapid apple decline. To the best of our knowledge, this is the first report of INA fungal strains isolated in Korea.

Species Transferability of Klebsiella pneumoniae Carbapenemase-2 Isolated from a High-Risk Clone of Escherichia coli ST410

  • Lee, Miyoung;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • 제30권7호
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    • pp.974-981
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    • 2020
  • Sequence type 410 (ST410) of Escherichia coli is an extraintestinal pathogen associated with multi drug resistance. In this study, we aimed to investigate the horizontal propagation pathway of a high-risk clone of E. coli ST410 that produces Klebsiella pneumoniae carbapenemase (KPC). blaKPC-encoding E. coli and K. pneumoniae isolates were evaluated, and complete sequencing and comparative analysis of blaKPC-encoding plasmids from E. coli and K. pneumoniae, antimicrobial susceptibility tests, polymerase chain reaction, multilocus sequence typing, and conjugal transfer of plasmids were performed. Whole-genome sequencing was performed for plasmids mediating KPC-2 production in E. coli and K. pneumoniae clinical isolates. Strains E. coli CPEc171209 and K. pneumoniae CPKp171210 were identified as ST410 and ST307, respectively. CPEc171209 harbored five plasmids belonging to serotype O8:H21, which is in the antimicrobial-resistant clade C4/H24. The CPKp171210 isolate harbored three plasmids. Both strains harbored various additional antimicrobial resistance genes. The IncX3 plasmid pECBHS_9_5 harbored blaKPC-2 within a truncated Tn4401a transposon, which also contains blaSHV-182 with duplicated conjugative elements. This plasmid displayed 100% identity with the IncX3 plasmid pKPBHS_10_3 from the K. pneumoniae CPKp171210 ST307 strain. The genes responsible for the conjugal transfer of the IncX3 plasmid included tra/trb clusters and pil genes coding the type IV pilus. ST410 can be transmitted between patients, posing an elevated risk in clinical settings. The emergence of a KPC-producing E. coli strain (ST410) is concerning because the blaKPC-2-bearing plasmids may carry treatment resistance across species barriers. Transgenic translocation occurs among carbapenem-resistant bacteria, which may spread rapidly via horizontal migration.

Genetic Organization of ascB-dapE Internalin Cluster Serves as a Potential Marker for Listeria monocytogenes Sublineages IIA, IIB, and IIC

  • Chen, Jianshun;Fang, Chun;Zhu, Ningyu;Lv, Yonghui;Cheng, Changyong;Bei, Yijiang;Zheng, Tianlun;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • 제22권5호
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    • pp.575-584
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    • 2012
  • Listeria monocytogenes is an important foodborne pathogen that comprises four genetic lineages: I, II, III, and IV. Of these, lineage II is frequently recovered from foods and environments and responsible for the increasing incidence of human listeriosis. In this study, the phylogenetic structure of lineage II was determined through sequencing analysis of the ascB-dapE internalin cluster. Fifteen sequence types proposed by multilocus sequence typing based on nine housekeeping genes were grouped into three distinct sublineages, IIA, IIB, and IIC. Organization of the ascB-dapE internalin cluster could serve as a molecular marker for these sublineages, with inlGHE, inlGC2DE, and inlC2DE for IIA, IIB, and IIC, respectively. These sublineages displayed specific genetic and phenotypic characteristics. IIA and IIC showed a higher frequency of recombination (${\rho}/{\theta}$). However, recombination events had greater effect (r/m) on IIB, leading to its high nucleotide diversity. Moreover, IIA and IIB harbored a wider range of internalin and stress-response genes, and possessed higher nisin tolerance, whereas IIC contained the largest portion of low-virulent strains owing to premature stop codons in inlA. The results of this study indicate that IIA, IIB, and IIC might occupy different ecological niches, and IIB might have a better adaptation to a broad range of environmental niches.

Genomic Analysis of the Carrot Bacterial Blight Pathogen Xanthomonas hortorum pv. carotae in Korea

  • Mi-Hyun Lee;Sung-Jun Hong;Dong Suk Park;Hyeonheui Ham;Hyun Gi Kong
    • The Plant Pathology Journal
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    • 제39권4호
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    • pp.409-416
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    • 2023
  • Bacterial leaf blight of carrots caused by Xanthomonas hortorum pv. carotae (Xhc) is an important worldwide seed-borne disease. In 2012 and 2013, symptoms similar to bacterial leaf blight were found in carrot farms in Jeju Island, Korea. The phenotypic characteristics of the Korean isolation strains were similar to the type strain of Xhc. Pathogenicity showed symptoms on the 14th day after inoculation on carrot plants. Identification by genetic method was multi-position sequencing of the isolated strain JJ2001 was performed using four genes (danK, gyrB, fyuA, and rpoD). The isolated strain was confirmed to be most similar to Xhc M081. Furthermore, in order to analyze the genetic characteristics of the isolated strain, whole genome analysis was performed through the next-generation sequencing method. The draft genome size of JJ2001 is 5,443,372 bp, which contains 63.57% of G + C and has 4,547 open reading frames. Specifically, the classification of pathovar can be confirmed to be similar to that of the host lineage. Plant pathogenic factors and determinants of the majority of the secretion system are conserved in strain JJ2001. This genetic information enables detailed comparative analysis in the pathovar stage of pathogenic bacteria. Furthermore, these findings provide basic data for the distribution and diagnosis of Xanthomonas hortorum pv. carotae, a major plant pathogen that infects carrots in Korea.

Characterization of Nivalenol-Producing Fusarium culmorum Isolates Obtained from the Air at a Rice Paddy Field in Korea

  • Kim, Da-Woon;Kim, Gi-Yong;Kim, Hee-Kyoung;Kim, Jueun;Jeon, Sun Jeong;Lee, Chul Won;Lee, Hyang Burm;Yun, Sung-Hwan
    • The Plant Pathology Journal
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    • 제32권3호
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    • pp.182-189
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    • 2016
  • Together with the Fusarium graminearum species complex, F. culmorum is a major member of the causal agents of Fusarium head blight on cereals such as wheat, barley and corn. It causes significant yield and quality losses and results in the contamination of grain with mycotoxins that are harmful to humans and animals. In Korea, F. culmorum is listed as a quarantine fungal species since it has yet to be found in the country. In this paper, we report that two isolates (J1 and J2) of F. culmorum were collected from the air at a rice paddy field in Korea. Species identification was confirmed by phylogenetic analysis using multilocus sequence data derived from five genes encoding translation elongation factor, histone H3, phosphate permease, a reductase, and an ammonia ligase and by morphological comparison with reference strains. Both diagnostic PCR and chemical analysis confirmed that these F. culmorum isolates had the capacity to produce nivalenol, the trichothecene mycotoxin, in rice substrate. In addition, both isolates were pathogenic on wheat heads and corn stalks. This is the first report on the occurrence of F. culmorum in Korea.