• 제목/요약/키워드: multi-locus sequence analysis

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Comparative Analysis of Salmonella enterica subsp. enterica Serovar Thompson Isolates associated with Outbreaks Using PFGE and wgMLST

  • Youngho Koh;Yunyoung Bae;Min-Jung Lee;Yu-Si Lee;Dong-Hyun Kang;Soon Han Kim
    • Journal of Microbiology and Biotechnology
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    • 제32권12호
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    • pp.1605-1614
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    • 2022
  • The strains associated with foodborne Salmonella enterica Thompson outbreaks in Korea have not been identified. Therefore, we characterized S. Thompson strains isolated from chocolate cakes linked to foodborne outbreaks in Korea. A total of 56 strains were isolated from preserved cake products, products in the supply chain distribution, the manufacturer's apparatus, and egg white liquid products used for cream preparation. Subsequently, serological typing, pathogenic gene-targeted polymerase chain reaction (PCR), pulsed-field gel electrophoresis (PFGE), and whole-genome multi-locus sequence typing (wgMLST) were performed to characterize these isolates. The antigen formula of all isolates was 7:k:1,5, namely Salmonella enterica subsp. enterica Serovar Thompson. All 56 isolates harbored invA, his, hin, and stn, and were negative for sefA and spvC based on gene-targeted PCR analyses. Based on PFGE results, these isolates were classified into one group based on the same SP6X01.011 pattern with 100% similarity. We selected 19 strains based on the region and sample type, which were subjected to wgMLST. Although the examined strains showed 100% similarity, they were classified into seven clusters based on allelic differences. According to our findings, the cause of these outbreaks was chocolate cake manufactured with egg white liquid contaminated with the same Salmonella Thompson. Additionally, comparative analysis of wgMLST on domestic isolates of S. Thompson from the three outbreaks showed genetic similarities of over 99.6%. Based on the results, the PFGE and wgMLST combination can provide highly resolved phylogeny and reliable evidence during Salmonella outbreak investigations.

Diversity of the Bambusicolous Fungus Apiospora in Korea: Discovery of New Apiospora Species

  • Sun Lul Kwon;Minseo Cho;Young Min Lee;Hanbyul Lee;Changmu Kim;Gyu-Hyeok Kim;Jae-Jin Kim
    • Mycobiology
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    • 제50권5호
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    • pp.302-316
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    • 2022
  • Many Apiospora species have been isolated from bamboo plants - to date, 34 bambusicolous Apiospora species have been recorded. They are known as saprophytes, endophytes, and plant pathogens. In this study, 242 bambusicolous Apiospora were isolated from various bamboo materials (branches, culms, leaves, roots, and shoots) and examined using DNA sequence similarity based on the internal transcribed spacer, 28S large subunit ribosomal RNA gene, translation elongation factor 1-alpha, and beta-tubulin regions. Nine Apiospora species (Ap. arundinis, Ap. camelliae-sinensis, Ap. hysterina, Ap. lageniformis sp. nov., Ap. paraphaeosperma, Ap. pseudohyphopodii sp. nov., Ap. rasikravindrae, Ap. saccharicola, and Ap. sargassi) were identified via molecular analysis. Moreover, the highest diversity of Apiospora was found in culms, and the most abundant species was Ap. arundinis. Among the nine Apiospora species, two (Ap. hysterina and Ap. paraphaeosperma) were unrecorded in Korea, and the other two species (Ap. lageniformis sp. nov. and Ap. pseudohyphopodii sp. nov.) were potentially novel species. Here, we describe the diversity of bambusicolous Apiospora species in bamboo organs, construct a multi-locus phylogenetic tree, and delineate morphological features of new bambusicolous Apiospora in Korea.

Development of a High-Resolution Multi-Locus Microsatellite Typing Method for Colletotrichum gloeosporioides

  • Mehta, Nikita;Hagen, Ferry;Aamir, Sadaf;Singh, Sanjay K.;Baghela, Abhishek
    • Mycobiology
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    • 제45권4호
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    • pp.401-408
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    • 2017
  • Colletotrichum gloeosporioides is an economically important fungal pathogen causing substantial yield losses indifferent host plants. To understand the genetic diversity and molecular epidemiology of this fungus, we have developed a novel, high-resolution multi-locus microsatellite typing (MLMT) method. Bioinformatic analysis of C. gloeosporioides unannotated genome sequence yielded eight potential microsatellite loci, of which five, CG1 $(GT)_n$, CG2 $(GT1)_n$, CG3 $(TC)_n$, CG4 $(CT)_n$, and CG5 $(CT1)_n$ were selected for further study based on their universal amplification potential, reproducibility, and repeat number polymorphism. The selected microsatellites were used to analyze 31 strains of C. gloeosporioides isolated from 20 different host plants from India. All microsatellite loci were found to be polymorphic, and the approximate fragment sizes of microsatellite loci CG1, CG2, CG3, CG4, and CG5 were in ranges of 213-241, 197-227, 231-265, 209-275, and 132-188, respectively. Among the 31 isolates, 55 different genotypes were identified. The Simpson's index of diversity (D) values for the individual locus ranged from 0.79 to 0.92, with the D value of all combined five microsatellite loci being 0.99. Microsatellite data analysis revealed that isolates from Ocimum sanctum, Capsicum annuum (chili pepper), and Mangifera indica (mango) formed distinct clusters, therefore exhibited some level of correlation between certain genotypes and host. The developed MLMT method would be a powerful tool for studying the genetic diversity and any possible genotype-host correlation in C. gloeosporioides.

임상 검체에서 분리된 Candida albicans의 MLST를 이용한 유전적 변이 특성 (Genetic Variations of Candida albicans Isolated from Clinical Specimens Using Multi-locus Sequence Typing Analysis)

  • 김희진;김상하;김성현;유영빈;김영권
    • 대한임상검사과학회지
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    • 제50권3호
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    • pp.331-336
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    • 2018
  • 본 연구에서는 국내 대전 소재 3차 병원 임상에서 분리된 Candida albicans 40 균주를 대상으로 균주 분리원에 따라 7종류의 housekeeping gene에 대한 염기서열 변이를 확인하고 MLST 분석을 통해 균주간의 계통학적 연관성에 대한 연구를 수행하여 다음과 같은 결과를 얻었다. Phylogenetic tree 분석 결과 sub-cluster 1로 central line blood (2), others (5), sputum (4), urine (7)을 포함해 총 18개가 분류되었으며, sub-cluster 2로는 central line blood (1), others (5), peripheral blood (6), sputum (1), urine (1)을 포함해 총 14개가 분류되어 동일 채취 부위에서 분리된 균주는 유전학적으로 유사성을 가지고 있을 가능성이 있음을 확인하였다. 앞으로 분리지역과 상병 등에 따른 C. albicans 유전자들의 변이 추세에 대한 자료의 축적과 임상 검체에 따른 계통학적 관계를 추정하여 감염병 연구 및 역학적 감시체계 구축에 도움이 될 것으로 생각된다.

Genetic Variations of Trichophyton rubrum Clinical Isolates from Korea

  • Yoon, Nam-Sup;Kim, Hyunjung;Park, Sung-Bae;Park, Min;Kim, Sunghyun;Kim, Young-Kwon
    • 대한의생명과학회지
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    • 제24권3호
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    • pp.221-229
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    • 2018
  • Trichophyton rubrum is one of the well-known pathogenic fungi and causes dermatophytosis and cutaneous mycosis in human world widely. However, there are not an available sequence type (ST) classification methods and previous studies for T. rubrum until now. Therefore, currently, molecular biological tools using their DNA sequences are used for genotype identification and classification. In the present study, in order to characterize the genetic diversity and the phylogenetic relation of T. rubrum clinical isolates, five different housekeeping genes, such as actin (ACT), calmodulin (CAL), RNA polymerase II (RPB2), superoxide dismutase 2 (SOD2), and ${\beta}$-tubulin (BT2) were analyzed using by multilocus sequence typing (MLST). Also, DNA sequence analysis was performed to examine the differences between the sequences of Trichophyton strains and the identified genetic variations sequence. As a result, most of the sequences were shown to have highly matched rates in their housekeeping genes. However, genetic variations were found on three different positions of ${\beta}$-tubulin gene and were shown to have changed from $C{\rightarrow}G$ (1766), $G{\rightarrow}T$ (1876), and $C{\rightarrow}A$ (1886). To confirm the association with T. rubrum inheritance, a phylogenetic tree analysis was performed. It was classified as four clusters, but there was little significant correlation. Even so, MLST analysis is believed to be helpful for determining the genetic variations of T. rubrum in cases where there is more large-scale data accumulation. In conclusion, the present study demonstrated the first MLST analysis of T. rubrum in Korea and explored the possibility that MLST could be a useful tool for studying the epidemiology and evolution of T. rubrum through further studies.

The First Korean Case of Nontuberculous Mycobacterial Lung Disease Caused by Mycobacterium abscessus Subspecies bolletii in a Patient with Bronchiectasis

  • Jeong, Byeong-Ho;Kim, Su-Young;Jeon, Kyeongman;Huh, Hee Jae;Ki, Chang-Seok;Lee, Nam Yong;Shin, Sung Jae;Koh, Won-Jung
    • Tuberculosis and Respiratory Diseases
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    • 제76권1호
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    • pp.30-33
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    • 2014
  • We report the first Korean case of lung diseases caused by Mycobacterium abscessus subsp. bolletii in a previously healthy male, except for a previous history of pulmonary tuberculosis and bronchiectasis. All serial isolates are identified as M. abscessus subsp. bolletii by multi-locus sequence analysis based on the hsp65, rpoB, and 16S rRNA fragments. At the genetic level, the isolate has the erm(41) gene with a T28 sequevar, associated with clarithromycin resistance, and no rrl mutation. The isolate is resistant to clarithromycin. Although the symptoms and radiographic findings have improved after combination of antibiotics, the follow-up sputum cultures are persistently positive.

New Finding of Golovinomyces salviae Powdery Mildew on Glechoma longituba (Lamiaceae), Besides Its Original Host Salvia spp.

  • In-Young Choi;Lamiya Abasova;Joon-Ho Choi;Young-Joon Choi;Hyeon-Dong Shin
    • 한국균학회지
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    • 제51권3호
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    • pp.239-243
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    • 2023
  • The Golovinomyces biocellatus complex is known to consist of powdery mildew from the Golovinomyces genus, associated with host plants from the Lamiaceae family. Recent molecular phylogenetic analyses have resolved the taxonomic composition of this complex, and Golovinomyces biocellatus sensu stricto is considered to be a pathogen of Glechoma species, globally. However, this paper presents a new finding of Golovinomyces salviae on Glechoma longituba, besides its original host species of Salvia. This information was inferred by molecular phylogenetic analyses from the multi-locus nucleotide sequence dataset of intergeneric spacer (IGS), internal transcribed spacer (ITS), large subunit (LSU) of rDNA, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Further, the asexual morphology of this fungus is described and illustrated.

First Report of Xenoroussoella triseptata Isolated from Soil in Korea

  • Jung-Joo Ryu;Seung-Yeol Lee;In-Kyu Kang;Leonid N. Ten;Hee-Young Jung
    • 한국균학회지
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    • 제50권3호
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    • pp.195-204
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    • 2022
  • A fungal strain, designated KNUF-20-NI009, was isolated from soil collected from Gunsan-si, Jeollabuk-do, Korea. The isolate showed cultural features typical of the genus Xenoroussoella. Colonies cultivated on malt extract agar were olivaceous-brown to pale olivaceous-white at the margins, with undersides of dark olivaceous to olivaceous-brown and a white margin. The conidia, with a size range of 2.7-5.1×1.6-3.3 ㎛ ($\bar{x}=3.6\times2.6{\mu}m$, n=50), were globoid to ellipsoid in shape, hyaline when immature, becoming light brown to golden-brown when mature, and characterized by 1 or 2 guttules. Multi-locus sequence analysis based on a combined dataset of internal transcribed spacer regions (ITS), large subunit rDNA (LSU), small subunit rDNA (SSU), translation elongation factor 1-alpha (TEF1α), and RNA polymerase II largest subunit (RPB2) sequences revealed KNUF-20-NI009 to be a strain of Xenoroussoella triseptata. This is the first report of this species in Korea.

국내에서 분리된 Fusarium sambucinum 종복합체 균주의 재동정 및 문헌 고찰 (Re-identification of Fusarium sambucinum Species Complex Strains in Korea and Their Literature Review)

  • 최윤희;;최효림;이지수;이다슬;홍승범
    • 식물병연구
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    • 제29권2호
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    • pp.118-129
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    • 2023
  • Fusarium sambucinum 종 복합체는 식물에 심각한 질병을 일으키는 분류군이다. 국내에서는 F. sambucinum 종 복합체가 일으키는 식물병에 대한 많은 연구가 보고되었으나, 한국식물 병명목록에는 2종(F. graminearum, F. sambucinum)이 14개의 기주에 식물병을 일으키는 정보만이 등록되어 있다. F. sambucinum 종복합체의 다양성 및 병원성을 확인하기 위해 농업미생물은행(Korean Agricultural Culture Collection)에 보존된 57 균주의 F. sambucinum 종 복합체를 다중 유전자 염기서열 분석(multi-locus sequence typing)을 바탕으로 7종(F. asiaticum, F. graminearum, F. vorosii, F. meridionale, F. bootii, F. kyushuense, F. armeniacum)으로 재동정하였다. 이전의 보고와 이번 연구의 결과에 따라, F. sambucinum 종 복합체 중 5종(F. asiatum, F. graminearum, F. vorosii, F. armeniacum, F. sambucinum)이 24개의 기주에 병원성을 보였으며, 3종(F. meridionale, F. bootii, F. kyushuense)의 병원성은 명확하지 않았다.

Molecular Phylogeny and Morphology Reveal the Underestimated Diversity of Mortierella (Mortierellales) in Korea

  • Lee, Jae-Sung;Nam, Bora;Lee, Hyang Burm;Choi, Young-Joon
    • 한국균학회지
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    • 제46권4호
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    • pp.375-382
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    • 2018
  • Members of the genus Mortierella (Mortierellales) are filamentous fungi, which are found on nearly all substrates, but more frequently in soil. Till date, 7 species of Mortierella have been reported in Korea, but being a ubiquitous group with high species diversity in temperate zones, this number is still low. During a survey of fungal biodiversity in Korea, we collected many isolates of Mortierella, and through morphological and molecular phylogenetic analyses, identified them to be 3 previously unrecorded species, namely, M. chienii, M. epicladia, and M. gamsii. A total of 10 Mortierella species in Korea, including the 3 species reported in the present study, are widely distributed in 5 out of 7 phylogenetic groups of this genus. This indicates that the diversity of Mortierella was so far underestimated in Korea. Multi-locus sequence analysis is required to provide a more reliable backbone for some uncertain phylogenetic groupings and to more clearly define a species of Mortierella, which would encourage deeper research in the diversity and ecological roles of Mortierella and allied genera.