• 제목/요약/키워드: Internal transcribed spacer rDNA

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PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp.

  • Min, Byung-Re;Lee, Young-Mi;Choi, Yong-Keel
    • Journal of Microbiology
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    • 제38권2호
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    • pp.66-73
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    • 2000
  • To investigate the genetic relationship among 12 species belonging to the Fusarium section Martiella, Dlaminia, Gibbosum, Arthrosporiella, Liseola and Elegans, the internal transcribed spacer(ITS) regions of ribosomal DNA (rDNA) were amplified with primer pITS1 and pITS4 using the polymerase chain reaction(PCR). After the amplified products were digested with 7 restriction enzymes, restriction fragment length polymorphism (RFLP) patterns were analyzed. The partial nucleotide sequences of the ITS region were determined and compared. Little variation was observed in the size of the amplified product having sizes of 550bp or 570bp. Based on the RFLP analysis, the 12 species studied were divided into 5 RFLP types. In particular, strains belonging to the section Martiella were separated into three RFLP types. Interestingly, the RFLP type of F. solani f. sp. piperis was identical with that of isolates belonging to the section Elegans. In the dendrogram derived from RFLP analysis of the ITS region, the Fusarium spp. examined were divided into two major groups. In general, section Martiella excluding F. solani f. sp. piperis showed relatively low similarity with the other section. The dendrogram based on the sequencing analysis of the ITS2 region also gave the same results as that of the RFLP analysis. As expected, 5.8S, a coding region, was highly conserved, whereas the ITS2 region was more variable and informative. The difference in the ITS2 region between the length of F. solani and its formae speciales excluding F. solani f. sp. piperis and that of other species was caused by the insertion/deletion of nucleotides in positions 143-148 and 179-192.

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Prevalence and Phylogenetic Analyses of Trichuris suis in Pigs in Hunan Province, Subtropical China

  • Tan, Lei;Wang, Aibing;Yi, Jing;Liu, Yisong;Li, Jiayu;Liu, Wei
    • Parasites, Hosts and Diseases
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    • 제56권5호
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    • pp.495-500
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    • 2018
  • Trichuris suis infection in pigs is ubiquitous in intensive and extensive farms, which causes potential threat to human health. The objective of this research was to investigate the prevalence of T. suis in pigs in Hunan province. Total 2,267 fresh fecal samples distributed in 28 pig farms from 7 different administrative regions (Hunan province) were evaluated for the existence of T. suis eggs using saturated NaCl floating method. The average infection rate of T. suis in pigs was 8.91% in Hunan province. To determine genetic variation of the gained T. suis isolates in the present study, the internal transcribed spacer (ITS) regions from nuclear ribosomal DNA (rDNA) of 7 T. suis isolates were cloned and analyzed. Nucleotide diversities were 1.0-3.5% and 0-3.8% for ITS-1 and ITS-2, respectively. Phylogenetic analyses indicated that all isolates collected in the present study and T. suis available in Genbank generated a monophyletic clade. The present investigation revealed high infection rates of T. suis in pigs in Hunan province, which shed light on making effective measures to prevent and control T. suis infection in pigs in Hunan province.

한국 자생 난초 뿌리에서 분리한 미기록 내생균 5종 (Five Species of Endophytic Fungi Isolated from Roots of Native Orchid Plants from Korea)

  • 이봉형;엄안흠
    • 한국균학회지
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    • 제45권4호
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    • pp.355-361
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    • 2017
  • 본 연구는 전라북도 진도에서 자란(Bletilla striata), 강원도 정선군 함백산에서 감자난초(Oreorchis patens)와 은대난초(Cephalanthera longibracteata)의 뿌리에서 내생균을 분리하고 이들의 형태 및 분자생물학적 방법을 통해 동정하였다. 각 균주의 형태학적 특징을 분석하기 위해 포자의 모양과 특징을 관찰하였고, 분자생물학적 동정을 위해 ITS1F와 ITS4 primer를 이용하여 internal transcribed spacer (ITS) 및large subunit (LSU) rDNA영역을 분석하였다. 그 결과 Phialocephala bamuru, Coniochaeta mutabilis, Phialophora foetens, Calonectria Canadensis, Neonectria ramulariae 5종의 국내 미기록 내생균이 동정되어 이를 보고하고자 한다.

구상나무의 잎에서 분리한 두 종의 국내 미기록 내생균 (Two Unreported Species of Endophytic Fungi Isolated from Leaves of Abies koreana in Korea)

  • 박혁;김동여;엄안흠
    • 한국균학회지
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    • 제46권1호
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    • pp.22-27
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    • 2018
  • 제주도 한라산 구상나무의 잎에서 내생균을 분리하여 다양성을 확인하는 과정에서 두 종의 국내 미기록 균주를 확인하였다. 분리된 균주는 형태적 특징과 internal transcribed spacer (ITS) 지역, large subunit (LSU) rDNA 지역 염기서열의 계통분석을 통해 동정하였다. 확인된 두 미기록종 균주는 Coleophoma parafusiformis와 Coniochaeta ligniaria이며, 두 균주에 대한 형태적 특징과 염기서열의 분석 결과를 제시하였다.

눈향나무의 잎에서 분리된 4종의 국내 미기록 내생균 (First Report on Four Novel Endophytic Fungal Species Isolated from Leaves of Juniperus chinensis var. sargentii in Korea)

  • 박혁;엄안흠
    • 한국균학회지
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    • 제46권2호
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    • pp.114-121
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    • 2018
  • 한라산에 서식하는 눈향나무의 침엽에서 내생균을 분리하였다. 분리된 균주들은 형태적 특성과 internal transcribed spacer 영역, large subunit rDNA 영역, translation elongation factor 영역 및 ${\beta}$-tublin 영역 염기서열의 계통분석을 이용하여 동정하였다. 그 결과 국내 미기록 내생균인 Allantophomopsis lycopodina, Epicoleosporium ramularioides, Kabatina juniperi, Seiridium podocarpi 4종의 균주를 확인하였다. 4종의 미기록 균주에 대한 형태적 특성 및 계통분석의 결과에 대해 기술하였다.

DNA 바코딩과 고해상 융해곡선분석에 기반한 인삼속 식물의 종 판별 (Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species)

  • 방경환;김영창;임지영;김장욱;이정우;김동휘;김기홍;조익현
    • 한국약용작물학회지
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    • 제23권6호
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    • pp.439-445
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    • 2015
  • Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

제주도에 서식하는 목본 식물의 잎에서 분리한 미기록 내생균 (Identification of Unrecorded Endophytic Fungi Isolated from Leaves of Woody Plants in Jejudo, Korea)

  • 이봉형;김동여;박혁;어주경;이향범;엄안흠
    • 한국균학회지
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    • 제44권4호
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    • pp.252-258
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    • 2016
  • 본 연구는 제주도 사려니숲과 동백동산에서 편백나무(Chamaecyparis obtusa), 삼나무(Cryptomeria japonica), 비자나무(Torreya nucifera), 꽝꽝나무(Ilex crenata), 동백나무(Camellia japonica) 등 5종의 목본 식물의 잎에서 내생균을 분리하고, 형태 및 분자생물학적 특징 통해 동정하였다. 이를 위해 primer인 ITS1F와 ITS4를 이용하여 internal transcribed spacer (ITS) rDNA영역과 primer LR0R과 LR16을 이용하여 large subunit rDNA 영역을 그리고 primer Bt2a과 Bt2b를 이용하여 ${\beta}$-tubulin 영역의 염기서열을 동시에 분석하였다. 그 결과 Mycosphaerella aleuritidis, Neofusicoccum eucalyptorum, Neofusicoccum parvum, Phyllosticta citrichinensis, Phyllosticta cryptomeriae, Phomopsis cotoneastri, Sphaerulina rhododendricola, Guignardia mangiferae, Lophodermium jiangnanense, Lophodermium minus 등 10종의 미기록 내생균을 동정하였고, 이를 보고하고자 한다.

Soft Rot on Cucumis melo var. makuwa Caused by Rhizopus oryzae

  • Kwon, Jin-Hyeuk;Kim, Jin-Woo;Lee, Yong-Hwan;Shim, Hong-Sik
    • Mycobiology
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    • 제38권4호
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    • pp.336-338
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    • 2010
  • Rhizopus oryzae is reported for the first time on Cucumis melo L. var. makuwa Makino. A detailed description of this Korean specimen is given, along with its rDNA internal transcribed spacer sequence. On the basis of mycological characteristics and molecular data, the fungus was identified as R. oryzae Went & Prinsen Geerligs.

Characterization of Rhizodermea veluwensis Isolated from the Roots of Rhododendron mucronulatum in Korea

  • Park, Hyeok;Eom, Ahn-Heum
    • 한국균학회지
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    • 제45권2호
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    • pp.102-106
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    • 2017
  • A fungal strain was isolated from surface-sterilized roots of Rhododendron mucronulatum, a plant species belonging to the Ericaceae family, collected from Mt. Minjujisan, Korea. This fungal strain was identified as Rhizodermea veluwensis based on its morphological characteristics and based on phylogenetic analysis of its internal transcribed spacer regions and large-subunit rDNA. R. veluwensis has not been previously reported in Korea, and for the first time, we report and describe it herein.

Rapid and Accurate Species-Specific Detection of Phytophthora infestans Through Analysis of ITS Regions in Its rDNA

  • Kim, Kyoung-Su;Lee, Youn-Su
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.651-655
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    • 2000
  • Polymerase chain reaction (PCR) was used to specifically detect Phytophthora infestans by analyzing the sequences of the ribosomal internal transcribed spacer regions (ITS) in the rDNA of the Phytophthora species. Based on the sequence data, PISP-1 together with the ITS3 primer were used to detect p. infestans. A single ca. 450 bp segment was observed in P. infestans, but not in the other fungal or bacterial isolates. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using these species-specific primers, a unique band was obtained within annealing temperatures of $55^{\circ}C$-$61^{\circ}C$ and template DNA levels of 10 pg-100 ng.

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