• 제목/요약/키워드: URP-PCR

검색결과 44건 처리시간 0.038초

Analysis of Genetic Relatedness in Alternaria species Producing Host Specific Toxins by PCR Polymorphism

  • Kang, Hee-Wan;Lee, Byung-Ryun;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • 제19권5호
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    • pp.221-226
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    • 2003
  • Twenty universal rice primers (URPs) were used to detect PCR polymorphisms in 25 isolates of six different Alternaria species producing host specific toxins (HST). Eight URPs could be used to reveal PCR polymorphisms of Alternaria isolates at the intra- and inter-species levels. Specific URP-PCR polymorphic bands that are different from those of the other Alternaria spp. were observed on A. gaisen and A. longipes isolates. Unweighted pair-group method with arithmetic mean (UPGMA) cluster analysis using 94 URP polymorphic bands revealed three clustered groups (A. gaisen group, A. mati complex group, and A. logipes group).

Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.120.2-120
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    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

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Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • 한국동물위생학회지
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    • 제40권1호
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

PCR다형성 밴드 유래 DNA probe에 의한 Erwinia carotovora subsp. carotovora 특이적 검출 (Specific Detection of Erwinia carotovora subsp. carotovora by DNA Probe Selected from PCR Polymorphic Bands)

  • 강희완;고승주;권순우
    • 한국식물병리학회지
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    • 제14권2호
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    • pp.164-170
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    • 1998
  • This study was carried out to develop DNA probe for specific detection of Erwinia carotovora subsp. carotovora. Universal rice primer (URP, 20 mer) developed from repetitive sequence of rice was applied for producing PCR DNA fingerprints of Erwinis spp. In E. carotovora subsp. carotovora strains, primer URP2F amplyfied polymorphic bands which are distinguisable from other Erwinia spp. A PCR band of 0.6 kb selected from PCr polymorphic bands of E. carotovora subsp. carotovora strains was cloned and evaluated as a diagnostic DNA probe. Among 28 bacterial strains including 22 Erwinia spp, the probe (pECC2F) only hybridized to total DNAs from e. carotovora subsp. carotovora strains and E. carotovora subsp. wasabiae, but sizes of hybridized bands were different between these subspecies, 10.0 kb and 3.5 kb respectively. In dot blot assays using probe pECC2F, as few as 103 colony forming units (CFU) of E. carotovora subsp. carotovora could be detected in a suspension containing about 1$\times$103 CFU of soil bacteria.

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가지과 작물에서 분리한 Alternaria 속 균의 형태적, 분자생물학적 특징 (Morphological and Molecular Characterization of Alternaria Isolates from Solanaceous Crops)

  • 유승헌;조혜선;김병련;박명수
    • 한국균학회지
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    • 제31권2호
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    • pp.103-113
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    • 2003
  • 국내의 가지과 작물에서 분리한 Alternaria 25 균주를 공시하여 분생포자의 형태적 특징을 조사하였고 A. solani와 A. tomaotphilad의 대표균주와 비교하였다. 분리균주 중 형태적으로 구별되는 몇 균주들과 대표균주를 공시하여 감자, 토마토, 가지, 고추에 대한 병원성 검정을 실시하였다. 대표균주를 포함한 Alternaria 17개 균주를 공시하여 ITS 영역과 histone h3 유전자의 염기서열분석, URP (universal rice primer)에 의한 핵산지문분석을 실시하였다. Alternaria 균주들은 분생포자의 형태적 특징에 의하여 A. tomaotophila(ATO), A. solani(ASO) 및 미동정의 Alternaria sp.(ASP)의 group으로 나눌 수 있었고 A. solani(ASO)는 분생포자 부리(beak)의 특징에 의하여 다시 ASO(1)과 ASO(2)의 2 type으로 나눌 수 있었다. ATO와 ASO 균주들은 실험에 사용한 감자, 토마토, 가지, 고추에 모두 병원성이 있었고 ATO 균주는 특히 토마토에 강한 병원성이 있었으나, ASP 균주는 감자에만 병원성이 있었다. 이 연구에서 사용한 molecular marker중 ITS와 histon H3 유전자의 염기서열 분석은 4개의 형태적 group을 명확히 구분할 수 없었지만, URP-PCR 핵산지문분석법은 이들 group을 명확히 구분할 수 있었다. 분생포자의 형태, 병원성검정, 분자생물학적 특징을 종합할 때 토마토 겹무늬병에 관여하는 A. tomatophila는 감자겹둥근무늬병에 관여하는 A. solani와 뚜렷이 구분할 수 있었다. 또한 감자에서 분리한 Alternaria sp.(ASP)는 A. solani(ASO)와 형태적으로 유사하였지만 분생포자의 폭이 두껍고 부리(beak)가 작다는 점에서 ASO와 구별되었으며 가지과 작물에서 보고된 바 없는 신종으로 추정되었다.

느티만가닥버섯 균주의 형태 및 유전적 유연관계 분석 (Analysis of Morphological and Genetic Relationships amomg Isolates of the Artificially Cultivated Mushroom, Hypsizygusmarmoreus)

  • 김민경
    • 한국균학회지
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    • 제48권3호
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    • pp.313-323
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    • 2020
  • Hypsizygus marmoreus의 형태적 특성과 유전적 유연관계를 조사하기 위해 한국 및 다른 국가에서 111개의 균주를 수집하였다. RAPD (Random amplified polymorphic DNA) 및 ITS rDNA 염기서열 분석을 이용하여 H. marmoreus 균주간의 유전적관계를 확인하였다. URP-PCR을 이용한 RAPD 분석결과, H. marmoreus의 모든 균주는 크게 3개의 그룹으로 분류되었으나, 90% 이상의 높은 유사성을 보였다. 또한 형태적 및 지리적으로 분류가 가능하였으나 갈색 균주 및 흰색 균주는 구별되지 않았다. 따라서 형태적, 지리적 차이가 있는 16개의 균주를 선별하여, 640 bp 길이의 ITS 염기서열을 정렬하고 비교하였다. ITS 염기서열의 유사성은 공시균주와 GenBank의 H. marmoreus 균주 사이에서 94.8-99.1 %였다. 수집지역과 형태에 차이가 있었지만, 실험에 사용된 공시균주는 모두 H. marmoreus에 가까운 유연관계를 형성하였다.

Morphology, Pathogenicity and Molecular analysis of Alternaria Isolates from Solanaceous Crops (oral)

  • Cho, H.S.;Park, M.S.;Kim, B.R.;Yu, S.H.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.112.3-113
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    • 2003
  • More than 30 isolates of Alternaria were obtained from various solanaceous crops in Korea. For all isolates, morphological characteristics of the conidia were determined and compared with those of representative isolates of A. solani and A. tomatophila. Pathogenicity test was performed to Potato, tomato, egg plant and red Pepper and molecular characteristics of them including the representative isolates were determined using sequence analyses of ITS rDNA and histone H3 gene, and URP-PCR analysis. Based on morphological characteristics, the isolates from the solanaceous crops were grouped as identical or very similar to either A. tomatophila(ATO), A. solani(ASO), and unidentified Altemaria sp.(ASP). Among the molecular markers used in this study, the URP-PCR analysis was found to be appropriate for taxonomic resolution of these species. Based on the conidial morphology, pathogenicity test and molecular characteristics, A. tomatophila(early blight of tomato) could be distinguished from A. solani(early blight of potato), and the Alternaria sp.(ASP) from potato, which was closely related to A. solani in conidial morphology, was considered as a new species.

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DNA Profiles of Trichoderma spp. in Korea

  • Park, Dong-Suk;Kang, Hee-Wan;Park, Young-Jin;Lee, Mi-Hee;Lee, Byoung-Moo;Hahn, Jang-Ho;Go, Seung-Joo
    • Mycobiology
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    • 제32권1호
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    • pp.24-34
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    • 2004
  • Molecular approaches, internal transcribed spacer(ITS) sequences of ribosomal DNA, and Universal Rice Primer Polymerase Chain Reaction(URP-PCR) were used to investigate the genetic diversity, taxonomic complexity, and relationships of Trichoderma species in mushroom farms. Forty-one isolates of 13 Trichoderma spp. were used in this study and clustered into eight groups. The DNA fingerprint patterns and ITS1 region sequence alignment data showed similar results, but not in some species, such as T. virens, T. atroviride, T. harzianum, and T. aureoviride. Results of this study have proven that the morphology-based taxonomic system has some limitations in terms of classification. The data obtained in this study would be a good index for classifying indistinguishable Trichoderma strains.