• Title/Summary/Keyword: URP-PCR

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Genetic Diversity of Paecilomyces japonica and Cordypces militaris Strains by URP-PCR Fingerprinting (URP-PCR핵산지문에 의한 눈꽃동충하초 (Paecilomyces japonica.)와 번데기동충하초(Cordypces militaris) 유전적 다양성분석)

  • Kim, Jong-Kun;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.180-184
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    • 2011
  • This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.

Genetic Divesity Analysis of Fungal Species by Universal Rice Primer (URP)-PCR (Universal Rice Primer (URP)-PCR에 의한 곰팡이 종의 유전적 다양성 검정)

  • Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.2
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    • pp.78-85
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    • 2012
  • URP primers that were derived from repetitive DNA sequence of rice weedy rice have been applied for producing PCR polymorphisms in different fungal species. URP-PCR protocol employed stringent PCR with high annealing temperature over $55^{\circ}C$ throughout the thermo-cycling reaction, giving high reproducibility. Under the PCR condition, Each single URP primer produced characteristic fingerprints from diverse genomes of different fungal species, indicating its universal applicability. URP-PCR has been accessed for applicability to various fungal species with 33 genus, 142 species and 1,489 isolates. Numerous related papers have demonstrated that URP-PCR profiles of fungal species are very useful for identifying fungal species at intra and inter species levels. The results were reviewed in this paper.

Genetic Diversity of Agrobacterium vitis Strains in Korea (국내 포도나무 혹병(Agrobacterium vitis) 균주의 유전적 다양성)

  • Kim, Jong-Kun;Choi, Jae-Eul;Kang, Hee-Wan
    • Research in Plant Disease
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    • v.13 no.3
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    • pp.137-144
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    • 2007
  • Fifty nine strains of Agrobacterium vitis, the causal agent of crown-gall disease on grapevine, originating from different geographical regions and 16 grapevine cultivars including 35 Kyoho cultivar of Korea, were characterized by PCR polymorphic analysis using Universal Rice Primer(URP). Of 12 URP primers, primers URP1F, URP2R, URP2F, and URP4R, URP17R were available for detecting PCR polymorphic bands among the A. vitis strains. PCR polymorphic bands produced by primers URP2F and URP17R were profiled to 12 strain types. A. vitis strains originated from Kyoho cultivar of grapevine showed relatively simple genetic diversify of the four PCR types, while the A. vitis strains originated from other grapevine cultivars and type culture strains showed various genetic diversity with 8 types. Unweighted Pair-Group Method with Arithmetic mean(UPGMA) cluster analysis using the URP-PCR polymorphic bands showed 59.4. vitis strains are genetically clustered into large seven groups.

Genetic Diversity of Pleurotus eringii Strains in Korea Based on Morphological Characteristics and PCR Polymorphism (형태적 특성과 PCR다형성 분석에 의한 국내 큰느타리버섯 계통의 유전적 다양성 분석)

  • Jeon, Sun-Jeong;Kim, Jong-Kun;Kim, Gum-Hee;Chi, Jeong-Hyun;Seo, Geon-Sik;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.19-27
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    • 2009
  • This study was conducted to investigate genetic characteristics of 25 Pleurotus eringii strains that have been released in Korea based on cultural, morphological features and PCR fingerprints. Strains PER-007 and PER-012 showed distinct cultural characteristics in growth rate, morphological characteristics of mycelial colony and fruiting bodies when compared to those of other strains. Strain PER-007 did not form primordium initiation in sawdust medium and PER-012 also showed different phenotypes on fruiting bodies. Eleven URP primers were used to detect PCR polymophic bands in P. eringii strains. Primers URP1F, URP2R, URP2F, URP4R, URP6R, URP9F and URP17R were selected as useful primers for amplifying PCR polymorphic bands in P. eringii strains. The genetic similarity index was calculated by using PCR polymorphic bands amplified by eight URP primers among the 25 strains. The P. eringii strains were grouped by four distinct clusters on the UPGMA analysis. The genetic similarity values ranged from 100% to 76% were observed in three major groups, suggesting close genetic relatedness of them. Exceptionally, PER-007 and PER-017 were involved in outgroup.

Genetic Analysis of Cultivars in Pleurotus spp. of Korea by URP-PCR Polymorphism (URP-PCR 다형성에 의한 국내 느타리버섯 품종의 유전적 특성 분석)

  • Kim, Jong-Kun;Lim, Seon-Hwa;Lee, Dae-Sung;Chi, Jeong-Hyun;Seo, Geon-Sik;Ju, Young-Cheol;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.35 no.2
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    • pp.61-67
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    • 2007
  • Twelve URP primers were used to assess genetic characteristics of oyster mushroom including 59 Pleurotsu ostreatus cultivars, two of P. florida cultivars, one P. sajor-caju cultivars, one P. abalonus cultivar and two P. eryngii cultivars registered in Korea. Six URP primers produced PCR polymorphic bands within and between the Pleurotus species. Primer URP2F produced distinct cultivar specific PCR polymorphic bands that profiled to 15 cultivar types. PCR polymorphic bands amplified by URP2F, URP6R, URP4R and URP2R were used for UPGMA cluster analysis. Fifty nine cultivars of Pleurotus ostreatus are genetically clustered into 5 groups, showing genetic similarity over 70% among them and P. abalonus. P. eryngii and P. sajor-caju, were involved in outside groups.

Genetic Diversity of Penicillium isolates Isolated from Pears with Postharvest Decay in Storage (수확 후 저장 배에서 분리한 Penicillium속 균의 유전적 다양성)

  • Han, Do-Suk;Hong, Sung-Kee;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.11-18
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    • 2012
  • This study was carried out to identify the genetic diversity of Penicillium isolates that were isolated from pears with postharvest decay in storage. URP-PCR was used to detect DNA diversity of 84 Penicillium isolates. Based on URP-PCR profiles, 18 Penicillium isolates were selected and their PCR polymorphic bands were produced by additional primers URP1F, URP2R, URP2F, and URP4R. UPGMA cluster analysis using the polymorphic bands showed four clustered groups and futhermore cultural and morphological features characterized the 18 Penicillium isolates. Group 1 was dominant, which occupies 70% in the four clustered groups and identified as P. expansum based on ITS sequence and morphological features.

Genomic Differentiation Among Oyster Mushroom Cultivars Released in Korea by URP-PCR Fingerprinting

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;You, Chang-Hyun;Lee, Byoung-Moo;Eun, Moo-Yong;Go, Seong-Joo
    • Mycobiology
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    • v.29 no.2
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    • pp.85-89
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    • 2001
  • URP primers of 20 mer derived from repetitive sequence of rice were used to assess genetic variation of oyster mushroom consisting of 10 cultivars of Pleurotus ostreatus, two cultivars of P. florida and two cultivars of P. sajor-caju which were registered in Korea. URP2F and URP38F primers produced cultivar-specific PCR polymorphic bands in the Pleurotus species. UPGMA cluster analysis using the URP-PCR data showed that 14 Pleurotus cultivars are genetically clustered into large three groups. The URP-PCR data provided important information for more efficient breeding strategies of Pleurotus cultivars.

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Molecular Characterization of Small-Spored Alternaria Species (소형의 포자를 형성하는 Alternaria 균류의 분자생물학적 특징)

  • Kim, Byung-Ryun;Park, Myung-Soo;Cho, Hye-Sun;Yu, Seung-Hun
    • Research in Plant Disease
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    • v.11 no.1
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    • pp.56-65
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    • 2005
  • To establish taxonomic system of morphologically similar species of small-spored Alternaria, phylogenetic analysis of internal transcribed spacer (ITS 1, ITS 2 and 5.8S rDNA) and mitochondrial small subunit (mt SSU) rDNA sequences and URP-PCR fingerprinting analysis from 11 species ofAlternaria were performed. Phylogenetic analysis of ITS and mt SSU rDNA sequences revealed that 10 out of 11 species of the smallspored Alternaria were phylogenetically identical with a bootstrap value of 100%. A. infectoria only was phylogenetically differentiated from the other species. The results suggest that the 10 small-spored Alternaria species are very closely related evolutionally and the markers can not be used for differentiation of the smallspored Alternaria species. URP-PCR fingerprinting analysis from eleven species of smallspored Alternaria using 10 URP primers showed that it was possible to differentiate the species, although genetic similarities were found among the species. The Alternaria sp. from common pokeweed could be distinguished from other species by URP-PCR analysis, and it was considered as a new species. A. infectoria could be easily distinguished from the other 10 species by phylogenetic analysis of ITS and mt SSU rDNA sequences and the URPPCR fingerprinting analysis.

Genetic Differentiation of Pseudomonas syringae Pathovar tomato from Other P. syringae Pathovars using REP-PCR and URP-PCR

  • Cho, Min-Seok;Park, Dong-Suk;Yun, Yeo-Hong;Kim, Seong-Hwan;Shim, Myung-Yong;Choi, Chang-Won;Kim, Young-Shick
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.60-67
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    • 2012
  • For the genetic differentiation of $Pseudomonas$ $syringae$ pathovar $tomato$, a total of 51 $P.$ $syringae$ pv. strains infecting 33 different host plants were analyzed using repetitive element PCR(REP-PCR) and universal rice primer PCR(URP-PCR). The entire DNA fingerprint profiles were analyzed using unweighted pair-group method with arithmetic averages (UPGMA). The 51 $P.$ $syringae$ pv. strains could be divided into five clusters based on 65% similarity by Rep-PCR using BOX, ERIC, and REP primers. $P.$ $syringae$ pv. $tomato$ cluster was well separated from other 31 $P.$ $syringae$ pathovars. $P.$ $syringae$ pv. $tomato$ cluster included only $P.$ $syringae$ pv. $maculicola$ and $P.$ $syringae$ pv. $tomato$. $P.$ $syringae$ pv. $tomato$ strains could be divided into two genetic groups. Meanwhile, the Pseudomonas pv. strains could be divided into four clusters based on 63% similarity by URP-PCR using 2F, 9F, and 17R primers. $P.$ $syringae$ pv. $tomato$ cluster was also well separated from 30 other $P.$ $syringae$ pathovars. In this case, $P.$ $syringae$ pv. $tomato$ cluster included $P.$ $syringae$ pv. $maculicola$, $P.$ $syringae$ pv. $berberidi$, and $P.$ $syringae$ pv. $tomato$. $P.$ $syringae$ pv. $tomato$ strains was also separated into two genetic groups by URP-PCR analysis. Overall, our work revealed that $P.$ $syringae$ pv. $tomato$ can be genetically differentiated from other $P.$ $syringae$ pathovars by the DNA fingerprint profiles of REP-PCR and URP-PCR. We first report that there are two genetically diverged groups in $P.$ $syringae$ pv. $tomato$ strains.

PCR Based Detection of Phellinus linteus using Specific Primers Generated from Universal Rice Primer(URP) Derived PCR Polymorphic Band

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;Lee, Byoung-Moo;Cho, Soo-Muk;Kim, Ki-Tae;Seo, Geon-Sik;Go, Seung-Joo
    • Mycobiology
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    • v.30 no.4
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    • pp.202-207
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    • 2002
  • This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.