• Title/Summary/Keyword: RAPD markers

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Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.130-130
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    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

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Genetic Variation and Identification of RAPD Markers from Some Garlic Cultivars in Korea and Mongolia (한국과 몽고 일부 재배마늘의 유전적 변이와 재배종 특이적 RAPD 마커의 탐색)

  • Bae, Seong-Kuk;Jung, Eun-A;Kwon, Soon-Tae
    • Korean Journal of Plant Resources
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    • v.23 no.5
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    • pp.458-464
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    • 2010
  • Twelve garlic cultivars collected from Korea and Mongolia were evaluated genetic similarity and diversity by RAPD method using oligo-nucleotide random primers. Genomic DNA isolated from twelve garlic cultivars were amplified by polymerase chain reaction using 143 primers, and 55 primers showed polymorphic DNA bands. Among a total of 187 bands amplified by 55 primers, 128 polymorphic bands were subjected to analysis for genetic relationship of garlic cultivars. Garlic cultivars were classified into three groups, such as group-I corresponded to Euiseong, Seosan, Samchuk and Yecheon-A, Yechun-B, Euiseong-norang, Jeongsun, Namdo, Yookback and Danyang cultivars, and group-II to Mongolia and group-III to Daeseo cultivars. Thirty DNA bands showing unique specificity to the specific cultivars are likely to be useful for identification of garlic local cultivars as DNA markers.

Spatial Autocorrelation Analysis of Carex humilis on Mt. Giri by RAPD (RAPD에 의한 지리산 내 산거울 집단의 공간적 상관관계 분석)

  • Lee, Bok-Kyu;Lee, Byeong-Ryong;Huh, Man-Kyu
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1287-1293
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    • 2010
  • The spatial distribution of alleles and geographical distances of a Carex humilis population on Mt. Giri in Korea were studied. A total of 102 DNA fragments (bands) were found among 107 plants. Among these 102 bands, 48 (47.1%) bands were polymorphic. In a simple variability of subpopulations by the percentage of polymorphic bands, distances I and V exhibited the lowest variation (16.7%). Distance VIII showed the highest variation (22.6%). The total genetic diversity (H) was 0.076 across species. Class VIII had the highest H (0.093), while class I had the lowest (0.063). Genetic similarity of individuals was found among subpopulations at up to a scale of 60 m distance, and this was partly due to a combination of alleles. Within the Mt. Giri population, a strong spatial structure was observed for RAPD markers, indicating a very low amount of migration among subpopulations and that the distribution of individual genotypes of a given population was clumped. The present study demonstrated that analysis of RAPD markers could be successfully used to study the spatial and genetic structures of C. humilis.

Fundamental Study for RAPD-PR Analysis in the Silkworm, Bombyx mori (누에 RAPD-PCR 분석을 위한 기초연구)

  • 황재삼;이진성
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.7-12
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    • 1996
  • Reproducible the random amplified polymorphic DNAs(RAPDs) patterns were obtained in the two silkworm strains(J111, Galwon) by adjusting concentration optimized of Taq DNA polymerase(one unit), dNTP(200$\mu$M), MgCl2(1.5mM) and template DNA(30ng). In addition, anealing temperature ranging 35$^{\circ}C$ to 42$^{\circ}C$ by the adjusted condition was investigated and fixed at 35$^{\circ}C$ in this study. Variation among individuals and between male and female of Jam 113 strain was not authorized. DNA polymorhpisms among silkworms were authorized by five RAPD markers using OPM04 random primer. Using the primer showing polymorhpims between parents(J111, Galwon) in thirty three individuals, RAPD-PCR for F2 analysis was performed and segregated 3 : 1 in the F2 population. Consequently, RAPDs detected in the parents were obtained as genetic markers, which can be used for construction of genetic map for this industrially particular insect, silkworm Bombyx mori

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Randomly Amplified Polymorphic DNA Analysis of Listeria Species Isolated from Foods in Korea (국내 식품으로부터 분리한 Listeria Species의 RAPD 분석)

  • 최영춘;박부길;이택수;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.606-614
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    • 2000
  • This study was carried out for comparing Listeria strains developing genetic markers for Listeroa strains using Listeria sp. genetic markers using Randomly Amlymorphic DNA (RAPD) analysis method. Five of RAPD promers (OPA-01, OP-26-01, OP-26-02, OPB-01, OP-26-10) showed the distinctive polymorphism among Kisteria sp. isolated from domestic foods. RAPD-PCR with five arbitrary primers produced 76 DNA polymorphism. Among them, OPA-01 and OP-26-01 primers produced about 1.5kb and 0.7 kb amplified DNA fragments for all the Listeric relationships of Listeria sp. using NTSYS program were grouped into 7 clusters and showed 0.54 to 0.93 similarity among strains. Especially, No. 3 and No. 20 isolates showed the genetically most similar relationship by 0.94, and No. 7 and No. 24, or No. 7 and N0. 45 isolates showed the least similarty by 0.54 From these results, RAPD analysis method deemed to be successfully applied the classification and genetic analysis for Listeria sp. isolates.

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Genetic Variation of Coreoleuciscus splendidus Populations from Four Major Rivers in Korea as Assessed by RAPD PCR (RAPD PCR에 의한 4대강 쉬리 Coreoleuciscus splendidus 개체군들의 유전변이 분석)

  • Song, Ha-Yoon;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.21 no.2
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    • pp.129-133
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    • 2009
  • Random Amplified Polymorphic DNA (RAPD) analysis was used to investigate the genetic variations of Coreoleuciscus splendidus within and among the West Korea Subdistrict populations (in Han and Geum Rivers) and the South Korea Subdistrict populations (in Seomjin and Nakdong Rivers). Twelve random primers were employed to generate RAPD markers. All primers were produced to identify specific RAPD markers between the West and South Korea Subdistrict populations. Analyses of genetic similarity and distance among the West and South Korea Subdistrict populations of C. splendidus also revealed similar results, with low genetic similarity (0.49~0.53) and high distance value (0.63~0.71). UPGMA dendrogram based on genetic distance was also similar in results. Therefore, the West Korea Subdistrict populations and the South Korea Subdistrict populations vary in genetic structure, and C. splendidus in the South Korea Subdistrict may represent a different species.

Identification of Potential Species-Specific Marker in Several Fish Species by RAPD Using Universal Rice Primers (Universal Rice Primer (URP)-RAPD 방법에 의한 어류 종 특이 marker의 동정)

  • KIM Woo-Jin;KIM Kyung-Kil;LEE Jeong-Ho;PARK Doo-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.3
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    • pp.317-320
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    • 2003
  • Morphologically similar fish species were subjected to the random amplified polymorphic DNA (RAPD) analysis using universal rice primer (URP). The fish species tested were sea basses (Lateolabrax japonicus and L. maculatus), eels (Anguilla japonica, A. bicolor bicolor, A. rostrata, and A. anguilla), and flounders (Limanda yokohamae and L. herzensteinin). Highly reproducible RAPD patterns were observed with several potential species-specific markers. The results indicate that RAPD technique using URP is useful for distinguishing fish psecies in a rapid manner.

Discrimination of the Genus Leontopodium Species (Gentianales: Asteraceae) Based on RAPD

  • Jeon, Mi Gyeong;Choi, Kang Jun;Kim, Ji Young
    • Journal of Forest and Environmental Science
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    • v.31 no.1
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    • pp.68-71
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    • 2015
  • Korean L. leiolepis of the genus Leontopodium could be discriminate from the foreign L. alpinum using random amplified polymorphic DNA (RAPD). Among the 12 URP markers used for the detection, the URP-5 marker and the URP-7 marker detected polymorphic DNA bands, ranging from 400-1000 bp in the size of amplified DNA fragments.

Genetic Divergence and Relationship among Abalone Species by RAPD Analysis (RAPD 분석을 이용한 전복류의 유전적 차이 및 유연관계)

  • Park, Choul-Ji;Kim, Hyun-Chul;Noh, Jae-Koo;Lee, Jeong-Ho;Myeong, Jeong-In
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.346-350
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    • 2008
  • RAPD analysis was examined to estimate the availability as a genetic marker. The availability was evaluated in terms of genetic divergence and relationships among Haliotis discus hannai, H. rufescens, H. rubra and H. midae in both hemispheres of the world. In results, RAPD analysis showed a clear genetic divergence between every pair of species. However, genetic relationships among the four species estimated by RAPD analysis unreflected to geographical distribution and morphological characteristics. In conclusion, RAPD is suitable genetic markers for estimates of genetic divergence and differences among abalone species.

Genetic Variation in the Selected Populations of Hovenia dulcis var. koreana Nakai. Based on RAPD Analysis

  • Kim Sea-Hyun;Han Jin-Gyu;Chung Hun-Gwan;Cho Yoon-Jin;Park Hyung-Soon
    • Plant Resources
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    • v.8 no.3
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    • pp.293-299
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    • 2005
  • This study used RAPD markers to assume genetic diversity and variation in selected populations of Hovenia dulcis var. koreana. Ratio of polymorphic RAPD markers were 93.4% in selected populations of Hovenia dulcis Thunb., difference of genetic structure among populations and within populations showed 16.45%, 83.55%, respectively in amount of total genetic variation of 4 populations. Total gene diversity($H_T$) that show genetic diversity appeared 0.313 and coefficient of gene differentiation($G_{ST}$) that compare genetic differentiation of populations appeared 0.1645, analysis of AMOVA for variation among populations and within populations was significantly different (P<0.001). Genetic diversity of whole populations showed that 12.44% difference among population and 87.56% difference within populations. As a result, difference within populations was larger than difference among populations in genetic diversity. Nei's genetic distance and cluster analysis appeared that mean genetic distance among populations was 0.076, thus dividing two main groups and geographic relationship did not show in populations.

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