• Title/Summary/Keyword: Polymorphic markers

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Genetic Relationships of Four Korean Oysters Based on RAPD and Nuclear rDNA ITS Sequence Analyses

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee-Jeong;Jung, Hyung-Taek
    • The Korean Journal of Malacology
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    • v.25 no.1
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    • pp.41-49
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    • 2009
  • Random amplified polymorphic DNA (RAPD) marker and sequence analyses of the internal transcribed spacer (ITS) region of ribosomal DNA were used to assess phylogenetic relationships of four Korean oyster species. The average number of species-specific markers identified from five universal rice primers (URPs) by RAPD-PCR was 1.8 for Crassostrea gigas, 3.2 for C. nippona, 3.6 for C. ariakensis, and 4.6 for Ostrea denselamellosa. The length of the ITS (ITS1-5.8S-ITS2) region ranged from 1,001 to 1,206 bp (ITS1, 426-518 bp; 5.8S, 157 bp; and ITS2, 418-536 bp), while the GC content ranged from 55.5-61.1% (ITS1, 56.8-61.8%; 5.8S, 56-57.3%; and ITS2, 54.1-62.2%). A phylogenetic analysis of the oysters based on our RAPD, ITS1, and ITS2 sequence data revealed a close relationship between C. gigas and C. nippona and a distant relationship between the genera Crassostrea and Ostrea. Our results indicated that RAPD and ITS sequence analysis was a useful tool for the elucidation of phylogenetic relationships and for the selection of species-specific markers in Korean oysters.

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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.

Genetic diversity analysis of Glycyrrhiza uralensis using 8 novel polymorphic microsatellite markers

  • Um, Yurry;Jin, Mei-Lan;Lee, Yi;Hur, Mok;Cha, Seon Woo;Jung, Chan Sik;Kim, Seong Min;Lee, Jeong-Hoon
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.174-180
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    • 2016
  • Licorice plant (Glycyrrhiza spp.) is an important herb, but the major portion of the national demand is imported to Korea because the domestic production base is vulnerable. We performed basic molecular breeding research for domestic cultivation and production. All publicly available G. uralensis EST sequences, which totaled 56,089, were assembled into 4,821 unigenes and examined for microsatellites. Eight polymorphic microsatellite loci were identified and 16 G. uralensis and 6 G. glabra accessions, which were collected from different locations, were genotyped using the microsatellites. Genetic diversity within the accessions was estimated by construction of a dendrogram. The dendrogram was clustered into two groups. The results showed that there is a correlative genetic relationship between species. The microsatellite markers were found to be useful for diversity analysis as they are able to successfully distinguish the Glycyrrhiza accessions.

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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Genotyping of Agaricus bisporus Strains by PCR Fingerprints

  • Min, KyongJin;Oh, YounLee;Kang, HeeWan
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.41-41
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    • 2014
  • Agaricus bisporus, commonly known as the button mushroom, is the most widely cultivated species of edible fungi. Low frequency of recombination ratio and homokaryotic or monokaryotic spore on meiotic basidia form obstacles for breeding programs. Since the first hybrid varieties for white button mushrooms were released in Europe, new varieties released afterwards were either identical of very similar to these first hybrids on morphologies. Therefore, different DNA markers have been used to define unique varieties of A. bisporus strains. Aim of this study is to assess the genetic diversity of different A. bisporus strains in Korea. Twelve UFP (Universal fungal primer, JK BioTech. Ltd), 12 simple sequence repeat (ISSR) and 30 SSR primers were used to assess genetic diversity of monokaryotic and dikaryotic Agaricus bisporus strains including other 19 Agaricus spp. Of them, four UFP, four SSR primers, $(GA)_8T$, $(AG)_8YC$, $(GA)_8C$ and $(CTC)_6$ and seven SSR markers produced PCR polymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted for UPGMA cluster analysis. Forty five strains of A. bisporus are genetically clustered into 6 groups, showing coefficient similarity from 0.75 to 0.9 among them. In addition, genetic variations of monokaryotic and dikaryotic Agaricus bisporus strains were partially detected by PCR technologies of this study. The varieties, Saea, saedo, Saejeong and Saeyeon that have recently been developed in Korea were involved in the same group with closely genetic relationship of coefficient similarity over 0.96, whereas, other strains were genetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese.

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Identification and Genetic Diversity of Korean Tomato Cultivars by RAPD Markers (한국 내 토마토 재재종의 RAPD에 의한 동정과 유전적 다양성)

  • Huh, Man-Kyu;Youn, Sun-Joo;Kang, Sun-Chul
    • Journal of Life Science
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    • v.21 no.1
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    • pp.15-21
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    • 2011
  • Cultivated tomato, Lycopersicum esculentum, is a very important crop. We selected 36 cultivars and studied them for identification and polymorphism by employing random amplified DNA (RAPD) analysis with 80 oligonucleotide primers. Of the 80 primers, 36 primers (45.0%) were polymorphic. Detection of polymorphism in cultivated tomato opens up the possibility of development of its molecular map by judicious selection of genotypes. Molecular markers can also be used for cultivar identification and protection of the plant breeder's intellectual property rights (plant breeders' rights, PBRs). As an example, DNA polymorphism using OPC-13 primer that did not produce the OPC-13-01 band was only found in Junk Pink and Ailsa Craighp cultivars. OPA-12-03 and OPB-15-07 were fragments specific to the TK-70 cultivar and were absent in other cultivars. DNA polymorphism in cultivated tomato in this study was correlated with a type of inflorescence, although some cultivars had exceptions. These approaches will be useful for developing marker-assisted selection tools for genetic enhancement of the tomato plant for desirable traits.

Molecular Authentication of Scrophularia herbs by PCR-RFLP Based on rpl-5 Region of Mitochondrial DNA (현삼속 식물의 종판별을 위한 Mitochondrial DNA의 염기서열 및 PCR-RFLP 분석)

  • Lee, Jeong-Hoon;Jo, Ick-Hyun;Lee, Jei-Wan;Park, Chun-Geun;Bang, Kyong-Hwan;Kim, Hong-Sig;Park, Chung-Berm
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.173-179
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    • 2010
  • This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.

Development and characterization of nine microsatellite loci from the Korean hare (Lepus coreanus) and genetic diversity in South Korea

  • Kim, Sang-In;An, Jung-Hwa;Choi, Sung-Kyoung;Lee, Yun-Sun;Park, Han-Chan;Kimura, Junpei;Kim, Kyung-Seok;Min, Mi-Sook;Lee, Hang
    • Animal cells and systems
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    • v.16 no.3
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    • pp.230-236
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    • 2012
  • The Korean hare, Lepus coreanus, is an important mammal in ecosystem food chains, and is distributed across the entire Korean peninsula and northeastern China. Polymorphic microsatellite loci were developed using the biotinenrichment technique for use in population genetics studies. Five trinucleotide and four dinucleotide microsatellite loci were selected and tested on 22 Korean hare specimens collected from Gangwon Province and Gyeongsangbuk Province in South Korea. The number of alleles across the two sampling regions ranged from three to nine with a mean of 6.1. Mean observed and expected heterozygosities and polymorphic information content were 0.540, 0.627 and 0.579, respectively. Only one locus, Lc06, showed departure from Hardy-Weinberg equilibrium after applying the Bonferroni correction. Four microsatellites, Lc01, Lc03, Lc12, and Lc19, satisfied the criteria to serve as a core set of markers recommended for population genetics studies. These new microsatellite markers will be widely applicable to future genetic studies for management and conservation of the Korean hare and related species, including assessment of the genetic diversity and population structure of L. coreanus.

Linkage Analysis of both RAPD and I-SSR Markers using Haploid Genome from a Single Tree of Pinus densiflora S. et Z. (소나무 단일(單一) 모수(母樹)의 반수체(半數體) 게놈을 이용(利用)한 RAPD 및 I-SSR 표식자(標識子)의 연관분석(連關分析))

  • Hong, Yong-Pyo;Chung, Jae-Min;Kim, Yong-Yul;Jang, Suk-Seong
    • Journal of Korean Society of Forest Science
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    • v.89 no.4
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    • pp.536-542
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    • 2000
  • A linkage map for Japanese red pine (Pinus densiflora) was constructed on the basis of two DNA marker systems of random amplified polymorphic DNAs (RAPDs) and inter-simple sequence repeats (I-SSR). Haploid genomic DNAs were extracted from megagametophyte tissues of 96 individual seeds in a single tree. A total of 98 DNA markers including 52 RAPD markers amplified by 25 primers and 46 I-SSR markers amplified by 18 primers were verified as Mendelian loci showing 1 : 1 segregation in 96 megagametophytes which were ${\chi}^2$-tested at 5% significance level. Of them, 63 segregating loci turned out to be linked into 20 linkage groups by the two-point analysis. However, 35 loci (17 RAPD and 18 I-SSR) of the 98 segregating loci did not coalesced into any linkage groups at a LOD of 3.0. The linked 63 loci were separated by an average distance of about 25.5 cM, which were spanned 1097.8 cM as a whole. The minimum and maximum map distances of the linkage groups were 4.3 cM and 54.9 cM, respectively. Incorporation of I-SSR loi into linkage map of RAPD loci resulted in extended and partially more saturated linkage blocks.

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Development and Application of PCR-based Markers for the Discrimination of Bang-Poong and Related Species (방풍류의 감별을 위한 분자마커의 탐색과 활용)

  • Hong, Seong-Mi;Lee, Mi-Young;Koh, Jae-Chul;Ko, Byoung-Soeb
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.1-6
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    • 2004
  • Bang-Poong and related species are an important herbal medicine. However, it is difficult to determine the commercial dry material through anatomical and chemotaxonomical characteristics. Here, we used a PCR-based technique for an accurate discrimination of Bang-Poong and related species. With the RAPD primers, 215 RAPDSs(random amplified polymorphic DNAs) were obtained, and 98% of them showed polymorphic patterns. RAPDs from the four primers were appropriate for the discrimination of S. divaricata $(T_{URCZ{\cdot}})\;S_{CHISKIN}$, those from the six primers for P. japonicum $T_{HUNBERG}$, those from the four primers for P. terebinthaceum $F_{ISHER}$, and those from the six primers for G. littoralis Fr. $S_{CHMIDT}$. The specific bands from the primer 425 were obtained and used to develop SCAR (sequence characterized amplified region) markers, based on the sequence information of the RAPD markers. The SCAR primers generated a 215 bp fragment specific to Peucedanum terebinthaceum $F_{ISHER}$, and a 177 bp and a 300 bp fragment specific to G. littoralis Fr. $S_{CHMIDT}$. As a result, the three SCAR markers were able to discriminate from two Bang-Poong related species.