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한국 토종오리의 개체 식별 및 품종 구분을 위한 Microsatellite 마커 탐색 (Investigation of Microsatellite Markers for Traceability and Individual Discrimination of Korean Native Ducks)

  • 서동원;술타나;최누리;김연수;진실;허강녕;진선덕;이준헌
    • 한국가금학회지
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    • 제42권1호
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    • pp.1-8
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    • 2015
  • 최근, 한국의 소비자들이 건강에 대한 관심이 증가하면서 단일불포화 지방산이 풍부해 건강에 긍정적인 영향을 줄 수 있는 오리고기의 수요가 급격하게 증가하고 있다. 하지만 대부분의 종 오리는 수입에 의존하고 있는 실정이기 때문에, 토종오리의 개발 및 보급이 필요한 실정이며, 이는 종자주권의 확립 및 농가 소득 증대에도 매우 필요한 일이라 할 수 있다. 따라서 본 연구에서는 24개의 microsatellite 마커를 확보하였으며, 이들 마커의 대립유전자수는 1~16개, 이형접합도는 0~0.865, 다형성은 0~0.841로 확인되었다. 이들 마커를 이용하여 임의 집단에서 동일개체 출현빈도를 계산한 결과는 임의 집단 $1.64{\times}10^{-16}$, 전형매 집단 $2.60{\times}10^{-7}$, 반형매 집단 $1.30{\times}10^{-12}$으로 높은 개체식별률과 친자확인도를 확인할 수 있었다. 하지만 이들 마커를 이용한 계통분석 결과, 토종오리와 실용오리 집단을 정확하게 구분하기에는 어려운 것으로 확인되었다. 따라서 추가연구를 통해 토종오리의 순종화 및 더 정확한 토종오리와 실용오리 집단 구분이 가능한 마커 개발이 필요할 것으로 사료된다.

Genetic diversity of Halla horses using microsatellite markers

  • Seo, Joo-Hee;Park, Kyung-Do;Lee, Hak-Kyo;Kong, Hong-Sik
    • Journal of Animal Science and Technology
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    • 제58권11호
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    • pp.40.1-40.5
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    • 2016
  • Background: Currently about 26,000 horses are breeding in Korea and 57.2% (14,776 horses) of them are breeding in Jeju island. According to the statistics published in 2010, the horses breeding in Jeju island are subdivided into Jeju horse (6.1%), Thoroughbred (18.8%) and Halla horse (75.1%). Halla horses are defined as a crossbreed between Jeju and Thoroughbred horses and are used for horse racing, horse riding and horse meat production. However, little research has been conducted on Halla horses because of the perception of crossbreed and people's weighted interest toward Jeju horses. Method: Using 17 Microsatellite (MS) Markers recommended by International Society for Animal Genetics (ISAG), genomic DNAs were extracted from the hair roots of 3,880 Halla horses breeding in Korea and genetic diversity was identified by genotyping after PCR was performed. Results and conclusion: In average, 10.41 alleles (from 6 alleles in HTG7 to 17 alleles in ASB17) were identified after the analysis using 17 MS Markers. The mean value of $H_{obs}$ was 0.749 with a range from 0.612(HMS1) to 0. 857(ASB2). Also, it was found that $H_{\exp}$ and PIC values were lowest in HMS1 (0.607 and 0.548, respectively), and highest in LEX3(0.859 and 0.843, respectively), and the mean value of $H_{\exp}$ was 0.760 and that of PIC was 0.728. 17 MS markers used in this studies were considered as appropriate markers for the polymorphism analysis of Halla horses. The frequency for the appearance of identical individuals was $5.90{\times}10^{-20}$ when assumed as random mating population and when assumed as half-sib and full-sib population, frequencies were $4.08{\times}10^{-15}$ and $3.56{\times}10^{-8}$, respectively. Based on these results, the 17 MS markers can be used adequately for the Individual Identification and Parentage Verification of Halla horses. Remarkably, allele M and Q of ASB23 marker, G of HMS2 marker, H and L of HTG6 marker, L of HTG7 marker, E of LEX3 marker were the specific alleles unique to Halla horses.

RAPD마커를 이용한 황기의 유전적 다양성 및 기원판별 (Genetic Diversity and Discrimination of Astragalus Membranaceus Bunge and A. Membranaceus var. Mogholicus Using RAPD Markers)

  • 방경환;허만규;조준형
    • 동의생리병리학회지
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    • 제18권3호
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    • pp.825-829
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    • 2004
  • This study was carried out to differentiate the origins of Astragalus membranaceus Bunge and A. membranaceus Bunge var. mogholicus Nakai. To identify the variation of the RAPD patterns between domestic and foreign Astragalus species, 40 random primers were applied to ten accessions of A. membranaceus and six accessions of A. membranaceus var. mogholicus genomic DNA, respectively, Ten primers of 40 primers could be used to discriminate the origins and 33 polymorph isms among 44 scored DNA fragments (33 fragments are specific for A. membranaceus and A. membranaceus var. mogholicus) were generated using these primers, 75.0 % of which were polymorphic. Especially, three primers of ten primers, OPA17, OPA11 and OPB11, were useful to differentiate between domestic and foreign Astragalus species. RAPD data from the 10 primers were used for cluster analysis and cluster analysis of RAPD markers showed that the two groups are distinct genetically. Consequently, RAPD analysis was a useful method to discriminate between A. membranaceus and A. membranaceus var. mogholicus.

Evaluation of Genetic Differentiation of Albizia lucida Populations from Eastern Region of the Indian Sub-continent by ISSR Markers

  • Aparajita, Subhashree;Rout, G.R.
    • Journal of Forest and Environmental Science
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    • 제24권1호
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    • pp.27-34
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    • 2008
  • Level and distribution of genetic diversity in seven populations of Albizia lucida Benth. in eastern region of the Indian sub-continent were estimated using ISSR markers. Relatively higher level of genetic diversity within populations was observed in seven populations of A. lucida (mean of 0.38). From the result of AMOVA, majority of genetic diversity was allocated within populations (96.2%) resulting in a moderate degree of population differentiation. The observed distribution pattern of I-SSR variant among the populations was coincided with the typical pattern of long-lived woody tree species. Genetic relationships among the populations, reconstructed by UPGMA method, revealed two genetic groups. The population of Anugul and Bargarh turned out to be the most closely related despite a distance location between them. These formations will be of great value in the development of conservation plans for species exhibiting high levels of genetic differentiation due to fragmentation, such as indication of conservation unit size, which populations should be chosen as priority in conservation plans and which samples should be introduced in areas with a low number of individuals of A. lucida.

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Seed Purity Test and Genetic Diversity Evaluation Using RAPD Markers in Radish (Raphanus sativus L.)

  • Huh, Man-Kyu;Choi, Joo-Soo
    • 한국작물학회지
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    • 제54권4호
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    • pp.346-350
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    • 2009
  • The cultivated radish (Raphanus sativus L.) is a major vegetable crop in the world wide and fast-growing species that grows inhabitats of six continents. It is very important to determine hybrid seed purity in the production of hybrid Brassica vegetable seeds to avoid unacceptable contamination with self-inbred (sib) seeds. The use of random amplified polymorphic DNA (RAPD) markers for evaluating seed purity in $F_2$-hybrid radish cultivars demonstrated. One hundred eighty seeds from the F1 male and female harvest were subsequently screened for seed purity using 13 primers. The 13 primers result in 17 cultivar-specific bands and 23 variable RAPD bands scored for cultivar. RAPD analysis of hybrid seeds from the harvest revealed 128 seeds tested except underdevelopment and decayed seeds were sibs. Especially, $F_2$ hybrids of radish, OPC13, OPD20 were presented clear hybrid bands. It maintains higher than average level of genetic diversity compared with their correspondent parents. RAPD amplification of DNA extracted from germinated individuals from the female harvest reveal that 10 of 208 seeds tested were self-inbred (4.8%). RAPD analysis of hybrid seeds from the male harvest revealed 7 of the 208 seeds tested were sibs (3.4%). The RAPD may lead to a better insight in to the hybrid seed purity.

Miniature 말의 성(sex) 결정과 친자감정 (Sex Determination and Parentage Testing In Miniature Horses)

  • 조길재;조병욱
    • 생명과학회지
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    • 제15권1호
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    • pp.45-48
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    • 2005
  • 서울대공원에서 사육중인 Miniature horse 10두의 혈통정립을 목적으로 PCR에 의한 성별 판정 및 16개 microsatellite marker를 사용하여 친자감정을 실시하였다. 성별 판정에서 430 bp의 SRY band가 관찰된 7두는 숫말로 판정되었고, 친자감정에서는 멘델의 유전양식에 부합된 3두가 친자관계가 확인되었다. 향후 Miniature horse의 혈통보존 및 관리에 유용한 자료가 될 것으로 사료된다.

RAPD 방법을 이용한 한국 야생쑥 6종간의 유전적 유연관계 분석 (Analysis of Genetic Polymorphism Among Six Korean Wild Artemisia spp. by Using RAPD Method)

  • 표현진;최관삼
    • 농업과학연구
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    • 제23권1호
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    • pp.99-107
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    • 1996
  • Eighteen nuclear probes were used to examine RFLP(restriction fragment length polymorphism) between six species of Artemisia spp. of Korea. Total DNA from six different species of Artemisia was separately cut with three restrict enzymes. The PstI enzyme was showed to reduce the variation of polymorphisms than the other two enzymes(EcoRl and BamHI). The genetic variation of polymorphism was similar between the Dhewegiki-ssug and Cham-ssug. RAPD analysis was applied to the same six species of Artemisia spp. in order to assess the degree of DNA polymorphism within the Artemisia genus. Six species of Artemisia were evaluated for variation using a set of 11 random 10-mer primers. Nine out of the eleven primers revealed scorable polymorphisms between six species of Artemisia spp. Genetic distances between each of the species were calculated and cluster analysis was used to generate a dendrogram showing phylogenetic relationships between them This result indicates that molecular markers will be more usable in intraspecific study of Artemisia spp. than isoenzyme markers.

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Genetic Analysis of Three River Populations of Catla catla (HAMILTON) Using Randomly Amplified Polymorphic DNA Markers

  • Islam, M.S.;Ahmed, A.S.I.;Azam, M.S.;Alam, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권4호
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    • pp.453-457
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    • 2005
  • The genetic variations in three major river populations viz. the Halda, the Jamuna and the Padma of the Indian major carp, Catla catla were analyzed by Random Amplified Polymorphic DNA (RAPD) markers. Four decamer primers were used for amplifying DNA of 10 individuals from each population. The proportion of polymorphic loci and the gene diversity estimates were 59.4 and 0.20 for the Halda, 37.5 and 0.14 for the Jamuna and 46.9 and 0.16 for the Padma populations respectively indicating the existence of a relatively high level of genetic variation in the Halda river population. The inter-population similarity indices, gene flow and genetic distance values indicated that the Jamuna-Padma population pair of catla was genetically closer than the Halda-Jamuna and the Halda-Padma population pairs in compliance with the geographical distances among them. The coefficient of gene differentiation ($G_{ST}$=0.13) reflects some degree of genetic differentiation among three populations of catla studied. The data suggest that the RAPD technique could be used to discriminate different river populations of catla.

SoEM: a novel PCR-free biodiversity assessment method based on small-organelles enriched metagenomics

  • Jo, Jihoon;Lee, Hyun-Gwan;Kim, Kwang Young;Park, Chungoo
    • ALGAE
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    • 제34권1호
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    • pp.57-70
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    • 2019
  • DNA metabarcoding is currently used for large-scale taxonomic identification to understand the community composition in various marine ecosystems. However, before being widely used in this emerging field, this experimental and analytic approach still has several technical challenges to overcome, such as polymerase chain reaction (PCR) bias, and lack of well-established metabarcoding markers, a task which is difficult but not impossible to achieve. In this study, we present an adapted PCR-free small-organelles enriched metagenomics (SoEM) method for marine biodiversity assessment. To avoid PCR bias and random artefacts, we extracted target DNA sequences without PCR amplification from marine environmental samples enriched with small organelles including mitochondria and plastids because their genome sequences provide a valuable source of molecular markers for phylogenetic analysis. To experimentally enrich small organelles, we performed subcellular fractionation using modified differential centrifugation for marine environmental DNA samples. To validate our SoEM method, two marine environmental samples from the coastal waters were tested the taxonomic capturing capacity against that of traditional DNA metabarcoding method. Results showed that, regardless of taxonomic levels, at least 3-fold greater numbers of taxa were identified in our SoEM method, compared to those identified by the conventional multi-locus DNA metabarcoding method. The SoEM method is thus effective and accurate for identifying taxonomic diversity and presents a useful alternative approach for evaluating biodiversity in the marine environment.

Intraspecific genetic variation in Corynandra chelidonii (Angiosperms: Cleomaceae) as revealed by SCoT, ISSR and RAPD analyses

  • Sirangi, Subash;Jogam, Phanikanth;Nemali, Gandhi;Ajmeera, Ragan;Abbagani, Sadanandam;Raju, Vatsavaya S.
    • Journal of Plant Biotechnology
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    • 제47권4호
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    • pp.289-297
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    • 2020
  • The genetic diversity of two subpopulations of Corynandra chelidonii, one of terrestrial and the other of aquatic environments, was measured with molecular markers, such as start codon targeted (SCoT), inter simple sequence repeats (ISSR), and random amplification of polymorphic DNA (RAPD). The traditional morphological traits such as habitat, habit, leaf morphology, the colour of the sepals and petals, number of stamens, and seed morphology formed the base for their realization as two varieties, C. chelidonii var. pallae and C. chelidonii var. chelidonii. The polymorphism between the two variants was 100% with the primers SCoT-2 and OPA-1 and 4, while maximum polymorphism was detected with ISSR-2, SCoT-3, and OPA-3. The study used, for the first time, more than one molecular marker to assess the genetic variation underscoring the morphological variation in Corynandra chelidonii (L.f.) Cochrane & Iltis. The study justifies the recognition of the two subpopulations of Corynandra chelidonii from aquatic and terrestrial environments as two distinct varieties, C. chelidonii var. pallae (Reddy & Raju) V.S.Raju and C. chelidonii var. chelidonii, respectively, based on the traditional taxonomic evidence.