• Title/Summary/Keyword: 분자 마커

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Construction of a Genetic Map using the SSR Markers Derived from "Wonwhang" of Pyrus pyrifolia (배 '원황'(Pyrus pyrifolia) 유전체 해독에 기반한 SSR 마커 개발 및 유전자 지도 작성)

  • Lee, Ji Yun;Seo, Mi-Suk;Won, So Youn;Lim, Kyoung Ah;Shin, Il Sheob;Choi, Dongsu;Kim, Jung Sun
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.434-441
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    • 2018
  • High-density genetic linkage mapping is critical for undertaking marker-assisted selection and confirming quantitative trait loci, as well as helping to build pseudomolecules of genomes. We constructed a genetic map using 94 $F_1$ populations generated from the interspecific cross between Korean cultivar "Wonwhang" (Pyrus pyrifolia, NCBI BioSample SAMN05196235) and European cultivar "Bartlett" (Pyrus communis). We designed a total of 24,267 SSR markers based on the genome sequences of "Wonwhang" for this. To select the markers that are linked to the traits important in pear breeding programs, SSR-containing genomic sequences were subjected to nucleotide sequence homology searches, which resulted in 510 SSR markers with high similarity to genes encoding proteins with putative functions such as transcription factors, resistance proteins, flowering time, and regulatory genes. Of these, 70 markers showed polymorphisms in parents and segregating populations and were used to construct a genetic linkage map, together with the unpublished 579 SNPs obtained from genotyping by sequencing analysis. The genetic linkage map covered 3,784.2 cM and the average distance between adjacent markers was 5.8 cM. Seventy SSR markers were distributed across 17 chromosomes with more than one locus.

Construction of a Microsatellite DNA Profile Database for Pear Cultivars and Germplasm (배 품종 및 유전자원에 대한 Microsatellite DNA 프로파일 데이터베이스 구축)

  • Hong, Jee-Hwa;Shim, Eun-Jo;Kwon, Yong-Sham
    • Horticultural Science & Technology
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    • v.35 no.1
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    • pp.98-107
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    • 2017
  • A DNA profile database was constructed to investigate the genetic relatedness of 72 germplasm samples of Pyrus and related cultivars using microsatellite markers. Three P. pyrifolia, four P. commus, and one P. betulifolia cultivars with different morphological traits were screened using 387 pairs of microsatellite primers. A core set of 11 primer pairs was selected to obtain 133 polymorphic amplified fragments meeting three criteria: high polymorphism information contents (PIC), high repeatability, and distinct allele patterns. The number of alleles per locus ranged between 4 and 22. Average PIC was 0.743 (range: 0.557 - 0.879). Cluster analysis using the unweighted pair - group method with arithmetical average (UPGMA) separated the 72 pear cultivars and germplasm samples into four major groups: Chinese, European pears, and a cluster of 55 Asian pears that could be reclassify into two subcluster, I - $1^{st}$ and II - $2^{nd}$, according to pedigree information. Almost all of the cultivars were discriminated by 11 microsatellite marker genotypes. The microsatellite DNA profile database may be utilized as tool to verify distinctness, uniformity, and stability between candidate cultivar, and to verify in the distinctness of existing cultivars.

Tag-SNP selection and online database construction for haplotype-based marker development in tomato (유전자 단위 haplotype을 대변하는 토마토 Tag-SNP 선발 및 웹 데이터베이스 구축)

  • Jeong, Hye-ri;Lee, Bo-Mi;Lee, Bong-Woo;Oh, Jae-Eun;Lee, Jeong-Hee;Kim, Ji-Eun;Jo, Sung-Hwan
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.218-226
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    • 2020
  • This report describes methods for selecting informative single nucleotide polymorphisms (SNPs), and the development of an online Solanaceae genome database, using 234 tomato resequencing data entries deposited in the NCBI SRA database. The 126 accessions of Solanum lycopersicum, 68 accessions of Solanum lycopersicum var. cerasiforme, and 33 accessions of Solanum pimpinellifolium, which are frequently used for breeding, and some wild-species tomato accessions were included in the analysis. To select tag-SNPs, we identified 29,504,960 SNPs in 234 tomatoes and then separated the SNPs in the genic and intergenic regions according to gene annotation. All tag-SNP were selected from non-synonymous SNPs among the SNPs present in the gene region and, as a result, we obtained tag-SNP from 13,845 genes. When there were no non-synonymous SNPs in the gene, the genes were selected from synonymous SNPs. The total number of tag-SNPs selected was 27,539. To increase the usefulness of the information, a Solanaceae genome database website, TGsol (http://tgsol. seeders.co.kr/), was constructed to allow users to search for detailed information on resources, SNPs, haplotype, and tag-SNPs. The user can search the tag-SNP and flanking sequences for each gene by searching for a gene name or gene position through the genome browser. This website can be used to efficiently search for genes related to traits or to develop molecular markers.

Construction of DNA Profile Data Base of Strawberry Cultivars Using Microsatellite Markers (Microsatellite 마커를 이용한 딸기 품종의 DNA Profile Database 구축)

  • Hong, Jee-Hwa;Choi, Keun-Jin;Kwon, Yong-Sham
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.853-863
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    • 2014
  • This study was carried out to construct a DNA profile database of 100 strawberry cultivars using microsatellite markers. Two hundred seventy four microsatellite primer pairs were screened with a set of 21 strawberry cultivars with different morphological traits. Twenty five primer pairs were selected because they produced reliable and reproducible fingerprints. These primer pairs were used to develop DNA profiles of 100 strawberry cultivars. Three to thirteen alleles were detected by each marker with an average of 7.50. The average polymorphism information content varied from 0.331 to 841 (average 0.706). Cluster analysis showed that the 100 cultivars were divided into 7 major groups reflecting geographic origin and pedigree information. Moreover, most of the cultivars could be discriminated by marker genotypes. These markers will be useful as a tool for the protection of plant breeders' intellectual property rights in addition to providing the means to intervene seed disputes relating to variety authentication.

Use of Microsatellite Markers Derived from Genomic and Expressed Sequence Tag (EST) Data to Identify Commercial Watermelon Cultivars (수박 시판 품종의 식별을 위한 Genomic과 Expressed Sequence Tag (EST)에서 유래된 Microsatellite Marker의 이용)

  • Kwon, Yong-Sham;Hong, Jee-Hwa;Kim, Du-Hyun;Kim, Do-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.737-750
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    • 2015
  • This study was carried out to construct a DNA profile database for 102 watermelon cultivars through the comparison of polymorphism level and genetic relatedness using genomic microsatellite (gMS) and expressed sequence tag (EST)-microsatellite (eMS) markers. Sixteen gMS and 10 eMS primers showed hyper-variability and were able to represent the genetic variation within 102 watermelon cultivars. With gMS markers, an average of 3.63 alleles per marker were detected with a polymorphism information content (PIC) value of 0.479, whereas with eMS markers, the average number of alleles per marker was 2.50 and the PIC value was 0.425, indicating that eMS detects a lower polymorphism level compared to gMS. Cluster analysis and Jaccard's genetic distance coefficients using the unweighted pair group method with arithmetic average (UPGMA) based on the gMS, eMS, and combined data sets showed that 102 commercial watermelon cultivars could be categorized into 6 to 8 major groups corresponding to phenotypic traits. Moreover, this method was sufficient to identify 78 out of 102 cultivars. Correlation analysis with Mantel tests for those clusters using 3 data sets showed high correlation ($r{\geq}0.80$). Therefore, the microsatellite markers used in this study may serve as a useful tool for germplasm evaluation, genetic purity assessment, and fingerprinting of watermelon cultivars.

Divergence Analysis of 16S rRNA and rpoB Gene Sequences Revealed from the Harmful Cyanobacterium Microcystis aeruginosa (유해 남조세균 Microcystis aeruginosa의 16S rRNA 및 rpoB 유전자 염기서열 변이 분석)

  • Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.296-302
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    • 2010
  • Microcystis (Cyanobacteria, Chroococcales) is one of the green tide-causing organisms in freshwaters, and some species produce microcystin that is hepatotoxin. In the aspects of freshwater quality controls and health concerns, therefore it is necessary to manage the harmful organisms. In the present study, RNA polymerase beta subunit (rpoB) gene sequences of Microcystis were determined and characterized in order to use a potential marker for the molecular detections of the species. Microcystis rpoB showed high divergences of DNA similarity and genetic distances when compared with those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.05). Parsimony analyses showed the rpoB gene evolves more than 2-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each M. aeruginosa strain more clearly compared with a 16S rRNA tree. This study found that the order Chroococcales, including Microcystis, has approximately two rRNA operons and single copy of the rpoB gene in their chromosomes. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the detection of Microcystis.

A New Record of Euonymus hamiltonianus Wall. in Dokdo: Using Molecular Markers of Nuclear and Plastid DNA (독도의 미기록 식물 참빗살나무: 핵과 엽록체 DNA의 분자마커 이용)

  • Song, Im-Geun;Park, Seon-Joo
    • Korean Journal of Environmental Biology
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    • v.32 no.1
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    • pp.88-94
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    • 2014
  • The present scientific scenario shows that, the long-distance dispersal of any species plays a major role in population as well as the flow of individual between populations. However, newly recorded plant at Dokdo was confirmed by using morphological and molecular markers of nrDNA and cpDNA. This scientific study has identified the plant species was as a Euonymus hamiltonianus Wall. (Celastraceae). And after careful studies, the laboratory results shows that the distribution of tree species at Dokdo as an Oceanic Island is a meaningful thing because of the inflow of plant in adverse condition of environment such as a shallow soil layers and steep terrain. So it is necessary to continuous monitor for the inflow of non-indigenous plant into Dokdo.

Identification of specific SNP molecular marker from Cudrania tricuspidata using DNA sequences of chloroplast TrnL-F region (구지뽕 나무의 엽록체 TrnL-F 영역 염기서열 분석을 통한 특이적 SNP 분자마커의 확인)

  • Lee, Soo Jin;Shin, Yong-Wook;Kim, Yun-Hee;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.135-141
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    • 2017
  • Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. For conservation and germplasm utilization of the plant, it is imperative to obtaining information regarding the genetic diversity of the plant populations. Although C. tricuspidata is an important medicinal plant registered in South Korea, no molecular markers are currently available to distinguish Korean-specific ecotypes from other ecotypes of different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via the amplification refractory mutation system (ARMS)-PCR analyses. Molecular authentication of twelve C. tricuspidata ecotypes from different regions was performed, using DNA sequences in the trnL-F chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific C. tricuspidata ecotypes from different regions.

Development of specific SNP molecular marker from Thistle using DNA sequences of ITS region (엉겅퀴의 ITS 영역 염기서열 분석을 통한 특이적 SNP 분자마커의 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.102-109
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    • 2018
  • Thistle is a perennial plant that is widely used for medicinal purposes. Information on the genetic diversity of thistle populations are great important for their conservation and germ plasmic utilization. Although thistle is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish them from other similar species from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from the nuclear ribosomal DNA internal transcribed spacer (ITS) regions of genomic sequences to identify distinct Korean-specific thistle species via an amplification refractory mutation system (ARMS)-PCR and high resolution melting (HRM) curve analyses. We performed molecular authentication of four different kinds of thistle species from different regions using DNA sequences in the ITS intergenic region. We also developed a quantitative PCR assay using species-specific ITS primers, which allowed us to estimate the ratio of Korean-specific thistle species using varying ratios of mixed genomic DNA templates from the two species. The SNP markers developed in this study are useful for rapidly identifying specific thistle species from different countries.