• Title/Summary/Keyword: SNP markers

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Molecular markers based on chloroplast and nuclear ribosomal DNA regions which distinguish Korean-specific ecotypes of the medicinal plant Cudrania tricuspidata Bureau

  • Lee, Soo Jin;Shin, Yong-Wook;Kim, Yun-Hee;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.235-242
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    • 2017
  • Cudrania tricuspidata Bureau is a widely-used, medicinal, perennial and woody plant. Obtaining information about the genetic diversity of plant populations is highly important with regard toconservation and germplasm utilization. Although C. tricuspidata is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish Korean-specific ecotypes from other ecotypes from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from the chloroplast and nuclear genomic sequences, which serve to to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR and high resolution melting (HRM) curve analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the maturaseK (MatK) chloroplast intergenic region and nuclear ribosomal DNA internal transcribed spacer (ITS) regions. The SNP markers developed in this study are useful for rapidly identifying specific C. tricuspidata ecotypes from different regions.

Beagle dogs parentage testing by using 22 ISAG microsatellite markers

  • Ji, Hye-jung;Kim, Eun-hee;Lee, Kyoung-kap;Kang, Tae-young;Lee, Joo-myoung;Shin, Hyoung-doo;Kim, Lyoung-hyo;Yun, Young-min
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.457-460
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    • 2007
  • The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

Screening of Rice Blast Resistance Genes from Aromatic Rice Germplasms with SNP Markers

  • Kim, Jeong-Soon;Ahn, Sang-Nag;Kim, Chung-Kon;Shim, Chang-Ki
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.70-79
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    • 2010
  • Rice blast is one of the serious devastating diseases. This study was carried out to determine the genetic diversities of blast resistance (R) genes form 86 accessions of aromatic rice with eight SNP markers, z4792, zt4792, z60510, zt6057, k6415, k6411, k39575 and t256, which showed the close-set linkage to 6 major genes, Piz, Piz-t, Pik, Pik-m, Pik-p, and Pit. Four accessions of indica type, Mayataung, Yekywin Yinkya Hmwe, Basmati9-93, and Basmati5854, showed the positive amplicons of six major genes. Among 86 accessions, 83 accessions were detected both or one of Piz and Piz-t genes. Seventy three accessions contained the Piz gene with z4792 marker. In addition, 30 and 71 accessions possessed Piz-t gene with zt4792 and zt6057 markers, respectively. Ten accessions showed the positive bands for the Piz-t gene with both zt4792 and zt6057 markers. Only one accession, Khau Nua Keo, was not amplified for both Piz and Piz-t gene. But japonica type, Gerdeh, possessed only Piz gene between Piz and Piz-t. Fifty two accessions showed the three of Pik multiple genes and Pit gene. Four accessions, Iari7447, Daebunhyangdo2, Shiyayuuine, and Basmati 6129 possessed a Pik-p gene. Especially, Pit gene on chromosome 1 was detected with t256 marker in all of 83 accessions, exception of A-2, one accession of japonica type.

Screening of the Dominant Rice Blast Resistance Genes with PCR-based SNP and CAPS Marker in Aromatic Rice Germplasm

  • Kim, Jeong-Soon;Ahn, Sang-Nag;Hong, Sung-Jun;Kwon, Jin-Hyeuk;Kim, Yeong-Ki;Jee, Hyeong-Jin;Shim, Chang-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.4
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    • pp.329-341
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    • 2011
  • The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and closest linkage to seven major rice blast resistance (R) genes, Pia, Pib, Pii, Pi5 (Pi3), Pita (Pita-2), and Pi9 (t). The allele specific PCR markers assay genotype of SCAR and STS markers was applied to estimate the presence or absence of PCR amplicons detected with a pair of PCR markers. One indica accession, Basmati (IT211194), showed the positive amplicons of five major rice blast resistance genes, Pia, Pi5 (Pi3), Pib, Pi-ta (Pi-ta2), and Pik-5 (Pish). Among 48 accessions of the PCR amplicons detected with yca72 marker, only five accessions were identified to Pia gene on chromosome 11. The Pib gene was estimated with the NSb marker and was detected in 65 of 84 accessions. This study showed that nine of 84 accessions contained the Pii gene and owned Pi5 (Pi3) in 42 of 84 accessions by JJ817 and JJ113-T markers, which is coclosest with Pii on chromosome 9. Only six accessions were detected two alleles of the Pita or Pita-2 genes. Three of accessions were identified as the Pi9 (t) gene locus.

Development of HRM Markers Based on Identification of SNPs from Next-Generation Sequencing of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link (오이풀, 흰오이풀, 긴오이풀의 NGS 기반 유전체 서열의 완전 해독 및 차세대 염기서열 재분석으로 탐색된 SNP 기반 HRM 분자표지 개발)

  • Sim, Mi-Ok;Jang, Ji Hun;Jung, Ho-Kyung;Hwang, Taeyeon;Kim, Sunyoung;Cho, Hyun-Woo
    • The Korea Journal of Herbology
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    • v.34 no.6
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    • pp.91-97
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    • 2019
  • Objective : To establish a reliable tool between for the distinction of original plants of Sanguisorbae Radix, we analyzed the complete chloroplast genome sequence of Sanguisorbae Radix and identified single nucleotide polymorphisms (SNPs). Materials and methods : The chloroplast genome sequence of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link obtained using next-generation sequencing technology were described and compared with those of other species to develop specific markers. Candidate genetic markers were identified to distinguish species from the chloroplast sequences of each species using Modified Phred Phrap Consed and CLC Genomics Workbench programs. Results : The structure of the chloroplast genome of each sample that had been assembled and verified was circular, and the length was about 155 kbp. Through comparative analysis of the chloroplast sequences, we found 220 nucleotides, 158 SNPs, and 62 Indel (insertion and/or deletion), to distinguish Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link. Finally, 15 specific SNP genetic markers were selected for the verification at positions. Avaliable primers for the dried herb, which is used as medicine, were used to develop the PCR amplification product of Sanguisorbae Radix to assess the applicability of PCR analysis. Conclusion : In this study, we found that Fendel-qPCR analysis based on the chloroplast DNA sequences can be an efficient tool for discrimination of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link.

Novel SNP in the coding region of the FTO gene is associated with marbling score in Hanwoo (Korean cattle)

  • Chung, Eui-Ryong
    • Journal of Animal Science and Technology
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    • v.56 no.8
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    • pp.27.1-27.6
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    • 2014
  • The fat mass and obesity associated (FTO) gene plays an important role in the regulation of energy homeostasis, fat deposition and obesity. For this reason, the FTO gene is a physiological and functional candidate gene for carcass and meat quality traits in beef cattle. The objectives of this study were to identify SNPs in the exonic regions of FTO gene and to evaluate the association of these SNPs with carcass traits in Hanwoo (Korean cattle). In this study, we newly identified two exonic SNPs in Hanwoo population. The g.125550A > T SNP was located in exon 3 and the g.175675C > T SNP was located in exon 6. Genotyping of the two SNP markers was carried out using PCR-RFLP analysis in Hanwoo steers to evaluate their association with carcass traits. As a result, g.125550A > T SNP genotype was significantly associated with effects on marbling score. Animals with the AA and TT homozygous genotypes had a significantly higher marbling score (p < 0.001) than those with AT heterozygous genotype, and this was significant after Bonferroni correction of the significance threshold (p = 0.003). Dominance effect was also observed for the marbling score (P < 0.05) with higher marbling score of homozygous animals. However, no significant associations with meat quality traits were observed for the g.175675C > T SNP. Our results suggest that the exonic SNP g.125550A > T in the FTO gene may be used as a DNA marker for the selection of Hanwoo with higher marbling.

A Whole Genome Association Study to Detect Single Nucleotide Polymorphisms for Carcass Traits in Hanwoo Populations

  • Lee, Y.-M.;Han, C.-M.;Li, Yi;Lee, J.-J.;Kim, L.H.;Kim, J.-H.;Kim, D.-I.;Lee, S.-S.;Park, B.-L.;Shin, H.-D.;Kim, K.-S.;Kim, N.-S.;Kim, Jong-Joo
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.417-424
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    • 2010
  • The purpose of this study was to detect significant SNPs for carcass quality traits using DNA chips of high SNP density in Hanwoo populations. Carcass data of two hundred and eighty nine steers sired by 30 Korean proven sires were collected from two regions; the Hanwoo Improvement Center of National Agricultural Cooperative Federation in Seosan, Chungnam province and the commercial farms in Gyeongbuk province. The steers in Seosan were born between spring and fall of 2006 and those in Gyeonbuk between falls of 2004 and 2005. The former steers were slaughtered at approximately 24 months, while the latter steers were fed six months longer before slaughter. Among the 55,074 SNPs in the Illumina bovine 50K chip, a total of 32,756 available SNPs were selected for whole genome association study. After adjusting for the effects of sire, region and slaughter age, phenotypes were regressed on each SNP using a simple linear regression model. For the significance threshold, 0.1% point-wise p value from F distribution was used for each SNP test. Among the significant SNPs for a trait, the best set of SNP markers were selected using a stepwise regression procedure, and inclusion and exclusion of each SNP out of the model was determined at the p<0.001 level. A total of 118 SNPs were detected; 15, 20, 22, 28, 20, and 13 SNPs for final weight before slaughter, carcass weight, backfat thickness, weight index, longissimus dorsi muscle area, and marbling score, respectively. Among the significant SNPs, the best set of 44 SNPs was determined by stepwise regression procedures with 7, 9, 6, 9, 7, and 6 SNPs for the respective traits. Each set of SNPs per trait explained 20-40% of phenotypic variance. The number of detected SNPs per trait was not great in whole genome association tests, suggesting additional phenotype and genotype data are required to get more power to detect the trait-related SNPs with high accuracy for estimation of the SNP effect. These SNP markers could be applied to commercial Hanwoo populations via marker-assisted selection to verify the SNP effects and to improve genetic potentials in successive generations of the Hanwoo populations.

Development of specific single nucleotide polymorphism molecular markers for Angelica gigas Nakai (ITS 영역의 HRM 분석을 통한 참당귀(Angelica gigas Nakai)의 특이적 SNP 분자표지 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Han, Eun-Hee;Shin, Yong-Wook;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.2
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    • pp.71-76
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    • 2021
  • Angelica is a perennial plant used widely for medicinal purposes. Information on the genetic diversity of Angelica populations is important for their conservation and germplasm utilization. Although Angelica is an important medicinal plant genus registered in South Korea, no molecular markers are currently available to distinguish individual species from other similar species in different countries, in particular, China and Japan. In this study, we developed single nucleotide polymorphism (SNP) markers derived from internal transcribed spacer regions of the nuclear ribosomal DNA to identify a distinct domestic species, Angelica gigas Nakai, via a high-resolution melting (HRM) curve analyses. We also performed HRM curve analysis of intentionally mixed genomic DNA samples from five Angelica species. Finally, we investigated A. gigas Nakai and A. sinensis using varying ratios of mixed genomic DNA templates. The SNP markers developed in this study are useful for rapidly identifying A. gigas species from different countries.

A Study on KASP Analysis Based on SNP to Rapidly Identify Caviar-Producing Sturgeon Species (캐비어를 생산하는 철갑상어의 신속 종판별을 위한 SNP 기반 KASP 분석에 관한 연구)

  • Sun Hee Lee;Bo Reum Park;Hyung Il Kim;Sooyeul Cho;Kyung-Hun Son
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.209-220
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    • 2024
  • Cavier is graded as Beluga, Osetra, and Sevruga based on the species of sturgeon (Acipenser sinensis). In this study, we developed an analytical method for determining the grade of black caviar using DNA barcodes and KASP markers. To identify the sturgeon species, ten black caviar samples were collected, and a reference DNA barcode library was developed using five genes (namely, 16S ribosomal RNA, cytochrome b, cytochrome c oxidase subunit I, cytochrome c oxidase subunit II, and NADH dehydrogenase subunit 5 genes). To develop the KASP markers, we selected 11 markers that could distinguish between the five grades of black caviar. As a result, specific markers for each of the targeted caviars were clustered into FAM-positive sections. DNA barcoding and the KASP assay revealed that one Beluga, six Osetra, and three Sevruga were identified among the ten caviar samples. Moreover, we found that the sturgeon species were mislabeled in two products. Here, we aimed to develop a KASP assay based on SNP that allows rapid and easy identification of caviar grade. These methods are expected to contribute to preventing the distribution of illegal products.