• Title/Summary/Keyword: SSR markers

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Genetic Analysis of Polymorphic DNA Markers in Cucumber (오이 다형성 마커를 이용한 유전분석)

  • Lee, Sun-Young;Chung, Sang-Min
    • Journal of Life Science
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    • v.21 no.3
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    • pp.468-472
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    • 2011
  • DNA marker is a powerful tool for plant genetics and breeding. In this study, 995 SSR markers were employed with chilling resistant cucumber, known as 'NC76', and chilling susceptible cucumber, known as 'GY14'. Using 2% agarose gel electrophoresis, 145 SSR markers were identified as length variation markers between 'NC76' and 'GY14'. The SSR markers that showed no length polymorphism were then screened using high resolution melting analysis technique and additional 30 polymorphic SSR markers were identified. As a preliminary evaluation for mapping, 20 markers among these 175 markers were employed to a $F_2$ population of 'NC76' x 'GY14' cross. Linkage analysis revealed 13 markers that joined into six linkage groups and seven markers that remained unlinked. This result indicates that these 175 markers could be used for construction of a genetic map using a cross between 'NC76' and 'GY14' for further investigation in developing markers related to resistance to chilling in cucumbers.

Whole genome re-sequencing and development of SSR markers in oriental melon (참외 전장유전체 염기서열 분석 및 SSR 마커 개발)

  • Song, Woon-Ho;Chung, Sang-Min
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.71-78
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    • 2019
  • The objective of this study was to use 'Danta PR', NGS (Next Generation Sequencing) technology for genome resequencing to develop polymorphic makers between Chinese oriental melon, 'Hyangseo 1' and Korean oriental melon. From the resequencing data that covered about 81 times of the genome size, 104,357 of SSR motifs and Indel, and 1,092,436 of SNPs were identified. 299 SSR and 307 Indel markers were chosen to cover each chromosome with 25 markers. These markers were subsequently used to identify genotypes of 'Danta PR' BC1 (F1 x 'Danta PR') population and a genetic linkage map was constructed. SSR, Indel, and SNPs identified in this study would be useful as a breeding tool to develop new oriental melon varieties.

Identification of Lettuce Germplasms and Commercial Cultivars Using SSR Markers Developed from EST (EST로부터 개발된 SSR 마커를 이용한 상추 유전자원 및 유통품종의 식별)

  • Hong, Jee-Hwa;Kwon, Yong-Sham;Choi, Keun-Jin;Mishra, Raghvendra Kumar;Kim, Doo Hwan
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.772-781
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    • 2013
  • The objective of this study was to develop simple sequence repeat (SSR) markers from expressed sequence tags (EST) of lettuce (Lactuca sativa) and identify 9 germplasms from 3 wild species of lettuce and 61 commercial cultivars using the developed EST-SSR markers. A total of 81,330 lettuce ESTs from NCBI databases were used to search for SSR and 4,229 SSR loci were identified. The highest proportion (59.12%, 2500) was represented by trinucleotide, followed by dinucleotide (29.70%, 1256) and hexanucleotide (6.62%, 280) among SSR repeat motifs. Totally 474 EST-SSR primers were developed from EST and a random set of 267 primers was used to assess the genetic diversity among 9 germplasms and 61 cultivars. Out of 267 primers, 47 EST-SSR markers showed polymorphism between 7 cultivars. Twenty-six EST-SSR markers among 47 EST-SSR markers showed high polymorphism, reproducibility, and band clearance. The relationship between 26 markers genotypes and 70 accessions was analyzed. Totally 127 polymorphic amplified fragments were obtained by 26 EST-SSR markers and two to nine SSR alleles were detected for each locus with an average of 4.88 alleles per locus. Average polymorphism information content was 0.542, ranging from 0.269 to 0.768. Genetic distance of clusters ranged from 0.05 to 0.94 between 70 accessions and dendrogram at a similarity of 0.34 gave 7 main clusters. Analysis of genetic diversity revealed by these 26 EST-SSR markers showed that the 9 germplasms and 61 commercial cultivars were discriminated by marker genotypes. These newly developed EST-SSR markers will be useful for cultivar identification and distinctness, uniformity and stability test of lettuce.

Construction of Linkage Map Using RAPD and SSR Markers in Soybean (Glycine max)

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    • Korean Journal of Plant Resources
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    • v.10 no.3
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    • pp.241-246
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    • 1997
  • Linkage maps based on molecular markers are valuable tools in plant breeding and genetic studies. A population of 76 RI lines from the mating of A3733 and PI437.088 was evaluated with Random Amplified Polymorphic DNA(RAPD) and Simple Sequence Repeats (SSR) markers to create soybean molecular linkage map, 302 RAPD and 21 SSR markers were genetically linked and formed forty linkage groups. These linkage groups spanned a genetic distance of 1,775 cM. The average distance between markers was 5.5 cM.

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Development of SSR markers for classification of Flammulina velutipes strains (팽이버섯 (Flammulina velutipes) 계통의 분류를 위한 SSR 마커개발)

  • Woo, Sung-I;Seo, Kyoung-In;Jang, Kab yeul;Kong, Won-Sik
    • Journal of Mushroom
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    • v.15 no.2
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    • pp.78-83
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    • 2017
  • Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From the SSR-enriched library, 490 white colonies were randomly selected and sequenced. Among the 490 sequenced clones, 85 (17.35%) were redundant. Among the remaining 405 unique clones, 201 (49.6%) contained microsatellite sequences. We used 12 primer pairs that produced reproducible polymorphic bands for four diverse strains, and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles, and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed ($H_O$) and expected ($H_E$) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, whereas for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among the 32 Flammulina velutipes strains on the basis of SSR data were investigated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from an SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenetic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

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.

Combined Genome Mapping of RFLP-AFLP-SSR in Pepper

  • Lee, Je Min;Kim, Byung-Dong
    • Genomics & Informatics
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    • v.1 no.2
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    • pp.108-112
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    • 2003
  • We have constructed a molecular linkage map of pepper (Capsicum spp.) in an interspecific $F_2$ population of 107 plants with 320 RFLP, 136 AFLP, and 46 SSR markers. The resulting linkage map consists of 15 linkage groups covering 1,720 cM with an average map distance of 3.7 cM between framework markers. Most RFLP markers ($80\%$) were pepper-derived clones and these markers were evenly distributed all over the genome. Genes for defense and biosynthesis of carotenoids and capsaicinoids were mapped on this linkage map. By using 30 primer combinations, AFLP markers were generated in the $F_2$ population. For development of SSR markers in Capsicum, microsatellites were isolated from two small-insert genomic libraries and the GenBank database. This combined map provides a starting point for high-resolution QTL analysis, gene isolation, and molecular breeding.

Genetically Independent Tetranucleotide to Hexanucleotide Core Motif SSR Markers for Identifying Lentinula edodes Cultivars

  • Saito, Teruaki;Sakuta, Genki;Kobayashi, Hitoshi;Ouchi, Kenji;Inatomi, Satoshi
    • Mycobiology
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    • v.47 no.4
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    • pp.466-472
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    • 2019
  • For the purpose of protecting the rights of Lentinula edodes breeders, we developed a new simple sequence repeat (SSR) marker set consisting only of genetically independent tetranucleotide or longer core motifs. Using available genome sequences for five L. edodes strains, we designed primers for 13 SSR markers that amplified polymorphic sequences in 20 L. edodes cultivars. We evaluated the independence of every possible marker pair based on genotype data. Consequently, eight genetically independent markers were selected. The polymorphic information content values of the markers ranged from 0.269 to 0.764, with an average of 0.409. The markers could distinguish among 20 L. edodes cultivars and produced highly repeatable and reproducible results. The markers developed in this study will enable the precise identification of L. edodes cultivars, and may be useful for protecting breeders' rights.

Analysis of Korean japonica rice cultivars using molecular markers associated with blast resistance genes

  • Suh, Jung-Pil;Roh, Jae-Hwan;Cho, Young-Chan;Han, Seong-Sook;Jeon, Yong-Hee;Kang, Kyung-Ho;Kim, Yeon-Gyu
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.215-222
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    • 2008
  • Fifty-two Korean japonica rice cultivars were analyzed for leaf blast resistance and genotyped with 4 STS and 26 SSR markers flanking the specific chromosome sites linked with blast resistance genes. In our analysis of resistance genes in 52 japonica cultivars using STS markers tightly linked to Pib, Pita, Pi5(t) and Pi9(t), the blast nursery reaction of the cultivars possessing the each four major genes were not identical to that of the differential lines. Eight of the 26 SSR markers were associated with resistant phenotypes against the isolates of blast nursery as well as the specific Korean blast isolates, 90-008 (KI-1113), 03-177 (KJ-105). These markers were linked to Pit, Pish, Pib, Pi5(t), Piz, Pia, Pik, Pi18, Pita and Pi25(t) resistance gene loci. Three of the eight SSR markers, MRG5836, RM224 and RM7102 only showed significantly associated with the phenotypes of blast nursery test for two consecutive years. These three SSR markers also could distinguish between resistant and susceptible japonica cultivars. These results demonstrate the usefulness of marker-assisted selection and genotypic monitoring for blast resistance of rice in blast breeding programs.

A Genetic Linkage Map of Soybean with RFLP, RAPD, SSR and Morphological Markers

  • Kim, Hong-Sik;Lee, Suk-Ha;Lee, Yeong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.2
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    • pp.123-127
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    • 2000
  • The objective of this study was to develop a linkage map of soybean under the genetic background of Korean soybean. A set of 89 F/sub 5/ lines was developed from a cross between 'Pureunkong', which was released for soy-bean sprout, and 'Jinpumkong 2', which had no beany taste in seed due to lack of lipoxygenase 1, 2, and 3. A linkage map was constructed for this population with a set of 113 genetic markers including 7 restriction fragment length polymorphism (RFLP) markers, 79 randomly amplified polymorphic DNA (RAPD) markers, 24 simple sequence repeat(SSR) markers, and 3 morphological markers. The map defined approximately 807.4 cM of the soybean genome comprising 25 linkage groups with 98 polymorphic markers. Fifteen markers remained unlinked. Seventeen linkage groups identified here could be assigned to the respective 13 linkage groups in the USDA soybean genetic map. RFLP and SSR markers segregated at only single genetic loci. Fourteen of the 25 linkage groups contained at least one SSR marker locus. Map positions of most of the SSR loci and their linkages with RFLP markers were consistent with previous reports of the USDA soybean linkage groups. For RAPD, banding patterns of 13 decamer primers showed independent segregations at two or more marker loci for each primer. Only the segregation at op Y07 locus was expressed with codominant manner among all RAPD loci. As the soybean genetic map in our study is more updated, molecular approaches of agronomically important genes would be useful to improve Korean soybean improvement.

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