• Title/Summary/Keyword: Simple Sequence Repeat

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In vitro micropropagation of M.26 (Malus pumila Mill) apple rootstock and assessment of the genetic diversity of proliferated plantlets using simple sequence repeat markers (사과 대목 M.26 (Malus pumila Mill)의 기내 대량번식 및 simple sequence repeat 마커를 이용한 증식된 식물체의 유전적 다양성 평가)

  • Cho, Kang Hee;Han, Bong Hee;Han, Jeom Hwa;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan;Kim, Mi Young;Kim, Myung-Su
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.382-391
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    • 2018
  • The objective of this study was to determine the most effective medium condition of shoot proliferation and root formation for the efficient in vitro micropropagation of M.26 (Malus pumila Mill). Simple sequence repeat (SSR) markers were used to analyze the genetic diversity of micro-propagated and greenhouse grown M.26. Shoot proliferation was carried out in MS (Murashige and Skoog) containing benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$) and thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$). The highest number of shoots (10.67 shoots per explant) was induced by adding BA at a concentration $1.0mg{\cdot}L^{-1}$. TDZ treatments caused higher hyperhydricity rate in cultured explants than in BA treatments. There was no significant effect of both BA and auxin on shoot proliferation, and the optimum proliferation medium for M.26 was MS medium containing $1.0mg{\cdot}L^{-1}$ BA. To find a suitable medium composition for shoot rooting, we tested different concentrations indole-3-butyric acid (IBA) and ${\alpha}$-naphthaleneacetic acid ($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS medium (1/4-1), sucrose ($0{\sim}30g{\cdot}L^{-1}$). The shoots showed good rooting on half-strength MS medium containing $1.0mg{\cdot}L^{-1}$ IBA and $15-20g{\cdot}L^{-1}$ sucrose. The rooting rate (100%), number of roots (10.45 ~ 13.60 roots per explant), root length (7.41 ~ 8.33 cm), and shoot length (4.93 ~ 5.38 cm) were good on this medium. Fifteen SSR primers were detected in a total of 30 alleles in 20 micro-propagated plantlets, all SSR profiles from micro-propagated plantlets were monomorphic and similar to greenhouse grown control plantlet M.26 plant. The results indicated that M.26 micro-propagated plantlets were genetically stable.

Identification of DNA Variations Using AFLP and SSR Markers in Soybean Somaclonal Variants

  • Lee, Suk-Ha;Jung, Hyun-Soo;Kyujung Van;Kim, Moon-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.69-72
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    • 2004
  • Somaclonal variation, defined as phenotypic and genetic variations among regenerated plants from a parental plant, could be caused by changes in chromosome structure, single gene mutation, cytoplasm genetic mutation, insertion of transposable elements, and DNA methylation during plant regeneration. The objective of this study was to evaluate DNA variations among somaclonal variants from the cotyledonary node culture in soybean. A total of 61 soybean somaclones including seven $\textrm{R}_1$ lines and seven $\textrm{R}_2$ lines from Iksannamulkong as well as 27 $\textrm{R}_1$ lines and 20 $\textrm{R}_2$ lines from Jinju 1 were regenerated by organogenesis from the soybean cotyledonary node culture system. Field evaluation revealed no phenotypic difference in major agronomic traits between somaclonal variants and their wild types. AFLP and SSR analyses were performed to detect variations at the DNA level among somaclonal variants of two varieties. Based on AFLP analysis using 36 primer sets, 17 of 892 bands were polymorphic between Iksannamulkong and its somaclonal variants and 11 of 887 bands were polymorphic between Jinju 1 and its somaclonal variants, indicating the presence of DNA sequence change during plant regeneration. Using 36 SSR markers, two polymorphic SSR markers were detected between Iksannamulkong and its somaclonal variants. Sequence comparison amplified with the primers flanking Satt545 showed four additional stretches of ATT repeat in the variant. This suggests that variation at the DNA level between somaclonal variants and their wild types could provide basis for inducing mutation via plant regeneration and broadening crop genetic diversity.

Genetic Variation of Rice Populations Estimated Using nrDNA ITS Region Sequence

  • Wang, Dong;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.249-255
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    • 2014
  • The rice belonging to Oryza sativa is not only has significant economic importance, for it is the major source of nutrition for about 3 billion all around the world. But also plays a vital role as a model organism, because it has a number of advantages to be a model plant, such as efficient transformation system and small genome size. Many methods and techniques have been conducted to attempt to distinguish different Oryza sativa species, such as amplified fragment length polymorphism (AFLP), random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and so on. However, studies using sequence analysis of internal transcribed spacer (ITS), a region of ribosomal RNA has not been reported until now. This study was undertaken with an aim to understand the phylogenetic relationships among sixteen isolates of Oryza sativa collected from abroad and fifteen isolates collected from Korea, using ribosomal RNA (rRNA) internal transcribed spacer (ITS) sequences to compare the phylogeny relationships among different Oryza sativa species. The size variation obtained among sequenced nuclear ribosomal DNA (nrDNA) ITS region ranged from 515bp to 1000bp. The highest interspecific genetic distance (GD) was found between Sfejare 45 (FR12) and Anapuruna (FR15). Taebong isolate showed the least dissimilarity of the ITS region sequence with other thirty isolates. This consequence will help us further understanding molecular diversification in intra-species population and their phylogenetic analysis.

Identification of New Microsatellite Markers in Panax ginseng

  • Kim, Joonki;Jo, Beom Ho;Lee, Kyoung Lyong;Yoon, Eui-Soo;Ryu, Gi Hyung;Chung, Ki Wha
    • Molecules and Cells
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    • v.24 no.1
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    • pp.60-68
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    • 2007
  • Microsatellites, also called simple sequence repeats (SSR), are very useful molecular genetic markers commonly used in crop breeding, species identification and linkage analysis. In the present study, we constructed a microsatellite-enriched genomic library of Panax ginseng, and identified 251 novel microsatellite sequences. Tri-nt repeat units were the most abundant (46.6%), followed by di-nt repeats (35.5%). The $(AG)_n$ motif was most common (23.1%), followed by the $(AAC)_n$ motif (22.3%). From the genotyping of 94 microsatellites using marker-specific primer sets, we identified 11 intraspecific polymorphic markers as well as 14 possible interspecific polymorphic markers differing between P. ginseng and P. quinquefolius. The exact allele structures of the polymorphic markers were determined and the alleles were named. This study represents the first report of the bulk isolation of microsatellites by screening a microsatellite-enriched genomic library in P. ginseng. The microsatellite markers could be useful for linkage analysis, genetic breeding and authentication of Panax species.

Genetic diversity and population structure of rice accessions from South Asia using SSR markers

  • Cui, Hao;Moe, Kyaw Thu;Chung, Jong-Wook;Cho, Young-Il;Lee, Gi-An;Park, Yong-Jin
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.11-22
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    • 2010
  • The population structure of a domesticated species is influenced by the natural history of the populations of its pre-domesticated ancestors, as well as by the breeding system and complexity of breeding practices implemented by humans. In the genetic and population structure analysis of 122 South Asia collections using 29 simple sequence repeat (SSR) markers, 362 alleles were detected, with an average of 12.5 per locus. The average expected heterozygosity and polymorphism information content (PIC) for each SSR locus were 0.74 and 0.72,respectively. The model-based structure analysis revealed the presence of three clusters with the 91.8% (shared > 75%) membership, with 8.2% showing admixture. The genetic distances of Clusters 1-3 were 0.55, 0.56, and 0.68, respectively. Polymorphic information content followed the same trend (Cluster 3 had the highest value and Cluster 1 had smallest value), with genetic distances for each cluster of 0.52, 0.52, and 0.65, respectively. This result could be used for supporting rice breeding programs in South Asia countries.

The complete chloroplast genome of Diarthron linifolium (Thymelaeaceae), a species found on a limestone outcrop in eastern Asia

  • KIM, Sang-Tae;OH, Sang-Hun;PARK, Jongsun
    • Korean Journal of Plant Taxonomy
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    • v.51 no.4
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    • pp.345-352
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    • 2021
  • Diarthron linifolium Turcz. is an annual herb usually found in sandy soil or limestone areas. Plants in the genus Diarthron are known to have toxic chemicals that may, however, be potentially useful as an anticancer treatment. Diarthron linifolium is a unique species among the species of the genus distributed in Korea. Here, we determine the genetic variation of D. linifolium collected in Korea with a full chloroplast genome and investigate its evolutionary status by means of a phylogenetic analysis. The chloroplast genome of Korean D. linifolium has a total length of 172,644 bp with four subregions; 86,158 bp of large single copy and 2,858 bp of small single copy (SSC) regions are separated by 41,814 bp of inverted repeat (IR) regions. We found that the SSC region of D. linifolium is considerably short but that IRs are relatively long in comparison with other chloroplast genomes. Various simple sequence repeats were identified, and our nucleotide diversity analysis suggested potential marker regions near ndhF. The phylogenetic analysis indicated that D. linifolium from Korea is a sister to the group of Daphne species.

Analysis of Microsatellite Patterns in the Genome of Red Sea Cucumber (홍해삼 유전체 분석에 의한 microsatellite의 분포도 연구)

  • Lee, Tae Wook;Kim, Sam Woong;Kim, Jung Sun;Chi, Won-Jae;Bang, Woo Young;Kim, Jang Hyeon;Yang, Chul Woong;Bang, Kyu Ho;Gal, Sang Wan
    • Journal of Life Science
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    • v.32 no.9
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    • pp.690-697
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    • 2022
  • This study was conducted to analyze genome of red sea cucumber and to use it as basic data for the development of genetic markers for red sea cucumber. Microsatellite marker analysis of Ulleungdo_normal and Ulleungdo_native red sea cucumbers revealed that dinucleotide simple sequence repeats (SSRs) had the highest ratio, at 81.3~81.4%, and the number of the detected SSRs tended to decrease as the number of repeating sequence units in SSRs increased. In general, microsatellites with between 5 and 10 iterations were most common. As the size of the SSR repeating sequence units increased, the SSR iterations gradually decreased. The di-, tri-, and tetra-nucleotides in SSRs were detected in the highest numbers as (AT)5, (AAT)5, and (AAAT)5, respectively. (CG) and (CCG) had very low frequencies compared to the numbers of other repeating SSR units. The numbers of di-and tri-nucleotide repeats were up to 35 and 32, respectively, and then increased discontinuously up to 44 and 43 repeats, respectively. Tetra-, penta-, and hexa-nucleotides in SSRs occurred in numbers up to 25, 21 and 14, respectively. This analysis of red sea cucumber indicated that it maintains its own repetition sequence and repetition number; therefore, we suggest that using it as basic data for molecular marker will be possible in future research.

Evaluation of horticultural traits and genetic relationship in melon germplasm (멜론 유전자원의 원예형질 특성 및 유연관계 분석)

  • Jung, Jaemin;Choi, Sunghwan;Oh, Juyeol;Kim, Nahui;Kim, Daeun;Son, Beunggu;Park, Younghoon
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.401-408
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    • 2015
  • Horticultural traits and genetic relationship were evaluated for 83 melon (Cucumis melo L.) cultivars. Survey of a total of 36 characteristics for seedling, leaf, stem, flower, fruit, and seed and subsequent multiple analysis of variance (MANOVA) were conducted. Principal component analysis (PCA) showed that 8 principle components including fruit weight, fruit length, fruit diameter, cotyledon length, seed diameter, and seed length accounted for 76.3% of the total variance. Cluster analysis of the 83 melon cultivars using average linkage method resulted in 5 clusters at coefficient of 0.7. Cluster I consisted of cultivars with high values for fruit-related traits, Cluster II for soluble solid content, and Cluster V for high ripening rate. Genotyping of the 83 cultivars was conducted using 15 expressed-sequence tagged-simple sequence repeat (EST-SSR) from the Cucurbit Genomics Initiative (ICuGI) database. Analysis of genetic relatedness by UPGMA resulted in 6 clusters. Mantel test indicated that correlation between morphological and genetic distance was very low (r = -0.11).

Development of an ISSR-Derived SCAR Marker in Korean Ginseng Cultivars (Panax ginseng C. A. Meyer)

  • Lee, Jei-Wan;Kim, Young-Chang;Jo, Ick-Hyun;Seo, A-Yeon;Lee, Jeong-Hoon;Kim, Ok-Tae;Hyun, Dong-Yun;Cha, Seon-Woo;Bang, Kyong-Hwan;Cho, Joon-Hyeong
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.52-59
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    • 2011
  • Recently, new ginseng cultivars having superior agricultural traits have been developed in Korea. For newly developed plant cultivars, the identification of distinctiveness is very important factors not only in plant cultivar management but also in breeding programs. Thus, eighty-five inter simple sequence repeat (ISSR) primers were applied to detect polymorphisms among six major Korean ginseng cultivars and two foreign ginsengs. A total of 197 polymorphic bands with an average 5.8 polymorphic bands and 2.9 banding patterns per assay unit across six Korean ginseng cultivars and foreign ginsengs from 236 amplified ISSR loci with an average 6.9 loci per assay unit were generated by 34 out of 85 ISSR primers. Three species of Panax ginseng including the Korean ginseng cultivars, P. quinquefolius, and P. notoginseng, could be readily discriminated using most tested primers. UBC-821, UBC-868, and UBC-878 generated polymorphic bands among the six Korean ginseng cultivars, and could distinguish them from foreign ginsengs. Sequence characterized amplified region (SCAR) marker system was introduced in order to increase the reproducibility of the polymorphism. One SCAR marker, PgI821C650, was successfully converted from the randomly amplified polymorphism by UBC-821. It showed the expected dominant polymorphism among ginseng samples. In addition, the specific polymorphism for Sunwon was generated by treating Taq I restriction enzyme to polymerase chain reaction products of PgI821C650. These results will serve as useful DNA markers for identification of Korean ginseng, especially Sunwon cultivar, seed management, and molecular breeding program supplemented with marker-assisted selection.

Analysis of the Genome Sequence of Strain GiC-126 of Gloeostereum incarnatum with Genetic Linkage Map

  • Jiang, Wan-Zhu;Yao, Fang-Jie;Fang, Ming;Lu, Li-Xin;Zhang, You-Min;Wang, Peng;Meng, Jing-Jing;Lu, Jia;Ma, Xiao-Xu;He, Qi;Shao, Kai-Sheng;Khan, Asif Ali;Wei, Yun-Hui
    • Mycobiology
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    • v.49 no.4
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    • pp.406-420
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    • 2021
  • Gloeostereum incarnatum has edible and medicinal value and was first cultivated and domesticated in China. We sequenced the G. incarnatum monokaryotic strain GiC-126 on an Illumina HiSeq X Ten system and obtained a 34.52-Mb genome assembly sequence that encoded 16,895 predicted genes. We combined the GiC-126 genome with the published genome of G. incarnatum strain CCMJ2665 to construct a genetic linkage map (GiC-126 genome) that had 10 linkage groups (LGs), and the 15 assembly sequences of CCMJ2665 were integrated into 8 LGs. We identified 1912 simple sequence repeat (SSR) loci and detected 700 genes containing 768 SSRs in the genome; 65 and 100 of them were annotated with gene ontology (GO) terms and KEGG pathways, respectively. Carbohydrate-active enzymes (CAZymes) were identified in 20 fungal genomes and annotated; among them, 144 CAZymes were annotated in the GiC-126 genome. The A mating-type locus (MAT-A) of G. incarnatum was located on scaffold885 at 38.9 cM of LG1 and was flanked by two homeodomain (HD1) genes, mip and beta-fg. Fourteen segregation distortion markers were detected in the genetic linkage map, all of which were skewed toward the parent GiC-126. They formed three segregation distortion regions (SDR1-SDR3), and 22 predictive genes were found in scaffold1920 where three segregation distortion markers were located in SDR1. In this study, we corrected and updated the genomic information of G. incarnatum. Our results will provide a theoretical basis for fine gene mapping, functional gene cloning, and genetic breeding the follow-up of G. incarnatum.