• Genomics & Informatics
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• v.9 no.4
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• pp.189-193
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• 2011

Simple sequence repeats (SSRs) are ubiquitous short tandem duplications found within eukaryotic genomes. Their length variability and abundance throughout the genome has led them to be widely used as molecular markers for crop-breeding programs, facilitating the use of marker-assisted selection as well as estimation of genetic population structure. Here, we report a software application, "SSR-Primer Generator " for SSR discovery, SSR-primer design, and homology-based search of in silico amplicons from a DNA sequence dataset. On submission of multiple FASTA-format DNA sequences, those analyses are batch processed in a Java runtime environment (JRE) platform, in a pipeline, and the resulting data are visualized in HTML tabular format. This application will be a useful tool for reducing the time and costs associated with the development and application of SSR markers.

• Genomics & Informatics
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• v.19 no.3
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• pp.33.1-33.10
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• 2021

Eucalyptus is one of the major plantation species with wide variety of industrial uses. Polymorphic and informative simple sequence repeats (SSRs) have broad range of applications in genetic analysis. In this study, two individuals of Eucalyptus tereticornis (ET217 and ET86), one individual each from E. camaldulensis (EC17) and E. grandis (EG9) were subjected to whole genome resequencing. Low coverage (10×) genome sequencing was used to find polymorphic SSRs between the individuals. Average number of SSR loci identified was 95,513 and the density of SSRs per Mb was from 157.39 in EG9 to 155.08 in EC17. Among all the SSRs detected, the most abundant repeat motifs were di-nucleotide (59.6%-62.5%), followed by tri- (23.7%-27.2%), tetra- (5.2%-5.6%), penta- (5.0%-5.3%), and hexa-nucleotide (2.7%-2.9%). The predominant SSR motif units were AG/CT and AAG/TTC. Computational genome analysis predicted the SSR length variations between the individuals and identified the gene functions of SSR containing sequences. Selected subset of polymorphic markers was validated in a full-sib family of eucalypts. Additionally, genome-wide characterization of single nucleotide polymorphisms, InDels and transcriptional regulators were carried out. These variations will find their utility in genome-wide association studies as well as understanding of molecular mechanisms involved in key economic traits. The genomic resources generated in this study would provide an impetus to integrate genomics in marker-trait associations and breeding of tropical eucalypts.

• 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.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.243-263
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• 2017

This study is to establish the varietal discrimination based on DNA profiling of different varieties of rice. We examined the genetic distance among Korean rice varieties using allele frequencies and a genetic diversity analysis with Simple Sequence Repeats (SSRs) markers. The analysis of the genetic diversity and genetic relationships of 243 Korean rice varieties was varied out using 20 SSRs markers. A total of 268 alleles were detected, ranging from 6 to 32, with an average of 13.45 alleles per locus, and and average of gene diversity (GD) of 0.556. Seven SSR markers were selected as key markers for discrimination among the Korean rice varieties. Concerning the results, 243 varieties (100%) were discriminated among by using acrylamide gel and fragment analyzer-based markers. In conclusion, this study provides useful basic data that can be utilized concerning Korean rice varieties breeding and development. In addition, we will have to manage and conserve as a valuable genetic resource, without losing the diversity of Korean rice varieties.

• Molecules and Cells
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• v.22 no.3
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• pp.300-307
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• 2006

Brassica rapa ssp. pekinensis (Chinese cabbage) is an economically important crop and a model plant for studies on polyploidization and phenotypic evolution. To gain an insight into the structure of the B. rapa genome we analyzed 12,017 BAC-end sequences for the presence of transposable elements (TEs), SSRs, centromeric satellite repeats and genes, and similarity to the closely related genome of Arabidopsis thaliana. TEs were estimated to occupy 14% of the genome, with 12.3% of the genome represented by retrotransposons. It was estimated that the B. rapa genome contains 43,000 genes, 1.6 times greater than the genome of A. thaliana. A number of centromeric satellite sequences, representing variations of a 176-bp consensus sequence, were identified. This sequence has undergone rapid evolution within the B. rapa genome and has diverged among the related species of Brassicaceae. A study of SSRs demonstrated a non-random distribution with a greater abundance within predicted intergenic regions. Our results provide an initial characterization of the genome of B. rapa and provide the basis for detailed analysis through whole-genome sequencing.

• 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.

• Horticulture, Environment, and Biotechnology : HEB
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• v.59 no.6
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• pp.889-897
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• 2018

Currently, there is a lack of genetic markers capable of effectively detecting polymorphisms in Clematis. Therefore, we developed new markers to investigate inter- and intraspecific diversity in Clematis. Based on the complete chloroplast genome of Clematis terniflora, simple sequence repeats were explored and primer pairs were designed for all ten adequate repeat regions (cpSSRs), which were tested in 43 individuals of 11 Clematis species. In addition, the nuclear ITS region was sequenced in 11 Clematis species. Seven cpSSR loci were found to be polymorphic in the genus and serve as markers that can distinguish different species and be used in different genetic analyses, including cultivar identification to assist the breeding of new ornamental cultivars.

• Molecules and Cells
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• v.25 no.4
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• pp.467-478
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• 2008

Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

• Journal of Korean Society of Forest Science
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• v.98 no.5
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• pp.568-575
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• 2009

Korean wild and forest cultivated ginseng has long been accepted as high medicinal values compared to field cultivated ginseng. Owing to the high price of Korean wild ginseng, Chinese wild and forest cultivated ginseng were smuggled and sold as Korean wild and forest cultivated ginseng. Therefore, an efficient method is required to distinguish Korean ginseng from Chinese ginseng. Microsatellites, simple sequence repeats (SSRs), are highly polymorphic loci present in DNA that consist of repeating units of base pairs. Thus SSR markers are highly advantageous for detection of small genetic variances of intra-species. In the present study, we constructed a microsatellite-enriched genomic library from South Korean wild Panax ginseng. After sequence analysis of 992 randomly picked positive colonies, 126 (12.7%) of the colonies were found to contain microsatellite sequences, and 38 primer pairs were designed. By polymorphism assessment using 36 primer pairs, 4 primers (PG409, PG450, PG491, and PG582) were shown to be polymorphic to distinguish the South Korean ginseng from the Chinese ginseng. These 4 microsatellite markers will provide powerful tools to authenticate South Korean ginseng from Chinese ginseng.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.44 no.2
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• pp.112-116
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• 1999

Genetic diversity of 31 rice varieties including 25 japonica and 6 indica varieties was evaluated using a combination of 19 microsatellite or simple sequence repeats (SSRs) and 28 random decamer oligonucle-otide primers. All 19 microsatellite primer sets representing 19 loci in the rice genome showed polymorphisms among the 31 varieties and revealed 91 alleles with an average of 4.80 bands per primer. Also all 28 random decamer primers used were informative and generated 114 non-redundant bands with a mean of 4.07 bands. Microsatellite markers detected higher number of alleles than random primers .although the mean difference was not statistically significant. A cluster analysis based on Nei＇s genetic distances calculated from the 205 bands resolved the 31 varieties into two major groups that correspond to indica and japonica subspecies, which is consistent with the genealogical information. As few as six random decamer primers or a combination of one microsatellite and four random decamer primers were sufficient to uniquely differentiate all 31 varieties. These combinations would be potentially useful in rice variety protection and identification considering that 25 out of 31 varieties used in this study are japonica rices with high grain quality and have close make up.