• 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.

• Journal of Life Science
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• v.20 no.4
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• pp.632-638
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• 2010

The genetic diversity among 48 persimmon (Diospyros kaki Thunb.) accessions, indigenous in Korea and introduced from Japan and China, was evaluated by using simple sequence repeat (SSR) markers. From 20 SSR primer sets, a total of 114 polymorphic markers were detected among 12 pollination-constant non-astringent (PCNA), 13 pollination-variant non-astringent (PVNA), 15 pollination-variant astringent (PVA), and 8 pollination-constant astringent (PCA) cultivars. Analysis of pair-wise genetic similarity coefficient (Nei-Li) and unweighted pair-group method with arithmetic averaging (UPGMA) clustering revealed two main clusters and four subclusters for cluster I. The subclustering pattern was in accordance with the classification of persimmon cultivars based on the nature of astringency loss. Phenetic relationships among the subclusters showed a closer relatedness of the PCNA group with the PVNA group, and the PVA with the PCA group. Genetic similarity co-efficiency was 0.499 on average and the highest (0.954) similarity was observed between 'Cheongdo-Bansi' and 'Haman-Bansi'. The similarity was lowest (0.192) between 'Damopan'and 'Atago'. Identification of each cultivar with the execption of 'Cheongdo-Bansi' and 'Gyeongsan-Bansi' was possible based on the SSR fingerprints, suggesting that these SSR markers are a useful tool for protecting intellectual property on newly developed cultivars.

• Journal of Life Science
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• v.23 no.6
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• pp.721-727
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• 2013

The objective of this study was to evaluate the suitability of microsatellite markers for variety identification in 42 apple varieties. For microsatellite analysis, 305 primer pairs were screened in 8 varieties and twenty six primer pairs showed polymorphism with clear band pattern and repetitive reproducibility. A total of 165 polymorphic amplified fragments were obtained in 42 varieties using 26 markers. Two to twelve alleles were detected for each locus with an average of 6.4 alleles per locus. A value of polymorphism information content (PIC) ranged from 0.461 to 0.849 with an average of 0.665. A total of 165 marker loci were used to calculate Jaccard's distance coefficients using unweighted pair-group method with arithmetical average (UPGMA) cluster analysis. Genetic distance of cluster ranged from 0.27 to 1.00. Analysis of genetic relationship revealed that these 26 microsatellite marker sets discriminated a total of 41 varieties except for 1 variety among 42 varieties. These markers will be utilized as molecular data in variety identification of apple.

• Journal of Mushroom
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• v.15 no.4
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• pp.273-278
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• 2017

Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

• Food Science of Animal Resources
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• v.27 no.2
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• pp.150-156
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• 2007

This study was conducted to analyze the genetic characteristics of Korean Native Pigs(KNP), and to establish an individual identification system comprising many microsatellite markers located on different pig autosomes. Genotype data from 13 microsatellites typed in 446 animals was used to determine the validation of a method of individual identification in 4 KNP. A total of 112 alleles of the 13 microsatellites were detected and average heterozygosities(polymorphic information content) ranged from 0.286(0.423) to 0.686(0.796) in this study. Comparing the pattern of allele frequency among the KNP, Yorkshire, Landrace and Duroc breeds, there was specific differentiation between populations at multi-allelic loci. The cumulative power of discrimination(CPD) was 99.999% by including 10 microsatellite loci for the individual identification system. The probability that two different individuals incidentally have same genotype was estimated to be $0.36{\times}10^{-9}$. The system employing these 10 markers therefore proved to be applicable to the individual identification of KNP.

• The Korean Journal of Mycology
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• v.42 no.2
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• pp.159-164
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• 2014

For development of a method for differentiation of Pleurotus eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotyping results. PCR using each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in the range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pairgroup method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into two clusters. Cluster I and II were comprised of four and eight cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivars and strains with 21 alleles and a PIC value of 0.9090. These results might be useful in providing an efficient method for the identification of P. eryngii cultivars with separate PCR reactions.

• Journal of Korean Society of Forest Science
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• v.108 no.3
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• pp.302-310
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• 2019

Precise and fast identification of crop cultivars is essential for efficient breeding and plant breeders' rights. Traditional methods for identification of jujube cultivars are based on the evaluation of morphological characteristics. However, due to time constraints and environmental influences, it is difficult to distinguish cultivars using only morphological traits. In this study, we cloned fragments from improved inter simple sequence repeats (ISSR) analysis, and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific ISSR bands of jujube cultivars from Dalizao and Boeundaechu were purified, cloned, and sequenced. As a result, four clones labeled 827Dalizao550, 827Boeun750, 846Boeun700, and 847Dalizao850 were identified. In order to investigate whether they were specific for the jujube cultivar, four pairs of SCAR primers were then designed and polymerase chain reaction (PCR) amplifications were conducted to analyze 32 samples, including jujube and sour jujube. In the PCR amplification of the 827Dalizao550 SCAR marker, the specific bands with 550 bp were amplified in six samples (Dalizao, Sandonglizao, Dongzao, Yuanlin No. 2, Suanzao 2, Suanzao 4), but unexpected bands (490 bp) were amplified in the others. Moreover, in the PCR amplification of the 847Dalizao850 SCAR marker, the specific bands with 850 bp were found in three samples (Dalizao, Sandonglizao, and Dongzao) and 900 bp unexpected bands were amplified in five samples (Pozao, Suanzao 1, Suanzao 2, Suanzao 3, Suanzao 4). These results showed that newly developed markers could be useful as a fast and reliable tool to identify jujube cultivars. However, further identification of polymorphic information and the development of SCAR markers are required for the identification of more diverse cultivars.

• Journal of Life Science
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• v.33 no.8
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• pp.623-631
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• 2023

This study was conducted to develop microsatellite markers in Parapristipoma trilineatum using next-generation sequencing. A total of 402,244,934 reads were generated on the Illumina Hiseq X Ten System, yielding 60,738,985,034 bp of sequences. The de novo assembly resulted in 1,320,995 contigs. A total of 952,326 contigs (0.016%) including 151 microsatellite loci were derived from the 1,320,995 contigs longer than 640 bp. A total of 34 primer sets were designed from the 151 microsatellite loci. As a result, 15 microsatellite loci were chosen and used for assuming population genetic parameters in the wild and farmed populations. The mean number of effective alleles was 12, ranging from 6 to 25. The observed heterozygosity (H_O) and the expected heterozygosity (H_E) ranged between 0.530 and 0.873, with an average of 0.750, and from 0.647 to 0.895, with an average of 0.793, respectively. According to these results, the developed set of 15 microsatellite markers is expected to be useful for the analysis of genetic characteristics in the population of P. trilineatum in Korea. There are requirements now for further genetic information, fishery resource management, breeding guidelines, support with the selection of breeds and studies on the effects of release, all of which will improve species conservation, and through future research, we aim to offer genetic foundational data with that goal.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.151-159
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• 2002

Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n＝9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

• Korean journal of applied entomology
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• v.37 no.1
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• pp.31-37
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• 1998

Random amplified polymorphic DNA (RAPD) markers were used to analyze genetic similarity among 8 clones of apierous green peach aphid, two types (M. persicae Sulzer and M. nicotianae lack man) classified by their mo~hologi~cahla raters and host preference (Blackman, 1987), collected from tobacco plants. The genetic variation among these clones was evaluated by polymerase chain reaction amplification with 20 random primers. The higher GC contents of primers, the better in amplification efficiency of PCR reaction in general. The genetic similarities among eight aphid clones were analyzed from UPGMA (unweighted pair group average method) cluster analysis based on simple matching coefficient. The range of genetic similarity coefficients was 0.414 to 0.808. The most close relationship among the clones was similarity coefficient of 0.808 between the PG2 and the PG3 clone. The eight aphid clones analyzed were clustered into three groups by the genetic similarity coefficient. The first group, PG1, PG2, PG3 clone including in M. persicae type by their morphological characters and RED clone in M. nicotianae type was clustered at the genetic similarity coefficient of 0.643. The second group, GR1, GR2, BRN in M. nicotianae type was at the 0.636;and the third group was DBR clone in M. persicae type. The results did not indicate any correlation between m&-phological types (M. persicae and M. nicotianae) and RAPD polymorphism. We could not detect any obvious genetic relationships of the two morphological types of the green peach aphid collected from tobacco plants.