• Korean Journal of Agricultural Science
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• v.40 no.2
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• pp.115-121
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• 2013

Polymorphic bands of two fine-leaf zoysiagrass mutants (CNU 70-1, CNU 70-2) induced via a gamma-ray irradiation on seeds of Zoysia japonica were obtained by using randomly amplified polymorphic DNA (RAPD) primers. The genotype-specific fragments were then converted into PCR-based sequence characterized amplified region (SCAR) markers, which are now amenable to detecting them among other zoysiagrass species widely noticeable in Korea. The CNU 70-1-specific primer set amplified about 900 bp successfully, while the CNU 70-6 marker produced the expected 1,500 bp band, by which those markers were nominated by CNU 70-1_900 and CNU 70-6_1500 SCARs, respectively. The developed SCAR markers can be an applicable tool in sod industry where illegal appropriation hampers breeder's right and profits due to the turfgrass plant vegetatively propagating.

• Korean Journal of Plant Taxonomy
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• v.48 no.3
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• pp.195-200
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• 2018

Genetic assessments of rare and endangered species are among the first steps necessary to establish the proper management of natural populations. Transcriptome-derived single-sequence repeat markers were developed for the Korean endangered species Astilboides tabularis (Saxifragaceae) to assess its genetic diversity. A total of 96 candidate microsatellite loci were isolated based on transcriptome data using Illumina pair end sequencing. Of these, 26 were polymorphic, with one to five alleles per locus in 60 individuals from three populations of A. tabularis. The observed and expected heterozygosity per locus ranged from 0.000 to 0.950 and from 0.000 to 0.741, respectively. These polymorphic transcriptome-derived simple sequence repeat markers would be invaluable for future studies of population genetics and for ecological conservation of the endangered species A. tabularis.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.3
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• pp.316-322
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• 2013

In order to evaluate the genetic diversity and discrimination among five Korean native chicken lines, a total of 86 individuals were genotyped using 150 microsatellite (MS) markers, and 15 highly polymorphic MS markers were selected. Based on the highest value of the number of alleles, the expected heterozygosity (He) and polymorphic information content (PIC) for the selected markers ranged from 6 to 12, 0.466 to 0.852, 0.709 to 0.882 and 0.648 to 0.865, respectively. Using these markers, the calculated genetic distance (Fst), the heterozygote deficit among chicken lines (Fit) and the heterozygote deficit within chicken line (Fis) values ranged from 0.0309 to 0.2473, 0.0013 to 0.4513 and -0.1002 to 0.271, respectively. The expected probability of identity values in random individuals (PI), random half-sib ($PI_{half-sibs}$) and random sibs ($PI_{sibs}$) were estimated at $7.98{\times}10^{-29}$, $2.88{\times}10^{-20}$ and $1.25{\times}10^{-08}$, respectively, indicating that these markers can be used for traceability systems in Korean native chickens. The unrooted phylogenetic neighbor-joining (NJ) tree was constructed using 15 MS markers that clearly differentiated among the five native chicken lines. Also, the structure was estimated by the individual clustering with the K value of 5. The selected 15 MS markers were found to be useful for the conservation, breeding plan, and traceability system in Korean native chickens.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.798-806
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• 2013

Precise, fast, and cost-effective identification of crop cultivars is essential for plant breeder's rights. Traditional methods for identification of persimmon cultivars are based on the evaluation of sets of morphological characteristics. However, it is difficult to distinguish closely related cultivars using only morphological traits. This study was conducted to develop DNA markers for identification of the 32 persimmon cultivars in Korea and Japan. A total of 309 randomly amplified polymorphic DNA (RAPD) markers were identified using 40 different random primers. Various number of polymorphic bands ranged from 4 (OPP-08) to 14 (UBC159) were detected with an average of 7.7. Resulting 57 RAPD fragments were selected, and their sequences were determined for developing sequence characterized amplified region (SCAR) markers. As a result, 15 of 57 RAPD fragments were successfully converted to SCAR markers. Single polymorphic bands of the same size as or smaller than the RAPD fragments were amplified depending on SCAR markers. Among these markers, a combination of eight SCAR markers (PS225_200, PSN05_420, PSF13_523, PSN11_540, PS372_567, PS485_569, PSP08_635, and PS631_735) provided sufficient polymorphisms to identify 32 persimmon cultivars. These newly developed markers will be a fast and reliable tool to identify persimmon cultivars.

• Journal of Life Science
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• v.19 no.5
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• pp.639-643
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• 2009

High temperature stress might affect the yield and quality of Chinese cabbage. In order to develop cultivars resistant to high temperature stress, we developed polymorphic DNA markers for trehalose synthesis genes related to abiotic stress resistance. A total of 28 Brassica rapa ESTs homologous to trehalose synthesis genes of Arabidopsis were found from the NCBI database. The polymorphic DNA sequences were searched between Chinese cabbages - Chiifu, which is relatively susceptible to high temperature stress, and Kenshin, which is tolerant to high temperature stress. Among the 28 ESTs, we found 10 ESTs that have either insertion/deletion and/or single nucleotide polymorphism between the two cultivars. Those polymorphic sites were then targeted for the development of 10 PCR based markers. These molecular markers related to trehalose genes could be used not only to test their relationship with abiotic stress resistance in Chinese cabbage, but also the development of abiotic stress resistant cultivars using MAS.

• Journal of Crop Science and Biotechnology
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• v.11 no.1
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• pp.39-44
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• 2008

Korean cultivated bramble, which is known as Bokbunja-ddal-gi is regarded to be originated from Korea native Rubus coreanus. However, little scientific evidence and significant morphological differences between Korean cultivated bramble(KCB) and R. coreanus throw doubt on the ancestry of KCB. This study was carried out to obtain phylogenetic information on KCB by comparing its nuclear genomic background with those of R. coreanus, black(R. occidentalis) and red(R. idaeus) raspberry, blackberry(R. lanciniatus) and R. crataegifolius. A total of 99 random amplified polymorphic DNA(RAPD) markers were generated and used for phylogenetic analysis of 76 Rubus accessions. Accessions of each species were grouped into each distinct subclade by the RAPD markers at a similarity coefficient of about 0.59. The KCB subclade formed a clade with R. occidentalis and R. crataegifolius subclades at a similarity coefficient of 0.47. The R. coreanus subclade formed a clade with R. idaeus, R. lanciniatus and R. crataegifolius subclades at a similar similarity coefficient. Only one KCB accession from Hoengsung was included in R. coreanus subclade. The accession shows leaf and flower characteristics different from the rest of the KCB accessions. The phylogenetic relationship inferred from the RAPD markers suggests that the nuclear genomic background of KCB accessions which show morphological similarity to black raspberry is more closely related to black raspberry than to R. coreanus. This brings about the need for close scientific evaluations on the ancestry of KCB at both morphological and molecular levels.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.5
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• pp.622-626
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• 2006

Molecular characterization of Hallikar, the native cattle breed of Karnataka, was undertaken using 19 cattle specific, highly polymorphic microsatellite markers recommended by FAO. The genomic DNA was subjected to PCR amplification and alleles were resolved through six per cent denaturing PAGE with a 10 bp DNA ladder followed by silver staining. Genotyping of animals was done based on allele size. The number of alleles ranged from three to nine with allele sizes ranging from 102 bp to 294 bp. These alleles were distributed in the frequency range between 0.0306 and 0.8673 in the population. The mean observed number of alleles was $6.368{\pm}1.4225$. The mean observed and expected heterozygosities were $0.7515{\pm}0.1734$ and $0.7850{\pm}0.1381$, respectively. The high heterozygosity observed implies presence of higher genetic variability within Hallikar breed. The PIC (Polymorphism Information Content) values ranged from 0.2322 (ETH152) to 0.8654 (ETH225). The percentage of polymorphic loci obtained was 100 as all the 19 microsatellite markers were found to be polymorphic. Except for ETH152, all the other loci had high PIC values, indicating that these markers are highly informative for characterization of Hallikar breed. The population was tested for Hardy-Weinberg equilibrium at 19 microsatellite loci, and at 74 per cent of the loci the population was found to be in disequilibrium.

• Korean Journal of Plant Resources
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• v.24 no.6
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• pp.702-711
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• 2011

Buckwheat (Fagopyrum esculentum Moench), one of the minor crops grown in Korea belonging to the Polygonaceae family, is an annual crop widely cultivated in Asia, Europe, and America and has a character of outcrossing and self-incompatibility. The objective of this study was to analyze the genetic variability, phylogenetic relationships and population structure of buckwheat landraces of Korea using SSR markers. Ten microsatellite markers have been detected from a total of 79 alleles among the 179 buckwheat accessions were collected from Korea. The number of allele per marker locus ($N_A$) ranged from 2 (GB-FE-001, GB-FE-043 and GB-FE-055) to 31 (GB-FE-035) with an average of 7.9 alleles. GB-FE-035 was the most polymorphic with the highest PIC value 0.93. Major allele frequencies ($M_{AF}$) for the 10 polymorphic loci varied from 0.12 to 0.97 with a mean allele frequency of 0.57. The expected heterozygosity ($H_E$) values ranged from 0.05 to 0.94 with an average of 0.53. The observed heterozygosity ($H_O$) ranged from 0.06 to 0.92 with an average of 0.42. The overall polymorphic information contents (PIC) values ranged from 0.05 to 0.93 with an average of 0.48. The landrace accessions of buckwheat used in the present study were not distinctly grouped according to geographic distribution. The study concludes that the results revealed genetic differentiation was low according to the geographic region because of outcrossing and self-incompatibility. We reported that our analyses on the genetic diversity of common buckwheat cultivars of Korea were performed by using of microsatellite markers.

• Horticultural Science & Technology
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• v.16 no.1
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• pp.21-24
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• 1998

RAPD markers were analyzed in order to detect the genetic variation and diversity of the fifty-two melon lines. SDS extraction method produced more and purer DNA than CTAB method. RAPD reaction conditions were optimized as follows ; 10ng template DNA, 270nM primer, $200{\mu}M$ each of dATP, dCTP, dGTP and dTTP, $0.3{\mu}unit$ dynazyme and 10x buffer brought to $15{\mu}l$ final volume with distilled water. The adequate annealing temperature was $39^{\circ}C$ and forty cycles of amplification produced the best RAPD band patterns. Among a total of 123 bands from 12 random primers, 25 polymorphic bands(20%) were selected as reliable markers. The average number of polymorphic bands per primer was 2.1 among the 52 lines. Intragroup genetic relationship based on the marker difference was closer than intergroup genetic relationship. The 52 lines could be grouped into two major group (Korean landraces and melon lines) and then melon group subdivided into two subgroups (net melon lines and no-net melon). This result corresponded to morphological grouping. Eight RAPD markers separated the Korean landraces and melon groups and four RAPD markers separated net melon and no-net melon groups.

• International Journal of Industrial Entomology and Biomaterials
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• v.10 no.2
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• pp.79-86
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• 2005

Molecular markers have increasingly been used in plant genetic analysis, due to their obvious advantages over conventional phenotypic markers, as they are highly polymorphic, more in number, stable across different developmental stages, neutral to selection and least influenced by environmental factors. Among the PCR based marker techniques, ISSR is one of the simplest and widely used techniques, which involves amplification of DNA segment present at an amplifiable distance in between two identical microsatellite repeat regions oriented in opposite direction. Though ISSR markers are dominant like RAPD, they are more stable and reproducible. Because of these properties ISSR markers have recently been found using extensively for finger printing, pohylogenetic analysis, population structure analysis, varietal/line identification, genetic mapping, marker-assisted selection, etc. In mulberry (Morus spp.), ISSR markers were used for analyzing phylogenetic relationship among cultivated varieties, between tropical and temperate mulberry, for solving the vexed problem of identifying taxonomic positions of genotypes, for identifying markers associated with leaf yield attributing characters. As ISSR markers are one of the cheapest and easiest marker systems with high efficiency in generating polymorphism among closely related varieties, they would play a major role in mulberry genome analysis in the future.