• Title/Summary/Keyword: Simple Sequence Repeat

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Interspecific relationships of Korean Viola based on RAPD, ISSR and PCR-RFLP analyses (RAPD, ISSR과 PCR-RFLP를 이용한 한국산 제비꽃속(Viola)의 종간 유연관계)

  • Yoo, Ki-Oug;Lee, Woo-Tchul;Kwon, Oh-Keun
    • Korean Journal of Plant Taxonomy
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    • v.34 no.1
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    • pp.43-61
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    • 2004
  • Molecular taxonomic studies were conducted to evaluate interspecific relationships in Korean Viola 34 taxa including two Japanese populations using RAPD(randornly amplified polymorphic DNA), ISSR(inter simple sequence repeat) and PCR-RFLP(restriction fragment length polymorphism) analysis. Only six and four primers out of 40 arbitrary and 12 ISSR primers were screened for 34 taxa, and were revealed 70 (98.6%) and 28 (96.6%) polymorphic bands, respectively. Fifteen restriction endonucleases produced 80 restriction sites and size variations from the large single copy region of cpDNA, 16 (20%) of which were polymorphic. The separate analyses from the RAPD, ISSR and PCR-RFLP data were incongruent in the relationships among 34 taxa, but combined data was in accordance with previous infrageneric classification system based on morphological characters, especially the subsection and series level. Section Chamaemelanium placed between subsect. Patellares and Vagimtae of section Nomimium was not formed as a distinct group. Viola alb ida complex including three very closely related taxa was recognized independent group within subsect. Patellares in combined data tree. This result strongly suggested that they should be treated to series Pinmtae. RAPD analysis was very useful to clarify the interspecific relationships among the species of Korean Viola than ISSH and PCR-RFLP analyses.

Genetic diversity and geographic differentiation in the endangered Primula farinosa subsp. modesta, a subalpine endemic to Korea (한반도 아고산대 특산·희귀식물 설앵초의 유전적 다양성과 지리적 분화)

  • Chung, Jae-Min;Son, Sung-Won;Kim, Sang-Yong;Park, Gwang-Woo;Kim, Sung-Shik
    • Korean Journal of Plant Taxonomy
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    • v.43 no.3
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    • pp.236-243
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    • 2013
  • Many plant species in subalpine regions are under threat of extinction as a result of climate change. In this study, the genetic diversity and geographic differentiation of three regions and six populations of Primula farinosa subsp. modesta (Bisset & Moore) Pax in Korea were assessed using the ISSR (Inter Simple Sequence Repeat) marker. The average genetic diversity (P = 60.62, SI = 0.299, h = 0.190) was relatively lower than that of other long-lived perennials, even though it is a self-incompatible species. AMOVA analysis showed that 50% of the total genetic diversity was partitioned among regions and Bayesian cluster analysis showed some remarkable geographic trends that were structured into 2 or 3 regions, suggesting limited gene flow among regions. Considering the population fragmentation, low level genetic diversity, and high genetic differentiation, it is essential to establish in situ and ex situ conservation strategies for P. farinosa subsp. modesta.

Genetic diversity of Millettia japonica in Korea as revealed by ISSR analysis (ISSR 분석으로 살펴본 애기등의 유전적 다양성)

  • Kim, Na-Rae;Kim, Yong-In;Lee, Jung-Hoon;Kim, Young-Dong
    • Korean Journal of Plant Taxonomy
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    • v.43 no.4
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    • pp.267-273
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    • 2013
  • This study employed inter-simple-sequence repeat (ISSR) to assess genetic variation among 189 individuals representing 10 populations (nine in Korea and one in Japan) of Millettia japonica, which has recently been lifted from the endangered species of Korea. The calculated Shannon's information index value (I = 0.2689) of the species was appreciable and was higher than other endangered leguminous woody taxa. Gochang (I = 0.2968), Namhae (I = 0.2951), and Mt. Toham (I = 0.2823) populations showed relatively high genetic diversity, whereas the Kyushu (in Japan) population (I = 0.2487) exhibited the lowest. The results of an analysis of molecular variance indicated that 86.49% of the diversity was attributed to within populations, and 13.51% to differences among populations, suggesting that M. japonica populations do not have significant geographic differentiation and that the gene flow between populations exists to some extent (Nm = 1.8446). Continuous habitat monitoring should be conducted to conserve genetic diversity of M. japonica, particularly for those populations with relatively high genetic diversity. Selection of many individuals from the populations in Gochang, Namhae, and Mt. Toham is thought to be an appropriate strategy for ex situ conservation of M. japonica in Korea.

A Study on the Genetic Variations of Tricholoma matsutake Collected from Eleven Sites of Korea Using I-SSR PCR (I-SSR PCR을 이용한 한국의 11개 주요 산지에서 채집한 송이의 유전변이에 관한 연구)

  • Cho, Duck-Hyun;Lee, Kyung-Joon;Han, Sim-Hee
    • The Korean Journal of Mycology
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    • v.28 no.1
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    • pp.32-37
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    • 2000
  • The objectives of this study was to identify genetic variations of Tricholoma matsutake (S. Ito et Imai) Sing. growing in different geographic ranges in South Korea. Mushrooms were collected during fruiting seasons from 1994 to 1997 from 11 major sites which included four sites (Bonghwa, UIjin, Goryoung, and Chungdo) in Kyongbuk Province, three sites (Changnyung, Hadong, and Hamyang) in Kyongnam Province, two sites (Yangyang and Inje) in Kangwon Province, one site (Goisan) in Choongbuk Province, and one site (Namwon) in Chonbuk Province. Two mushrooms each from three to eight shiros in each sites were collected. Genetic characteristics were analyzed by Inter-Simple Sequence Repeat Polymerase Chain Reaction (I-SSR PCR) method using six primers. With a total of 131 DNA bands identified, Nei's genetic distance and UPGMA tree were constructed. It was estimated that genetic variations between sites amounted to 12.9%, while 87.1% of total variation was explained by variations among individuals within sites. The cluster analysis indicated that the eleven major sites were clustered into four groups, group I (Yangyang, Hamyang, Inje, Hadong and UIjin), group II (Changnyung, Namwon and Chungdo), group III (Goryoung), and group IV (Bonghwa and Goisan). It is concluded that matsutake mushrooms in South Korea have a considerable degree of genetic variations between major sites.

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Simple Sequence Repeat Markers Linked to Quantitative Trait Loci Controlling Seed Weight, Protein and Oil Contents in Soybean (콩에서 종실의 무게와 oil 및 단백질 함량을 조절하는 양적 형질 유전자좌와 연관된 simple sequence repeat marker)

  • Kim, Hyeun-Kyeung;Kang, Sung-Taeg;Choung, Myoung-Gun;Jung, Chan-Sik;Oh, Ki-Won;Baek, In-Youl;Son, Beung-Gu
    • Journal of Life Science
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    • v.16 no.6
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    • pp.949-954
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    • 2006
  • Soybean [Glycine max (L.) Merr.] is an important crop, accounting for 48% of the world market in oil crops. Improvement of the quality and quantity of soybean seed constituents is one of the most important objectives in soybean breeding. Protein content and seed size are important properties to determine the quality of tofu and soy sprouts respectively. The objective of this study was to identify quantitative trait loci (QTLs) that control seed weight, protein and oil content in soybean. The 117 $F_{2:10}$ recombinant inbred lines (RlL) developed from a cross of 'Keunolkong' and 'Shinpaldalkong' were used. Narrow-sense heritability estimates based on a plot mean on seed weight, protein and oil content were 0.8, 0.78 and 0.71, respectively. Four independent QTLs for seed weight were identified from linkage group (LG) F, I and K. Five QTL for protein content were located on LG D1b, E, H, I and L. Oil content was related with six QTLs located on LG D1b, E, G, I, J and N. Protein and oil content have three common QTLs on LG D1b, E and I. Thus, we identified major loci improving soybean seed quality.

A Database of Simple Sequence Repeat (SSR) Marker-Based DNA Profiles of Citrus and Related Cultivars and Germplasm (SSR Marker를 이용한 감귤속 품종 및 유전자원에 대한 DNA Profile Data Base 구축)

  • Hong, Jee-Hwa;Chae, Chi-Won;Choi, Keun-Jin;Kwon, Yong-Sham
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.142-153
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    • 2016
  • The present study investigated identification of cultivars through phylogenetic analysis of 108 Citrus varieties and related cultivars using simple sequence repeat (SSR) markers. Two hundred three SSR primer pairs were used to detect polymorphic markers among 8 Citrus cultivars consisting of 4 mandarins, 1 orange, 1 tangor, 1 tangelo, and 1 pumelo. Eighteen SSR primer pairs were reproducible and showed highly polymorphic alleles. These markers were applied to assess genetic variations of the 108 varieties. Each marker detected 5-14 alleles, with an average of 9.28. The polymorphism information content varied from 0.417 to 0.791 with an average of 0.706. Cluster analysis with SSR markers resulted in 13 major groups reflecting cultivar types and pedigree information. Twelve orange cultivars in the $I-1^{st}$ sub-cluster and 23 mandarin cultivars in the $II-1^{st}$ sub-cluster, respectively, were not discriminated using the SSR markers. This could be due to narrow genetic backgrounds originated through bud mutation or nucellars seedlings. The SSR profile database of Citrus cultivars will be useful as a tool for protection of plant breeders' intellectual property rights in addition to assessing genetic diversity in Citrus cultivars and germplasms.

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.

High-density genetic mapping using GBS in Chrysanthemum

  • Chung, Yong Suk;Cho, Jin Woong;Kim, Changsoo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.57-57
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    • 2017
  • Chrysanthemum is one of the most important floral crop in Korea produced about 7 billion dollars (1 billion for pot and 6 billion for cutting) in 2013. However, it is difficult to breed and to do genetic study because 1) it is highly self-incompatible, 2) it is outcrossing crop having heterozygotes, and 3) commercial cultvars are hexaploid (2n = 6x = 54). Although low-density genetic map and QTL study were reported, it is not enough to apply for the marker assisted selection and other genetic studies. Therefore, we are trying to make high-density genetic mapping using GBS with about 100 $F_1s$ of C. boreale that is oHohhfd diploid (2n = 2x = 18, about 2.8Gb) instead of commercial culitvars. Since Chrysanthemum is outcrossing, two-way pseudo-testcross model would be used to construct genetic map. Also, genotype-by-sequencing (GBS) would be utilized to generate sufficient number of markers and to maximize genomic representation in a cost effective manner. Those completed sequences would be analyzed with TASSEL-GBS pipeline. In order to reduce sequence error, only first 64 sequences, which have almost zero percent error, would be incorporated in the pipeline for the analysis. In addition, to reduce errors that is common in heterozygotes crops caused by low coverage, two rare cutters (NsiI and MseI) were used to increase sequence depth. Maskov algorithm would also used to deal with missing data. Further, sparsely placed markers on the physical map would be used as anchors to overcome problems caused by low coverage. For this purpose, were generated from transcriptome of Chrysanthemum using MISA program. Among those, 10 simple sequence repeat (SSR) markers, which are evenly distributed along each chromosome and polymorphic between two parents, would be selected.

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Development of a Simple Method to Determine the Mouse Strain from Which Cultured Cell Lines Originated

  • Yoshino, Kaori;Saijo, Kaoru;Noro, Chikako;Nakamura, Yukio
    • Interdisciplinary Bio Central
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    • v.2 no.4
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    • pp.14.1-14.9
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    • 2010
  • Misidentification of cultured cell lines results in the generation of erroneous scientific data. Hence, it is very important to identify and eliminate cell lines with a different origin from that being claimed. Various methods, such as karyotyping and isozyme analysis, can be used to detect inter-species misidentification. However, these methods have proved of little value for identifying intra-species misidentification, and it will only be through the development and application of molecular biological approaches that this will become practical. Recently, the profiling of microsatellite variants has been validated as a means of detecting gene polymorphisms and has proved to be a simple and reliable method for identifying individual cell lines. Currently, the human cell lines provided by cell banks around the world are routinely authenticated by microsatellite polymorphism profiling. Unfortunately, this practice has not been widely adopted for mouse cells lines. Here we show that the profiling of microsatellite variants can be also applied to distinguish the commonly used mouse inbred strains and to determine the strain of origin of cultured cell lines. We found that approximately 4.2% of mouse cell lines have been misidentified; this is a similar rate of misidentification as detected in human cell lines. Although this approach cannot detect intra-strain misidentification, the profiling of microsatellite variants should be routinely carried out for all mouse cell lines to eliminate inter-strain misidentification.

Diversity analysis of japonica rice using microsatellite markers (Microsatellite를 이용한 자포니카 벼의 다양성 분석)

  • Xiao, Luo;Balkunde, Sangshetty;Yang, Paul;Lee, Hyun-Sook;Ahn, Sang-Nag
    • Korean Journal of Agricultural Science
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    • v.39 no.1
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    • pp.9-15
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    • 2012
  • The study was conducted to evaluate the genetic similarity among commercial japonica rice varieties in Korea and China and to develop markers to differentiate between japonica cultivars developed in Korea and China. The genetic similarity and cluster of 38 accessions were analyzed using 47 SSR(simple sequence repeat) markers. The number of alleles by 47 SSR markers ranged from 2 to 9 with an average of 3.6. A total of 169 alleles were detected among these tested rice varieties. The PIC value varied from 0.05 to 0.79 with an average of 0.44. The Chinese japonica cultivars could be differentiated from the japonica cultivars in Korea by combining 2 SSR markers, RM223 and RM266. Cluster analysis showed that 38 tested varieties could be distinguished into japonica and indica based on the genetic distance.