• Title/Summary/Keyword: Simple Sequence Repeat

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Development of Multiplex Microsatellite Marker Set for Identification of Korean Potato Cultivars (국내 감자 품종 판별을 위한 다중 초위성체 마커 세트 개발)

  • Cho, Kwang-Soo;Won, Hong-Sik;Jeong, Hee-Jin;Cho, Ji-Hong;Park, Young-Eun;Hong, Su-Young
    • Horticultural Science & Technology
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    • v.29 no.4
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    • pp.366-373
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    • 2011
  • To analyze the genetic relationships among Korean potato cultivars and to develop cultivar identification method using DNA markers, we carried out genotyping using simple sequence repeats (SSR) analysis and developed multiplex-SSR set. Initially, we designed 92 SSR primer combinations reported previously and applied them to twenty four Korean potato cultivars. Among the 92 SSR markers, we selected 14 SSR markers based on polymorphism information contents (PIC) values. PIC values of the selected 14 markers ranged from 0.48 to 0.89 with an average of 0.76. PIC value of PSSR-29 was the lowest with 0.48 and PSSR-191 was the highest with 0.89. UPGMA clustering analysis based on genetic distances using 14 SSR markers classified 21 potato cultivars into 2 clusters. Cluster I and II included 16 and 5 cultivars, respectively. And 3 cultivars were not classified into major cluster group I and II. These 14 SSR markers generated a total of 121 alleles and the average number of alleles per SSR marker was 10.8 with a range from 3 to 34. Among the selected markers, we combined three SSR markers, PSSR-17, PSSR-24 and PSSR-24, as a multiplex-SSR set. This multiplex-SSR set used in the study can distinguish all the cultivars with one time PCR and PAGE (Polyacrylamide gel electrophoresis) analysis and PIC value of multiplex-SSR set was 0.95.

Inter Simple Sequence Repeats (ISSR) Marker Analysis of Genetic Diversity in Korean Phasianus colchicus karpowi and Genetic Relationships Among Subspecies of Phasianus spp. (Inter Simple Sequence Repeats (ISSR) 표지자를 이용한 한국꿩의 유전적 다양성 및 아종간의 유연관계 분석)

  • Yoon, Seong-Il
    • Korean Journal of Environmental Biology
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    • v.26 no.2
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    • pp.66-75
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    • 2008
  • The level of genetic diversity and genetic relationships among Korean ring-necked pheasant (Phasianus colchicus karpowi) habitat and subspecies have been investigated based on Inter Simple Sequence Repeat (ISSR) markers. Wild and domesticated Korean ring-necked pheasant, hybrids between domesticated Korean ring-necked and foreign subspecies, and four foreign subspecies; Chinese ring-necked (P. c. torquatus), Melanistic mutant (P. c. mut. tenebrosus), XL White (P. c. mut) and Southern green (P. c. versicolor) were used for comparison. On the basis of the results of AMOV A, 94.08% of genetic diversity in Korean ring-necked was allocated among individuals within habitat differences. Estimate of $\Phi$st, which represents the degree of genetic differentiation among habitats was 5.9%. Based on the dendrogram reconstructed by UPGMA, Yangpyung habitat of the eight habitats turned out to be distinct from others habitat. Interestingly, domesticated Korean ring-necked and hybrid mixture showed closer genetic relationship with four foreign subspecies than Korean ring-necked. As a consequence of AMOVA, 96.63% of genetic diversity in four foreign subspecies was allocated among individuals within subspecies. Estimate of $\Phi$st representing the degree of genetic differentiation among subspecies was 3.4%, which was lower than that among habitats of Korean ring-necked. The lower level of genetic difference among four foreign subspecies showed that these subspecies were genetically closer even though they were morphologically classified into four different subspecies. When seven habitats of Korean ring-necked pheasant and four foreign subspecies were divided into Korean and Foreign Pheasant Groups, respectively, more than 17% of genetic diversity was allocated between groups (about 4% among habitats/subspecies within groups). This observation implied that Korean ring-necked pheasant is genetically quite different from four foreign subspecies. On the basis of cluster analysis, three foreign subspecies (Chinese ring-necked pheasant, Melanistic mutant pheasant, and XL White pheasant) formed a distinct group with domesticated Korean ring-necked pheasant and hybrid mixture at 98% confidence interval.

Analysis of Genetic Polymorphism and Relationship of Korean Ginseng Cultivars and Breeding Lines using EST-SSR Marker (EST-SSR 마커를 이용한 인삼 품종과 육성계통의 유전적 다형성 및 유연관계 분석)

  • Bang, Kyong-Hwan;Seo, A-Yeon;Chung, Jong-Wook;Kim, Young-Chang;Jo, Ick-Hyun;Kim, Jang-Uk;Kim, Dong-Hwi;Cha, Seon-Woo;Cho, Yong-Gu;Kim, Hong-Sig
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.4
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    • pp.277-285
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    • 2012
  • In this study, Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Polymorphic and reproducible bands were produced by 14 primers out of total 30 primers used in this study. Fourteen EST-SSR loci generated a total of 123 bands. Amplified PCR products showed the highly reproducible banding patterns at 110~920 bp. The number of amplified bands for each EST-SSR primers ranged from 2 to 19 with a mean of 8.8 bands. P26 and P35 primers showed 13 and 12 banding patterns, respectively. The number of alleles for each EST-SSR locus ranged from 1.67 to 2.00 with a mean of 1.878 alleles. P34 and P60 primers showed the highest and the lowest genetic polymorphism, respectively. Cluster analysis based on genetic similarity estimated by EST-SSR markers classified Korean cultivars and breeding lines into 4 groups. Group included Gopoong and Chunpoong and 9 breeding lines (55%), group included 2 breeding lines (10%), group included 3 breeding lines (15%), group included Gumpoong and 3 breeding lines (20%). Consequently, the EST-SSR marker developed in this study may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.

Distribution and Frequency of SSR Motifs in the Chrysanthemum SSR-enriched Library through 454 Pyrosequencing Technology (국화 SSR-enriched library에서 SSR 반복염기의 분포 및 빈도)

  • Moe, Kyaw Thu;Ra, Sang-Bog;Lee, Gi-An;Lee, Myung-Chul;Park, Ha-Seung;Kim, Dong-Chan;Lee, Cheol-Hwi;Choi, Hyun-Gu;Jeon, Nak-Beom;Choi, Byung-Jun;Jung, Ji-Youn;Lee, Kyu-Min;Park, Yong-Jin
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.546-551
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    • 2011
  • Chrysanthemums, often called mums or chrysanths, belong to the genus Chrysanthemum, which includes about 30 species of perennial flowering plants in the family Asteraceae. We extracted DNA from Dendranthema grandiflorum ('Smileball') to construct a simple sequence repeat (SSR)-enriched library, using a modified biotin-streptavidin capture method. GS FLX (Genome Sequencer FLX System which provides the flexibility to perform the broad range of applications) sequencing (at the 1/8 run specification) resulted in 18.83 mega base pairs (Mbp) with an average read length of 280.06 bp. Sequence analyses of all SSR-containing clones revealed a predominance of di-nucleotide motifs (16,375, 61.5%) followed by tri-nucleotide motifs (6,616, 24.8%), tetra-nucleotide motifs (1,674, 6.3%), penta-nucleotide motifs (1,283, 4.8%), and hexa-nucleotide motifs (693, 2.6%). Among the di-nucleotide motifs, the AC/CA class was the most frequently identified (93.5% of all di-nucleotide types), followed by the GA/AG class (6.1%), the AT/TA class (0.4%), and the CG/GC class (0.03%). When we analyzed the distribution of different repeat motifs and their respective numbers of repeats, regardless of the motif class, of 100 SSR markers, we found a higher number of di-nucleotide motifs with 70 to 80 repeats; we also found two di-nucleotide motifs with 83 and 89 repeats, respectively, but their product lengths were within optimum size (297 and 300 bp). In future work, we will screen for polymorphisms of possible primer pairs. The results will provide a useful tool for assessing molecular diversity and investigating the population structure among and within Chrysanthemum species.

Genetic Diversity and Structure of the Korean Endemic Species, Coreanomecon hylomeconoides Nakai, as Revealed by ISSR markers (한국 특산식물 매미꽃(Coreanomecon hylomeconoides Nakai) 집단의 유전다양성 및 구조)

  • Son, Sung-Won;Chung, Jae-Min;Kim, Eun-Hye;Choi, Kyoung-Su;Park, SeonJoo
    • Korean Journal of Plant Resources
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    • v.26 no.2
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    • pp.310-319
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    • 2013
  • The genetic diversity and structure of eight populations of Coreanomecon hylomeconoides Nakai, an endemic Korean plant, were investigated using 50 ISSR loci from eight primers. The average percentage of polymorphic loci was 47.3%. The Shannon's index (SI=0.218) and gene diversity (h=0.142) were relatively lower than those of other long-lived perennials. The Sancheong (SI=0.233, h=0153), Gwangyang (SI=0.263, h=0.171), and Suncheon (SI=0.241, h=0.159) populations showed greater genetic diversity than the Namhae and Gwangju populations, which are on the edge of the distribution. Analysis of molecular variance (AMOVA) showed that 18% of the total variation could be attributed to differences among populations, and 82% to differences within populations, indicating moderate gene flow among adjacent populations. These results were supported by value of Nm (2.184). The UPGMA conducted using the genetic distance and Bayesian cluster analysis showed a remarkable geographic trend structured into east and west regions. Overall, the results indicate that the Sancheong and Gwangyang populations, which had a large population size and higher degree of genetic diversity, should be the focus of in situ conservation.

Genetic Diversity and Population Structure of Korean Mint Agastache rugosa (Fisch & Meyer) Kuntze (Lamiaceae) Using ISSR Markers

  • Kang, Man Jung;Sundan, Suresh;Lee, Gi An;Ko, Ho Cheol;Chung, Jong Wook;Huh, Yun Chan;Gwag, Jae Gyun;Oh, Se Jong;Kim, Yeon Gyu;Cho, Gyu Taek
    • Korean Journal of Plant Resources
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    • v.26 no.3
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    • pp.362-369
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    • 2013
  • Agastache rugosa, a member of the mint family (Labiatae), is a perennial herb widely distributed in East Asian countries. It is used in traditional medicine for the treatment of cholera, vomiting, and miasma. This study assessed the genetic diversity and population structures on 65 accessions of Korean mint A. rugosa germplasm based on inter simple sequence repeat (ISSR) markers. The selected nine ISSR primers produced reproducible polymorphic banding patterns. In total, 126 bands were scored; 119 (94.4%) were polymorphic. The number of bands generated per primer varied from 7 to 18. A minimum of seven bands was generated by primer 874, while a maximum of 18 bands was generated by the primer 844. Six primers (815, 826, 835, 844, 868, and 874) generated 100% polymorphic bands. This was supported by other parameters such as total gene diversity ($H_T$) values, which ranged from 0.112 to 0.330 with a mean of 0.218. The effective number of alleles ($N_E$) ranged from 1.174 to 1.486 with a mean value of 1.351. Nei's genetic diversity (H) mean value was 0.218, and Shannon's information index (I) mean value was 0.343. The high values for total gene diversity, effective number of alleles, Nei's genetic diversity, and Shannon's information index indicated substantial variations within the population. Cluster analysis showed characteristic grouping, which is not in accordance with their geographical affiliation. The implications of the results of this study in developing a strategy for the conservation and breeding of A. rugosa and other medicinal plant germplasm are discussed.

Genetic relationships of pear germplasms using simple sequence repeat marker (SSR 마커를 이용한 배 유전자원의 유연관계)

  • Chun, Jae An;Cho, Kang Hee;Kim, Se Hee;Lee, Han-Chan;Choi, In Myong;Park, Seo Jun
    • Journal of Plant Biotechnology
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    • v.43 no.4
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    • pp.466-472
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    • 2016
  • This study analyzed the genetic diversity of 115 pear germplasms using 15 SSR markers. Three to forty-one SSR alleles were detected for each locus with an average of 16 alleles per locus. The average availability of markers was 0.966. The average observed heterozygosity ($H_{obs}$) was 0.603 (range: 0.140 to 0.929). The average expected heterozygosity ($H_{exp}$) was 0.718 (range: 0.463 to 0.904). The average polymorphism information content (PIC) was 0.692 (range: 0.403 to 0.897). The genetic relationships of pear germplasms were classified into two major groups by geographic origins and genetic characteristics according to genetic distance. The first group was composed of European pear belonging to Pyrus communis. The second group consisted of P. pyrifolia, P. ussuriensis, P. bretschneideri, P. betulaefolia, P. calleryana, interspecific hybrids, and unclear germplasms. The results of this study suggest that genotype analysis of pear germplasms using SSR markers can identify the genetic diversity of germplasms, and can be used to provide basic information for pear breeding.

Isolation of Bacillus subtilis GS-2 Producing γ-PGA from Ghungkukjang Bean Paste and Identification of γ-PGA (청국장으로부터 분리한 Poly(γ-glutamic acid)를 생산하는 균주 Bacillus subtilis GS-2의 분리 및 γ-PGA의 확인)

  • Bang, Byung-Ho;Jeong, Eun-Ja;Rhee, Moon-Soo;Kim, Yong-Min;Yi, Dong-Heui
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.1-6
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    • 2011
  • ${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.

Genetic Diversity of Quercus gilva in Je-ju Island (제주도 개가시나무의 유전구조와 유전적 다양성)

  • Kim, Go-Un;Jang, Kyoung-Soo;Lim, Hyungwoo;Kim, Eun-Hye;Lee, Kye-Han
    • Journal of Korean Society of Forest Science
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    • v.107 no.2
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    • pp.151-157
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    • 2018
  • This study was to analyze the genetic diversity of Quercus gilva Blume growing in Jeju Island for developing a preservation strategy. We examined the genetic diversity and structure using 6 ISSR primers and investigated 67 polymorphic ISSR amplicons in 80 trees distributed among five populations. The average of proportion of polymorphic loci were 93%, the average level of Shannon's information index was 0.237, and Nei's genetic diversity was 0.156. According to the analysis of the molecular variance (AMOVA), $F_{st}$ was 0.169 indicating there was a genetic variation among five populations. 17% of the total variation was allocated among the five populations, while the other 83% of the total variation was in individual trees in each population. The result could be due to the uneven number of trees among the five populations. Based on these results, the preservation strategy could be developed, for examples, considering for designation as "forest genetic resources conservation area" about the habitat, monitoring continuously, fostering the growth of seedling, ex situ preservation of genetic resources, and comparing the differences of environmental and genetic characteristic with population in ex situ.

Identification of Quantitative Trait Loci Associated with Leaf Length. Width and Length/width Ratio in Two Recombinant Inbred Lines of Soybean (Glycine max L.) (두 집단의 재조합 근친교잡 계통 (RIL) 콩에서 엽장과 엽폭 및 장폭비와 관련된 양적헝질 유전자좌 분석)

  • Kim, Hyeun-Kyeung;Kang, Sung-Taeg
    • Journal of Life Science
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    • v.14 no.5
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    • pp.821-828
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    • 2004
  • The increasing apparent photosynthetic rate per leaf area may improve seed yield in soybean. Leaf area, length and width are related to the photosynthetic capability of the plant. In this study, two populations derived from the cross of Keunolkong, Shinpaldalkong and Iksanl0 were evaluated with simple sequence repeat (SSR) markers to identify length, width and length/width ratio of leaf. Leaf length/width ratio were significantly negative correlation with leaf width in K/S and K/I populations. In the K/S population, two minor QTLs for leaf length (LL) were found on LG Dlb+W and 1. Two QTLs on LG J and L were related to LL in K/I population. Two and three minor QTLs were identified in leaf width with total phenotypic variation of 13% and 18.04 in K/S and K/I populations, respectively. The leaf length/width ratio, two QTLs on LG I and L, and three QTLs on LG Cl, E and L were related to K/S and K/I populations, respectively. Thus it is assumed that the leaf traits are very much dependent on the genotype used and different breeding approach should be considered for the selection of favorite leaf traits in soybean breeding programs.