• Title/Summary/Keyword: Polymorphic markers

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Use of Simple Sequence Repeat (SSR) Markers for Variety Identification of Tomato (Lycopersicon esculentum) (Simple Sequence Repeat (SSR) Marker를 이용한 토마토 품종 식별)

  • Kwon, Yong-Sham;Park, Eun-Kyung;Bae, Kyung-Mi;Yi, Seung-In;Park, Soon-Gi;Cho, Il-Ho
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.289-295
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    • 2006
  • This study was carried out to evaluate the suitability of simple sequence repeat (SSR) markers for varietal identification and genetic diversity in 28 commercial tomato varieties. The relationship between marker genotypes and 28 varieties was analyzed. Of the 219 pairs of SSR primers screened against ten tomato varieties, 18 pairs were highly polymorphic with polymorphism information content (PIC) ranging from 0.467 to 0.800. Among the polymorphic loci, two to nine SSR alleles were detected for each locus with an average of 3.3 alleles per locus. Genetic distances were estimated according to Jaccard's methods based on the probability that the amplified fragment from one genotype would be present in another genotype. These varieties were categorized into cherry and classic fruit groups corresponding to varietal types and genetic distance of cluster ranging from 0.35 to 0.97. The phonogram discriminated all varieties by marker genotypes. The SSR markers proved to be useful variety identification and genetic resource analysis of tomato.

Identification of Monoecious and Dioecious Plants of. Schisandra nigra Using the RAPD Markers (RAPD 표지인자를 이용한 흑오미자의 자웅동주 및 자웅이주 식물의 동정)

  • 이효연;한효심;이갑연;한상섭;정재성
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.309-313
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    • 1998
  • RAPD (Random Amplified Polymorphic DNA) analysis was conducted to Schisandra nigra plants in order to select the specific markers for monoecious and dioecious individuals. RAPD results using eighty random 10-mer primers revealed that S. nigra had a different banding pattern from S. chinensis and Kadsura japonica. When DNA isolated from leaves of monoecious and dioecious plants were used as PCR template, only five primers, OPA-17, OPA-19, OPB-03, OPB-09 and OFB-16, showed polymorphic band patterns. No variation in banding profiles within male or female individuals was observed when these five primers were used whereas three monoecious plants (No 1, No 2 and No 3) showed different banding patterns one another, A 750 bp segment was amplified by primer OPB-3 from male individuals. On the other hand, two segments, 950 bp and 1690 bp, with OPA-19 and 700 bp of segment with OPB-3 were amplified in female individuals. These result indicate that the specific buds of male and female S. nigra could be used as genetic markers for the early discrimination of male and female individuals.

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Reverse Random Amplified Microsatellite Polymorphism Reveals Enhanced Polymorphisms in the 3' End of Simple Sequence Repeats in the Pepper Genome

  • Min, Woong-Ki;Han, Jung-Heon;Kang, Won-Hee;Lee, Heung-Ryul;Kim, Byung-Dong
    • Molecules and Cells
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    • v.26 no.3
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    • pp.250-257
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    • 2008
  • Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

Development of SCAR Markers for the Authentication of Acori Rhizoma Based on the Analysis of RAPD and Multiplex-PCR (RAPD 분석과 multiplex-PCR을 이용한 석창포 감별용 SCAR 마커 개발)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Yeong;Choo, Byung-Kil;Kim, Ho-Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.3
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    • pp.162-169
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    • 2011
  • The rhizomes and herbal medicines originating from Acorus gramineus, A. calamus, A. tatarinowii, and A. gramineus var. pusilus, show significant similarity, and the correct identification of species is very difficult. Random Amplified Polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop a reliable method for identification of these four species. Several distinct SCAR markers were developed from species-specific RAPD amplicons for each species. Furthermore, a useful molecular marker was established for multiplex-PCR, in order to the four species could be distinguished concurrently. These markers allow efficient and rapid identification of closely-related Acorus species and will be useful for standardization of herbal medicines.

Assessment of genetic diversity of Prangos fedtschenkoi (Apiaceae) and its conservation status based on ISSR markers

  • Mustafina, Feruza U.;Kim, Eun Hye;Son, Sung-Won;Turginov, Orzimat T.;Chang, Kae Sun;Choi, Kyung
    • Korean Journal of Plant Taxonomy
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    • v.47 no.1
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    • pp.11-22
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    • 2017
  • Prangos fedtschenkoi (Regel et Schmalh.) Korovin (Apiaceae) is an endemic species for mountainous Middle Asia, which is both a rare and useful plant. Organic extractions from this species are being used in pharmaceutics and cosmetology. In recent years, P. fedtschenkoi distribution area has considerably decreased, presumably, resulting from human activities such as agriculture, construction works, overgrazing and collection from wild for pharmaceutic purposes. Six populations were found in Uzbekistan and their genetic divergence and differentiation were studied with 10 inter-simple sequence repeat (ISSR) markers, selected out of 101. Totally 166 amplified ISSR fragments (loci) were revealed, of which 164 were polymorphic. Relatively moderate level of polymorphism was found at population level with polymorphic bands ranging from 27.71% to 47.59%. Mean P = 39.05%, $N_a=1.40$, $N_e=1.25$, S.I. = 0.21, and $H_e=0.14$ were revealed for all loci across six populations. AMOVA showed higher variation among populations (62%) than within them (38%). The Bayesian model determined 5 clusters, or genetic groups. The posteriori distribution of the Theta II estimator detected full model identifying high inbreeding, intensified by low gene flow (Nm = 0.3954). Mantel test confined population 6 as distinct cluster corresponding to geographic remoteness (R = 0.5137, $p{\leq}0.005$). Results were used as the bases for developing conserve measures to restore populations.

Confirmation of $F_1$ Hybridity Using RAPD Markers in Soybean

  • Chung, Jong-Il;Ko, Mi-Suk;Shim, Jung-Hyun;Kim, Seok-Hyeon;Kang, Jin-Ho
    • Plant Resources
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    • v.2 no.1
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    • pp.22-25
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    • 1999
  • Molecular markers are useful to confirm the hybridity of F1 plant derived from cross of two homozygous parents with similar morphological traits. RAPD markers were used to test F1 hybrid plant obtained from cross of two homozygous soybean (Glycine max) parents. Fl plant for cross I was made from the mating of Hobbit87 (female) and L63-1889 (male) and Fl plant for cross II was obtained from the mating of H1053 (female) and L63-1889 (male). Selfing plant per each cross was also obtained. Among 20 Operon primers used, OPA04 and OPA09 show polymorphism between cross I and II parent. Band in size 1Kb of OPA04 and 2.1Kb of OPA09 primer was polymorphic band. This fragment identified Fl hybrid plant and selfing plant in cross I and II. Female parent Hobbit87 in cross I and H1053 in cross II has no this fragment (recessive allele). However, male parent L63-1889 and Fl hybrid plant in cross I and II has this size of polymorphic band (dominant allele). This indicated that Fl hybrid and selfing plants were detected by RAPD marker before phenotypic marker would be used to identify Fl hybridity. Amplification products of selfing plant for cross I and II were completely same to the those of female parent. When mature, flower color of Fl hybrid plant in cross I and II was purple and flower color of selfing plant in cross I and II was white. Purple flower is dominant trait. Fl hybridity was successfully detected at very early growth stage using RAPD marker. Therefore, RAPD marker can be used broadly to confirm Fl hybridity in many crops.

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New Sources of Resistance and Identification of DNA Marker Loci for Sheath Blight Disease Caused by Rhizoctonia solani Kuhn, in Rice

  • Pachai, Poonguzhali;Ashish, Chauhan;Abinash, Kar;Shivaji, Lavale;Spurthi N., Nayak;S.K., Prashanthi
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.572-582
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    • 2022
  • Sheath blight disease caused by the necrotrophic, soilborne pathogen Rhizoctonia solani Kuhn, is the global threat to rice production. Lack of reliable stable resistance sources in rice germplasm pool for sheath blight has made resistance breeding a very difficult task. In the current study, 101 rice landraces were screened against R. solani under artificial epiphytotics and identified six moderately resistant landraces, Jigguvaratiga, Honasu, Jeer Sali, Jeeraga-2, BiliKagga, and Medini Sannabatta with relative lesion height (RLH) range of 21-30%. Landrace Jigguvaratiga with consistent and better level of resistance (21% RLH) than resistant check Tetep (RLH 28%) was used to develop mapping population. DNA markers associated with ShB resistance were identified in F2 mapping population developed from Jigguvaratiga × BPT5204 (susceptible variety) using bulk segregant analysis. Among 56 parental polymorphic markers, RM5556, RM6208, and RM7 were polymorphic between the bulks. Single marker analysis indicated the significant association of ShB with RM5556 and RM6208 with phenotypic variance (R2) of 28.29 and 20.06%, respectively. Co-segregation analysis confirmed the strong association of RM5556 and RM6208 located on chromosome 8 for ShB trait. This is the first report on association of RM6208 marker for ShB resistance. In silico analysis revealed that RM6208 loci resides the stearoyl ACP desaturases protein, which is involved in defense mechanism against plant pathogens. RM5556 loci resides a protein, with unknown function. The putative candidate genes or quantitative trait locus harbouring at the marker interval of RM5556 and RM6208 can be further used to develop ShB resistant varieties using molecular breeding approaches.

Determination of Genetic Diversity Using 15 Simple Sequence Repeats Markers in Long Term Selected Japanese Quail Lines

  • Karabag, Kemal;Balcioglu, Murat Soner;Karli, Taki;Alkan, Sezai
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.12
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    • pp.1696-1701
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    • 2016
  • Japanese quail is still used as a model for poultry research because of their usefulness as laying, meat, and laboratory animals. Microsatellite markers are the most widely used molecular markers, due to their relative ease of scoring and high levels of polymorphism. The objective of the research was to determine genetic diversity and population genetic structures of selected Japanese quail lines (high body weight 1 [HBW1], HBW2, low body weight [LBW], and layer [L]) throughout 15th generations and an unselected control (C). A total of 69 individuals from five quail lines were genotyped by fifteen microsatellite markers. When analyzed profiles of the markers the observed ($H_o$) and expected ($H_e$) heterozygosity ranged from 0.04 (GUJ0027) to 0.64 (GUJ0087) and 0.21 (GUJ0027) to 0.84 (GUJ0037), respectively. Also, $H_o$ and $H_e$ were separated from 0.30 (L and LBW) to 0.33 (C and HBW2) and from 0.52 (HBW2) to 0.58 (L and LBW), respectively. The mean polymorphic information content (PIC) ranged from 0.46 (HBW2) to 0.52 (L). Approximately half of the markers were informative ($PIC{\geq}0.50$). Genetic distances were calculated from 0.09 (HBW1 and HBW2) to 0.33 (C and L). Phylogenetic dendrogram showed that the quail lines were clearly defined by the microsatellite markers used here. Bayesian model-based clustering supported the results from the phylogenetic tree. These results reflect that the set of studied markers can be used effectively to capture the magnitude of genetic variability in selected Japanese quail lines. Also, to identify markers and alleles which are specific to the divergence lines, further generations of selection are required.

Development and Validation of Single Nucleotide Polymorphism (SNP) Markers from an Expressed Sequence Tag (EST) Database in Olive Flounder (Paralichthys olivaceus)

  • Kim, Jung Eun;Lee, Young Mee;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Kim, Kyung-Kil
    • Development and Reproduction
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    • v.18 no.4
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    • pp.275-286
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    • 2014
  • To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Identification of Korean Strawberry Cultivars using DNA markers (DNA 표지를 이용한 딸기 국내 육성 품종 판별)

  • Cho, Kang-Hee;Rho, Il Rae;Cho, Yong Seop;Park, Pue-Hee
    • Korean Journal of Breeding Science
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    • v.40 no.4
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    • pp.401-407
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    • 2008
  • This study was conducted to develop the DNA markers for identification of the strawberry cultivars in Korea and Japan. We developed fifteen cleaved amplified polymorphic sequence (CAPS) markers based on the Fragaria gene sequences. Among them six CAPS markers showed polymorphism exclusively in one cultivar. Five CAPS markers (ANR-MspI, ANR-BamHI, ACO-HinfI, DFR-AseI, FGT-MspI) provided enough polymorphism to identify eight Korean strawberry cultivars except for 'Maehyang' and 'Sunhong'. To complement the fifteen CAPS markers, we selected another fifteen sequence-related amplified polymorphism (SRAP) and one of them, me1/em5_460bp marker, made it possible to discriminate between 'Maehyang' and 'Sunhong'. Therefore, application of the five CAPS markers and one SRAP marker were sufficient to identify the nineteen Korean and Japanese strawberry cultivars. These markers could be used practically for cultivar identification of Korean and Japanese strawberry.