• Title/Summary/Keyword: SSR markers

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Determination of the Origin of Angelica Roots using Angelica gigas Chloroplast Based SSR Markers (엽록체기반 SSR marker를 이용한 당귀의 기원 판별)

  • Park, Sang Ik;Hwangbo, Kyeong;Gil, Jinsu;Chung, Hee;Kim, Ho Bang;Kim, Ok Tae;Kim, Seong Cheol;Koo, Sung Cheol;Um, Yurry;Lee, Yi
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.6
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    • pp.361-366
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    • 2017
  • Background: In the herbal medicinal industry, Angelica gigas Nakai, Angelica sinensis (Oliv.) Diels. and Angelica acutiloba (Siebold & Zucc.) Kitag. are often confused, because the roots of the three species can not be distinguished by their appearance. This confusion can cause serious side effects. In this study, we determined the origins of Angelica roots distributed in the Korean market using the simple sequence repeat (SSR) markers developed based on the A. gigas chloroplast DNA sequence. Methods and Results: We collected twenty seven A. gigas and three A. acutiloba samples from the Seoul, Daegu, and Cheongju herbal medicinal markets. Fifty sections of one collection were mixed and ground to make a powder, which was used for DNA extraction using the cetyl trimethylammonium bromide (CTAB) method. Chloroplast based SSR markers were applied to the DNA for the determination of the species. In addition, polymorphism was found in eight samples. The phylogenetic analysis showed that the A. gigas roots collected from herbal medicinal markets were clearly discriminated from A. sinensis and A. acutiloba even though they were grouped into four clusters. Conclusions: This study showed that chloroplast based SSR markers would help the discrimination of Angelica roots in the Korean herbal medicinal industry and the markers are useful to prevent confusion between Angelica roots.

Two-Year Estimates of Mating System in Seed Orchard of Pinus densiflora Revealed by cpSSR and nSSR Markers (안면도 소나무 채종원 교배양식 추정모수의 연간비교)

  • Kim, Young Mi;Hong, Yong Pyo;Park, Jae In
    • Journal of Korean Society of Forest Science
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    • v.104 no.4
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    • pp.578-587
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    • 2015
  • Nuclear SSR (nSSR) and chloroplast SSR (cpSSR) markers were analyzed to assess the parameters of mating system in seed orchard, such as outcrossing rates, the number of potential pollen contributors, paternal contribution rates, degree of pollen contamination, and biparental inbreeding ($t_m-t_s$). In 2006, 2007, seeds were collected from the seed orchard of Pinus densiflora, established in 1977 at Anmyeon island. Estimates of outcrossing rates ranged from 94.9 to 100% (mean 98.9%) in 2006 and from 91.2 to 100% (mean 97.7%) in 2007 on the basis of the analysis of cpSSR haplotypes and from 90.3 to 100% (mean 95.9%) in 2006 and from 81.6 to 100% (mean 95.3%) in 2007 on the basis of the analysis of nSSR genotypes. By cross checking of both DNA markers, mean cumulative outcrossing rates of 100% and 98.9% were estimated in each year. Mean contamination rates were estimated as 48.9% and 42.4%, respectively. On the basis of cpSSR haplotype observed in each seed, paternal contribution rates (the number of pollen contributors) were estimated as 0.458 (mean 16.2) in 2006 and 0.512 (mean 14.8) in 2007. In conclusion, considering pretty high level of outcrossing rates observed in a seed orchard, there might be little to be influenced by inbreeding depression for genetic potential of the seeds induced by selfing. Estimates of the mating system parameters obtained from the two reproductive years were not statistically different, which revealed stable genetic quality of seeds produced in different years. Observed results from this study may provide useful information for the management and establishment of the seed orchard of the progressive generation.

Analysis and mapping of the re-1 gene for reduced embryo size in rice

  • Kien, Trinh Hong;Oh, Ji Min;Yang, Paul;Hong, Soon Kwan;Ahn, Sang Nag
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.23-27
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    • 2010
  • The objective of this study was to map the gene for reduced embryo size in rice using DNA markers. The reduced embryo size mutant was induced from N-methyl-N-nitrosourea (MNU) treated Taichung 65. Genetic analysis revealed that the phenotype of the reduced embryo was controlled by a single recessive gene, designated as re-1. For mapping the gene controlling embryo size, an $F_2$ population was developed from a cross between the Korean Tongil-type, Milyang 23 (Oryza sativa ssp. indica) and the mutant. The ratio of $F_2$ seeds nearly fitted to 3:1 ratio, indicating that this phenotype was controlled by a single recessive gene. Bulked sergeant analysis was performed with SSR markers. The gene for the reduced embryo size was detected on chromosome 1. The gene was further mapped between two SSR markers, RM315 and RM265 on chromosome 1 (approximately 1.5 Mb interval). The linked markers will facilitate selection of this grain character in a breeding program and provide the foundation for positional cloning of this gene.

Genotype Fingerprinting, Differentiation and Association between Morphological Traits and SSR Loci of Soybean Landraces

  • Park, lk-Young
    • Plant Resources
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    • v.1 no.2
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    • pp.81-91
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    • 1998
  • Fifty-nine Korean soybean (Glycine max L. Merr.) landrace accessions were tested for genotype fingerprinting, differentiation and association between morphological traits and SSR profile. Using 8 SSR loci, 59 varieties were divided into 55 groups, and only 4 pairs of varieties were not uniquely identified. The resolving power of SSR for soybean genotyping was much higher than that of the morphological traits that were studied. Identification efficiency also differed among SSR loci. Those loci with higher numbers of alleles distinguished varieties more effectively. Genetic differentiation values of the soybean landraces varied from 0.57 to 0.82 with a mean of 0.68. The number of alleles detected by the 8 loci ranged from 3 to 8. and the effective number of alleles ranged from 2.3 to 5.1. In a study of the association of SSR alleles with morphological traits, some alleles seemed to be related with some specific morphological traits. Comparison of two kinds of dendrograms which were derived from SSR markers and quantitative traits indicated that the dendrograms were not consistent. Considering the correlation between single SSR locus and qualitative traits governed by major genes, the data suggest that alleles of microsatellite loci be more closely related to some traits determined by major genes than those determined by minor genes.

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Development of Microsatellite Markers and their Use in Genetic Diversity and Population Analysis in Eleutherococcus senticosus

  • Lee, Kyung Jun;An, Yong-Jin;Ham, Jin-Kwan;Ma, Kyung-Ho;Lee, Jung-Ro;Cho, Yang-Hee;Lee, Gi-An
    • Korean Journal of Plant Resources
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    • v.30 no.3
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    • pp.323-330
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    • 2017
  • Eleutherococcus senticosus (Siberian ginseng) is an important medicinal tree found in northeast Asia. In this study, we analyzed the genome-wide distribution of microsatellites in E. senticosus. By sequencing 711 clones from an SSR-enriched genomic DNA library, we obtained 12 polymorphic SSR markers, which also revealed successful amplicons in E. senticosus accessions. Using the developed SSR markers, we estimated genetic diversity and population structure among 131 E. senticosus accessions in Korea and China. The number of alleles ranged from 2 to 11, with an average of 7.4 alleles. The mean values of observed heterozygosity ($H_O$) and expected heterozygosity ($H_E$) were 0.59 and 0.56, respectively. The average polymorphism information content (PIC) was 0.51 in all 131 E. senticosus accessions. E. senticosus accessions in Korea and China showed a close genetic similarity. Significantly low pairwise genetic divergence was observed between the two regions, suggesting a relatively narrow level of genetic basis among E. senticosus accessions. Our results not only provide molecular tools for genetic studies in E. senticosus but are also helpful for conservation and E. senticosus breeding programs.

Genetic Diversity of Finger Millet (Eleusine coracana (L.) Gaertn.) Landraces Based on EST-SSR

  • Myung Chul Lee;Yu-Mi Choi;Myoung-Jae Shin;Hyemyeong Yoon;Seong-Hoon Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.46-46
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    • 2020
  • Finger millet is more nutritious than other and millets and widely cultivate in tropical regions of the world. Furthermore, it is more tolerant against biotic and abiotic stresses such as pest, drought and salt. For this reason, finger millet is one of the putative crops to introduce and cultivate on reclaimed land and prepare the global climate exchange in Korea. In present study, genetic diversity and structure of different populations of finger millet from Africa and South Asia was examined at molecular level using newly developed EST-Simple Sequence Repeat (EST-SSR) markers. In total, 46 primers produced 292 alleles in a size range of 100-500 bp and mean Polymorphism Information Content (PIC) and Marker Index (MI) were 0.372 and 1.04, respectively. 46 primers showed polymorphism and 21 primers were identified as having a PIC value above 0.5. Principal coordinates analysis and the dendrogram constructed out of combined data of both markers showed grouping of finger millet accessions to their respective area of collection. The 156 accessions were more classified into four groups, such as three groups of Africa collection and one group of Asia. Results of present study can be useful in identifying diverse accessions and management of this plant resource. Moreover, the novel SSR markers developed can be utilized for various genetic analyses in this species in future.

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Establishing a Core Collection of Proso Millet (Panicum miliaceum) Germplasm

  • Myung Chul Lee;Yu-Mi Choi;Myoung-Jae Shin;Hyemyeong Yoon;Kebede Taye Desta
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.47-47
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    • 2020
  • The Korean National Agrobiodiversity Center holds the more than 1300 accessions of proso millet, but a large portion of accessions are landrace of Korea that has very similar traits. To comprehend the maximum genetic diversity of this crop, a core collection with minimum number of accessions will facilitate easy access to genetic material. Here we assessed the genetic diversity and population structure in a germplasm collection of 830 accessions by employing EST-SSR markers and morphological traits. A total of 107 alleles were detected with an average allele number of 4.9 per locus among the 830 accessions based on 37 EST-SSR markers. The number of alleles per locus ranged from 2 to 7. Polymorphism information content and expected heterozygosity ranged from 0.06 to 0.68 (mean = 0.21) and 0.06 to 0.73 (mean = 0.23), respectively. The germplasm collection was separated into two groups based on population structure analysis, whereas principal coordinate analysis (PCoA) could not cluster accessions according to their geographic origin. Subsequently, a preliminarily developed core collection with a total of 141 accessions (17%) was selected from the whole set of germplasm by combining allelic variations of EST-SSR markers and eight different phenotypic traits. The core collection optimally represented the whole germplasm collection and displayed a similar level of PCoA value and genetic variation from the initial collection. The results obtained here provide a primary resource for further genetic analysis and establish a reference for further development of appropriate genetic breeding strategies.

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Evaluation of Germplasm and Development of SSR Markers for Marker-assisted Backcross in Tomato (분자마커 이용 여교잡 육종을 위한 토마토 유전자원 평가 및 SSR 마커 개발)

  • Hwang, Ji-Hyun;Kim, Hyuk-Jun;Chae, Young;Choi, Hak-Soon;Kim, Myung-Kwon;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.30 no.5
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    • pp.557-567
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    • 2012
  • This study was conducted to achieve basal information for the development of tomato cultivars with disease resistances through marker-assisted backcross (MAB). Ten inbred lines with TYLCV, late blight, bacterial wilt, or powdery mildew resistance and four adapted inbred lines with superior horticultural traits were collected, which can be useful as the donor parents and recurrent parents in MAB, respectively. Inbred lines collected were evaluated by molecular markers and bioassay for confirming their disease resistances. To develop DNA markers for selecting recurrent parent genome (background selection) in MAB, a total of 108 simple sequence repeat (SSR) primer sets (nine per chromosome at average) were selected from the tomato reference genetic maps posted on SOL Genomics Network. Genetic similarity and relationships among the inbred lines were assessed using a total of 303 polymorphic SSR markers. Similarity coefficient ranged from 0.33 to 0.80; the highest similarity coefficient (0.80) was found between bacterial wilt-resistant donor lines '10BA333' and '10BA424', and the lowest (0.33) between a late blight resistant-wild species L3708 (S. pimpinelliforium L.) and '10BA424'. UPGMA analysis grouped the inbred lines into three clusters based on the similarity coefficient 0.58. Most of the donor lines of the same resistance were closely related, indicating the possibility that these lines were developed using a common resistance source. Parent combinations (donor parent ${\times}$ recurrent parent) showing appropriate levels of genetic distance and SSR marker polymorphism for MAB were selected based on the dendrogram. These combinations included 'TYR1' ${\times}$ 'RPL1' for TYLCV, '10BA333' or '10BA424' ${\times}$ 'RPL2' for bacterial wilt, and 'KNU12' ${\times}$ 'AV107-4' or 'RPL2' for powdery mildew. For late blight, the wild species resistant line 'L3708' was distantly related to all recurrent parental lines, and a suitable parent combination for MAB was 'L3708' ${\times}$ 'AV107-4', which showed a similarity coefficient of 0.41 and 45 polymorphic SSR markers.

Determination and Application of Combined Genotype of Simple Sequence Repeats (SSR) DNA Marker for Cultivars of Cymbidium goeringii (춘란(Cymbidium goeringii) 품종에 대한 Simple Sequence Repeats (SSR) DNA 마커의 복합 유전자형 결정과 적용)

  • Lee, Dae-Gun;Koh, Jae-Chul;Chung, Ki-Wha
    • Horticultural Science & Technology
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    • v.30 no.3
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    • pp.278-285
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    • 2012
  • Cymbidium goeringii is one of the most important and popular species in the orchid family in north-east Asia. In the present study, we prepared multiplex PCR system, and used it for the genotyping of eight simple sequence repeats (SSR) markers (CG409, CG415, CG709, CG722, CG787, CG1023, CG1210, and CG1281) in subject with 40 samples of cultivated varieties. All the analyzed samples showed different combined genotypes. The average combined power of discrimination was very high value of $7.14{\times}10^{-10}$, and observed heterozygosity (Ho = 0.466) was similar with two wild populations of C. goeringii, which may indicate no or little occurrence of genetic change after collection from wild habitats. The present study also developed a two-dimensional barcode to express information of genotype results of eight SSR markers (SSR DNA ID). The discrimination power of DNA ID between two individuals will be statistically more than 99.999999%. The SSR DNA ID and two-dimensional barcode may be very usefully applied for the discrimination and maintence of cultivars of C. goeringii.

Isolation and characterization of EST-SSR markers for Astilboides tabularis (Saxifragaceae), endangered species in Korea

  • JUNG, Eui-Kwon;KANG, Dae-Hyun;YOO, Ki-Oug;KWAK, Myounghai;KIM, Young-Dong;KIM, Bo-Yun
    • 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.