• Title/Summary/Keyword: SSR markers

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SSR Analysis of Genetic Diversity and Nitrogen Use Efficiency Traits in Rice

  • Kim, Myung Ki;Oh, Myeong Kyu;Lee, Jeong Heui;Kim, Yeon Gyu;Lee, Young Tae;Kim, Kwang Ho;Ahn, Sang Nag
    • Korean Journal of Breeding Science
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    • v.40 no.2
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    • pp.119-127
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    • 2008
  • A total of 41 microsatellite markers were used with 29 genotypes to examine the relationship between SSR polymorphisms and N-use efficiency related traits with a goal to identify the putative QTLs related to these traits. These primers yielded a total of 183 alleles (average 4.46 alleles per primer), and polymorphism information content (PIC) values of the SSRs ranged from 0.119 to 0.805 with mean value of 0.425. Correlation coefficients were obtained among the four N-use efficiency traits in the 34 accessions and significant positive correlations of relative ratios between grain yield and harvest index (r=0.3404) and total dry matter (r=0.7976), while N uptake showed a moderate level of correlation with the ratios of the grain yield and total dry matter, respectively. 36.5% (15/41) SSR markers were monomorphic among the 25 japonica accessions out of the 29 accessions. Association between SSR genotypes and phenotypic performances from the total (29) or japonica (25) accessions was tested based on a single point analysis. Three putative QTL regions were detected for the ratio of grain yield. These include the chromosomal region containing the RM283 locus on chromosome 1 and RM25 on chromosome 8 (all and japonica accessions) and the region with the SSR marker, RM206 on chromosome 11 (the japonica accessions). For the total dry matter ratio, two chromosomal regions were identified as the putative QTL region. One is the region with the SSR marker, RM162 on chromosome 6 (all and japonica accessions) and the other was the one with the SSR marker RM25 on chromosome 8 (the japonica accessions). Among these markers, RM25 showed associations with both traits.

Cultivar Discrimination of Korean and Chinese Boxthorn (Lycium chinense Mill. and Lycium barbarum L.) using SSR Markers (SSR 마커를 이용한 한국산과 중국산 구기자의 품종 판별)

  • Chung, Jong-Wook;Lee, Gi-An;Lee, Sok-Su;Bang, Kyong-Hwan;Park, Chung-Berm;Park, Yong-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.6
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    • pp.445-451
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    • 2009
  • This study was undertaken to develop a technique of discrimination using SSR makers in boxthorn cultivars. Forty one boxthorn cultivars, which were collected from Korea and China, were evaluated by 10 SSR markers. Total of 61 alleles were detected, ranging from 3 to 13 with an average of 6.1 alleles per locus. The averages of gene diversity and PIC values were 0.482 and 0.428, with a range from 0.25 (GB-LCM-022 and GB-LCM-087) to 0.83 (GB-LCM-167) and from 0.24 (GB-LCM-022 and GB-LCM-087) to 0.81 (GB-LCM-167), respectively. Five markers out of 10 markers, GB-LCM-022, GB-LCM-075, GB-LCM-104, GB-LCM-167 and GB-LCM-217, were selected as key markers for discrimination in boxthorn cultivars. All of boxthorn cultivars were individually distinguished by the combination of five SSR markers.

Linkage Analysis of both RAPD and I-SSR Markers using Haploid Genome from a Single Tree of Pinus densiflora S. et Z. (소나무 단일(單一) 모수(母樹)의 반수체(半數體) 게놈을 이용(利用)한 RAPD 및 I-SSR 표식자(標識子)의 연관분석(連關分析))

  • Hong, Yong-Pyo;Chung, Jae-Min;Kim, Yong-Yul;Jang, Suk-Seong
    • Journal of Korean Society of Forest Science
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    • v.89 no.4
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    • pp.536-542
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    • 2000
  • A linkage map for Japanese red pine (Pinus densiflora) was constructed on the basis of two DNA marker systems of random amplified polymorphic DNAs (RAPDs) and inter-simple sequence repeats (I-SSR). Haploid genomic DNAs were extracted from megagametophyte tissues of 96 individual seeds in a single tree. A total of 98 DNA markers including 52 RAPD markers amplified by 25 primers and 46 I-SSR markers amplified by 18 primers were verified as Mendelian loci showing 1 : 1 segregation in 96 megagametophytes which were ${\chi}^2$-tested at 5% significance level. Of them, 63 segregating loci turned out to be linked into 20 linkage groups by the two-point analysis. However, 35 loci (17 RAPD and 18 I-SSR) of the 98 segregating loci did not coalesced into any linkage groups at a LOD of 3.0. The linked 63 loci were separated by an average distance of about 25.5 cM, which were spanned 1097.8 cM as a whole. The minimum and maximum map distances of the linkage groups were 4.3 cM and 54.9 cM, respectively. Incorporation of I-SSR loi into linkage map of RAPD loci resulted in extended and partially more saturated linkage blocks.

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Construction of an Integrated Pepper Map Using RFLP, SSR, CAPS, AFLP, WRKY, rRAMP, and BAC End Sequences

  • Lee, Heung-Ryul;Bae, Ik-Hyun;Park, Soung-Woo;Kim, Hyoun-Joung;Min, Woong-Ki;Han, Jung-Heon;Kim, Ki-Taek;Kim, Byung-Dong
    • Molecules and Cells
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    • v.27 no.1
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    • pp.21-37
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    • 2009
  • Map-based cloning to find genes of interest, marker-assisted selection (MAS), and marker-assisted breeding (MAB) all require good genetic maps with high reproducible markers. For map construction as well as chromosome assignment, development of single copy PCR-based markers and map integration process are necessary. In this study, the 132 markers (57 STS from BAC-end sequences, 13 STS from RFLP, and 62 SSR) were newly developed as single copy type PCR-based markers. They were used together with 1830 markers previously developed in our lab to construct an integrated map with the Joinmap 3.0 program. This integrated map contained 169 SSR, 354 RFLP, 23 STS from BAC-end sequences, 6 STS from RFLP, 152 AFLP, 51 WRKY, and 99 rRAMP markers on 12 chromosomes. The integrated map contained four genetic maps of two interspecific (Capsicum annuum 'TF68' and C. chinense 'Habanero') and two intraspecific (C. annuum 'CM334' and C. annuum 'Chilsungcho') populations of peppers. This constructed integrated map consisted of 805 markers (map distance of 1858 cM) in interspecific populations and 745 markers (map distance of 1892 cM) in intraspecific populations. The used pepper STS were first developed from end sequences of BAC clones from Capsicum annuum 'CM334'. This integrated map will provide useful information for construction of future pepper genetic maps and for assignment of linkage groups to pepper chromosomes.

Development of Molecular Markers for Xanthomonas axonopodis Resistance in Soybean

  • Kim Ki-Seung;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.429-433
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    • 2004
  • A single recessive gene, rxp, controls the bacterial leaf pustule (BLP) resistance in soybean and in our previous article, it has been mapped on linkage group (LG) D2 of molecular genetic map of soybean. A total of 130 recombinant inbred lines (RILs) from a cross between BLP-resistant SS2-2 and BLP-susceptible Jangyeobkong were used to identify molecular markers linked to rxp. Fifteen simple sequence repeat (SSR) markers on LG D2 were screened to construct a genetic map of rxp locus. Only four SSR markers, Satt135, Satt372, Satt448, and Satt486, showed parental polymorphisms. Using these markers, genetic scaffold map was constructed covering 26.2cM. Based on the single analysis of variance, Satt372 among these four SSR markers was the most significantly associated with the resistance to BLP. To develop new amplified fragment length polymorphism (AFLP) marker linked to the resistance gene, bulked segregant analysis (BSA) was employed. Resistance and susceptible bulks were made by pooling equal amount of genomic DNAs from ten of each in the segregating population. A total of 192 primer combinations were used to identify specific bands to the resistance, selecting three putative AFLP markers. These AFLP markers produced the fragment present in SS2-2 and the resistant bulk, and not in Jangyeobkong and the susceptible bulk. Linkage analysis revealed that McctEact97 $(P=0.0004,\;R^2=14.67\%)$ was more significant than Satt372, previously reported as the most closely linked marker.

Confirmation of Parentage of the Pear Cultivar 'Niitaka' (Pyrus pyrifolia) Based on Self-incompatibility Haplotypes and Genotyping with SSR Markers

  • Kim, Hoy-Taek;Nou, Ill-Sup
    • Horticultural Science & Technology
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    • v.34 no.3
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    • pp.453-460
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    • 2016
  • The parentage of the horticulturally important pear cultivar 'Niitaka' was confirmed by determining its S-genotypes based on the S-RNase and $PpSFBB^{-{\gamma}}$ genes, and genotyping using simple sequence repeat (SSR) markers. Previous reports suggested that the cultivars 'Amanogawa' and 'Imamuraaki' were the parents of 'Niitaka', although the cultivars 'Chojuro' and 'Shinchu' were also examined as candidate parents, along with two other cultivars. In the present study, the S-genotype of 'Niitaka' was determined to be $S^3S^9$. The $S^9$-RNase of 'Niitaka' was found to be likely inherited from the parent 'Amanogawa' ($S^1S^9$) and the $S^3$-RNase from 'Chojuro' ($S^3S^5$) or 'Shinchu' ($S^3S^5$). Based on the S-genotypes, the cultivar 'Imamuraaki' ($S^1S^6$) had no contribution to the parentage of 'Niitaka' ($S^3S^9$). A total of 67 polymorphic SSR markers were used to further confirm the parentage of 'Niitaka'. Discrepancies were found at several SSR loci between 'Niitaka' and the cultivars 'Imamuraaki' and 'Shinchu', whereas 'Niitaka' inherited alleles from 'Amanogawa' and 'Chojuro' at all SSR loci. Therefore, our findings established that 'Amanogawa' and 'Chojuro' are the parents of pear cultivar 'Niitaka', and not 'Imamuraaki' as previously reported.

Mating System in Natural Population of Pinus koraiensis at Mt. Seorak Based on Allozyme and cpSSR Markers (동위효소 표지와 cpSSR 표지를 이용한 설악산 잣나무 집단의 교배양식)

  • Hong, Yong-Pyo;Ahn, Ji-Young;Kim, Young-Mi;Hong, Kyung Nak;Yang, Byeong-Hoon
    • Journal of Korean Society of Forest Science
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    • v.102 no.2
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    • pp.264-271
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    • 2013
  • Mating system parameters were estimated in a natural population of Pinus koraiensis which was located at Gwongeumseong in Mt. Seorak, South Korea. The estimated parameters from allozyme were as follows: 0.882 of multilocus outcrossing rates($t_m$), 0.881 of singlelocus outcrossing rates($t_s$), 0.368 of correlated paternity($r_p$), and 2.7 of number of effective pollen contributors. The estimated parameters from cpSSR markers were as follows: 0.831 of average of outcrossing rates and 12.4 of the average number of effective pollen contributors. The average outcrossing rate from two genetic markers was 0.857, which was similar to those estimated in other conifer species. More number of potential pollen contributors was estimated from cpSSR marker analysis compared with that estimated from allozyme marker analysis. This result sugges$t_s$ that cpSSR markers may be more useful than allozyme markers for identifying potential pollen contributors in the analysis of mating system.

Development of EST-SSR markers for genetic diversity analysis in little millet (Panicum sumatrense) genetic resources

  • Lee, Myung-Chul;Choi, Yu-Mi;Lee, Sukyeung;Yoon, Hyemyeong;Oh, Sejong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.74-74
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    • 2018
  • Little millet (Panicum sumatrense) is well known for its salt and drought stress tolerance and high nutritional value, but very limited knowledge of genetic variation and genomic information is available. This study was to develop highly polymorphic EST-SSR markers based on cross-species transferability of derived SSRs from switchgrass EST databases and characterize newly developed EST - SSRs to better understand the genetic diversity of collected 37 germplasm accessions of little millet. A total of 779 primer pairs were designed from the 22,961 EST sequences of switchgrass (Pancium virgatum), of which 48 EST - SSR markers were developed based on the trials of transferability of these primers in little millet. The EST - SSR amplicons showed reproducible single band polymorphism and produced a total of 160 alleles with an average of 3.3 alleles per locus in 37 accessions of little millet. T he average values of expected and observed heterozygosities were 0.266 and 0.123, respectively. T he polymorphic information content (PIC) values were observed in range of 0.026 to 0.549 with an average of 0.240. The genetic relatedness among the little millet accessions was evaluated by neighbor-joining dendrogram, which grouped all accessions into two distinct groups. The validation thus demonstrated the utility of the switchgrass EST - SSR markers in assessing genomic relationships in little millet. T he findings from this study could be useful for designing strategies for the identification of diverse germplasm for conservation and future molecular breeding programs for little millet.

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Development and validation of SSR markers in Eleutherococcus senticosus

  • Lee, Kyung Jun;An, Yong-Jin;Ham, Jin-Kwan;Ma, Kyung-Ho;Lee, Jung-Ro;Cho, Yang-Hee;Lee, Gi-An
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.84-84
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    • 2017
  • Eleutherococcus senticosus (Siberian ginseng) is an important medicinal tree found in Russian taiga, and northern regions of Korea, Japan, and China. In this study, we analyzed the genome-wide distribution of microsatellites in E. senticosus using developed SSR markers. A total of 711 clones from an SSR-enriched genomic DNA library were sequenced, of which 47 clones (6.6%) were redundant. Of the 664 independent clones, only 12 polymorphic SSR markers were obtained, 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.

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High-Throughput Development of Polymorphic Simple Sequence Repeat Markers Using Two Whole Genome Sequence Data in Peucedanum japonicum

  • Lee, Junki;Joh, Ho Jun;Kim, Nam-Hoon;Lee, Sang-Choon;Jang, Woojong;Choi, Beom Soon;Yu, Yeisoo;Yang, Tae-Jin
    • Plant Breeding and Biotechnology
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    • v.5 no.2
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    • pp.134-142
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    • 2017
  • Resource plants are important and have strong potential for a variety of utilities as crops or pharmaceutical materials. However, most resource plants remain wild and thus their utility for breeding and biotechnology is limited. Molecular markers are useful to initiate genetic study and molecular breeding for these understudied resource plants. We collected various wild collections of Peucedanum japonicum which is indigenous resource plants utilized as oriental medicine and leafy vegetables in Korea. In this study, we produced two independent whole genome sequences (WGSs) from two collections and identified large scale polymorphic simple sequence repeat (pSSR) based on our pipeline to develop SSR markers based on comparison of two WGSs. We identified a total of 452 candidate pSSR contigs. To confirm the accuracy and utility of pSSR, we designed ten SSR primer pairs and successfully applied those to seven collections of P. japonicum. The WGS and pSSR candidates identified in this study will be useful resource for genetic research and breeding purpose for the valuable resource plant, P. japonicum.