• Title/Summary/Keyword: Marker Loci

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Development of SCAR Markers for Early Identification of Cytoplasmic Male Sterility Genotype in Chili Pepper (Capsicum annuum L.)

  • Kim, Dong Hwan;Kim, Byung-Dong
    • Molecules and Cells
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    • v.20 no.3
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    • pp.416-422
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    • 2005
  • We previously used Southern blot analysis to detect restriction-length polymorphisms between male fertile and cytoplasmic male sterile (CMS) cytoplasms at the coxII and atp6 loci of the mtDNA of Capsicum annuum L. Two copies of atp6 were found in each male fertile and CMS pepper lines. Interestingly, one of the copies of atp6 in CMS pepper was a 3'-truncated pseudogene. The open reading frame of the coxII gene was the same in the fertile (N-) and CMS (S-) lines. However, the nucleotide sequence in the S-cytoplasm diverged from that in the N-cytoplasm 41 bp downstream of the stop codon. To develop CMS-specific sequence-characterized amplified region (SCAR) markers, inverse PCR was performed to characterize the nucleotide sequences of the 5' and 3' flanking regions of mitochondrial atp6 and coxII from the cytoplasms of male fertile (N-) and CMS (S-) pepper plants. Based on these data, two CMS-specific SCAR markers, 607 and 708 bp long, were developed to distinguish N-cytoplasm from S-cytoplasm by PCR. The CMS-specific PCR bands were verified for 20 cultivars containing either N- or S-cytoplasm. PCR amplification of CMS-specific mitochondrial nucleotide sequences will allow quick and reliable identification of the cytoplasmic types of individual plants at the seedling stage, and assessment of the purity of $F_1$ seed lots. The strategy used in this report for identifying CMS-specific markers could be adopted for many other crops where CMS is used for F1 seed production.

DNAse 1 Hypersensitive Sites of Lung Specific Transcription Factor Gene (폐특이 전사조절 유전자의 DNAse 1 Hypersensitive Sites)

  • Lee, Yong-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.879-886
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    • 2000
  • Background : Thyroid Transcription Factor-1(TTF-1) acts as a tissue specific transcription factor in the regulation of lung specific gene expression and as morphogenic protein during lung organogenesis. Currently, there is very little information on the cis-acting sequences and transcription factors that direct the TTF-1 gene expression. DNAse 1 hypersensitive (DH) sites represent a marker for active or potentially active chromatin and are likely to be especially important in gene regulation, being associated with many DNA sequences that regulate gene expression. It is clear that DH regions correlate with genetic regulatory loci and binding for sequence-specific DNA-binding proteins. Methods : We have used DH site assays to identify putative distal regulatory elements in H441 lung adenocarcinoma cells, which express the TTF-1 gene and HeLa cells. Results : There are four DH sites 5' of the TTF-1 gene. These sites are located at base pair approximately +150, -450, -800, and -1500 from the start of transcription. Conclusion : These data suggest that there may be at least one intragenic site and regulatory region 5' prime to the promotor region.

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Analysis of Genetic Diversity and Population Structure of Buckwheat (Fagopyrum esculentum Moench) Landraces of Korea Using SSR Markers

  • Song, Jae-Young;Lee, Gi-An;Yoon, Mun-Sup;Ma, Kyung-Ho;Choi, Yu-Mi;Lee, Jung-Ro;Jung, Yeon-Ju;Park, Hong-Jae;Kim, Chung-Kon;Lee, Myung-Chul
    • Korean Journal of Plant Resources
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    • v.24 no.6
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    • pp.702-711
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    • 2011
  • Buckwheat (Fagopyrum esculentum Moench), one of the minor crops grown in Korea belonging to the Polygonaceae family, is an annual crop widely cultivated in Asia, Europe, and America and has a character of outcrossing and self-incompatibility. The objective of this study was to analyze the genetic variability, phylogenetic relationships and population structure of buckwheat landraces of Korea using SSR markers. Ten microsatellite markers have been detected from a total of 79 alleles among the 179 buckwheat accessions were collected from Korea. The number of allele per marker locus ($N_A$) ranged from 2 (GB-FE-001, GB-FE-043 and GB-FE-055) to 31 (GB-FE-035) with an average of 7.9 alleles. GB-FE-035 was the most polymorphic with the highest PIC value 0.93. Major allele frequencies ($M_{AF}$) for the 10 polymorphic loci varied from 0.12 to 0.97 with a mean allele frequency of 0.57. The expected heterozygosity ($H_E$) values ranged from 0.05 to 0.94 with an average of 0.53. The observed heterozygosity ($H_O$) ranged from 0.06 to 0.92 with an average of 0.42. The overall polymorphic information contents (PIC) values ranged from 0.05 to 0.93 with an average of 0.48. The landrace accessions of buckwheat used in the present study were not distinctly grouped according to geographic distribution. The study concludes that the results revealed genetic differentiation was low according to the geographic region because of outcrossing and self-incompatibility. We reported that our analyses on the genetic diversity of common buckwheat cultivars of Korea were performed by using of microsatellite markers.

Genetic Diversity and Population Genetic Structure of Cephalotaxus koreana in South Korea

  • Hong, Kyung Nak;Kim, Young Mi;Park, Yu Jin;Lee, Jei Wan
    • Korean Journal of Plant Resources
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    • v.27 no.6
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    • pp.660-670
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    • 2014
  • The Korean plum yew (Cephalotaxus koreana Nakai) is a shade-tolerant, coniferous shrub. The seeds have been used as a folk medicine in Korea, and an alkaloid extract (HTT) is known to have anticancer properties. We estimated the genetic diversity of 429 trees in 16 populations in South Korea using 194 polymorphic amplicons from seven combinations of AFLP primer-restriction enzymes. The average number of effective alleles and the percentage of polymorphic loci were 1.37 and 79.4%, respectively. Shannon's diversity index and the expected heterozygosity were 0.344 and 0.244, respectively. We divided 16 populations into four groups on the UPGMA dendrogram and the PCA biplot. The first two principal components explained 84% of the total genetic variation. Genetic differentiation between populations explained 14% of total genetic variation, and the remaining 86% came from difference between individuals within populations, as determined by an analysis of molecular variance (AMOVA). However, the genetic differentiation did not correlate with the geographic distance between populations from the Mantel test. The Bayesian statistics, which are comparable to Wright's $F_{ST}$ and Nei's $G_{ST}$, were ${\theta}^I=0.406$ and ${\theta}^{II}=0.172$, respectively. The population genetic diversity was slightly lower, and the strength of genetic differentiation was much weaker, than the average of those plants having similar life histories, as assessed using arbitrary marker systems. We discuss strategies for the genetic conservation of the plum yew in Korea.

Genic Vadadon and Speciation of Fishes of the Genus Moroco(Cyprinidae) (버들치속(잉어과) 어류의 유전적 변이 및 종분화)

  • 양서영;민미숙
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.75-83
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    • 1989
  • Surveys of electrophoretic variation in isozymes and general proteins encoded by 26 loci were conducted to assess species recognition and to estimate the degree of genic variation and species divergence for seven species of the genus Moroco inhabiting in Korea and Japan. Estimates of the average calculated heterozygosity per species of M semotilus, M sp., M percnurus, M lagowskii, M oxycephalus, M steindachneri and M jouyf are low: 0.021, 0.019, 0.051, 0.031, 0.023, 0.046, and 0.007, respectively, and observed heterozygosities are 0.038, 0.022, 0.060, 0.027, 0.025, 0.042, and 0.002, respectively. Allozyme analyses show these species to be distinct genetically with the lafter four species being more closely related one another than any one of them is to the rest of the species. However, these four species (M. lagowskii, M. oxycephalus, M. steindachneri and M jouyi), had unique genetic markers in each species to be recognized as valid species. These results contrast to the previous report of Chung et of. (1986) mainly due to their error in analyzing the isozyme pallems, particularly in MDH and PGI analyses. The genetic distances among M semotilus, M sp., and M percnurus are near the high end of the scale of such estimate for freshwater fish congeners. Based on estimated divergent time of these species of the genus Moroco (5 to 0.6 million years) it is assumed that they are speciated during late Pliocene to middle Pleistocene epoch prior to migration to Korean and Japanese waters through Paleo Amur River system.

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QTL Analysis of Soybean Seed Weight Using RAPD and SSR Markers

  • Chung, Jong-Il;Ko, Mi-Suk;Kang, Jin-Ho
    • Plant Resources
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    • v.3 no.3
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    • pp.184-193
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    • 2000
  • Soybean [Glycine max (L.) Merr.] seed weight is a important trait in cultivar development. Objective of this study was to identify and confirm quantitative trait loci (QTLs) for seed weight variation in the F2 and F2:3 generations. QTLs for seed weight were identified in F2 and F2:3 generations using interval mapping (MapMaker/QTL) and single-factor analysis of variance (ANOVA). In the F2 plant generation (i.e., F3 seed), three markers, OPL9a, OPM7a, and OPAC12 were significantly (P<0.01) associated with seed weight QTLs. In the F2:3 plant row generation (i.e., F4 seed), five markers, OPA9a, OPG19, OPL9b, OPP11, and Sat_085 were significantly (P<0.01) associated with seed weight QTLs. Two markers, OPL9a and OPL9b were significantly (P<0.05) associated with seed weight QTLs in both generations. Two QTLs on USDA soybean linkage group C1 and R were identified in both F2 and F2:3 generations using interval mapping. The linkage group C1 QTL explained 16% of the variation in seed weight in both generations, and the linkage group R QTL explained 39% and 41% of the variation for F2 and F2:3 generation, respectively. The linkage group C2 QTL identified in F2:3 generation explained 14.9% of variation. Linkage groups C1, C2 and R had previously been identified as harbouring seed size QTLs. The consistency of QTLs across generations and populations indicates that marker-assisted selection is possible in a soybean breeding program.

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A test of the hybrid origin of Korean endemic Sedum latiovalifolium (Crassulaceae) (한국특산 태백기린초(돌나물과)의 교잡 기원에 대한 검증)

  • Yoo, Young-Gi;Park, Ki-Ryong
    • Korean Journal of Plant Taxonomy
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    • v.46 no.4
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    • pp.378-391
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    • 2016
  • Eighteen morphological characters from 40 populations and ten isozyme loci from 35 populations of Sedum latiovalifolium and related species were examined to investigate the degree of morphological and genetic variation. The high-frequency marker alleles $MDH-2^a$ and $PGI-1^a$ in S. aizoon, S. kamtschaticum, and S. zokuriense did not appear in the populations of S. latiovalifolium. In addition, the high-frequency allele ($MDH-2^c$) in S. latiovalifolium appeared at a very low frequency in other subg. aizoon species. Thus, the isozyme data did not support a hybrid origin of S. latiovalifolium from S. aizoon with S. kamtschaticum. The results of an unweighted pair-group method using the arithmetic average method and a principal components analysis using morphological data also did not support a hybrid origin of S. latiovalifolium. However, our data strongly suggest that some individuals from the populations found in the Gumdaebong area were most likely hybrids due to introgression between S. latiovalifolium and S. kamtschaticum or S. aizoon and S. kamtschaticum.

Chromosome-specific polymorphic SSR markers in tropical eucalypt species using low coverage whole genome sequences: systematic characterization and validation

  • Patturaj, Maheswari;Munusamy, Aiswarya;Kannan, Nithishkumar;Kandasamy, Ulaganathan;Ramasamy, Yasodha
    • Genomics & Informatics
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    • v.19 no.3
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    • pp.33.1-33.10
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    • 2021
  • Eucalyptus is one of the major plantation species with wide variety of industrial uses. Polymorphic and informative simple sequence repeats (SSRs) have broad range of applications in genetic analysis. In this study, two individuals of Eucalyptus tereticornis (ET217 and ET86), one individual each from E. camaldulensis (EC17) and E. grandis (EG9) were subjected to whole genome resequencing. Low coverage (10×) genome sequencing was used to find polymorphic SSRs between the individuals. Average number of SSR loci identified was 95,513 and the density of SSRs per Mb was from 157.39 in EG9 to 155.08 in EC17. Among all the SSRs detected, the most abundant repeat motifs were di-nucleotide (59.6%-62.5%), followed by tri- (23.7%-27.2%), tetra- (5.2%-5.6%), penta- (5.0%-5.3%), and hexa-nucleotide (2.7%-2.9%). The predominant SSR motif units were AG/CT and AAG/TTC. Computational genome analysis predicted the SSR length variations between the individuals and identified the gene functions of SSR containing sequences. Selected subset of polymorphic markers was validated in a full-sib family of eucalypts. Additionally, genome-wide characterization of single nucleotide polymorphisms, InDels and transcriptional regulators were carried out. These variations will find their utility in genome-wide association studies as well as understanding of molecular mechanisms involved in key economic traits. The genomic resources generated in this study would provide an impetus to integrate genomics in marker-trait associations and breeding of tropical eucalypts.

Microsatellite Markers for Non-Invasive Examination of Individual Identity, Genetic Variation, and Population Differentiation in Two Populations of Korean Long-Tailed Goral (Naemorhedus caudatus)

  • Kim, Baek-Jun
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.3 no.4
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    • pp.191-198
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    • 2022
  • Natural habitats of the Korean long-tailed goral (Naemorhedus caudatus) have been fragmented by anthropogenic activities in South Korea in the last decades. Here, the individual identity, genetic variation, and population differentiation of the endangered species were examined via the multiple-tube approach using a non-invasive genotyping method. The average number of alleles was 3.16 alleles/locus for the total population. The Yanggu population (1.66) showed relatively lower average number of alleles than the Inje population (3.67). Of the total 19 alleles, only seven (36.8%) alleles were shared by the two populations. Using five polymorphic out of six loci, four and six different goral individuals from the captive Yanggu (n=24) and the wild Inje (n=28) population were identified, respectively. The allele distribution was not identical between the two populations (Fisher's exact test: P<0.01). A considerably low migration rate was detected between the two populations (no. of migrants after correction for size=0.294). Additionally, the F statistics results indicated significant population differentiation between them, however, quite low (FST=0.327, P<0.01). The posterior probabilities indicated that the two populations originated from a single panmictic population (P=0.959) and the assignment test results designated all individuals to both populations with nearly equal likelihood. These could be resulted from moderate population differentiation between the populations. No significant evidence supported recent population bottleneck in the total Korean goral population. This study could provide us with useful population genetic information for conservation and management of the endangered species.

Development of novel microsatellite markers to analyze the genetic structure of dog populations in Taiwan

  • Lai, Fang-Yu;Lin, Yu-Chen;Ding, Shih-Torng;Chang, Chi-Sheng;Chao, Wi-Lin;Wang, Pei-Hwa
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1314-1326
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    • 2022
  • Objective: Alongside the rise of animal-protection awareness in Taiwan, the public has been paying more attention to dog genetic deficiencies due to inbreeding in the pet market. The goal of this study was to isolate novel microsatellite markers for monitoring the genetic structure of domestic dog populations in Taiwan. Methods: A total of 113 DNA samples from three dog breeds-beagles (BEs), bichons (BIs), and schnauzers (SCs)-were used in subsequent polymorphic tests applying the 14 novel microsatellite markers that were isolated in this study. Results: The results showed that the high level of genetic diversity observed in these novel microsatellite markers provided strong discriminatory power. The estimated probability of identity (P(ID)) and the probability of identity among sibs (P(ID)sib) for the 14 novel microsatellite markers were 1.7×10-12 and 1.6×10-5, respectively. Furthermore, the power of exclusion for the 14 novel microsatellite markers was 99.98%. The neighbor-joining trees constructed among the three breeds indicated that the 14 sets of novel microsatellite markers were sufficient to correctly cluster the BEs, BIs, and SCs. The principal coordinate analysis plot showed that the dogs could be accurately separated by these 14 loci based on different breeds; moreover, the Beagles from different sources were also distinguished. The first, the second, and the third principal coordinates could be used to explain 44.15%, 26.35%, and 19.97% of the genetic variation. Conclusion: The results of this study could enable powerful monitoring of the genetic structure of domestic dog populations in Taiwan.