• Title/Summary/Keyword: molecular sequence data

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Development of EST-SSR Markers for Evaluation of Genetic Diversity and Population Structure in Finger Millet (Eleusine coracana (L.) Gaertn.)

  • Lee, Myung Chul;Choi, Yu-Mi;Hyun, Do-Yoon;Lee, Sukyeung;Kim, Jin-Hee;Oh, Sejong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.105-105
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    • 2018
  • Finger millet, Eleusine coracana Gaertn., is more nutritious than other cereals and millets and widely cultivate in tropical regions of the world. However, status of its genetic diversity remained concealed due to lack of research work in this species. In recent years, microsatellites have become the most used markers for studying population genetic diversity. 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 using a total of 1,927 ESTs of Eleusine coracana available in the NCBI database. 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 was 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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Phylogenetic Analysis by RFLP and Sequencing of Mitochondrial DNA in a Korean Population

  • Lee, Jin-Young;Kim, Heui-Soo;Ha, Bae-Jin;Park, Yeong-Hong
    • Archives of Pharmacal Research
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    • v.29 no.1
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    • pp.88-95
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    • 2006
  • Analysis of molecular nature of mitochondrial DNA (mtDNA) could be powerful marker for anthropological studies of modern populations. While population genetic studies on mtDNA have been reported for several ethnic groups, no such study has been documented for the Korean population. We surveyed mtDNA polymorphisms in the HVS I of noncoding D-loop region and its upstream region from 430 unrelated healthy Korean population by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing analysis. PCR product with 2,790 bp spanning the specific mtDNA region (mt13715-16504) was subjected to RFLP analysis using 6 restriction enzyme (Hinf I, Hae III, Alu I, Dde I, Mbo I, Rsa I). On the PAUP analysis of PCR-RFLP results, 38 mtDNA haplotypes (Hap 1-38) were detected in the Korean populations, which were classified into 11 haplogroups (Grp 1-11) of related haplotypes encompassing all 38 haplotypes. In comparison of sequencing data with Anderson's reference sequence, the transition type was more prevalent than the transversion type. Insertions or deletions were not found. In addition, three of the polymorphic sites (A16240C, A16351G, G16384A) in HVS-I region are determined newly. The polymorphic sites were distributed randomly in the region, though the frequency at each site was variable. Thus, this research might be required for the genealogical study of Orientals.

BcSNPdb: Bovine Coding Region Single Nucleotide Polymorphisms Located Proximal to Quantitative Trait Loci

  • Moon, Sun-Jin;Shin, Hyoung-Doo;Cheong, Hyun-Sub;Cho, Hye-Young;NamGoong, Sohg;Kim, Eun-Mi;Han, Chang-Su;Sung, Sam-Sun;Kim, Hee-Bal
    • BMB Reports
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    • v.40 no.1
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    • pp.95-99
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    • 2007
  • Bovine coding region single nucleotide polymorphisms located proximal to quantitative trait loci were identified to facilitate bovine QTL fine mapping research. A total of 692,763 bovine SNPs was extracted from 39,432 UniGene clusters, and 53,446 candidate SNPs were found to be a depth >3. In order to validate the in silico SNPs experimentally, 186 animals representing 14 breeds and 100 mixed breeds were analyzed. Genotyping of 40 randomly selected candidate SNPs revealed that 43% of these SNPs ranged in frequency from 0.009 to 0.498. To identify non-synonymous SNPs and to correct for possible frameshift errors in the ESTs at the predicted SNP positions, we designed a program that determines coding regions by protein-sequence referencing, and identified 17,735 nsSNPs. The SNPs and bovine quantitative traits loci informations were integrated into a bovine SNP data: BcSNPdb (http://snugenome.snu.ac.kr/BtcSNP/). Currently there are 43 different kinds of quantitative traits available. Thus, these SNPs would serve as valuable resources for exploiting genomic variation that influence economically and agriculturally important traits in cows.

Expression of a rice DREB1 gene, OsDREB1D, enhances cold and high-salt tolerance in transgenic Arabidopsis

  • Zhang, Yang;Chen, Chen;Jin, Xiao-Fen;Xiong, Ai-Sheng;Peng, Ri-He;Hong, Yi-Huan;Yao, Quan-Hong;Chen, Jian-Min
    • BMB Reports
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    • v.42 no.8
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    • pp.486-492
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    • 2009
  • OsDREB1D, a special DREB (dehydration responsive element binding protein) homologous gene, whose transcripts cannot be detected in rice (Oryza sativa L), either with or without stress treatments, was amplified from the rice genome DNA. The yeast one-hybrid assay revealed that OsDREB1D was able to form a complex with the dehydration responsive element/C-repeat motif. It can also bind with a sequence of LTRE (low temperature responsive element). To analyze the function of OsDREB1D, the gene was transformed and over-expressed in Arabidopsis thaliana cv. Columbia. Results indicated that the over-expression of OsDREB1D conferred cold and high-salt tolerance in transgenic plants, and that transgenic plants were also insensitive to ABA (abscisic acid). From these data, we deduced that this OsDREB1D gene functions similarly as other DREB transcription factors. The expression of OsDREB1D in rice may be controlled by a special mechanism for the redundancy of function.

Genetic Evolution and Characteristics of Ichthyotoxic Cochlodinium polykrikoides(Gymnodiniales, Dinophyceae) (어류치사성 Cochlodinium polykrikoides 적조생물의 유전적 진화 및 특성)

  • Cho, Eun-Seob;Jeong, Chang-Su
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1453-1463
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    • 2007
  • This study presents a molecular phylogenetic analysis of the harmful dinoflagellate Cochlodinium polykrikoides, by use of partial sequence of small subunit (SSU) rRNA gene from most of the major taxa(24 species) in dinoflagellates. The class Dinophyceae clade formed a strong monophyletic relationship with C. polykrikoides and several taxa. On the basis of deeper nodes, the phylogenetic relationships placed C. polykrikoides closer to the order Prorocentrales rather than to the order Gymnodiniales, which was supported by a strong bootstrap value (100%) in the analyses of Neighbor-Joining and Parsimony methods. There is strong support for C. polykrikoides being placed in the same branch as Gymnodiniaceae and being connected in a clade with Prororcentrum micans among Prorocentrales. Morphological data show that C. polykrikoides is well associated with the genus Gyrodinium; however, this species is genetically closer to Gymnodinium than to Gyrodinium. The placement of C. polykrikoides always formed an independent branch separated from other dinoflagellates. In conclusion, planktonic P. micans plays an important role as an ancestor of Gymnodinium, whereas C. polykrikoides appears to be used an intermediate position between P. micans and Gymnodinium based on evolution.

Re-evaluation of specimens recorded as Amanita virosa in Korea (한국에 기록된 독우산광대버섯의 재동정)

  • Jin, Yong-Ju;Yoo, Ki-Bum;Ahn, Geum Ran;Kim, Seong Hwan;Seok, Soon-Ja
    • The Korean Journal of Mycology
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    • v.45 no.1
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    • pp.14-22
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    • 2017
  • A molecular phylogenetic analysis of 40 fungal specimens that were collected from the Korean peninsula from 2000 to 2015 and recorded as Amanita virosa was performed using internal transcribed spacer sequence data. Results confirmed that Amanita oberwinklerana (14 specimens), Amanita rimosa (5), Amanita pallidorosea (20), and Amanita virosa (1) belong to section Phalloideae of subgenus Lepidella, and the morphological features of these specimens were re-examined. The former three species with deadly poisonous white mushrooms were not yet recorded in Korea. Because of their morphological similarities with A. virosa, they are frequently overlooked or misidentified in the field. All collections were deposited in the Herbarium Conservation Center of the National Institute of Agricultural Sciences.

Development of a High-Resolution Multi-Locus Microsatellite Typing Method for Colletotrichum gloeosporioides

  • Mehta, Nikita;Hagen, Ferry;Aamir, Sadaf;Singh, Sanjay K.;Baghela, Abhishek
    • Mycobiology
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    • v.45 no.4
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    • pp.401-408
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    • 2017
  • Colletotrichum gloeosporioides is an economically important fungal pathogen causing substantial yield losses indifferent host plants. To understand the genetic diversity and molecular epidemiology of this fungus, we have developed a novel, high-resolution multi-locus microsatellite typing (MLMT) method. Bioinformatic analysis of C. gloeosporioides unannotated genome sequence yielded eight potential microsatellite loci, of which five, CG1 $(GT)_n$, CG2 $(GT1)_n$, CG3 $(TC)_n$, CG4 $(CT)_n$, and CG5 $(CT1)_n$ were selected for further study based on their universal amplification potential, reproducibility, and repeat number polymorphism. The selected microsatellites were used to analyze 31 strains of C. gloeosporioides isolated from 20 different host plants from India. All microsatellite loci were found to be polymorphic, and the approximate fragment sizes of microsatellite loci CG1, CG2, CG3, CG4, and CG5 were in ranges of 213-241, 197-227, 231-265, 209-275, and 132-188, respectively. Among the 31 isolates, 55 different genotypes were identified. The Simpson's index of diversity (D) values for the individual locus ranged from 0.79 to 0.92, with the D value of all combined five microsatellite loci being 0.99. Microsatellite data analysis revealed that isolates from Ocimum sanctum, Capsicum annuum (chili pepper), and Mangifera indica (mango) formed distinct clusters, therefore exhibited some level of correlation between certain genotypes and host. The developed MLMT method would be a powerful tool for studying the genetic diversity and any possible genotype-host correlation in C. gloeosporioides.

FXRα Down-Regulates LXRα Signaling at the CETP Promoter via a Common Element

  • Park, Sung-Soo;Choi, Hojung;Kim, Seung-Jin;Kim, Ok Jin;Chae, Kwon-Seok;Kim, Eungseok
    • Molecules and Cells
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    • v.26 no.4
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    • pp.409-414
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    • 2008
  • The cholesteryl ester transfer protein (CETP), a key player in cholesterol metabolism, has been shown to promote the transfer of triglycerides from very low density lipoprotein (VLDL) and low density lipoprotein (LDL) to high density lipoprotein (HDL) in exchange for cholesterol ester. Here we demonstrate that farnesoid X receptor ${\alpha}$ ($FXR{\alpha}$; NR1H4) down-regulates CETP expression in HepG2 cells. A $FXR{\alpha}$ ligand, chenodeoxycholic acid (CDCA), suppressed basal mRNA levels of the CETP gene in HepG2 cells in a dose-dependent manner. Using gel shift and chromatin immunoprecipitation (ChIP) assays, we found that $FXR{\alpha}$ could bind to the liver X receptor ${\alpha}$ ( $LXR{\alpha}$; NR1H3) binding site (LXRE; DR4RE) located within the CETP 5' promoter region. $FXR{\alpha}$ suppressed $LXR{\alpha}$-induced DR4RE-luciferase activity and this effect was mediated by a binding competition between $FXR{\alpha}$ and $LXR{\alpha}$ for DR4RE. Furthermore, the addition of CDCA together with a $LXR{\alpha}$ ligand, GW3965, to HepG2 cells was shown to substantially decrease mRNA levels of hepatic CETP gene, which is typically induced by GW3965. Together, our data demonstrate that $FXR{\alpha}$ down-regulates CETP gene expression via binding to the DR4RE sequence within the CETP 5' promoter and this $FXR{\alpha}$ binding is essential for $FXR{\alpha}$ inhibition of $LXR{\alpha}$-induced CETP expression.

Mapping of the Reduced Culm Number Trait in Rice (Oryza sativa L.) rcn10(t) Mutant

  • Yeo, Un-Sang;Lee, Jong-Hee;Kim, Choon-Song;Jeon, Meong-Gi;Oh, Tae-Yong;Han, Chang-Deok;Shin, Mun-Sik;Oh, Byeong-Geun
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.223-227
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    • 2008
  • In rice, tillering is an important trait determining yield. To study tillering at the agricultural and molecular aspects, we have examined a spontaneous rice mutant that showed reduction in the number of culms. The mutant was derived from a $F^6$ line of the cross of Junambyeo*4 / IR72. It could produce, on average, 4 tillers per hill in the paddy field while wild-type plants usually have 15. Except the reduced culm numbers, they also show pale green phenotypes. The phenotypes of this mutant were co-segregated as the monogenic Mendelian ratio (${\chi}^b=0.002$, p=0.969). In order to locate a gene responsible for the rcn phenotype, the mutant with the japonica genetic background was crossed with Milyang21 of the indica background. Bulked segregant analysis was used for rapid determination of chromosomal location. Three SSR markers (RM551, RM8213, and RM16467) on chromosome 4 were genetically associated with the mutant phenotype. Each of the 217 $F_2$ plants was genotyped with simple sequence length polymorphisms. The data showed that RM16572 on chromosome 4 was the closest marker that showed perfect co-segregation among the $F_2$ population. We suggest the new rcn gene studied here name as $rcn10^t$ because there was no report which exhibit a rcn phenotype with a pleiotropic effect of pale green (chlorophyll deficiency), and mapped at same position on chromosome 4.

Conservation Biology of Endangered Plant Species in the National Parks of Korea with Special Reference to Iris dichotoma Pall. (Iridaceae)

  • So, Soonku;Myeong, Hyeon-Ho;Kim, Tae Geun;Oh, Jang-Geun;Kim, Ji-young;Choi, Dae-hoon;Yun, Ju-Ung;Kim, Byung-Bu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.32-32
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    • 2019
  • The aim of this study was to provide basic guidelines for conservation and management of endangered plants in the national parks of Korea. Iris dichotoma Pall. (Iridaceae), which is a popular garden plant, is considered a second-class endangered species by Korean government and it is listed as a EN (Endangered) species in Red Data Book of Korea. We analyzed ecological conditions of I. dichotoma habitats based on vegetation properties and soil characteristics. This species which is known to inhabit in grassland adjacent to the ocean of lowlands slope and its population was located at an elevation of 8 m to 11 m. In the study sites, the mean of soil organic matter, total nitrogen and soil pH were 6.16%, 0.234% and 5.39 respectively. Additionally, the genetic variation and structure of three populations were assessed using ISSR (Inter Simple Sequence Repeat) markers. The genetic diversity of I. dichotoma (P = 59.46%, H = 0.206, S = 0.310) at the species level was relatively high. Analysis of molecular variance (AMOVA) showed 82.1% of the total genetic diversity was occurred in within populations and 17.9% variation among populations. Lastly, we developed predicted distribution model based on climate and topographic factors by applying SDMs (Species Distribution Models). Consequently, current status of I. dichotoma habitats is limited with natural factors such as the increase of the coverage rate of the herbs due to ecological succession. Therefore, it is essential to establish in situ and ex situ conservation strategies for protecting natural habitats and to require exploring potential and alternative habitats for reintroduction.

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