• Title/Summary/Keyword: Phylogenetic analyses

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Isolation of an Agarolytic Bacteria, Cellvibrio mixtus SC-22 and The Enzymatic Properties (한천분해세균 Cellvibrio mixtus SC-22의 분리 및 효소적 특성)

  • Cha, Jeong-Ah;Kim, Yoo-Jin;Seo, Yung-Bum;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.157-162
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    • 2009
  • An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.

Diversity of Duodenal and Rectal Microbiota in Biopsy Tissues and Luminal Contents in Healthy Volunteers

  • Li, Gangping;Yang, Min;Zhou, Kan;Zhang, Lei;Tian, Lugao;Lv, Shangze;Jin, Yu;Qian, Wei;Xiong, Hanhua;Lin, Rong;Fu, Yu;Hou, Xiaohua
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1136-1145
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    • 2015
  • The diverse microbial communities that colonize distinct segments of the gastrointestinal tract are intimately related to aspects of physiology and the pathology of human health. However, most recent studies have focused on the rectal or fecal microbiota, and the microbial signature of the duodenum is poorly studied. In this study, we compared the microbiota in duodenal and rectal samples to illustrate the characteristic microbial signatures of the duodenum in healthy adults. Nine healthy volunteers donated biopsies and luminal contents from the duodenum and rectum. To determine the composition and diversity of the microbiota, 454-pyrosequencing of bacterial 16S rRNA was performed and multiple bioinformatics analyses were applied. The α-diversity and phylogenetic diversity of the microbiota in the duodenal samples were higher than those of the rectal samples. There was higher biodiversity among the microbiota isolated from rectal biopsies than feces. Proteobacteria were more highly represented in the duodenum than in the rectum, both in the biopsies and in the luminal contents from the healthy volunteers (38.7% versus 12.5%, 33.2% versus 5.0%, respectively). Acinetobacter and Prevotella were dominant in the duodenum, whereas Bacteroides and Prevotella were dominant in the rectum. Additionally, the percentage of OTUs shared in biopsy groups was far higher than in the luminal group (43.0% versus 26.8%) and a greater number of genera was shared among the biopsies than the luminal contents. Duodenal samples demonstrated greater biological diversity and possessed a unique microbial signature compared with the rectum. The mucosa-associated microbiota was more relatively conserved than luminal samples.

Isolation and Identification of Three Pseudomonas koreensis Strains with Anti-microbial Activities Producing Inducers of the Expression of Egr-1 Gene (Egr-1 유전자의 발현 유도물질을 생산하는 항균성 저 영양 세균의 분리 및 동정)

  • Yoon, Sang-Hong;Kim, Dong-Gwan;Lee, Young-Han;Shin, Soon-Young;Kwon, Soon-Woo;Lee, Chang-Muk;Kang, Han-Chul;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.119-125
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    • 2011
  • The Egr-1 gene is known to be a transcription factor for activating the expression of many tumor-repressing genes. In this study, three strains activating the promoter of the Egr-1 gene were selected, through the use of Egr-1 luciferase reporter assay and western blotting, from amongst approximately 3,800 oligotrophic bacteria isolated from the cultivated soils of various regions within Korea. These strains were identified as Pseudomonas koreensis on the basis of phylogenetic tree analysis of their 16S ribosomal DNA sequences and biochemical characteristics analyses using a variety of commercial kits (API 20NE, ID 32GN, API ZYM kits). In addition, we discovered that these strains produced anti-bacterial activity against Bacillus subtilis, Staphylococcus aureus and Listeria monocytogenes.

Systematic Study on the Aconitum alboviolaceum Complex (Ranunculaceae) in Korea (한국산 줄바꽃 종집단의 분류학적 연구)

  • Lee, Soo-Rang;Park, Chong-Wook
    • Korean Journal of Plant Taxonomy
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    • v.37 no.4
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    • pp.477-502
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    • 2007
  • The Aconitum alboviolaceum Kom. complex includes four controversial species described from Korea; A. albouiolaceum, A. pseudolaeue, A. longecassidatum, and A. quelpaertense. The main objective of this study was to determine the taxonomic identities and systematic relationships among the species in the A. albouiolaceum complex based on morphology, numerical analyses and DNA sequence analysis. In the present study, variations in the principal morphological characters and chloroplast DNA noncoding region sequences (psbA-trnH IGS, trnL intron, and trnL-trnF IGS) were examined for 95 individuals from 20 populations. Also, neighbor-joining analysis was adopted to infer their relationships. Morphological variation appeared to be considerably high but not to be related to geographic distribution. These morphological results suggest that reevaluation of key morphological characters are needed for the proper taxonomic treatment of the complex. The length of psbA-trnH IGS region ranged from 241 to 250 bp, that of the trnL intron from 526 to 532 bp, and that of the trnL-trnF IGS region from 466 to 472 by in all taxa. Nine haplotypes were recognized from the analysis. Seven populations shared more than two haplotypes, while other thirteen populations shared only one haplotype. In the phylogenetic analysis, the nine haplotypes formed four groups, separated A. sibiricum, one of the sister groups of the complex. There also was no distinct grouping pattern supporting the species and populations observed. These results suggest that introgression or speciation might have been involved in the formation of taxa of A. alboviolaceum complex.

Molecular Cloning, Characterization and Functional Analysis of a 2C-methyl-D-erythritol 2, 4-cyclodiphosphate Synthase Gene from Ginkgo biloba

  • Gao, Shi;Lin, Juan;Liu, Xuefen;Deng, Zhongxiang;Li, Yingjun;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.39 no.5
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    • pp.502-510
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    • 2006
  • 2C-methyl-D-erythritol 2, 4-cyclodiphosphate synthase (MECPS, EC: 4.6.1.12) is the fifth enzyme of the non-mevalonate terpenoid pathway for isopentenyl diphosphate biosynthesis and is involved in the methylerythritol phosphate (MEP) pathway for ginkgolide biosynthesis. The full-length mecps cDNA sequence (designated as Gbmecps) was cloned and characterized for the first time from gymnosperm plant species, Ginkgo biloba, using RACE (rapid amplification of cDNA ends) technique. The full-length cDNA of Gbmecps was 874 bp containing a 720 bp open reading frame (ORF) encoding a peptide of 239 amino acids with a calculated molecular mass of 26.03 kDa and an isoelectric point of 8.83. Comparative and bioinformatic analyses revealed that GbMECPS showed extensive homology with MECPSs from other species and contained conserved residues owned by the MECPS protein family. Phylogenetic analysis indicated that GbMECPS was more ancient than other plant MECPSs. Tissue expression pattern analysis indicated that GbMECPS expressed the highest in roots, followed by in leaves, and the lowest in seeds. The color complementation assay indicated that GbMECPS could accelerate the accumulation of $\beta$-carotene. The cloning, characterization and functional analysis of GbMECPS will be helpful to understand more about the role of MECPS involved in the ginkgolides biosynthesis at the molecular level.

Mitochondrial Cytochrome b Sequence Variations and Population Structure of Siberian Chipmunk (Tamias sibiricus) in Northeastern Asia and Population Substructure in South Korea

  • Lee, Mu-Yeong;Lissovsky, Andrey A.;Park, Sun-Kyung;Obolenskaya, Ekaterina V.;Dokuchaev, Nikolay E.;Zhang, Ya-Ping;Yu, Li;Kim, Young-Jun;Voloshina, Inna;Myslenkov, Alexander;Choi, Tae-Young;Min, Mi-Sook;Lee, Hang
    • Molecules and Cells
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    • v.26 no.6
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    • pp.566-575
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    • 2008
  • Twenty-five chipmunk species occur in the world, of which only the Siberian chipmunk, Tamias sibiricus, inhabits Asia. To investigate mitochondrial cytochrome b sequence variations and population structure of the Siberian chipmunk in northeastern Asia, we examined mitochondrial cytochrome b sequences (1140 bp) from 3 countries. Analyses of 41 individuals from South Korea and 33 individuals from Russia and northeast China resulted in 37 haplotypes and 27 haplotypes, respectively. There were no shared haplotypes between South Korea and Russia - northeast China. Phylogenetic trees and network analysis showed 2 major maternal lineages for haplotypes, referred to as the S and R lineages. Haplotype grouping in each cluster was nearly coincident with its geographic affinity. In particular, 3 distinct groups were found that mostly clustered in the northern, central and southern parts of South Korea. Nucleotide diversity of the S lineage was twice that of lineage R. The divergence between S and R lineages was estimated to be 2.98-0.98 Myr. During the ice age, there may have been at least 2 refuges in South Korea and Russia - northeast China. The sequence variation between the S and R lineages was 11.3% (K2P), which is indicative of specific recognition in rodents. These results suggest that T. sibiricus from South Korea could be considered a separate species. However, additional information, such as details of distribution, nuclear genes data or morphology, is required to strengthen this hypothesis.

Intra-, Inter-specific Variation of Korean Rana (Amphibia: Ranidae) Based on the Partial Sequence of Mitochondrial 16S rDNA (미토콘드리아 16S rDNA부분 염기서열을 이용한 한국산 개구리 속(Amphibia: Ranidae)의 종간, 종내 변이에 대한 연구)

  • 송재영;신정아;장민호;윤병수;정규회
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.66-74
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    • 2004
  • In order to clarify intra-and inter-specific variation of Korean Rana species, the partial DNA sequences of mitochondrial 16S rDNA gene were determined from 6 Korean and 1 Japanese Rana species, DNA sequences from Korean and Japanese species were comparison-analysed within, and also with the sequences from three species of Japanese brown frogs. DNA similarities were calculated as 91.3∼97.3% among brown frog (R. amurensis coreana, R. dybowskii and R. huanrenensis), as 96.11∼97.26% among pond frogs (R. nigromaculata and R. planeyi chosenica). Genetic distance of pond frog and wrinkle fyog (R. rugosa) were near than that of pond frog and brown frog. Two clusters were formed brown frogs and the other group by neigh-bor-joining and maximum-likelihood analysis, also the populations of R. nigromaculata were well distinguished between Korean peninsula and Korean island. But result from maximum-likelihood analysis slightly differed from neighbor-joining to cluster of R. rugosa. Further analyses for their population will be necessary to study the phylogenetic status.

Fusarium Wilt Caused by Fusarium oxysporum on Passionfruit in Korea (Fusarium oxysporum에 의한 패션프루트 시들음병)

  • Joa, Jae-Ho;Choi, In-Young;Choi, Min-Kyung;Heo, Byong-Soo;Jang, Jong-Ok;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.24 no.1
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    • pp.75-80
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    • 2018
  • From 2014 to 2016, Fusarium wilt disease was found on fassionfruit in Iksan and Jeju, Korea. Symptoms included wilting of foliage, drying and withering of leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelia with 35 mm after one week. Macroconidia were falcate to almost straight, thin-walled and usually 2-3 septate. Microconidia were usually formed on monophialides of the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $3-12{\times}2.5-6{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of two molecular markers, internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$, the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of F. oxysporum on fassionfruit in Korea.

Current status and prospects of the authentication of Angelica species (Angelica 속 식물의 종판별을 위한 연구현황 및 전망)

  • Gil, Jinsu;Park, Sang ik;Lee, Yi;Kim, Ho Bang;Kim, Seong-Cheol;Kim, Ok-Tae;Cha, Seon-Woo;Jung, Chan Sik;Um, Yurry
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.151-156
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    • 2016
  • Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, natural cosmetics, and pharmaceutical industries. However, names are different from each country and their phylogenetic origins are not clear. These lead consumers to be confused. In particular, when they are morphologically similar and distributed as dried roots, it is extremely difficult to differentiate their origins even by specialists. Recently, molecular markers have been extensively applied to identify the origin of many crops. In this review, we tried to overview the current research achievements for the development of suitable 'origin identification' regarding to the differentiation of Angelica species. Furthermore, more advanced techniques including amplification genome based marker analyses are also discussed for their practical applications in the authentication of particular medicinal plant in Angelica species.

Genetic diversity and divergence among Korean cattle breeds assessed using a BovineHD single-nucleotide polymorphism chip

  • Kim, Seungchang;Cheong, Hyun Sub;Shin, Hyoung Doo;Lee, Sung-Soo;Roh, Hee-Jong;Jeon, Da-Yeon;Cho, Chang-Yeon
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.11
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    • pp.1691-1699
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    • 2018
  • Objective: In Korea, there are three main cattle breeds, which are distinguished by coat color: Brown Hanwoo (BH), Brindle Hanwoo (BRH), and Jeju Black (JB). In this study, we sought to compare the genetic diversity and divergence among there Korean cattle breeds using a BovineHD chip genotyping array. Methods: Sample data were collected from 168 cattle in three populations of BH (48 cattle), BRH (96 cattle), and JB (24 cattle). The single-nucleotide polymorphism (SNP) genotyping was performed using the Illumina BovineHD SNP 777K Bead chip. Results: Heterozygosity, used as a measure of within-breed genetic diversity, was higher in BH (0.293) and BRH (0.296) than in JB (0.266). Linkage disequilibrium decay was more rapid in BH and BRH than in JB, reaching an average $r^2$ value of 0.2 before 26 kb in BH and BRH, whereas the corresponding value was reached before 32 kb in JB. Intra-population, interpopulation, and Fst analyses were used to identify candidate signatures of positive selection in the genome of a domestic Korean cattle population and 48, 11, and 11 loci were detected in the genomic region of the BRH breed, respectively. A Neighbor-Joining phylogenetic tree showed two main groups: a group comprising BH and BRH on one side and a group containing JB on the other. The runs of homozygosity analysis between Korean breeds indicated that the BRH and JB breeds have high inbreeding within breeds compared with BH. An analysis of differentiation based on a high-density SNP chip showed differences between Korean cattle breeds and the closeness of breeds corresponding to the geographic regions where they are evolving. Conclusion: Our results indicate that although the Korean cattle breeds have common features, they also show reliable breed diversity.