• Title/Summary/Keyword: 분자 계통분석

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Genetic identification of Aeromonas species using a housekeeping gene, rpoD, in cultured salmonid fishes in Gangwon-Do (강원도 양식 연어과 어류에서 분리된 에로모나스 종의 유전학적 동정)

  • Lim, Jongwon;Koo, Bonhyeong;Kim, Kwang Il;Jeong, Hyun Do;Hong, Suhee
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.79-88
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    • 2017
  • At the present, fish farms are suffering a lot of economic losses due to infectious diseases caused by various pathogens including aeromonad. Aeromonad is ubiquitous bacteria that causes infectious diseases. At least 26 species in the genus Aeromonas have been reported to cause fatal infections not only in salmonid fishes, but also in other freshwater and seawater fishes. Molecular techniques based on nucleic acid sequences of 16S rDNA and housekeeping genes can be used to identify the Aeromonas species. In this study, The genus Aeromonas was isolated from salmonid fishes of sixteen fish farms in Gangwon-Do, Korea and phylogenetically identified based on the sequences of 16S rDNA and housekeeping genes for Aeromonad, i.e. RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) or DNA gyrase subunit B (gyrB). Consequently, 96 strains were collected from Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), masou salmon (Oncorhynchus masou) and rainbow trout (Oncorhynchus mykiss), and 36 isolates were identified as the genus Aeromonas by 16S rDNA analysis. Thirty six Aeromonad isolates were further analysed based on rpoD or gyrB gene sequences and found Aeromonas salmonicida (24 isolates), A. sobria (10 isolates), A. media (1 isolates) and A. popoffii (1 isolates), indicating that A. salmonicida is a main infectious bacteria in Salmonid fishes in Gangwon-Do. It was also proved that the phylogenetic identification of Aeromonas species based on the sequences of housekeeping gene is more precise than the 16S rDNA sequence.

Analysis on the Microbial Community Structure of Red Granule in the Anaerobic Ammonium Oxidation Reactor (혐기성 암모늄 산화 반응기 내 붉은색 입상슬러지의 미생물 군집구조 분석)

  • Bae, Hyo-Kwan;Park, Kyung-Soon;Chung, Yun-Chul;Jung, Jin-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.10
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    • pp.1055-1064
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    • 2006
  • Extremely slow growing anammox(anaerobic ammonium oxidation) bacteria were cultivated using a combination of UASB(Upflow Anaerobic Sludge Blanket) reactor seeded with anaerobic granular sludge and carbon-fiber cultivating reactor. After 180 days of continuous cultivation, average nitrogen removal rate showed 0.54 kg $N/m^3-day$ when 0.6 kg $N/m^3-day$ of nitrogen loading was applied. The black granule was changed to brown and red granule as continuous operation, and the red granule was highly dependant on the high anammox activity. Microbial community structure of red granule in the UASB reactor was analyzed by molecular methods such as gene cloning, phylogenetic tree analysis, and FISH(Fluorescence In Situ Hybridization) method. As a result of gene cloning and phylogenetic tree analysis, 5 kinds of phylum were found to be Planctomycetes, Proteobacteria, Acidobacteria, Chlorobi and Chloroflexi. 13 clones were matched to anammox bacteria among 51 clones in the red anammox granule. In-silico test which used cloning information and FISH probe of the AMX368 was conducted to detect the presence of anammox bacteria in the red anammox granule. As a result of in-silico test only one clone was exactly matched to AMX368 but 11 clones was mutated one base among 18 bases representing all 12 clones are anammox bacteria. A filamentous Chloroflexi might be related to the granulation of anammox bacteria. As a result of FISH analysis, anammox bacteria was abundant in the red anammox granule.

Qualitative and Quantitative Analysis of Microbial Community Structure in the Sequencing Batch Reactor for Enriching ANAMMOX Consortium (연속회분식 반응기를 이용한 혐기성 암모늄 산화균 농후배양에서의 정성 및 정량적 미생물 군집구조 분석)

  • Bae, Hyo-Kwan;Jung, Jin-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.10
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    • pp.919-926
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    • 2009
  • Enrichment of anaerobic ammonium oxidation (ANAMMOX) bacteria is the essential step for operating full-scale ANAMMOX bioreactor because adding a significant amount of seeding sludge is required to stabilize the ANAMMOX reactor. In this study, the enrichment of ANAMMOX bacteria from an activated sludge using sequencing batch reactor was conducted and verified by analyzing changes in the microbial community structure. ANAMMOX bacteria were successfully enriched for 70 days and the substrate removal efficiencies showed 98.5% and 90.7% for $NH_4\;^+$ and $NO_2\;^-$ in the activity test, respectively. The phylogenetic trees of Planctomycetes phylum showed that the diverse microbial community structure of an activated sludge was remarkably simplified after the enrichment. All 36 clones, obtained after the enrichment, were affiliated with ANAMMOX bacteria of Candidatus Brocadia (36%) and Candidatus Anammoxoglobus (64%) genera. The quantification using real-time quantitative PCR (RTQ-PCR) revea ed that the 16S rDNA concentration of ANAMMOX bacteria was 74.8% compared to the granular ANAMMOX sludge obtained from an upflow ANAMMOX sludge bed reactor which had been operated for more than one year. The results of molecular analysis supported that the enriched sludge could be used as a seeding sludge for a full-scale ANAMMOX bioreactor.

Identification and phylogenetic analysis of the human endogenous retrovirus HERV-W pol in cDNA library of human fetal brain (인간태아의 뇌로부터 유래된 cDNA liberary에서 내생레트로바이러스 HERV-W pol 유전자의 동정과 계통)

  • Kim, Heui-Soo;Jeon, Seung-Heui;Yi, Joo-Mi;Kim, Tae-Hyung;Lee, Won-Ho
    • Journal of Life Science
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    • v.13 no.3
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    • pp.291-297
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    • 2003
  • A human endogenous retroviral family (HERV-W) has recently been described that is related to multiple sclerosis-associated retrovirus (MSRV) sequences that have been identified in particles recovered from monocyte cultures from patients with multiple sclerosis. Two pol fragments (HWP-FB10 and HWP-FBl2) of HERV-W family were identified and analysed by the PCR approach with cDNA library of human fetal brain. They showed 89 percent nucleotide sequence similarity with that of the HERV-W (accession no. AF009668). Deletion/insertion or point mutation in the coding region of the pol fragments from human fetal brain resulted in amino acid frameshift that induced a mutated protein. Phylogenetic analysis of the HERV-W family from GenBank database indicates that the HWP-FB10 is very closely related to the AC000064 derived from human chromosome 7q21-q22. Further studies on the genetic relationship with neighbouring genes and functional role of these new HERV-W pol sequences are indicated.

Use of Microsatellite Markers Derived from Genomic and Expressed Sequence Tag (EST) Data to Identify Commercial Watermelon Cultivars (수박 시판 품종의 식별을 위한 Genomic과 Expressed Sequence Tag (EST)에서 유래된 Microsatellite Marker의 이용)

  • Kwon, Yong-Sham;Hong, Jee-Hwa;Kim, Du-Hyun;Kim, Do-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.737-750
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    • 2015
  • This study was carried out to construct a DNA profile database for 102 watermelon cultivars through the comparison of polymorphism level and genetic relatedness using genomic microsatellite (gMS) and expressed sequence tag (EST)-microsatellite (eMS) markers. Sixteen gMS and 10 eMS primers showed hyper-variability and were able to represent the genetic variation within 102 watermelon cultivars. With gMS markers, an average of 3.63 alleles per marker were detected with a polymorphism information content (PIC) value of 0.479, whereas with eMS markers, the average number of alleles per marker was 2.50 and the PIC value was 0.425, indicating that eMS detects a lower polymorphism level compared to gMS. Cluster analysis and Jaccard's genetic distance coefficients using the unweighted pair group method with arithmetic average (UPGMA) based on the gMS, eMS, and combined data sets showed that 102 commercial watermelon cultivars could be categorized into 6 to 8 major groups corresponding to phenotypic traits. Moreover, this method was sufficient to identify 78 out of 102 cultivars. Correlation analysis with Mantel tests for those clusters using 3 data sets showed high correlation ($r{\geq}0.80$). Therefore, the microsatellite markers used in this study may serve as a useful tool for germplasm evaluation, genetic purity assessment, and fingerprinting of watermelon cultivars.

First Report of an Unrecorded Nematode-trapping Fungus, Arthrobotrys sinensis in Korea (국내 미기록 선충포식성 곰팡이 Arthrobotrys sinensis의 형태 및 분류)

  • Ha, Jihye;Kang, Heonil;Kang, Hangwon;Kim, Donggeun;Lee, Dongwoon;Kim, Yongchul;Choi, Insoo
    • Korean journal of applied entomology
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    • v.58 no.1
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    • pp.9-13
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    • 2019
  • Nematode-trapping fungi use various specialized traps to capture nematodes. A fungus that can capture nematodes in three dimensional adhesive networks was isolated from the soil around the root of Cucumis melo L. (Oriental melon) in Seongju, Korea. The conidiophores were found to be septate, hyaline, erect and $290-528(342.8){\mu}m$ high. It produces obovoid shape and 1-3 septate (commonly 2-septate) conidia with a size of $30.5{\times}20.3{\mu}m$. Molecular analysis of 5.8 S rDNA displayed 99% similarity to Arthrobotrys sinensis. On the basis of morphological, morphometric and molecular studies, the fungus was identified as A. sinensis. It is the first report in Korea which can be one of biological control resource of plant-parasitic nematode.

Construction of a DNA Profile Database for Commercial Cucumber (Cucumis sativus L.) Cultivars Using Microsatellite Marker (Microsatellite 마커를 이용한 오이 유통품종 DNA Profile Data Base 구축)

  • Kwon, Yong-Sham;Choi, Keun-Jin
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.344-351
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    • 2013
  • Microsatellite is one of the most suitable marker for cultivar identification as it has great discrimination power for cultivars with narrow genetic variation. The polymorphism level between 358 microsatellite primer pairs and 11 commercial cucumber cultivars was investigated. Thirty-one primer pairs showed high polymorphism within cucumber cultivars with different fruit types. These markers were applied for the constructing DNA profile data base of 110 commercial cucumber cultivars through multiplex PCR and fluorescence based automatic detection system. A total of 139 polymorphic amplified fragments were obtained by using 31 microsatellite markers. The average of PIC value was 0.610 ranging from 0.253 to 0.873. One hundred and thirty nine microsatellite loci were used to calculate Jaccard's distance coefficients for UPGMA cluster analysis. A clustering group of varieties, based on the results of microsatellite analysis, were categorized into plant shape and fruit type. Almost the cultivars were discriminated by marker genotypes. This information may be useful to compare through genetic relationship analysis between existing variety and candidate varieties in distinctive tests and protection of plant breeders' intellectual property rights through variety identification.

Morphological, Molecular and Pathogenic Characteristics of Botrytis elliptica and Botrytis cinerea Associated with Leaf Blight of Lily (백합 잎마름병에 관여하는 Botrytis elliptica와 Botrytis cinerea의 형태적, 분자적 특성과 병원성)

  • Kim, Won-Ki;Park, Myung-Soo;Hahm, Soo-Sang;Yu, Seung-Hun
    • The Korean Journal of Mycology
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    • v.35 no.2
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    • pp.68-75
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    • 2007
  • The seventy nine isolates of Botrytis spp. were obtained from leaf blight lesions of lily in Korea, Japan and Netherlands. Morphological and cultural characteristics of them were investigated and molecular characteristics of them were determined using sequence analysis of RNA polymerase II (RPB2) and heat-shock protein 60 (HSP60) gene. A selection of Botrytis isolates were evaluated for their pathogenicity to lily. Based on morphological and cultural characteristics, the Botrytis isolates were divided into two groups, and identified as B. elliptica (n = 54) and B. cinerea (n = 25). Based on analysis of RPB2 and HSP60 sequences, the Botrytis isolates were also divided into two groups and well supported morphological groupings. Spore suspensions of B, elliptica showed significant pathogenicity on lily leaves and flowers, however those of B. cinerea showed pathogenicity only on flowers but not on leaves. The latter showed pathogenicity on lily leaves only when spore suspensions amended with PDB were used as inocula.

Effect of Sequence Variation in Bovine Mitochondrial DNA D-Ioop Region on ~ilk Production for Hanwoo (한우 산유량에 미치는 Mitochondrial DNA D-loop영역의 염기서열 변이효과)

  • Kong, H.S.;Oh, J.D.;Lim, H.J.;Lee, H.K.;Jeon, G.J.;Yoon, D.H.;Jeon, G.J.;Choi, J.G.;Choi, Y.H.;Cho, B.W.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.729-734
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    • 2004
  • This study was performed to analyze the sequence variations of mtDNA D-loop and their effects on milk in Hanwoo(Korean cattle). The resulting sequences were compared with previously published sequences for other cattle breeds (GenBank JOI394). The Polymerase Chain Reaction was performed to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty polymorphic sites by nucleotide substitution were found in mtDNA D-loop region of Hanwoo. The frequencies of positions at 8, 169, 16042, 16051, 16057, 16093, 16119, 16122, 16209, 16255 and 16302 nt with high levels of sequence polymorphism were 0.150, 0.950, 0.085, 0.138, 0.106, 0.085, 0.138, 0.212, 0.085, 0.148 and 0.180, respectively. The substitution effect at 16119(p<0.1) and 16185(p< 0.05) nt was found significant on milk production. Polymorphism of mtDNA sequence in D-Ioop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.

Multi-locus Phylogeny Analysis of Korean Isolates of Phytophthora Species Based on Sequence of Ribosomal and Mitochondrial DNA (핵 및 미토콘드리아 DNA 염기서열을 이용한 국내 Phytophthora 속의 Multi-locus phylogeny 분석)

  • Seo, Mun-Won;Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.40-47
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    • 2010
  • To investigate genetic relationships either interspecies or intraspecies of 14 Korean Phytophthora species, sequence analyses of nuclear DNA (ypt gene and rDNA-IGS region) and mitochondrial DNA (Cox gene, $\beta$-tubuline gene, and EF1A gene) were performed. All of 14 Korean Phytophthora species clearly clustered into foreign isolates of each species. These Korean isolates in Phytophthora species also showed no correlation between molecular classification and morphological classification like as in case of foreigners. P. palmivora KACC 40167 reported previously from genetic groups of Phytophthora species in Korea was not consistent with the classification system, and therefore was required re-examination for the genetic group analysis. Korean isolates of P. drechsleri KACC 40195 showed very close relationship with P. cryptogea KACC 40161 above 94% bootstrap value in P. cryptogea-P. drechsleri complex group. Identification of these isolates is still unclear, because P. cryptogea and P. drechsleri were not differentiated in this study. On the other hand, it was required to unify species for these two species, since P. parasitica and P. nicotianae were clustered into a group on the level of 99 to 100% sequence homology. Comparing to the sequences of foreigners, Korean isolates were newly divided to ten groups in the phylogenic system. These results could be prepared useful informations to understand genetic diversity of Phytophthora species in Korea.