• Title/Summary/Keyword: 12S rDNA

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Molecular Systematics of the Genus Megoura (Hemiptera: Aphididae) Using Mitochondrial and Nuclear DNA Sequences

  • Kim, Hyojoong;Lee, Seunghwan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.510-522
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    • 2008
  • To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.

A Phylogenetic Study of Korean Rodents (Muridae, Sciuridae) Based on Mitochondrial and Nuclear DNA

  • Jung, Gi-La;Lee, Seo-Jin;Kim, Chuel-Kyu;Lee, Hang;Kim, Chang-Bae
    • Animal Systematics, Evolution and Diversity
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    • v.26 no.2
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    • pp.99-104
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    • 2010
  • The subfamily Murinae is a very controversial group concerning their phylogenetic relationship. Previous studies could not resolve phylogeny among four genera Apodemus, Micromys, Mus and Rattus of the Muridae. In the present study, eight rodent species resident in South Korea were collected and phylogenetically analyzed based on sequence data of five mitochondrial and nuclear DNA regions: 12S rRNA, cytochrome b gene (cyt b), cytochrome oxidase II (COII), control region of mitochondrial DNA, and a thyroglobulin (Tg) of nuclear DNA. According to the phylogeny of the concatenated data, M. musculus separated early in Murinae (ML 100%; BA 1.00 pp) and the genus Rattus grouped with the harvest mouse, M. minutes; these were separated from the genus Apodemus with relatively strong support (ML 74%; BA 0.76 pp). The Siberian chipmunk population was also examined using the five genes to obtain better resolution. The phylogeny for Korean rodents determined using the 12S rRNA, cyt b, COII and control regions discriminated the Siberian chipmunk populations from Korea, Russia, and China.

Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

  • Park, Joung-Eon;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Cha, Jae-Ho;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.29 no.3
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    • pp.121-131
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    • 2001
  • This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

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Genetic Stock Identification of Common Carp (Cyprinus carpio) by Detection of Intraspecific DNA Sequence Variation in the Mitochondrial 12S rRNA Gene (미토콘드리아 12S rRNA 유전자 변이 조사를 통한 잉어(Cyprinus carpio)의 유전학적 동정)

  • 남윤권;주수동;정창화;노충환;조재윤;김동수
    • Journal of Aquaculture
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    • v.10 no.4
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    • pp.403-407
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    • 1997
  • Intraspecific sequence variation was detected by polymerase chain reaction (PCR) and direct sequencing of a 350-nucleotide region of the mitochondrial 12S rRNA gene of two natural populations (Han River and Nakdong River) and one hatchery stock (Jinhae Inland Fisheries Institute) of local strain common carp, one Israeli strain of common carp stock from Pukyong National University (PKU), and one hybrid between Israeli strain of common carp female and local strain common carp male from PKU stock. There is little variation in 350 bases of the mitochondrial 12S rRNA gene sequences among 2 natural and 1 hatchery local strain common carp populatins, representing abut 7 to 20 nucleotide differences (less than 6%). The sequence of specimens from Han River was more similar to that from Nakdong River (identity=98.0%) than to that from Jinhae Inland Fisheries Institute (identity=96.3%). Sequence variation between Israeli strain and wild local strain common carp was higher than the variation within natural stocks. The level of variation was ranged from 15.7 to 17.7%. The hybrid showed very similar nucleotide4 sequence of 12S rRNA gene to the sequence of Israeli strain with the identity of 98.9%.

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Identification and Characterization of Hydrogen Peroxide-generating Lactobacillus fermentum CS12-1

  • Kang, Dae-Kyung;Oh, H.K.;Ham, J.-S.;Kim, J.G.;Yoon, C.H.;Ahn, Y.T.;Kim, H.U.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.90-95
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    • 2005
  • Lactic acid bacteria were isolated from silage, which produce high level of hydrogen peroxide in cell culture supernatant. The 16S rDNA sequences of the isolate matched perfectly with that of Lactobacillus fermentum (99.9%), examined by a 16S rDNA gene sequence analysis and similarity search using the GenBank database, thus named L. fermentum CS12-1. L. fermentum CS12-1 showed resistance to low pH and bile acid. The production of hydrogen peroxide by L. fermentum CS12-1 was confirmed by catalase treatment and high-performance liquid chromatography. L. fermentum CS12-1 accumulated hydrogen peroxide in culture broth as cells grew, and the highest concentration of hydrogen peroxide reached 3.5 mM at the late stationary growth phase. The cell-free supernatant of L. fermentum CS12-1 both before and after neutralization inhibited the growth of enterotoxigenic Escherichia coli K88 that causes diarrhea in piglets.

Physical Characteristics and Antagonistic Effect of Ampelomyces (Ampelomyces의 생리적 특성 및 길항능력)

  • Kim, Ji-Young;Lee, Wang-Hyu;Kim, Hyung-Moo
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.209-216
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    • 2009
  • During the period of June, 2005 to May, 2008, 44 host plants infected with powdery mildew were collected in the Jeon-ju and Jang-su districts of Jeonbuk province and in the Jang-sung district of Jeonnam province. The hyperparasites, Ampelomyces were confirmed in 12 plant species. Most of the pycnidium shapes of Ampelomyces were circular or oval shaped, and the sizes were different even within the same host plant, and also the color of pycnidium was ranged from light brown to dark brown. Ampelomyces species were isolated from 4 hosts including Impatiens balsamina L., Cucurbita pepo, Rudbeckia laciniata var. elatier and Youngia sonchifolia, and thus the most appropriate 12 Ampelomyces strains for the current experiment were selected. When analyzing the selected 12 strains' incubational and nutritional characteristics, the malt extract agar was the most appropriate media. When investigating the effect of osmotic pressure on the spore germination, 0.15M NaCl concentration was the optimum germination concentration. When the isolated Ampelomyces sp. was tested in-vitro, it was found to be effective to control in other plant pathogens, isolated Ampelomyces showed no pathogenicity to the plant. strains isolated . studied on rDNA ITS sequence analysis. The rDNA ITS sequence data of Ampelomyces sp. isolate BSLAH16 from Impatiens balsamina L. were analyzed and identified.

Molecular Identification and Morphological Description of Gymnothorax minor (Muraenidae, Anguilliformes) Leptocephali off Jeju Island (제주 근해에서 채집된 나망곰치, Gymnothorax minor (뱀장어목, 곰치과) 엽상자어의 분자동정 및 형태기재)

  • Lee, Seung-Jong;Ji, Hwan-Sung;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.25 no.2
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    • pp.59-64
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    • 2013
  • Two leptocephali (36.2~69.2 mm in total length) of the family Muraenidae were collected off Jeju Island, Korea. They were identified using 742 base pairs of mitochondrial DNA 12S rRNA sequences, and their morphological description were presented. The two leptocephali were identified to Gymnothorax minor based on genetic distance (d=0.000), being well consistent with that of G. minor adult collected off Tongyeong. Gymnothorax minor leptocephali are characterized by morphological combinations: total myomeres 137~139, predorsal myomeres 32, preanal myomeres 82~86, body depth in total length more than 10%.

Comparison of Phylogenetic Characteristics of Viable but Non-Culturable (VBNC) Bacterial Populations in the Pine and Quercus Forest Soil by 16S rDNA-ARDRA (16S rDNA-ARDRA법을 이용한 소나무림과 상수리나무림 토양 내 VBNC 세균군집의 계통학적 특성 비교)

  • Han Song-Ih;Kim Youn-Ji;Whang Kyung-Sook
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.116-124
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    • 2006
  • In this study was performed to analyze quantitatively the number of viable but non-culturable bacteria in the Pine and Quercus forest soil by improved direct viable count (DVC) and plate count (PC) methods. The number of living bacteria of Pine and Quercus forest soil by PC method were less then 1% of DVC method. This result showed that viable but non-culturable (VBNC) bacteria existed in the forest soil with high percentage. Diversity and structure of VBNC bacterial populations in forest soil were analyzed by direct extracting of DNA and 16S rDNA-ARDRA from Pine and Quercus forest soil. Each of them obtained 111 clones and 108 clones from Pine and Quercus forest soil. Thirty different RFLP types were detected from Pine forest soil and twenty-six different RFLP types were detected from Quercus forest soil by HeaIII. From ARDRA groups, dominant clones were selected for determining their phylogenetic characteristics based on 16S rDNA sequence. Based on the 16S rDNA sequences, dominant clones from ARDRA groups of Pine forest soil were classified into 7 major phylogenetic groups ${\alpha}$-proteobacteria (12 clones), ${\gamma}$-proteobacteria (3 clones), ${\delta}$-proteobacteria (1 clone), Flexibacter/Cytophaga (1 clone), Actinobacteria (4 clones), Acidobacteria (4 clones), Planctomycetes (5 clones). Also, dominant clones from ARDRA groups of Quercus forest soil were classified into 6 major phylogenetic groups : ${\alpha}$-proteobacte,ia (4clones), ${\gamma}$-proteobacteria (2 clones), Actinobacteria (10 clones), Acidobacteria (8 clones), Planctomycetes (1 clone), and Verrucomicobia (1 clone). Result of phylogeneric analysis of microbial community from Pine and Quercus forest soils were mostly confirmed at uncultured or unidentified bacteria, VBNC bacteria of over 99% existent in forest soil were confirmed variable composition of unknown micro-organism.

Identification of Bacteria by Sequence Analysis of 16S rRNA in Testes of Jeju Horses (제주마 고환내 세균의 16S rRNA 염기서열 분석을 이용한 동정)

  • Park, Yong-Sang;Kim, Nam-Young;Han, Sang-Hyun;Park, Nam-Geon;Ko, Moon-Suck;Cho, Won-Mo;Chae, Hyun-Seok;Cho, In-Chul;Cho, Sang-Rae;Woo, Jae-Hoon;Kang, Tae-Young
    • Journal of Veterinary Clinics
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    • v.31 no.1
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    • pp.36-39
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    • 2014
  • Many bacteria colonized in the horse semen affect quality of the sperm and some may cause infection in the mare reproductive tract and infertility of susceptible mare. This study was initiated to determine the prevalence of bacteria in testes of Jeju horses by determining rRNA sequence. The samples were swabed from the testes of nine Jeju horses (aged from 8 to 12 months after birth). Bacteria isolated from testes were identified by 16S rDNA sequencing. 1.6-kbp PCR products for 16S rRNA coding region were obtained using the universal primers. The PCR products were further purified and sequenced. Maximum similar species were found by BLAST search in the GenBank DNA database. BLAST results showed that the sequences were similar to those of Acinetobacter sp (A. schindleri, A. ursingii)., Bacillus cereus, Corynebacterium glutamicum, Escherichia coli, Gamma proteobacterium, Micrococcus luteus, Pseudomonas mendocina, Shigella sonnei, Sphingomonas sp., Staphylococcus sp (S. cohnii, S. saprophyticus, S. xylosus)., and Stenotrophomonas maltophilia. DNA sequences for 16S rRNA is provided useful informations for species identification of pathogenic microorganisms for the reproductive organs in horses.

Genetic Phylogeny among Three Species Red Seabream, Black Seabream and Rock Bream Based on Mitochondrial DNA Sequences

  • Kim, Mi-Jung;An, Hye-Suck;Kim, Kyung-Kil;Park, Jung-Youn
    • Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.171-178
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    • 2009
  • The Perciformes include approximately 40% of all bony fishes and are the largest order of vertebrates. This order includes some of the most economically relevant marine fishes, particularly the red seabream, black seabream and rock bream. A 409 bp fragment of the cytochrome b (cyt b) gene and 403 bp and 518 bp fragments of ribosomal RNA (12S and 16S rRNA, respectively) were sequenced from five populations of natural and cultured red seabreams, natural black seabream, and natural and cultured rock breams. The mitochondrial DNA sequences were utilized for the genetic identification and population structural analyses of these three species. Phylogenetic relationships of intra- and inter-species were elucidated using three types of molecular genetic markers from three species of the order Perciformes in Korea. We noted no significant differences in the intra-specific variation of the cyt b and rRNA genes in each population however, inter-specific divergences were greater than intra-specific variation. Inter-specific variation was induced more by transition than transversion type in the cyt b and rRNA genes. The cyt b gene and rRNA genes make it possible to determine the inter-species divergence. The rRNA genes have more conserved sequences than the cyt b gene. Therefore, these genes are expected to prove useful among species belonging to the different genera or families.