• Title/Summary/Keyword: Phylogenetic analyses

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Isolation of Two cDNAs Encoding a Putative Peptidohlycan Recognition Protein Gene from the Domestic Silkworm, Bombyx mori

  • Kim, Sang-Hyun;Lee, Heui-Sam;Kim, Jin-Won;Lee, Young-Sin;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.31-36
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    • 2002
  • Peptidohlycan recognition protein (PGRP) is one of the pattern recognition proteins in innate immunity of insect. We isolated differentially expressed two cDNAa, BTL-LPI and BTL-LP2, in the fat body of Bombyx mori larvae injected with bacteria by subtractive hybridization method. These two clones showed amino acid sequence divergence of 30.4%. In the comparison with other insect PGRP genes, BTL-LP2 showed 48.8% and 45.2% of sequence homology to the known PGRP genes of Bombyx mori and Tricoplusia ni, respectively, and BTL-LP2 was 31.8% and 30.9% , respectively. Phylogenetic analysis showed relatively close relationship of the BTL-LP2 to the known insect PGRP, unlike BTL-LPI, which was equidistant both to insect and mammals, suggesting a divergent relationships of the two newly cloned B. mori PGRP genes. Northern blot analyses confirmed an induction of the expression of BTL-LP2 by the bacterial infection in the Int body of B. mori, suggesting the involvement of the gene in the insect immunity.

Morphological and molecular identification of Alaria paradisea (Phaeophyceae, Laminariales) from the Kurile Islands

  • Klimova, Anna V.;Klochkova, Nina G.;Klochkova, Tatyana A.;Kim, Gwang Hoon
    • ALGAE
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    • v.33 no.1
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    • pp.37-48
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    • 2018
  • Alaria is the second largest genus of the Laminariales, which is distributed far into the northern Pacific and Atlantic oceans. Due to its high morphological plasticity, over 100 specific and sub-specific names have been used in Alaria, this has been tailored down to the present 17 species through morphological revision and molecular phylogenetic analysis. Endemic species of Alaria from Russian Far East have not been thoroughly revised since their original description, and few of them were confirmed using molecular data until recently. We carried out morphological and molecular studies on A. paradisea which is an endemic species distributed on the Kurile Islands, first described by Miyabe and Nagai in 1932 as Pleuropterum paradiseum. The range of morphological variability and its distribution was re-evaluated using the type specimen as well as other specimens. Analyses of partial mitochondrial cytochrome c oxidase subunit 1 and nuclear-encoded internal transcribed spacer sequences showed that A. paradisea nested within the genus Alaria, but differs morphologically from any other Alaria species in having additional sporophylls with a central midrib (${\beta}-sporophylls$). Our results showed that A. paradisea clearly belongs to the genus Alaria based on DNA data, although the key morphological character that was used to include this species to the genus Pleuropterum, ${\beta}-sporophylls$, is stable and distinguishes it from other Alaria species.

Germ Cell Development during Spermatogenesis and Taxonomic Values of Sperm Morphology in Septifer (Mytilisepta) virgatus (Bivalvia: Mytilidae)

  • Kim, Jin-Hee;Kim, Sung-Han
    • Development and Reproduction
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    • v.15 no.3
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    • pp.239-247
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    • 2011
  • Spermatogenesis and taxonomic values of mature sperm morphology of in male Septifer (Mytilisepta) virgatus were investigated by transmission electron microscope observations. The morphologies of the sperm nucleus and the acrosome of this species are the cylinder shape and cone shape, respectively. Spermatozoa are approximately 45-50 ${\mu}m$ in length including a sperm nucleus (about 1.26 ${\mu}m$ long), an acrosome (about 0.99 ${\mu}m$ long), and tail flagellum (about 45-47 ${\mu}m$). Several electron-dense proacrosomal vesicles become later the definitive acrosomal vesicle by the fusion of several Golgi-derived vesicles. The acrosome of this species has two regions of differing electron density: there is a thin, outer electron-dense opaque region (part) at the anterior end, behind which is a thicker, more electron-lucent region (part). In genus Septifer in Mytilidae, an axial rod does not find and also a mid-central line hole does not appear in the sperm nucleus. However, in genus Mytilus in Mytilidae, in subclass Pteriomorphia, an axial rod and a mid-central line hole appeared in the sperm nucleus. These morphological differences of the acrosome and sperm nucleus between the genuses Septifer and Mytilus can be used for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are five, as seen in subclass Pteriomorphia.

Identification and Characterization of the Antifungal Substances of a Novel Streptomyces cavourensis NA4

  • Pan, Hua-Qi;Yu, Su-Ya;Song, Chun-Feng;Wang, Nan;Hua, Hui-Ming;Hu, Jiang-Chun;Wang, Shu-Jin
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.353-357
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    • 2015
  • A new actinomycete strain NA4 was isolated from a deep-sea sediment collected from the South China Sea and showed promising antifungal activities against soilborne fungal pathogens. It was identified as Streptomyces cavourensis by morphological, physiological, and phylogenetic analyses based on its 16S rRNA gene sequence. The main antifungal components were isolated and identified from the fermentation culture as bafilomycins B1 and C1. These compounds exhibited significant antifungal activities and a broad antifungal spectrum. The results suggest that the Streptomyces cavourensis NA4 and bafilomycins B1 and C1 could be used as potential biocontrol agents for soilborne fungal diseases of plants.

Fumarate Reductase-Producing Enterococci Reduce Methane Production in Rumen Fermentation In Vitro

  • Kim, Seon-Ho;Mamuad, Lovelia L.;Kim, Dong-Woon;Kim, Soo-Ki;Lee, Sang-Suk
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.558-566
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    • 2016
  • Biotic agents such as fumarate-reducing bacteria can be used for controlling methane (CH4) production in the rumen. Fumarate-reducing bacteria convert fumarate to succinate by fumarate reductase, ultimately leading to the production of propionate. Fumarate-reducing bacteria in the genus Enterococcus were isolated from rumen fluid samples from slaughtered Korean native goats. The enterococci were identified as Enterococcus faecalis SROD5 and E. faecium SROD by phylogenetic analyses of 16S rRNA gene sequences. The fumarate reductase activities of the SROD5 and SROD strains were 42.13 and 37.05 mM NADH oxidized/min/mg of cellular nitrogen (N), respectively. Supplementation of rumen fermentation in vitro with the SROD5 and SROD strains produced significantly higher propionate, butyrate, and total volatile fatty acid (VFA) concentrations than controls at 12 h; VFA concentrations tended to increase after 24 h of incubation. The generated CH4 concentration was significantly lower in the SROD5 and SROD treatment groups after 24 h of incubation. These findings indicate that E. faecium SROD has potential as a direct-fed microbial additive for increasing total VFAs while decreasing CH4 production in rumen fermentation in vitro.

Isolation and Characterization of a Xylanolytic Bacterium, Bacillus sp. MX47 (Xylanase 생산균 Bacillus sp. MX47의 분리 및 동정)

  • Chi, Won-Jae;Park, Da Yeon;Park, Jae-Seon;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.419-423
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    • 2012
  • A xylanolytic bacterial strain, MX47, was isolated from rotting plant matter in soil. The strain was aerobic and gram positive, and grew between pH 6.0 and 11.0. Cells were susceptible to thiostrepton and chloramphenicol. The major fatty acids (>3%) comprised 64.55% of iso-$C_{15:0}$, 22.76% of anteiso-$C_{15:0}$, and 3.92% of iso-$C_{17:0}$. The G/C content of the DNA was 44.15 mol%. The predominant respiratory quinone was menaquinone 7 (MK-7). Searches for 16S rRNA gene sequence similarity as well as phylogenetic analyses strongly suggested that the strain should be classified to the genus Bacillus. However, its biochemical characteristics, including acid production and enzyme activities, are different from those of other Bacillus strains in the same clade, and therefore, we propose the name Bacillus sp. MX47.

Sensitive, Accurate PCR Assays for Detecting Harmful Dinoflagellate Cochlodinium polykrikoides Using a Specific Oligonucleotide Primer Set

  • Kim Chang-Hoon;Park Gi-Hong;Kim Keun-Yong
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.122-129
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    • 2004
  • Harmful Cochlodinium polykrikoides is a notorious harmful algal bloom (HAB) species that is causing mass mortality of farmed fish along the Korean coast with increasing frequency. We analyzed the sequence of the large subunit (LSD) rDNA D1-D3 region of C. polykrikoides and conducted phylogenetic analyses using Bayesian inference of phylogeny and the maximum likelihood method. The molecular phylogeny showed that C. polykrikoides had the genetic relationship to Amphidinium and Gymnodinium species supported only by the relatively high posterior probabilities of Bayesian inference. Based on the LSU rDNA sequence data of diverse dinoflagellate taxa, we designed the C. polykrikoides-specific PCR primer set, CPOLY01 and CPOLY02 and developed PCR detection assays for its sensitive, accurate HAB monitoring. CPOLY01 and CPOLY02 specifically amplified C. polykrikoides and did not cross-react with any dinoflagellates tested in this study or environmental water samples. The effective annealing temperature $(T_{p})$ of CPOLY01 and CPOLY02 was $67^{\circ}C$. At this temperature, the conventional and nested PCR assays were sensitive over a wide range of C. polykrikoides cell numbers with detection limits of 0.05 and 0.0001 cells/reaction, respectively.

A Newly Isolated Rhizopus microsporus var. chinensis Capable of Secreting Amyloytic Enzymes with Raw-Starch-Digesting Activity

  • Li, Yu-Na;Shi, Gui-Yang;Wang, Wu;Wang, Zheng-Xiang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.383-390
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    • 2010
  • A newly isolated active producer of raw-starch-digesting amyloytic enzymes, Rhizopus microsporus var. chinensis CICIM-CU F0088, was screened and identified by morphological characteristics and molecular phylogenetic analyses. This fungus was isolated from the soil of Chinese glue pudding mill, and produced high levels of amylolytic activity under solid-state fermentation with supplementation of starch and wheat bran. Results of thin-layer chromatography showed there are two kinds of amyloytic enzymes formed by this strain, including one $\alpha$-amylase and two glucoamylases. It was found in the electron microscope experiments that the two glucoamylases can digest raw corn starch and have an optimal temperature of $70^{\circ}C$. These results signified that amyloytic enzymes secreted by strain Rhizopus microsporus var. chinensis CICIM-CU F0088 were types of thermostable amyloytic enzymes and able to digest raw corn starch.

Sequence Validation for the Identification of the White-Rot Fungi Bjerkandera in Public Sequence Databases

  • Jung, Paul Eunil;Fong, Jonathan J.;Park, Myung Soo;Oh, Seung-Yoon;Kim, Changmu;Lim, Young Woon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1301-1307
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    • 2014
  • White-rot fungi of the genus Bjerkandera are cosmopolitan and have shown potential for industrial application and bioremediation. When distinguishing morphological characters are no longer present (e.g., cultures or dried specimen fragments), characterizing true sequences of Bjerkandera is crucial for accurate identification and application of the species. To build a framework for molecular identification of Bjerkandera, we carefully identified specimens of B. adusta and B. fumosa from Korea based on morphological characters, followed by sequencing the internal transcribed spacer region and 28S nuclear ribosomal large subunit. The phylogenetic analysis of Korean Bjerkandera specimens showed clear genetic differentiation between the two species. Using this phylogeny as a framework, we examined the identification accuracy of sequences available in GenBank. Analyses revealed that many Bjerkandera sequences in the database are either misidentified or unidentified. This study provides robust reference sequences for sequence-based identification of Bjerkandera, and further demonstrates the presence and dangers of incorrect sequences in GenBank.

DNA Barcode Examination of Bryozoa (Class: Gymnolaemata) in Korean Seawater

  • Lee, Hyun-Jung;Kwan, Ye-Seul;Kong, So-Ra;Min, Bum-Sik;Seo, Ji-Eun;Won, Yong-Jin
    • Animal Systematics, Evolution and Diversity
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    • v.27 no.2
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    • pp.159-163
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    • 2011
  • DNA barcoding of Bryozoa or "moss animals" has hardly advanced and lacks reference sequences for correct species identification. To date only a small number of cytochrome c oxidase subunit I (COI) sequences from 82 bryozoan species have been deposited in the National Center for Biotechnology Information (NCBI) GenBank and Barcode of Life Data Systems (BOLD). We here report COI data from 53 individual samples of 29 bryozoan species collected from Korean seawater. To our knowledge this is the single largest gathering of COI barcode data of bryozoans to date. The average genetic divergence was estimated as 23.3% among species of the same genus, 25% among genera of the same family, and 1.7% at intraspecific level with a few rare exceptions having a large difference, indicating a possibility of presence of cryptic species. Our data show that COI is a very appropriate marker for species identification of bryozoans, but does not provide enough phylogenetic information at higher taxonomic ranks. Greater effort involving larger taxon sampling for the barcode analyses is needed for bryozoan taxonomy.