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Morphology and molecular characterization of the epiphytic dinoflagellate Amphidinium massartii, isolated from the temperate waters off Jeju Island, Korea

  • Lee, Kyung Ha;Jeong, Hae Jin;Park, Kila;Kang, Nam Seon;Yoo, Yeong Du;Lee, Moo Joon;Lee, Jin-Woo;Lee, Soojin;Kim, Taekyung;Kim, Hyung Seop;Noh, Jae Hoon
    • ALGAE
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    • v.28 no.3
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    • pp.213-231
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    • 2013
  • Amphidinium massartii Biecheler is an epiphytic and toxic dinoflagellate. Prior to the present study, A. massartii has been reported in the waters off the Mediterranean, Australian, USA, and Canadian coasts. We isolated Amphidinium cells from the coastal waters of Jeju Island, Korea and their morphology and rDNA sequences indicated that they were A. massartii. Herein, we report for the first time the occurrence of A. massartii in the waters of the temperate region in the northwestern Pacific Ocean. The large subunit (LSU) rDNA sequences of the Korean strains were 0.7% different from those of an Australian strain of A. massartii CS-259, the closest species, but were 4.1-5.8% different from those of the other Australian strains and the USA strains of A. massartii and from those of Amphidinium sp. HG115 that was isolated from subtropical Okinawan waters. In phylogenetic trees based on LSU, internal transcribed spacer, small subunit rDNA, and cytochrome b sequences, the Korean strains belonged to the A. massartii clade, which was clearly divergent from the A. carterae clade. The morphology of the Korean A. massartii strains was similar to that of the originally described French strain and recently described Australian strain. However, we report for the first time here that scales were observed on the surface of the flagella. In conclusion, the Korean A. massartii strains have unique rDNA sequences, even though they have a very similar morphology to that of previously reported strains. This report extends the known range of this dinoflagellate to the temperate waters of the northwestern Pacific Ocean.

Evolution of sea Urchin Strongylocentrotus intermedius Based on DNA Sequences of a Mitochondrial Gene, Cytochrome c Oxidase Subunit I (미토콘드리아 유전자, 치토그롬 옥시다제(subunit I)의 염기서열을 이용한 새치성게(Strongylocentrotus intermedius)의 진화과정 분석)

  • Lee, Youn-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.5 no.2
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    • pp.157-168
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    • 2000
  • Sea urchin S. intermedius occurring in the Korean east coast is a cold water species that belongs to the family Strongylocentrotidae of Echinoidea. Although it is known that there are nine species in the family, species identification criteria, phylogenetic relationships, time and process of evolution of the family members have not been uncovered clearly. In the present study, I tried to find some clues to such problems for S. intermedius by means of DNA sequences. For this, cytochrome c oxidase subunit I (COI), one of the mitochondrial genes that evolve fast and follow maternal inheritance was analyzed. DNA was extracted from the female gonad of S. intermedius, a segment of COI gene amplified by polymerase chain reaction (PCR), and finally a total of 1077 base pair sequence of COI obtained by cloning and sequencing the PCR product. The sequence was compared with homologous genes of other sea urchins and echinoderm species. Phylogenetic trees of the COI gene segment revealed that S. intenedius is a sister species of S. purpuratus which lives along the east coast of the Paciflc. With reference to the fossil records of sea urchins and genetic distances in the molecular phylogenies, it is estimated that the two species were separated about 0.89 million years ago when the earth temperature fluctuated significantly. The current disjunct distribution patterns of the two species and the climate change of the earth at the time of separation suggest that speciation might have occurred by vicariance. The COI gene sequence obtained here now can be used as a molecular character which discerns S. intermedius from the other sea urchin species of Strongylocentrotidae.

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Phylogeny of Korean Rhus spp. Based on ITS and rbcL Sequences (ITS 및 rbcL 염기서열에 근거한 한국 자생 옻나무속의 계통분류)

  • Lee, Won-Kyung;Kim, Myong-Jo;Heo, Kweon
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.1
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    • pp.60-66
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    • 2004
  • This study was carried out to confirm the phylogenetic relationships in Korean Rhus species. Sequences from internal transcribed spacers (ITS) of nuclear ribosomal DNA and rbcL gene of chloroplast DNA were determined. Cotinus coggygria was selected as outgroup because it is closest allied with Rhus in Anacardiaceae. Also, ingroup was limited as six Korean Rhus species. ITS 1 sequences in six species of Rhus and one species of Cotinus ranged from 246 to 253 bp and ITS 2 sequences from 234 to 244 bp. Concerning the G+C content of the studied taxa, ITS 1 sequences ranged from 58.0 to 68.13% and ITS 2 from 59.75 to 68.46%. On the other hand, rbcL sequences were same size in the all species examined by 1,428 bp. G+C contents of rbcL sequences were ranged from 43.56 to 43.77% which means there are nearly no different from interspecies each other. Phylogenetic tree strongly supports the colse relationships between R. succedanea and R. sylvestris. Rhus javanica and Cotinus coggygria were also closely allied with each other in ITS and rbcL trees. Therefore, R. javanica was regarded as most primitive species among the Korean Rhus species. ITS 1 region of nuclear ribosomal DNA was suggested as very useful taxonomical marker for genus Rhus.

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.

Molecular Phylogenetic Study of Anemone pendulisepala (Ranunculaceae) (태백바람꽃(Anemone pendulisepala, Ranunculaceae)의 분자계통학적 검토)

  • Lee, Chang Shook;Lee, Nam Sook;Yeau, Sung Hee
    • Korean Journal of Plant Taxonomy
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    • v.36 no.4
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    • pp.263-277
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    • 2006
  • Anemone pendulisepala, recently described as a new species, is distributed in sympatry with A. reflexa, A. amurensis, and A. raddeana at Mt. taebeark and Mt. Baekdu. Anemone pendulisepala was previously proposed to be a hybrid species between A. reflexa and A. amurensis becaue it displavs overlapping features with them in involucre shape, petiole length, sepal apex and xylem shape, To verify the taxonomic status and to examine the hybridity of A. pendulisepala, sequences of ITS region of nuclear ribosomal DNA and the psba-trnH, rps16 and trnLF region of cpDNA from 36 accessions of 5 taxa including outgroup were analyzed. In maximum parsimony tree based on ITS sequences, A. pendulisepala had the same sequences of A. reflexa and was clustered with monophyletic A. amurensis, and then A. raddeana. Anemone pendulisepala was distinguished from the other taxa by having four base insertion in rps16 region, two species-specific bases and insertion in trnLF region. In the phylogenetic trees of combined cpDNA, A. pendulisepala showed monophyly with the bootstrap 100%. Anemone pendulisepala exhibited no polymorphism and shared no sequences with putative parental or related taxa examined in this study. Molecular data suggest that A. pendulisepala should be a distinct species, and no evidence of the hybridization was detectcd.

Genome-Wide Analysis of Type VI System Clusters and Effectors in Burkholderia Species

  • Nguyen, Thao Thi;Lee, Hyun-Hee;Park, Inmyoung;Seo, Young-Su
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.11-22
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    • 2018
  • Type VI secretion system (T6SS) has been discovered in a variety of gram-negative bacteria as a versatile weapon to stimulate the killing of eukaryotic cells or prokaryotic competitors. Type VI secretion effectors (T6SEs) are well known as key virulence factors for important pathogenic bacteria. In many Burkholderia species, T6SS has evolved as the most complicated secretion pathway with distinguished types to translocate diverse T6SEs, suggesting their essential roles in this genus. Here we attempted to detect and characterize T6SSs and potential T6SEs in target genomes of plant-associated and environmental Burkholderia species based on computational analyses. In total, 66 potential functional T6SS clusters were found in 30 target Burkholderia bacterial genomes, of which 33% possess three or four clusters. The core proteins in each cluster were specified and phylogenetic trees of three components (i.e., TssC, TssD, TssL) were constructed to elucidate the relationship among the identified T6SS clusters. Next, we identified 322 potential T6SEs in the target genomes based on homology searches and explored the important domains conserved in effector candidates. In addition, using the screening approach based on the profile hidden Markov model (pHMM) of T6SEs that possess markers for type VI effectors (MIX motif) (MIX T6SEs), 57 revealed proteins that were not included in training datasets were recognized as novel MIX T6SE candidates from the Burkholderia species. This approach could be useful to identify potential T6SEs from other bacterial genomes.

Comparison of the Genetic Relationships and Osteological Aspects in Six Branchiostegid Fish Species (Perciformes)

  • Ryu, Jung-Hwa;Kim, Jin-Koo;Park, Jung-Youn
    • Animal cells and systems
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    • v.13 no.3
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    • pp.323-329
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    • 2009
  • We analyzed partial sequences of cytochrome b (cyt-b), a mitochondrial DNA (mtDNA) gene, to determine the genetic relationships between six horsehead fish species: Branchiostegus japonicus, Branchiostegus albus, Branchiostegus auratus, Branchiostegus argentatus, Branchiostegus wardi, and an unidentified Branchiostegus species. The specimens were collected in Korea, China, Japan, and Vietnam. We compared their molecular phylogenetic relationships inferred from mtDNA cyt-b sequences with an osteological analysis. The unidentified species, B. sp., was similar to B. albus in terms of the lack of triangular silver-white dot at the posterior region of eyes (vs. large one present in B. japonicus), but was also similar to B. japonicus in terms of the presence of a straight-shaped first hemal spine (vs. a curve-shaped hemal spine in B. albus). Analysis of the mtDNA cyt-b sequences indicated that the smallest estimated sequence divergence was between the B. japonicus and B. sp. (0.70-0.94%), whereas the largest difference was between B. auratus and B. argentatus (23.06-23.36%). Both the maximum parsimony and maximum likelihood trees showed that the B. sp. was closely clustered with B. japonicus, and that B. auratus was most distant from the other species. When comparing the osteological characters, UPGMA tree showed that the B. japonicus and B. sp. were the most closely clustered species, and B. auratus was the most distantly clustered fish relative to the other species. The shape of the nasal, otolith and first hemal spine was informative for distinguishing B. auratus from the other species. These osteological differences were consistent with the differences in mtDNA.

Cloning and Characterization of a new tobamovirus infecting Hibiscus rosa-sinensis

  • Srinivasan, L.K.G.;Wong, S.M.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.125.3-126
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    • 2003
  • A near full-length sequence of a new tobamovirus infecting Hibiscus rosa-sinensis L. was determined. The genome consists of 58 nucleotides (nt) 5' UTR, followed by a 4.9 kb ORF which methyl transferase helicase domain (128 kDa), readthrough protein RNA dependent RNA polymerase (RdRp) 185 kDa and a 52 kDa protein. The 128 kDa protein had a maximum homology of 51.4 % to TMGMV and amino acids (an) were 54.3 % identical to TMV- vulgare strain. The 185 kDa RdRp had a maximum homology of 53.5% to TMV-Ob and KGMMV-Y and a 59.6% homology at the an level to CGMMV-SH. The MP gene encodes 282 aa and its theoretical molecular weight is 30.4 kDa. The nt and an sequence identities of MP ranged from 38.8% to 43.9% and 30.9% to 37.9%, respectively. The CP gene encodes 163 residues and with a theoretical molecular weight of 18.2 kDa The (nt) and aa sequences of the CP were 46.9 % to 51.6% and 45.3% to 57.1% identical to other tobamoviruses, respectively. The predicted virion origin of assembly (OAS) was located in the CP gene. Phylogenetic trees generated based on the nt and as sequences of RdRp, MP and CP genes indicated that this new virus clustered with subgroup II tobamoviruses. Although the CP ORF of this virus shared a high nt and aa sequence identity with Sunn-hemp mosaic virus (SHMV), Western analysis showed that it is serologically unrelated to SHMV. We propose the name Hibiscus virus S (HVS) for this Singapore isolate. This is the first report on a near full-length sequence of a Tobamovirus that infects hibiscus.

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Report on 31 unrecorded bacterial species in Korea that belong to the phylum Actinobacteria

  • Choi, Jung-Hye;Cha, Ju-Hee;Bae, Jin-Woo;Cho, Jang-Cheon;Chun, Jongsik;Im, Wan-Taek;Jahng, Kwang Yeop;Jeon, Che Ok;Joh, Kiseong;Kim, Seung Bum;Seong, Chi Nam;Yoon, Jung-Hoon;Cha, Chang-Jun
    • Journal of Species Research
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    • v.5 no.1
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    • pp.1-13
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    • 2016
  • To discover and characterize indigenous species in Korea, a total of 31 bacterial strains that belong to the phylum Actinobacteria were isolated from various niches in Korea. Each strain showed the high sequence similarity (>99.1%) with the closest bacterial species, forming a robust phylogenetic clade. These strains have not been previously recorded in Korea. According to the recently updated taxonomy of the phylum Actinobacteria based upon 16S rRNA trees, we report 25 genera of 13 families within 5 orders of the class Actinobacteria as actinobacterial species found in Korea. Cellular morphology, Gram staining, basic biochemical characteristics are described in the species description.

Neobenedenia girellae infection of aquarium-raised snubnose pompano (Trachinotus blochii) in Korea

  • Nam, U-Hwa;Seo, Hyun-Joon;Hwang, Ilson;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.33 no.1
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    • pp.15-21
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    • 2020
  • We found skin flukes in snubnose pompano (Trachinotus blochii) from a public aquarium and attempted clear identification of them to the species level by morphology and molecular analyses. Skin flukes were collected from snubnose pompano showing dyspnea, anorexia and mild hemorrhage on the skin. All the fish samples (n=2) were infected with the flukes on the skin, gill and eyes, covered with excessive mucus. The isolated worms were transferred for making slide specimen and PCR amplification targeting 18S rDNA, 28S rDNA, mitochondrial cytochrome c oxidase subunit 1 (mt cox1) and cytochrome b (Cytb) genes for further analyses. Morphology and measurements data of our slide specimen coincided with those of Neobenedenia girellae. The sequence data of 2 genes (28S rDNA and Cytb) and the phylogenetic trees revealed that our specimen consistently belonged to the N. girellae clade. For 18S rDNA and mt cox1 genes, there was no sequence of either of these 2 Neobenedenia species from the type host available in GenBank. This is the first record of N. girellae in snubnose pompano, but it is still unclear if the snubnose pompano is a natural host for N. girellae or not because N. girellae is known to have an unusual broad host range and the host-switching can occur particularly in captive conditions such as aquarium or aquaculture facilities.