• Title/Summary/Keyword: ITs rDNA

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Cloning of Heat Shock Protein 70 and Its Expression Profile under an Increase of Water Temperature in Rhynchocypris kumgangensis (금강모치(Rhynchocypris kumgangensis)에서 heat shock protein 70의 클로닝과 수온상승에 의한 발현 변화 분석)

  • Im, Jisu;Ghil, Sungho
    • Journal of Korean Society on Water Environment
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    • v.29 no.2
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    • pp.232-238
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    • 2013
  • Water temperature is key factor influencing growth and reproduction of fish and its increase give rise to various physiological changes including gene expression. Heat shock protein (Hsp), one of the molecular chaperones, is highly conserved throughout evolution and its expression is induced by various stressors such as temperature, oxidative, physical and chemical stresses. Here, we isolated partial cDNA clones encoding 70-kDa Hsp (Hsp70) and $\beta$-actin using reverse transcriptase-PCR (RT-PCR) from gut of Rhynchocypris kumgangensis, a Korean indigenous species and cold-water fish, and investigated expression profiles of Hsp70 under an increase of water temperature using $\beta$-actin as an internal control for RT-PCR. Cloned Hsp70 cDNA of R. kumgangensis showed homology to Ctenopharyngodon idella (96%), Hypophthalmichthys molitrix (96%), Danio rerio (93%) and Oncorhynchus mykiss (81%) Hsp70. Cloned $\beta$-actin cDNA of R. kumgangensis showed homology to D. rerio (98%), H. molitrix (97%), C. idella (97%) and O. mykiss (90%) $\beta$-actin. Both mRNA of Hsp70 and $\beta$-actin were expressed in gut, brain, and liver in R. kumgangensis. Futhermore, expression of Hsp70, in brain, was highly augmented by an increase of water temperature. These results suggest that Hsp70 mRNA expression level in brain can be used as a biological molecular marker to represent physiological stress against an increase of water temperature.

Sclerotium Rot of Pulsatilla koreana Nakai Caused by Sclerotium rolfsii (Sclerotium rolfsii에 의한 할미꽃 흰비단병)

  • Kwon, Jin-Hyeuk;Hwang, Gap-Chun;Lee, Seol-Bi;Choi, Yong-Jo;Lee, Sang-Dae;Nguyen, Le-Minh
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.334-336
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    • 2015
  • Sclerotium rot on Pulsatilla koreana was observed in the exhibition field of Jinju Agriculture Technology Center in July 2013. The infected plants showed water-soaked, blighted and rotted symptoms. White mycelial mats spread over lesions, and then numerous sclerotia were formed on flower stalk and flower stem near the soil line. The sclerotia were globoid in shape, 1-3 mm in size and white to brown in color. The optimum temperature for mycelial growth and sclerotia formation on PDA was $30^{\circ}C$ and the hyphal width was $4-8{\mu}m$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. For molecular identification, the complete ITS rDNA sequence of the causal fungus was sequenced and analyzed. On the basis of mycological characteristics, ITS rDNA sequence analysis, and pathogenicity to host plants, this fungus was identified as Sclerotium rolfsii Saccardo. This is the first report of sclerotium rot on Pulsatilla koreana caused by S. rolfsii in Korea.

Isolation of Streptomyces sp. YU100 Producing Extracellular Phospholipase D

  • Lim, Si-Kyu;Choi, Jae-Woong;Lee, Eun-Tag;Khang, Yong-Ho;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.71-76
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    • 2002
  • Soil samples were screened for actinomycete strains capable of producing phospholipase D, and a strain, Streptomyces sp. YU100, showing a high transphosphatidylation activity was isolated. This strain secreted phospholipase D in a culture broth after 12 h of cultivation, and its productivity continued to increase for 36 h of fermentation. In addition, its transphosphatidylation rate of phosphatidylcholine to phosphatidylserine was almost $68\%$ within 1 h. The morphological and chemotaxonomical characteristics showed that this strain could be classified as a number of the Streptomycetaceae family, particularly due to the spiral form of its spore chain consisting of 60-70 smooth spores $(0.75{\times}1.0{\mu}m$) on an aerial mycelium, FA-2c type of fatty acid profile in the cell wall, and LL-DAP component in the cell wall peptidoglycan. A phylogenetic analysis of the 16S rDNA provided a clue that the strain YU100 was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyes sp. ASB27, S. peucetius JCM9920, and S. griseus ATCC10137. A dendrogram based on the 16S rDNA sequences also showed a phylogenetic relationship between the strain YU100 and these strains. However, the strain YU100 has not yet been assigned to a particular species, because of absence of any other classified species with a high matching score.

Taxonomic Revision of Notohymena gangwonensis (Protozoa: Ciliophora), with Notes on Its Cortical Granules and Scanning Electron Micrographs

  • Moon, Ji Hye;Kim, Kang-San;Chae, Kyu Seok;Min, Gi-Sik;Jung, Jae-Ho
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.2
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    • pp.113-122
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    • 2020
  • From a moss sample, we isolated and identified Notohymena gangwonensis Kim et al., 2019 based on morphological and molecular data. The moss and type population has completely identical 18S rRNA (nuclear small subunit ribosomal RNA) gene sequences and both are highly similar in morphological and morphometric attributes, except for the diameter and arrangement of the cortical granules. Thus, we reexamined the type materials(i.e., micrographs and gDNA) and resulted in finding mistakes made by the authors of the species. Based on these data and supporting materials newly obtained (i.e., internal transcribed spacer [ITS] 1, ITS2, 5.8S, and partial 28S rDNA sequences, and scanning electron micrographs), we provide improved diagnosis of the species to clarify its identity. In addition, a key for Notohymena species is provided.

Morphological and molecular characterization of the genus Coolia (Dinophyceae) from Bahía de La Paz, southwest Gulf of California

  • Morquecho, Lourdes;Garate-Lizarraga, Ismael;Gu, Haifeng
    • ALGAE
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    • v.37 no.3
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    • pp.185-204
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    • 2022
  • The genus Coolia A. Meunier 1919 has a global distribution and is a common member of epiphytic dinoflagellate assemblages in neritic ecosystems. Coolia monotis is the type species of the genus and was the only known species for 76 years. Over the past few decades, molecular characterization has unveiled two species complexes that group morphologically very similar species, so their limits are often unclear. To provide new knowledge on the biogeography and species composition of the genus Coolia, 16 strains were isolated from Bahía de La Paz, Gulf of California. The species were identified by applying morphological and molecular approaches. The morphometric characteristics of all isolated Coolia species were consistent with the original taxa descriptions. Phylogenetic analyses (large subunit [LSU] rDNA D1 / D2 and internal transcribed spacer [ITS] 1 / 5.8S / ITS2) revealed a species assemblage comprising Coolia malayensis, C. palmyrensis, C. tropicalis, and the C. cf. canariensis lineage. This is the first report of Coolia palmyrensis and C. cf. canariensis in Mexico and C. tropicalis in the Gulf of California. Our results strengthen the biogeographical understanding of these potentially harmful epiphytic dinoflagellate species.

Classification and Characterization of Bacteriophages of Lactobacillus casei -Analysis of Restriction Patterns of Phage DNA- (Lactobacillus casei bacteriophage의 분류 및 특성에 관한 연구 - Phage DNA의 제한효소 절편 비교 분석-)

  • 김영창;강현삼
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.115-121
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    • 1985
  • Five representative virulent phages (J1, TK93, K1, PD5, and CP1) and one temperate phage (.phi.1043) of Lactobacillus casei were compared to each other by analyzing the agarose gel electrophoretic patterns of restriction enzyme-digested phage DNAs. Nucleic acids of all the tested phages were double stranded DNA. DNAs of J1, TK93, K1, and ${\phi}$ 1043 phages had a size of about 42kb, but the size of PD5 and CP1 DNAs was avout 140kb. J1, TK93, K1, PD5, CP1, and ${\phi}$ 1043 DNAs were digested to 13, 13, 11, 14, 14, and 12 fragments by EcoR1, respectively, and showed its characteristec restriction patterns. Cohesive ends were present in J1, TK93, and ${\phi}$ 1043, but were absent in K1, PD5, and CP1. Restriction maps of J1 and TK93 DNAs showed nearly complete homology and their evolutionary relationship based upon the restriction analysis was discussed.

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Morphological and genetic characterization and the nationwide distribution of the phototrophic dinoflagellate Scrippsiella lachrymosa in the Korean waters

  • Lee, Sung Yeon;Jeong, Hae Jin;You, Ji Hyun;Kim, So Jin
    • ALGAE
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    • v.33 no.1
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    • pp.21-35
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    • 2018
  • The phototrophic dinoflagellate genus Scrippsiella is known to have a worldwide distribution. Here, we report for the first time, the occurrence of Scrippsiella lachrymosa in Korean waters. Unlike the other stains of S. lachrymosa whose cultures had been established from cysts in the sediments, the clonal culture of the Korean strain of S. lachrymosa was established from motile cells. When the sulcal plates of S. lachrymosa, which have not been fully described to date, were carefully examined using scanning electron microscopy, the Korean strain of S. lachrymosa clearly exhibited the anterior sulcal plate (s.a.), right sulcal plate (s.d.), left sulcal plate (s.s.), median sulcal plate (s.m.), and posterior sulcal plate (s.p.). When properly aligned, the large subunit (LSU) rDNA sequence of the Korean strain of S. lachrymosa was ca. 1% different from those of two Norwegian strains of S. lachrymosa, the only strains for which LSU sequences have been reported. The internal transcribed spacer (ITS) rDNA sequence of the Korean strain of S. lachrymosa was also ca. 1% different from those of the Scottish and Chinese strains and 3% different from those of the Canadian, German, Greek, and Portuguese strains. Thus, the Korean S. lachrymosa strain has unique LSU and ITS sequences. The abundances of S. lachrymosa in the waters of 28 stations, located in the East, West, and South Sea of Korea, were quantified in four seasons from January 2016 to October 2017, using quantitative real-time polymerase chain reaction method and newly designed specific primer-probe sets. Its abundances were >$0.1cells\;mL^{-1}$ at eight stations in January and March 2016 and March 2017, and its highest abundance in Korean waters was $26cells\;mL^{-1}$. Thus, S. lachrymosa has a nationwide distribution in Korean waters as motile cells.

Phylogenetic Analysis of Schizonepeta Spike on the Basis of DNA Sequences (염기서열을 이용한 한약재 형개의 기원 및 유연관계 분석)

  • Jigden, Baigalmaa;Kim, Myung-Kyum;Noh, Jong-Hun;Sun, Hua;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.1
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    • pp.46-53
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    • 2009
  • Schizonepeta spike (Korean name "Hyung-Gae") has been used for oriental medicinal purposes in Korea, China and Japan. In this study, twenty six "Hyung-Gae" samples were collected including nine certified Schizonepeta tenuifolia plants, and seventeen commercially marketed "Hyung-Gae" products. Chloroplast trnL-F and rDNA ITS regions of the "Hyung-Gae" samples were sequenced and used to identify whether the samples were genuine S. tenuifolia or not. As the result, the trnL-F and ITS sequences of all the "Hyung-Gae" samples were shown to be identical and it was proven that commercially available medicinal products "Hyung-Gae" are genuine S. tenuifolia. Phylogenetic tree of S. tenuifolia using the trnL-F sequences was constructed and compared with phylogenetic tree using ITS of rDNA region sequences. In these tree, S. tenuifolia was affiliated in the family Lamiaceae. It is proven that trnL-F and ITS phylogenetic trees are useful to study taxonomic position of S. tenuifolia.

Occurrence of Gray Mold Caused by Botrytis cinerea on Common Fig in Korea (Botrytis cinerea에 의한 무화과 잿빛곰팡이병)

  • Cheong, Seong-Soo;Choi, In-Young;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.38-41
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    • 2013
  • In 2010 and 2011, gray mold was found on common fig (Ficus carica) fruit grown at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. Gray mold symptoms on common fig fruit mainly occurred after harvest season until December. The typical symptom included brown water-soaked rot and fruit decay. The diseased fruit was covered by gray to brown colored conidiophore and conidia. The conidiophores were tree shape and measured $15-33{\times}2{\mu}m$. Conidia on conidiophore were ellipsoidal or lemon shape, colorless, single cell, and measured $7.3-14.6{\times}6.8-11.1{\mu}m$. The nucleotide sequences of the rDNA ITS region obtained from the pure culture of the gray mold on common fig were 100% similar to the sequences of the GenBank accession number HQ171052, EU519210, HQ171053, FN812726, HM849615, and EU563120 of B. cinerea isolates. In phylogenetic tree, the representative isolate was placed within same clade of B. cinerea. Based on the morphological characteristics and analysis of rDNA ITS sequence data, the fungus was identified as B. cinerea.

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.