• Title/Summary/Keyword: internal transcribed spacer 2 (ITS2)

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Detection of Laminariaceae Species Based on PCR by Family-specific ITS Primers

  • Choi, Chang-Geun;Kim, Jong-Myoung
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.157-162
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    • 2012
  • To analyze nucleotide sequence encoding internal transcribed spacer (ITS) regions specific to the Laminariaceae family, genomic DNA was isolated from six brown algae species distributed along the east coast of Korea. These included three species from the Laminariaceae family (Agarum clathratum Dumortier, Costaria costata [C. Agardh] Saunders, and Saccharina japonica Areschoug) and two species from the Alariaceae family (Undaria pinnatifida [Harvey] Suringer and Ecklonia cava Kjellman), both in the order Laminariales, and one species from the family Sargassaceae in the order Fucales (Sargassum serratifolium). Based on a sequence analysis of ITS-1 and ITS-2 for A. clathratum, C. costata, and E. cava, oligonucleotides were designed from the regions that showed sequence conservation in Laminariaceae. Following polymerase chain reaction using three sets of primers, amplification of ITS-1 and ITS-2 was detected in reactions using genomic DNA isolated from the species belonging to Laminariaceae, but not from the species belonging to the other families. The results indicate that this method can be used for the detection and identification of Laminariaceae species.

Variation of nuclear and mitochondrial DNAs in Korean and Chinese isolates of Clonorchis sinensis

  • Lee, Soo-Ung;Huh, Sun
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.145-148
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    • 2004
  • We compared the DNA sequence difference of isolates of Clonorchis sinensis from one Korean (Kimhae) and two Chinese areas (Guangxi and Shenyang), The sequences of nuclear rDNA (18S, internal transcribed spacer 1 and 2: ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1: cox1) were compared. A very few intraspecific nucleotide substitution of the 18S, ITS1, ITS2 and cox1 was found among three isolates of C. sinensis and a few nucleotide insertion and deletion of ITS1 were detected. The 18S, ITS1, ITS2 and cox1 sequences were highly conserved among three isolates. These findings indicated that the Korean and two Chinese isolates are similar at the DNA sequence level.

Isolation and Identification of Wild Yeasts from Soils of Fields in Daejeon Metropolitan City and Chungcheongnam-do, Korea (대전광역시와 충청남도 밭 토양으로부터 야생효모의 분리 및 동정)

  • Han, Sang-Min;Han, Jae-Won;Bae, Sang-Min;Park, Won-Jong;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.1-7
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    • 2016
  • This study focused on isolation and identification of wild yeasts from soils in fields near mountains and elucidation of its yeast distribution. Several kinds of yeasts were isolated from various soils of Daejeon metropolitan city and Chungcheongnam-do in Korea and identified by BLAST search of nucleotide sequences of internal transcribed spacer (ITS) region including 5.8S rRNA and D1/D2 region of 26S rDNA. Ninety-seven strains of 20 species from 61 soil samples were isolated, of which Cryptococcus podzolicus (11 strains), Debaryomyces hansenii (6 strains), and Trichosporon asahii (6 strains) were dominant species.

Identification of Unrecorded Endophytic Fungi Isolated from Leaves of Woody Plants in Jejudo, Korea (제주도에 서식하는 목본 식물의 잎에서 분리한 미기록 내생균)

  • Lee, Bong-Hyung;Kim, Dong-Yeo;Park, Hyeok;Eo, Ju-Kyeong;Lee, Hyang Burm;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.252-258
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    • 2016
  • In this study, endophytic fungi were isolated from the leaves of five species of woody plants in Jeju, Korea, namely Chamaecyparis obtusa, Cryptomeria japonica, Torreya nucifera, Ilex crenata, and Camellia japonica. The isolated fungal endophytes were identified based on their morphological and molecular characteristics including a sequence analysis of the internal transcribed spacer and 26S regions of rDNA and ${\beta}$-tubulin genes. Ten species of fungal endophytes have not been previously reported in Korea, namely Mycosphaerella aleuritidis, Neofusicoccum eucalyptorum, Neofusicoccum parvum, Phyllosticta citrichinensis, Phyllosticta cryptomeriae, Phomopsis cotoneastri, Sphaerulina rhododendricola, Guignardia mangiferae, Lophodermium jiangnanense, and Lophodermium minus.

Notes on Five Unrecorded Endophytic Fungi Isolated from Coniferous Leaves and Orchid Roots in Korea (한국에 서식하는 침엽수의 잎과 난초과 식물의 뿌리에서 분리한 5종의 국내 미기록 내생균)

  • Park, Hyeok;Lee, Bong-Hyung;Bae, Yu-Ra;Kim, Dong-Yeo;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.365-370
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    • 2016
  • We collected leaves of Pinus koraiensis and Thuja koraiensis and roots of Bletilla striata from various sites in Korea. The leaf and root samples were surface-sterilized and endophytic fungi were isolated. Fungal isolates were identified based on their morphological characteristics and a phylogenetic analysis of internal transcribed spacer regions, large subunit regions, and the ${\beta}$-tubulin gene. Consequently, we identified five species of endophytic fungi, namely Colletotrichum simmondsii, Fusarium sterilihyphosum, Diatrypella pulvinata, Ochroconis globalis, and Sphaeria chrysosperma. These species have not been previously reported in Korea and we report them here with descriptions and illustrations.

Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species (DNA 바코딩과 고해상 융해곡선분석에 기반한 인삼속 식물의 종 판별)

  • Bang, Kyong Hwan;Kim, Young Chang;Lim, Ji Young;Kim, Jang Uk;Lee, Jung Woo;Kim, Dong Hwi;Kim, Kee Hong;Jo, Ick Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.6
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    • pp.439-445
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    • 2015
  • Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

Diversity of Fungal Endophytes in Various Tissues of Panax ginseng Meyer Cultivated in Korea

  • Park, Young-Hwan;Lee, Soon-Gu;Ahn, Doek-Jong;Kwon, Tae-Ryong;Park, Sang-Un;Lim, Hyoun-Sub;Bae, Han-Hong
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.211-217
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    • 2012
  • Endophytic fungi were isolated from various tissues (root, stem, petiole, leaf, and flower stalk) of 3- and 4-year-old ginseng plants (Panax ginseng Meyer) cultivated in Korea. The isolated endophytic fungi were identified based on the sequence analysis of the internal transcribed spacer (ITS), 1-5.8-ITS 2. A morphological characterization was also conducted using microscopic observations. According to the identification, 127 fungal isolates were assigned to 27 taxa. The genera of Phoma, Alternaria and Colletotrichum were the most frequent isolates, followed by Fusarium, Entrophospora and Xylaria. Although 19 of the 27 taxa were identified at the species level, the remainder were classified at the genus level (6 isolates), phylum level (Ascomycota, 1 isolate), and unknown fungal species (1 isolate). Endophytic fungi of 13 and 19 species were isolated from 3- and 4-year-old ginseng plants, respectively, and Phoma radicina and Fusarium solani were the most frequently isolated species colonizing the tissues of the 3- and 4-year-old ginseng plants, respectively. The colonization frequency (CF%) was dependant on the age and tissue examined: the CFs of the roots and stems in the 3-year-old ginseng were higher than the CF of tissues in the 4-year-old plants. In contrast, higher CFs were observed in the leaves and petioles of 4-year-old plants, and endophytic fungi in the flower stalks were only detected in the 4-year-old plants. In conclusion, we detected diverse endophytic fungi in ginseng plants, which were distributed differently depending on the age and tissue examined.

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

  • Kamaruddin, Mudyawati;Tokoro, Masaharu;Rahman, Md. Moshiur;Arayama, Shunsuke;Hidayati, Anggi P.N.;Syafruddin, Din;Asih, Puji B.S.;Yoshikawa, Hisao;Kawahara, Ei
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.471-478
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    • 2014
  • Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

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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New Records of Fungi Isolated from Indoor Air of Greenhouse Used for Shiitake Cultivation in Korea (표고 재배사 실내 공기에서 분리한 국내 미기록 진균)

  • Kwon, Hyuk Woo;Yun, Yeo Hong;Kim, Jun Young;Kim, Seong Hwan;Ko, Han Kyu
    • The Korean Journal of Mycology
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    • v.43 no.1
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    • pp.58-63
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    • 2015
  • Mold contamination is one of the detrimental factors affecting sawdust media-based shiitake cultivation in greenhouses. During mold monitoring of indoor air of greenhouses, several fungi were isolated. Among them, Aspergillus pulverulentus and Cosmospora butyri were found to be new in Korea and Lecanicillium psalliotae and L. antillanum were known in Korea without taxonomic validation. In this study the morphological characteristics and phylogenetic analysis based on the internal transcribed spacer (ITS) rDNA region or ${\beta}$-tubulin gene of the four identified species were described.