• Title/Summary/Keyword: ITS rDNA sequences

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Phylogenetic relationships of genera Polyporus on the basis of ITS region sequences (rDNA의 ITS 부위 염기서열 분석에 의한 겨울우산버섯(Polyporus)속 균주의 유전적인 유연관계 분석)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Kong, Won S.
    • Journal of Mushroom
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    • v.10 no.1
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    • pp.37-43
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    • 2012
  • This study was carried to identify a correct species and asses genetic diversity within the same species of Polyporus spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Polyporus spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that three strains were different species and four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Polyporus clustered into five distinct group, most of which correlated with species-groups identified by RAPD method. Four isolates included strain PM02 showed high similarity with P. arcularius, four isolates included strain PM03 high similarity with P. alveolaris, three isolates included strain PM01 high similarity with P. tuberaster, and PM 06 and PM04 high similarity with P. brumalis and P. squamossus. Isolates were collected in the United States(PM10, PM11) was identified as P. alveolarius and P. arcularius. RAPD analysis of genetic polymorphisms of genus Polyporus showed a very different band patterns. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 6 isolates of Polyporus spp. may be need to reclassify or eliminate from preserved catalogue.

New Records of Three Tintinnopsis Species and Redescription of One Antetintinnopsis Species (Protozoa: Ciliophora) from Coastal Waters of Korea

  • Ji Hye Moon;Jae-Ho Jung
    • Animal Systematics, Evolution and Diversity
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    • v.40 no.1
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    • pp.1-15
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    • 2024
  • Our study aimed to investigate the diversity of tintinnid species in Korea by collecting samples from coastal waters. As a result, we identified and redescribed three newly recorded species of the genus Tintinnopsis Stein, 1867 and one previously recorded species of the genus Antetintinnopsis Wang et al., 2021 in Korea. The loricae morphology and molecular phylogeny based on the 18S rDNA sequences of these four were analyzed. Tintinnopsis kiaochowensis Yin, 1956 is characterized by having an irregular collar with spiral turns and an obconical-shaped bowl. Tintinnopsis orientalis Kofoid and Campbell, 1929 is characterized by the inverted bell-shaped lorica with size of 121-140×86-94 ㎛. Tintinnopsis parvula Jörgensen, 1912 is characterized by its narrower collar than bowl and acute angle of the bowl (39-75°). The recorded species, Antetintinnopsis gracilis (Kofoid and Campbell, 1929) Wang et al., 2021 is characterized by a cylindrical collar narrower than the bowl width and with a size of 98-131×37-46 ㎛.

Molecular Authentication and Phylogenetic Analysis of Plant Species for Breeae and Cirsii Herba based on DNA barcodes (DNA 바코드 분석을 통한 소계(小薊) 및 대계(大薊) 기원식물 감별과 종간 유연관계 분석)

  • Moon, Byeong Cheol;Lee, Young Mi;Ji, Yunui;Choi, Goya;Chun, Jin Mi;Kim, Ho Kyoung
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.75-84
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    • 2013
  • Objectives : The origin of Breeae Herba (So-gye) and Cirsii Herba (Dae-gye) is differently prescribed in Korean and Chinese modern pharmacopoeia. Since the similar morphological characteristics and chaotic plant names, moreover, the aerial part of Carduus crispus have been used as the Cirsii Herba. To develop a reliable method for correct identification of these herbal medicines and to evaluate the genetic relationship of these closely related plant species, we analyzed sequences of DNA barcode regions. Methods : Thirty-one samples of 6 medicinal plants (B. segeta, B. setosa, C. japonicum var. maackii, C. setidens, C. chanroenicum, and C. crispus) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL) were analyzed after amplification using appropriate primers reported in previous studies. The nucleotides of species-specific authentic marker and phylogenetic relations were estimated based on the entire sequences of DNA barcodes by the analysis of ClastalW and UPGMA, respectively. Results : In comparative analysis of DNA barcode sequences, we obtained specific nucleotides to discriminate the medicinal plant of Breeae/Cirsii Herba in species level and evaluated the phylogenetic relationship of these species. Futhermore, we identified distinct marker nucleotides enough to authenticate respective species. These sequence differences at corresponding positions were avaliable genetic markers to determine the botanical origins of Breeae Herbal as well as Cirsii Herba. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by providing of definitive information that can identify each plant species and distinguish from unauthentic adulterants and substitutes.

Monophyly of the Family Desmoscolecidae (Nematoda, Demoscolecida) and Its Phylogenetic Position Inferred from 18S rDNA Sequences

  • Hwang, Ui Wook;Choi, Eun Hwa;Kim, Dong Sung;Decraemer, Wilfrida;Chang, Cheon Young
    • Molecules and Cells
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    • v.27 no.5
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    • pp.515-523
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    • 2009
  • To infer the monophyletic origin and phylogenetic relationships of the order Desmoscolecida, a unique and puzzling group of mainly free-living marine nematodes, we newly determined nearly complete 18S rDNA sequences for six marine desmoscolecid nematodes belonging to four genera (Desmoscolex, Greeffiella, Tricoma and Paratricoma). Based on the present data and those of 72 nematode species previously reported, the first molecular phylogenetic analysis focusing on Desmoscolecida was done by using neighbor joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) methods. All four resultant trees consistently and strongly supported that the family Desmoscolecidae forms a monophyletic group with very high node confidence values. The monophyletic clade of desmocolecid nematodes was placed as a sister group of the clade including some members of Monhysterida and Araeolaimida, Cyartonema elegans (Cyartonematidae) and Terschellingia Iongicaudata (Linhomoeidae) in all the analyses. However, the present phylogenetic trees do not show any direct attraction between the families Desmoscolecidae and Cyartonematidae. Within the monophyletic clade of the family Desmoscolecidae in all of the present phylogenetic trees, there were consistently observed two distinct subgroups which correspond to the subfamilies Desmoscolecinae [Greeffiella sp. + Desmoscolex sp.] and Tricominae [Paratricoma sp. + Tricoma sp].

Interspecific Distinguishability of Veiled Lady Mushrooms (Dictyophora spp.) Based on rDNA-ITS Analysis (rDNA-ITS 분석에 의한 망태버섯속균(Dictyophora spp.)의 종간 구분 가능성)

  • Cheong, Jong-Chun;Lee, Myung-Chul;Kim, Bum-Gi;Park, Dong-Seok;Hong, Sung-Beom;Park, Jeong-Sik
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.1-7
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    • 2004
  • To establish the phylogenetic relationships of Dictyophora spp., rDNA-ITS regions of 11 strains of veiled lady mushroom collected from various countries were amplified and sequenced. It was observed that the 11 strains were divided into four groups based on PCR band patterns of each ITS region cleaved by eight different restriction enzymes in cleaved amplified polymorphic sequence analysis (CAPS). The phylogenic relationship of each group by cleaved amplified polymorphic sequence (CAPS) analysis matches well with previously reported morphological phylogeny, such as 5 strains of D. indusiata, 4 strains of D. echinovolvata, and a strain of Phallus rugulosus. Sequence analysis using the cluster V methods showed more detail classification than CAPS analysis. The 5.8S region showed two point nucleotide base exchanges from G to A according to four groups, and four groups were subdivided by sequence variation of ITS I and ITS II regions. But sequence variation of Phallus rugulosus was not showed in full ITS region. This study further delineates the taxonomic level at which ITS sequences, in comparison to ribosomal gene sequence, are most useful in systematics and other mushroom study.

Species Diversity and Seasonal Changes of Dominant Ulva Species (Ulvales, Ulvophyceae) in Mikawa Bay, Japan, Deduced from ITS2 rDNA Region Sequences

  • Kawai, Hiroshi;Shimada, Satoshi;Hanyuda, Takeaki;Suzuki, Teruaki;Gamagori City Office, Gamagori City Office
    • ALGAE
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    • v.22 no.3
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    • pp.221-228
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    • 2007
  • Frequent occurrences of green tides caused by Ulva species (Ulvales, Ulvophyceae) associated with eutrophication along enclosed coasts are currently causing environmental problems in coastal ecosystems. In addition, increasing intercontinental introductions of coastal marine organisms, including Ulva, are also a serious issue. However, due to the considerable morphological plasticity of this genus, the taxonomy of Ulva species based on morphological studies is problematic. Therefore, in order to elucidate the species diversity and seasonal changes of the dominant Ulva species in Mikawa Bay, central Honshu, Japan, we made seasonal collections of Ulva species at seven localities, and identified the dominant species using the ITS2 rDNA region sequences. We identified the following nine taxa as common Ulva species in the area: 1) Ulva pertusa Kjellman; 2) U. ohnoi Hiraoka et Shimada; 3) U. linza L.; 4) U. californica Wille; 5) U. flexuosa Wulfen; 6) U. fasciata Delile; 7) U. compressa L.; 8) U. armoricana Dion et al.; 9) U. scandinavica Bliding. Among the species, U. pertusa was most common and dominant from spring to summer, and U. ohnoi from autumn to winter. Ulva californica and U. scandinavica have not been reported before from Japan.

Vegetative Growth and Phylogenetic Relationship of Commercially Cultivated Strains of Pleurotus eryngii based on ITS sequence and RAPD

  • Alam, Nuhu;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • v.37 no.4
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    • pp.258-266
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    • 2009
  • Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

Isolation, Identification and Characterization of a Antidementia Acetylcholinesterase Inhibitor-Producing $Yarrowia$ $lipolytica$ S-3

  • Kang, Min-Gu;Yoon, Min-Ho;Choi, Young-Jun;Lee, Jong-Soo
    • Mycobiology
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    • v.40 no.1
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    • pp.42-46
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    • 2012
  • This report describes the isolation and identification of a potent acetylcholinesterase (AChE) inhibitor-producing yeasts. Of 731 species of yeast strain, the S-3 strain was selected as a potent producer of AChE inhibitor. The selected S-3 strain was investigated for its microbiological characteristics. The S-3 strain was found to be short-oval yeast that did not form an ascospore. The strain formed a pseudomycelium and grew in yeast malt medium containing 50% glucose and 10% ethanol. Finally, the S-3 strain was identified by its physiological characteristics and 26S ribosomal DNA sequences as $Yarrowia$ $lipolytica$ S-3.

Molecular Identification of Asian Isolates of Medicinal Mushroom Hericium erinaceum by Phylogenetic Analysis of Nuclear ITS rDNA

  • Park, Hyuk-Gu;Ko, Han-Gyu;Kim, Seong-Hwan;Park, Won-Mok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.816-821
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    • 2004
  • A reliable molecular phylogenetic method to identify Hericium erinaceum, the most industrially valuable species in the Hericium genus, was established. Sequencing and phylogenetic analyses of the PCR-amplified ITS and 5.8S rDNA from Hericium fungi, including 6 species and 23 isolates, showed that variation in nucleotide sequences and size exists in both ITS1 and ITS2 regions, but not in the 5.8S region. These two ITS regions provided different levels of information on the relationship of H. erinaceum to other Hericium species. Based on the ITS1 sequence, both the parsimony and neighbor joining trees clearly distinguished Asian H. erinaceum isolates from other Hericium species and isolates. The intraspecific divergence of the ITS2 region was suitable to dissect the Asian H. erinaceum isolates into a few groups.

Development of Rapid Molecular Detection Marker for Colletotrichum spp. in Leaf and Fruit Tissues of Sweet Persimmon

  • Iee, Sang-Pyo;Lee, Youn-Su
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.989-992
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    • 2002
  • Sweet persimmon (Diospyros kaki Thunb.) is widely cultivated in the southern part of Korea and its cultivation is increasing. However, anthracnose disease caused by Colletotricuhum species is one of the major hinderances to the cultivation and production of sweet persimmon. Therefore, in the current study, PCR was used to specifically detect Colletotrichum spp., based on the sequences of the ITS II regions in the rDNA. Using the sequence data, CO-1 was designated to detect Colletotrichum together the with ITS 4 primer. The result showed that a single segment of ca. 500 bp was observed only in Colletotrichum, but not in any other fungal and bacterial isolates. The annealing temperatures and template DNA quantites were also investigated to identify optimal conditions for detection. Using these species-specific primers, a unique band was obtained at annealing temperatures ranging from $55^{\circ}C\;and\;61^{\circ}C$ and template DNA levels from 10 pg- $10{\mu}g$.