• Title/Summary/Keyword: Internal transcribed spacer rDNA

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First Report of Three Didymella Species Isolated from Freshwater Ecosystem in Korea (담수환경에서 발굴된 Didymella속 3종의 국내 최초 보고)

  • Mun, Hye Yeon;Goh, Jaeduk;Oh, Yoosun;Jeong, Ae-Ran;Chung, Namil
    • The Korean Journal of Mycology
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    • v.46 no.1
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    • pp.1-8
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    • 2018
  • Three fungal isolates, NNIBRFG108, 1139, and 1480, were isolated from freshwater environments; NNIBRFG108 from plant litter in Samcheok, Gangwon and NNIBRFG1139 and 1480 from the soil in Jeju & Gimcheon, Gyeongbuk, Korea. Based on the morphological characteristics and phylogenetic analysis of internal tanscribed spacer (ITS), 28S rDNA region, and ${\beta}$-tubulin gene, NNIBRFG108, NNIBRFG1139, and NNIBRFG1480 isolates were confirmed as Didymella segeticola, D. ellipsoidea, and D. aeria, respectively. Neither species has previously been described in Korea.

Taxonomic status of Goodyera rosulacea (Orchidaceae): molecular evidence based on ITS and trnL sequences (로젯사철란(Goodyera rosulacea: Orchidaceae)의 분류학적 위치: ITS와 trnL 염기서열에 의한 분자적 증거)

  • Lee, Chang Shook;Eom, Sang Mi;Lee, Nam Sook
    • Korean Journal of Plant Taxonomy
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    • v.36 no.3
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    • pp.189-207
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    • 2006
  • Goodyera rosulacea, which is morphologically similar to G. repens, is described recently as a new species based on its distinct morphological characters such as rosette-formed leaves, short rhizome and habitat. To verify the taxonomic identity of G. rosulacea and its taxonomic relationship within Korean Goodyera taxa, sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA and the trnL region of cpDNA from 24 accessions including 1 outgroup accession were analyzed. Aligned sequences were analyzed using maximum parsimony and distance method, and the taxonomic identity and the taxonomic relationships among the related taxa were estimated by the existence of private marker gene and the phylogenetic tree of the aligned sequences. Molecular data indicate that G. rosulacea gas several private marker genes and shows monophyly in phylogenetic trees of both ITS and trnL sequences. the pairwise distance between G. rosulacea and the orher taxa of Korean Goodyera was 3.49-6.68% for ITS region and 5.05-9.53% for trnL region, indicating that G. rosulacea could be treated as an independent species. Therefore, our molecular data support the taxonomic of G. rosulacea as a distinct species of Korea. In phylogenetic trees, G. rosulacea formed same clade with G. repens, which has similar morphological characters with G. rosulacea, and showed the lowest pairwise distance with G. repens among Korean Goodyera taxa. These molecular data sugguested that G. rosulacea and G. repens are closely related taxa.

Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

  • Alam, Nuhu;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Tae-Soo;Lee, U-Youn
    • Mycobiology
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    • v.38 no.1
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    • pp.17-25
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    • 2010
  • The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

Phylogenetic Analysis of the Entomopathogenic Fungal Species and Taxonomical Positions of Their Commercial Products (동충하초의 계통분류 및 시판동충하초의 분류학적 위치)

  • 김순한;이영자;김인복;김미경;한정아;홍무기;이순호;이재동
    • Journal of Life Science
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    • v.13 no.4
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    • pp.400-411
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    • 2003
  • This study was carried out to identify the phylogenetic relationship and to know the distribution of the entomopathogenic fungi by comparing the DNA sequences of internal transcribed spacer regions (ITS1 and ITS2) and 5.8S ribosomal DNA (rDNA) repeat unit. The entomopathogenic fungi had their specific sequences in ITS1 and 2 regions depending on species. The comparison of the ITS sequences of standard strains indicated that the sequences ITS1 were more variable than those of ITS2. It seems that Paecilomyces tenuipes, Isaria japonicus and P. japonicus are the same species but called as different names because of very similar sequences, and unidentified Paecilomyces sp. KACC 40220 and KACC 40656 showed identical sequences to P. tenuipes. Thirty six strains of the commercial products of entomopathogenic fungi used in this study were divided into four groups by the phylogenetic analysis based on 5.85 rDNA and ITS regions. We found twenty-three strains were P. tenuipes / japonica, eleven strains were C. militaris, and other two strains were Beauveria bassiana and C. multiaxialis, respectively.

Analysis of Phylogenetic Relationship of 30 Cultivars of Korean Mulberry (Rosales: Moraceae) in Korea

  • Kwon, O-Chul;Kim, Hyun-Bok;Sung, Gyoo-Byung;Kim, Yong-Soon;Ju, Wan-Taek
    • International Journal of Industrial Entomology and Biomaterials
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    • v.37 no.2
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    • pp.82-89
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    • 2018
  • This study was carried out to understand phylogenetic relationships of the 30 mulberry cultivars converved in Korea based on the ITS rDNA region, and they were compared to 40 reference sequences from GenBank. The size and the G+C content of the ITS rDNA gene regions from the 30 Korean mulberry cultivars and 40 reference sequences varied from 612-630 bp and 58.19-61.62%, respectively. Based on the results of the comparative phylogenetic analysis of the ITS rDNA regions of the 30 Korean mulberry cultivars and 40 reference sequences, they were divided into three groups (Group 1, 2, and 3) and two subgroups (Group 1A and 1B within Group 1). The sequence lengths of the Korean mulberry cultivar numbers 1-26 and 27-30 were 615 bp and 616 bp, respectively. At 205 bp location of ITS1 rDNA region, the cultivar numbers 1-26 contain the nucleotide thymine but the cultivar numbers 27-30 contain the nucleotide adenine. In addition, the insertion of the nucleotide adenine at 206 bp location was found only in the four Korean mulberry cultivars (numbers 27-30). Based on these sequence information and phylogenetic result, the 30 Korean mulberry cultivars were identified as M. alba and M. australis. This study will contribute to the construction of genetic database constructions and accurate variety identifications for unidentified mulberry varieties in Korea.

Identification 4 kinds of Muxiang using Multiplex PCR (Multiplex PCR을 이용한 4 종류 목향(木香)의 감별)

  • Doh, Eui Jeong;Lee, Guemsan;Ju, Young-Sung;Oh, Seung Eun
    • The Korea Journal of Herbology
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    • v.29 no.3
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    • pp.19-26
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    • 2014
  • Objectives : Aucklandiae Radix (Muxiang) one of important herbal medicines in oriental medicine, is defined as the dried root of Aucklandia lappa (Asteraceae). Owing to the similarities in the morphology and name, Inulae Radix (Tu-Muxiang) and Vladimiriae Radix (Chuan-Muxiang) as well as Aristolochiae Radix (Qing-Muxiang) originated from other medicinal plants are often used as substitutes and/or adulterants of Aucklandiae Radix. Therefore, a reliable authentication of these herbal medicines is necessarily for the public health and prevention of misuse. Methods : 32 samples of medicinal plants supplying Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix were collected in Korea and China. The ITS (Internal transcribed spacer) nucleotide sequences of samples were determined. The PCR primers to amply DNA marker of each herbal medicine were designed basing on the specific ITS regions showing differences in the sequences among medicinal plants. Results : Primer set Al R/IS F designed in this work amplified 220 bp PCR product only in samples of Aucklandiae Radix. In contrast, primer set Ih F/IS R, Vs R/IS F, and AcR F1/Ac R amplified 250 bp product, 356 bp prouct, and 516 bp product respectively to identify Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. Conclusions : The primers designed basing on the nucleotide sequences of ITS regions appearing differenced in the sequences among medicinal plants amplified the DNA markers for the identification of Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. These herbal medicines were more efficiently identified by multiplex PCR method using all primers in a single PCR process.

Comparative Analysis of Acanthopanax senticosus Harms from Korea, China and Russia Based on the ITS Sequences of Nuclear Ribosomal DNA (ITS 염기서열분석에 의한 한국산, 중국산 및 러시아산 가시오갈피의 유연관계 분석)

  • Han Hyo-Shim;Kim Doo-Young;Lee Kab-Yeon;Park Wan-Geun;Cho In-Kyung;Jung Jae-Sung
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.54-58
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    • 2006
  • The genetic analyses of Acanthopanax senticosus Harms from Korea, China and Russia, were made by comparing the internal transcribed spacer (ITS) sequences of the nuclear ribosomal DNA. The ITS region of A. senticosus was amplified by polymerase chain reaction (PCR) using the universal primers and then directly sequenced. The length of the ITS region including 162 bp 5.85 rRNA gene ranged from 608 bp (for Korean and Chinese) to 611 bp (for Russian). The G+C content of ITS region were 60.20% for Korean and Chinese plants and 60.06% for Russian plants. Sequence comparisons indicated that ITS regions of A. senticosus from Korea and China were identical, whereas the ITS sequence of A. senticosus from Russia showed 99.2% homology with the plants from Korea. Variation in sequences were attributable to 5 bp substitution such as transversion or insertion events. These results suggested that A. senticosus Harms from Korea and China were closely related in phylogenetic relationship compared to Russian. In addition, A. senticosus Harms were more similar to Kalopanax pictus than A. sessiliflorus in their ITS sequences.

Identification of Acremonium acutatum and Trichothecium roseum isolated from Grape with White Stain Symptom in Korea

  • Oh, Soh-Young;Nam, Ki-Woong;Yoon, Deok-Hoon
    • Mycobiology
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    • v.42 no.3
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    • pp.269-273
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    • 2014
  • During 2010 and 2012 grape harvest seasons in Gyeonggi-do, Korea, a white stain symptom was observed on the harvested grape fruits in 'Campbell-Early' and 'Kyoho' varieties. In samples collected from the infected vine, two different strains of pathogenic fungi have been found and identified as Acremonium acutatum and Trichothecium roseum based on fungal morphology and nucleotide sequence of internal transcribed spacer (ITS) and supported by the phylogenetic analysis of the rDNA-ITS region. The DNA homologies of the isolated strains were 99.8% and 99.6% identical with T. roseum (IFB-22133) and A. acutatum (CBS682.71), respectively. In the pathogenicity test, the spores of A. acutatum and T. roseum sprayed on the grapes caused white stain symptoms on the fruits in two weeks after the artificial inoculation, which is similar to observations in the field. To our knowledge, this is the first report of white stain symptoms caused by A. acutatum and T. roseum on the grapes in Korea.

Diversity, Phylogeny, and Host-Specialization of Hyaloperonospora Species in Korea

  • Lee, Jae Sung;Lee, Hyang Burm;Shin, Hyeon-Dong;Choi, Young-Joon
    • Mycobiology
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    • v.45 no.3
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    • pp.139-149
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    • 2017
  • The genus Hyaloperonospora (Peronosporaceae; Oomycota) is an obligate biotrophic group that causes downy mildew disease on the Brassicaceae and allied families of Brassicales, including many economically relevant crops, such as broccoli, cabbage, radish, rape, and wasabi. To investigate the diversity of Hyaloperonospora species in northeast Asia, we performed a morphological analysis for the dried herbarium specimens collected in Korea, along with molecular phylogenetic inferences based on internal transcribed spacer rDNA and cox2 mtDNA sequences. It was confirmed that 14 species of Hyaloperonospora exist in Korea. Of these, three species, previously classified under the genus Peronospora, were combined to Hyaloperonospora: H. arabidis-glabrae comb. nov. (ex Arabis glabra), H. nasturtii-montani comb. nov. (ex Rorippa indica), and H. nasturtii-palustris comb. nov. (ex Rorippa palustris). In addition, finding two potentially new species specific to northeast Asian plants is noteworthy in support of the view that the species abundance of Hyaloperonospora has been underestimated hitherto.

Taxonomy and Phylogeny of Peronospora Species (Oomycota) Parasitic to Stellaria and Pseudostellaria in Korea, with the Introduction of Peronospora casparyi sp. nov.

  • Lee, Jae Sung;Shin, Hyeon-Dong;Lee, Hyang Burm;Choi, Young-Joon
    • Mycobiology
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    • v.45 no.4
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    • pp.263-269
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    • 2017
  • The genus Peronospora, an obligate biotrophic group belonging to Oomycota, causes serious damage to a variety of wild and ornamental plants, as well as cultivated crops, such as beet, rose, spinach, and tobacco. To investigate the diversity of Peronospora species parasitic to Stellaria and Pseudostellaria (Caryophyllaceae) plants in Korea, we performed a morphological analysis on dried herbarium specimens and molecular phylogenetic inferences based on internal transcribed spacer rDNA and cox2 mitochondrial DNA sequences. As a result, it was confirmed that there are four species of Peronospora parasitic to specific species of Stellaria and Pseudostellaria, all of which were hitherto unrecorded in Korea: P. alsinearum (ex Stellaria media), P. stellariae-aquaticae (ex Stellaria aquatica), P. stellariae-uliginosae (ex Stellaria alsine), and P. pseudostellariae (ex Pseudostellaria palibiniana). In addition, Peronospora specimens parasitic to Pseudostellaria davidii differed morphologically from P. pseudostellariae owing to the large and ellipsoidal conidia; this morphological discrepancy was also validated by the high genetic divergence between the two species. Peronospora casparyi sp. nov. is described and illustrated here.