• Title/Summary/Keyword: Internal transcribed spacer rDNA

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Identification of Puccinia iridis on Iris domestica in Korea (범부채에서 녹병균 Puccinia iridis의 동정)

  • Choi, In-Young;Choi, Young-Joon;Kim, Jin-Young;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.89-94
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    • 2019
  • A rust fungus on Iris domestica (syn. Belamcanda chinensis) from China was previously identified as Puccinia iridis. Accordingly, the identity of the rust fungus on the same host species in Korea needs to be reexamned. Morphological characteristics of the Korean materials matched with P. iridis. Molecular phylogenetic analyses based on internal transcribed spacer and large subunit rDNA sequences of two representative materials confirmed the identification by high sequence similarities of 100% and 99% with the reference sequences available in NCBI GenBank database. Phylogenetic tree inferred from neighbor-joining method proved them to be clustered in P. iridis group. Thus, the rust fungus on I. domestica in Korea was determined as P. iridis. On the other hand, a previous record that Puccinia belamcandae is another rust agent on I. domestica in Korea should be reconfirmed in future.

Monitoring of Fasciola Species Contamination in Water Dropwort by COX1 Mitochondrial and ITS-2 rDNA Sequencing Analysis

  • Choi, In-Wook;Kim, Hwang-Yong;Quan, Juan-Hua;Ryu, Jae-Gee;Sun, Rubing;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.53 no.5
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    • pp.641-645
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    • 2015
  • Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.

Phylogenetic Study of Ganoderma spp. Based on the DNA Sequences in ITS II Region (ITS II 영역의 DNA 염기서열 분석에 의한 불로초(Ganoderma)속의 계통분류학적 고찰)

  • Park, Dong-Suk;Go, Seung-Joo;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.39-43
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    • 1999
  • The internal transcribed spacer II regions (ITS II) of the ribosomal DNA gene repeat from Ganoderma spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 9 species including Ganoderma lucidum, G. tsugae, G. pfeifferi, G. resinaceum, G. australe-applanatum, G. oregonense, G. neo-japonicum, G. applanatum and Inonotus xeranticus as an out-group were compared. The spacer regions of them were $247{\sim}257$ nucleotides in length and contained partial sequences of 5.8S and 25S gene. The reciprocal homologies of each ITS II sequence of the species were in the range of $70{\sim}100%$ except outgroup species, I. xeranticus. According to the analysis of ITS II sequences, Ganoderma spp. constructed 5 clusters. Ganoderma lucidum isolates were to be divided into two groups. One group was consisted of isolates from South Korea. The other group comprised isolates from UK. G. lucidum isolates belonging to the group I were closely related with G. tsugae. These results suggested that G. lucidum from Korea should be G. tsugae, otherwise G. tsugae was to be synonym of G. lucidum.

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Phylogenetic Relationships of Coprinoid Taxa and an Agaric-like Gastroid Taxon Based on the Sequences of Internal Transcribed Spacer (ITS) Regions (ITS 영역의 염기서열을 이용한 먹물버섯류 및 주름버섯 유사 복균류와의 계통학적 유연관계)

  • Park, Dong-Suk;Go, Seung-Joo;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.406-411
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    • 1999
  • The phylogenetic relationships of black-spored basidioid taxa (mainly coprinoid taxa) and an agaric-like gastroid taxon were studied. The sequences of internal transcribed spacer regions (ITS) partially including 17S, 25S and 5.8S from 14 species (Coprinus comatus, C. atramentarius, C. micaceus, C. cinereus, C. disseminatus, C. rhizophorus, C. radians, C. echinosporus, Psathyrella candolleana, Podaxis pistillaris, Conocybe lactea, Bolbitius demangei, Agaricus balzei, and Stropharia rugosoannulata) were compared. The reciprocal homologies of ITS sequences among these species were in the range of $38.7{\sim}77.2%$. Black-spored taxa were classified into four clusters. Cluster I comprised C. micaceus, C. radians, and C. disseminata. Cluster II is consisted of C. cinereus, C. echinosporus, C. rhizophorus, and C. atramentarius. On the other hand, C. comatus is in cluster III with Agaricus balzei and Podaxis pistillaris even though this species is belonging to the section Coprinus in morphological aspect. Psathyrelloid taxon is included in cluster II. The question of the origin of secotioid (agaric-like) fungi has been taken, though largely on theoritical suggestions as to whether gastroid taxa give rise to agarics with secotioid taxa as intermediates or whether secotioid taxa are evolutionary novelities arising from many distinct groups of agarics. In this relationship, it was shown that secotioid taxon evolved from within agaric species.

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Phylogenetic Relationships of Genera Coprinus and Psathyrella on the Basis of ITS Region Sequences (먹물버섯속(Coprinus)과 눈물버섯속(Psathyrella)의 ITS 영역 염기서열에 의한 계통학적 유연관계 분석)

  • Park, Dong-Suk;Go, Seung-Joo;Kim, Yang-Sup;Seok, Soon-Ja;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.274-279
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    • 1999
  • The internal transcribed spacer regions(ITS) of the ribosomal DNA gene repeat from Coprinus and Psathyrella spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 11 species including Coprinus comatus, C. atramentarius, C. micaceus, C. cinereus, C. rhizophorus, C. radians, C. echinosporus, C. disseminatus, Psathyrella candolleana, P. spadiceogrisea and Stropharia rugosoannulata were compared. The spacer region I and II were $258{\sim}301\;bp\;and\;253{\sim}275\;bp$ in length respectively and partially contained 17S, 5.8S and 25S. The reciprocal homologies of ITS sequences among these strains were in the range of $43.9{\sim}96.0%$. According to the analysis of ITS sequences, Coprinus and Psathyrella spp. were classified into three clusters. Cluster I consisted of Coprinus lagopus, C. cinereus, C. echinosporus, C. rhizophorus, and C. atramentarius. Cluster II comprised C. micaceus, C. radians, C. disseminatus, Psathyrella candolleana, and P. spadiceogrisea. On the other hand C. comatus is in Cluster III with Stropharia rugosoannulata even though this species is belonging to the section Coprinus in morphological aspect. These results suggest that taxonomic position of Psathyrella would better be inculded in genus Coprinus. Coprinus comatus, the type species of Coprinus, gives a doubt on monophyletic evolution and is assumed to be paraphyletic or polyphyletic.

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Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences (nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별-)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Seo, Hyeong-Seok;Lee, A-Young;Chun, Jin-Mi;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Taxonomical Classification and Species-specific Detection of Genus Some Phellinus using Phylotype (Phylotype에 의한 수종의 Phellinus속의 분류체계 확립 및 종간구별을 위한 신속동정법 개발)

  • Kim, Cheng-Yun;Lee, Jae-Yun;Kim, Gi-Young;Lee, Ki-Won;Park, Jae-Min;Kim, Mun-Ok;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.121-128
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    • 2003
  • This study was carried out to identify the phylogenetic relationship of Phellinus species and to know its distribution by comparing the DNA sequences of internal transcribed spacer regions(ITS1 and IST2) and 5.8S ribosomal DNA (rDNA) repeat unit. The Phellinus species had their specific sequences in IST1 and 2 regions depending on suedes. The comparison of the ITS sequences of standard strains indicated that the sequences of ITS1 were more variable than those of ITS2. Nine strains of the commercial products of Phellinus species used in this study were identified as P. lintues, P. baumii, P. igniarius, and P. pini. Most of commercial species were P. pini and P. baumii, and P. gilvus was not found. Also, P. linteus was only found in form of mycelial culture rather than fruiting body. Moreover, the species-specific primers were designed based on ITS sequence data. Each species-specific primers were bound in P. lintues(ITSF-PL2R), P. baumii(PB1F-ITS4R), P. igniarius(IF1-IR3), P. pini(PF1-PR3), and P. gilvus(GF2-GR4), respectively. These primer sets would be useful fer the detection of specific-species among unidentified Phellinus species rapidly.

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.

Isolation of Yeasts from Wild Flowers in Gyonggi-do Province and Jeju Island in Korea and the Production of Anti-gout Xanthine Oxidase Inhibitor (경기도와 제주도 야생화들로부터 효모의 분리, 동정 및 항통풍성 Xanthine oxidase 저해물질의 생산)

  • Hyun, Se Hee;Mun, Hye Yeon;Lee, Hyang Burm;Kim, Ha-Kun;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.383-390
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    • 2013
  • Several yeasts were isolated from flowers found in Gyonggi-do Province and Jeju island in Korea. They were then identified by a comparison of their PCR-amplified D1/D2 regions of 26S rDNA, internal transcribed spacer 1 and 2 inclusive of 5.8S rDNA, using the BLAST database. A total of fifty four yeast strains were isolated from wild flowers in Gyonggi-do and the genus Pseudozyma was noted as being dominant. A total of thirty two strains were isolated from Songaksan and Seongsan-ilchulbong in Jeju island and Sporobolomyces ruberrimus was seen to be dominant. The anti-gout xanthine oxidase inhibitory activities of the culture broths and cell-free extracts from eighty six yeast strains were then determined. The cell-free extracts of Pseudozyma hubeiensis 228-S-1 exhibited the highest xanthine oxidase inhibitory activity of 19.6%. The XOD inhibitor was also maximally produced when Pseudozyma hubeiensis 228-S-1 was cultured at $30^{\circ}C$ for 36h in YEPD medium.

Morphological and cultural characteristics of a novel Phellinus linteus KACC93057P (신규 목질진흙버섯 KACC93057P의 배양적, 형태적 특성)

  • Min, Gyeong-Jin;Kwak, A-Min;Seok, Sun-Ja;Kang, Hee-Wan
    • Journal of Mushroom
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    • v.14 no.3
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    • pp.75-80
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    • 2016
  • Morphological and culture characteristics of the novel Phellinus linteus variant KACC93057P collected in Korea were characterized in this study. The surface of the was angular, sessile, tough-woody, concentrically zonate, and dark brown in color. Basidiocarppores were circular, with 5-7 pores per mm. The hyphal system was dimitic, and basidiospores were ellipsoid or oval, $4.5-6{\times}4-5$, exhibiting characteristics typical of P. linteus. The optimum temperature for mycelial growth was $25-30^{\circ}C$, and optimal pH for growth was 5-7. The mycelial growth rate of P. linteus KACC93057P was faster than that of other P. baumii isolates. On growth medium, KACC93057P formed aerial mycelia with density higher than that of other isolates. The internal transcribed spacer (ITS)-ribosomal DNA sequences were closely related to the sequences P. linteus complex.