• 제목/요약/키워드: ITS1-5.8S rDNA sequences.

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Internal Transcribed Spacer와 5.8S ribosomal DNA의 염기서열 분석에 의한 Agaricus blazei와 근연종에 대한 계통분류학적인 연구 (Phylogenetic Analysis of Agaricus blazei and Related Taxa by Comparing the Sequences of Internal Transcribed Spacers and 5.8S rDNA)

  • 김기영;하명규;이태호;이재동
    • 미생물학회지
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    • 제35권3호
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    • pp.180-184
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    • 1999
  • Agaricus 속의 계통분류학적 유연관계를 알아보기 위하여 Internal transcribed spacers 1,2, 와 5.8S rDNA 에 해당하는 부위를 PCR 기법으로 증폭하여 염기서열을 비교 조사하였다. 본 실험에 사용되어진 aGARICUS 속 5균주에 대한 염기서열을 분석한 결과, 이들 속은 크게 두 개의 군으로 분류되었다. 첫 번째 분류군은 aGARICUS BLAZEI atcc 76739, 현재 우리나라에서 배양중인 Agaricus blazei, agaricus arvensis IMSNU 32049, 그리고 Agaricus campestris VPI-OKM 25665로 이루어 졌다. 특히, Agaricus blazei ATCC 76739와 현재 우리나라 농가에서 배양중인 Agaricus blazeisms ITS 부위에서 5개의 염기서열에서 변이가 발견되었다. 이러한 변이는 지리적 또는 배양상의 변이로 인하여 특정염기서열에서 변이가 발생한 것으로 간주되므로 이들은 상호 동일종일 것으로 추정된다. 또한, Agaricus arvensis IMSNU 32049 와 Agaricus ampestris VPI-LKM 25665는 Agaricus blazei 하위 분류군을 형성하였다. 두 번째 분류군은 Agaricus bisporus CH 3004와 Agaricus pocillator DUKE-J 173 으로 이루어졌다.

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Characterization of Sclerotinia sclerotiorum Isolated from Paprika

  • Jeon, Young-Jae;Kwon, Hyuk-Woo;Nam, Ji-Sun;Kim, Seong-Hwan
    • Mycobiology
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    • 제34권3호
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    • pp.154-157
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    • 2006
  • A fungal isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Sclerotinia sclerotiorum based on its ability of sclerotium formation, physiological and molecular properties. When the isolate was grown on potato dextrose agar, oatmeal agar, and malt extract agar, it grew most well on PDA. Optimal temperature and pH for its growth were $25^{\circ}C$ and pH 7, respectively. The fungal isolate produced sclerotia on PDA within 10 days, and the color and shape of the sclerotia were similar to those of S. sclerotiorum. The ITS rDNA regions including ITS1 and ITS2 and 5.8S sequences were amplified using ITS1F and ITS4 primers from the genomic DNAs of the paprika isolate and other known pathogenic S. sclerotiorum isolated from different crops in Korea, and their nucleotide sequences were determined. Sequence comparison analysis showed the ITS rDNA of the paprika isolate shares 100% sequence identity with those of S. sclerotiorum isolated from red pepper, lettuce and a S. sclerotiorum isolate registered in GenBank DNA database. Neighbor joining analysis based on the ITS rDNA sequence revealed the paprika isolate has very close phylogenetic relationships with known Sclerotinia sclerotiorum isolates. This is the first report that S. sclerotiorum has been found associated with paprika rot in paprika growing countries.

Phylogenetic Analyses of Nuclear rDNA ITS Sequences of Korean Allium L. Subgenus Rhizirideum(Alliaceae)

  • Lee, Nam-Sook
    • Animal cells and systems
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    • 제5권4호
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    • pp.283-290
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    • 2001
  • Phylogenetic relationships among the Korean taxa of the genus Allium subgenus Rhizirideum and some related taxa were assessed on the basis of in sequences of nuclear ribosomal DNA. Twenty-eight accessions of the genus Allium L. consisting of subgenera Rhizirideum (19 taxa), Allium (5 taxa) and Amerallium (one taxon) were analyzed. The variation in the ITS region was informative at the levels of section except for sect. Reticulato- bulbosa which is known to be of multiple origin. The ITS 2 region was longer than the ITS 1 region, and all of the investigated Allium taxa were the same in length in the 5.8S region except for A. monanthum. Allium cyaneum var. cyaneum was the shortest (635 bp) and A. victorialis the longest (646 bp) among the investigated Korean taxa. The three morphologically similar taxa, A. thunbergii, A. sacculiferum that has been included in A. thunbergii, and A. deltoid- fistulosum, had the same ITS lengths of 641 bp, but were clearly distinguished in the phylogenetic analysis of their ITS sequences.

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Mycobacteria에 대해 항균력을 나타내는 엉겅퀴의 분류를 위한 ITS1, 5.8S rRNA, ITS2의 염기서열 분석 (Identification of a Carduus spp. Showing Anti-Mycobacterial Activity by DNA Sequence Analysis of Its ITS1, 5.8S rRNA and ITS2)

  • 배영민
    • 생명과학회지
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    • 제20권4호
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    • pp.578-583
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    • 2010
  • 세균 및 진균류의 증식을 억제하는 능력이 있는 것으로 보고된 누로와 대계의 추출물을 사용하여 Mycobacterium smegmatis 및 Mycobacterium fortuitum의 증식을 억제하는 능력이 있는지를 시험하였다. 그 결과, 누로의 추출물에서는 증식억제능을 발견할 수 없었으나, 대계의 추출물에서는 뚜렷한 증식억제능이 관찰되었다. 따라서 본 연구에 사용된 대계(엉겅퀴)에 대한 분류학적 또는 진화적 분석을 수행하기 위하여 genomic DNA를 추출한 후, ITS1, 5.8S rRNA 유전자 및 ITS2를 포함하는 부분을 PCR로 증폭시켰다. PCR 산물의 염기서열을 분석한 결과, 733-bp의 염기서열이 얻어졌고, 이것을 GenBank에 등록하였다(accession number GU188570). 이렇게 얻어진 염기서열을 사용하여 BLAST analysis를 수행한 결과, 염기서열이 일치하는 생물체는 아직까지 GenBank에 보고된 적이 없고, 가장 가까운 식물들로는 귀화식물로서 전국적으로 분포하는 Carduus crispus (지느러미엉겅퀴) 및 현재까지 국내에 자생하는 것으로 보고된 적이 없는 Carduus defloratus로서 각각 3개씩의 염기가 다른 것으로 나타났다.

Phylogenetic Relationship among Several Korean Coastal Red Tide Dinoflagellates Based on their rDNA Internal Transcribed Spacer Sequences

  • Cho, Eun-Seob;Kim, Gi-Yong;Park, Hyung-Sik;Nam, Byung-Hyouk;Lee, Jae-Dong
    • Journal of Life Science
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    • 제11권2호
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    • pp.74-80
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    • 2001
  • The nucleotide sequences of the internal transcribed spacer regions (ITS1 and ITS2) of ribosomal DNA (rDNA), and the 5.85 rRNA gene, have been determined for 13 strains of dinoflagellates in order to analyze the phylo-genetic relationship. The DNA sequences contained considerable variation in the ITS regions, but little in the 5.85 rDNA. In addition, the ITS1 was more variable than the ITS2 in all species examined. The nucleotide length of this region varied from 519 bp to 596 bp depending on the taxa. The investigated taxa were divided into three large groups based on the ITS length, i. e., a group with short ITS region (A. fraterculus and Alexandrium sp.), a with ITS region group (P. micans, P. minimum and P. triestinum) and a with ITS region group (G. impudicum, C. polykrikoides, G. sanguineum, G. catenatum and H. triquetra). The relationship between nucleotide length of ITS1 and that of ITS2 was negative, whereas G+C content and nucleotide length showed positive correlation. In phylogenetic analyses producing NJ trees, the topology was similar cluster and clearly divided the taxa into three groups based on 5.8S rDNA that were similar to those based on morphological characteristics. In particular, G. impudicum was more closely related to G. catenatum than to C. polykrikoides using phylogenetic analysis. From this study, we chew that the length of ITS region contributes to discriminate Korean harmful algal species and ITS analysis is a useful method for resolving the systematic relationships of dinoflagellates.

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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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    • 제38권1호
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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.

Molecular Identification of Gyrodinium impudicum and Gymnodinium sanguineum by Comparing the Sequences of the Internal Transcribed Spacers 1, 2 and 5.8S Ribosomal DNA

  • Kim Gi Young;Ha Myoung-Gyu;Cho Eun Seob;Lee Tae-Ho;Lee Sang Jun;Lee Jae-Dong
    • Fisheries and Aquatic Sciences
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    • 제2권1호
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    • pp.66-77
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    • 1999
  • The sequences coding for the 5.8S rDNA and the internal transcribed spacers (ITS1 and ITS 2) from the isolates of nine isolates of Gyrodinium impudicum and two isolates of Gymnodinium sanguineum species were amplified, sequenced and compared with the previously known Alexandrium species and Gymnodinium catenatum. The genetic distance analyses based on the sequence alignment indicated that Gymnodinium catenatum and Gyrodinium impudicum species were some related, Alexandrium species was distant. G. catenatum and G. sanguineum were quite separate, but these two species belonged to the same genus. G. impudicum and G. catenatum forming the closet cluster showed some variation in the alignment of ITS regions. The length of ITS1 varied more than that of ITS2 and the length of ITS1 and ITS2 was different for each G. impudicum, Gymnodinium and Alexandrium species. Also, the length of ITS1 was shorter than that of ITS2. However, on the sequences of G. sanguineum, the length of ITS1 was longer about 23 nucleotides than that of ITS2. The phylogenetic analysis and rDNA similarity of G. impudicum and G. catenatum $(59\%)$ is higher than the that of G. catenatum and G. sanguineum $(55\%)$. It was thought that the phylogenetic analysis and the genetic distance revealed that G. impudicum and G. catenatum were clearly different species and G. impudicum may belong to the genus of Gymnodinium.

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Ansanella granifera gen. et sp. nov. (Dinophyceae), a new dinoflagellate from the coastal waters of Korea

  • Jeong, Hae Jin;Jang, Se Hyeon;Moestrup, Ojvind;Kang, Nam Seon;Lee, Sung Yeon;Potvin, Eric;Noh, Jae Hoon
    • ALGAE
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    • 제29권2호
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    • pp.75-99
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    • 2014
  • A small dinoflagellate, Ansanella granifera gen. et sp. nov., was isolated from estuarine and marine waters, and examined by light microscopy, scanning electron microscopy, and transmission electron microscopy. In addition, the identity of the sequences (3,663-bp product) of the small subunit (SSU), internal transcribed spacer (ITS) region (ITS1, 5.8S, ITS2), and D1-D3 large subunit (LSU) rDNA were determined. This newly isolated, thin-walled dinoflagellate has a type E eyespot and a single elongated apical vesicle, and it is closely related to species belonging to the family Suessiaceae. A. granifera has 10-14 horizontal rows of amphiesmal vesicles, comparable to Biecheleria spp. and Biecheleriopsis adriatica, but greater in number than in other species of the family Suessiaceae. Unlike Biecheleria spp. and B. adriatica, A. granifera has grana-like thylakoids. Further, A. granifera lacks a nuclear fibrous connective, which is present in B. adriatica. B. adriatica and A. granifera also show a morphological difference in the shape of the margin of the cingulum. In A. granifera, the cingular margin formed a zigzag line, and in B. adriatica a straight line, especially on the dorsal side of the cell. The episome is conical with a round apex, whereas the hyposome is trapezoidal. Cells growing photosynthetically are $10.0-15.0{\mu}m$ long and $8.5-12.4{\mu}m$ wide. The cingulum is descending, the two ends displaced about its own width. Cells of A. granifera contain 5-8 peripheral chloroplasts, stalked pyrenoids, and a pusule system, but lack nuclear envelope chambers, a nuclear fibrous connective, lamellar body, rhizocysts, and a peduncle. The main accessory pigment is peridinin. The SSU, ITS regions, and D1-D3 LSU rDNA sequences differ by 1.2-7.4%, >8.8%, and >2.5%, respectively, from those of the other known genera in the order Suessiales. Moreover, the SSU rDNA sequence differed by 1-2% from that of the three most closely related species, Polarella glacialis, Pelagodinium bei, and Protodinium simplex. In addition, the ITS1-5.8S-ITS2 rDNA sequence differed by 16-19% from that of the three most closely related species, Gymnodinium corii, Pr. simplex, and Pel. bei, and the LSU rDNA sequence differed by 3-4% from that of the three most closely related species, Protodinium sp. CCMP419, B. adriatica, and Gymnodinium sp. CCMP425. A. granifera had a 51-base pair fragment in domain D2 of the large subunit of ribosomal DNA, which is absent in the genus Biecheleria. In the phylogenetic tree based on the SSU and LSU sequences, A. granifera is located in the large clade of the family Suessiaceae, but it forms an independent clade.

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

  • 김성윤;이재윤;김기영;이기원;박재민;김문옥;이태호;이재동
    • 한국균학회지
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    • 제31권3호
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    • pp.121-128
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    • 2003
  • 국내 유통균주에 대한 Phellinus속의 분류체계를 확립하고 종간구별을 위한 신속동정법을 개발하기 위해 계통학적 정보를 지니고 있는 ITS1, 5.8S rRNA및 ITS2부위의 염기서열을 밝히고 ITS1과 ITS2 부위의 다양한 염기서열을 이용하여 종 특이적인 유전자 탐침을 개발하였다. 본 연구에서 조사된 바에 의하면, 상황버섯으로 시판되고 있는 국내유통균주는 총 9균주로서 P. pini가 4종, P. baumii가 3종, P. igniarius와 P. linteus가 각각 1종으로 확인되었으며 P. gilvus은 조사되지 알았다. 그 중 P. pini는 대부분 중국에서 수입된 제품이었고 P. igniarius를 포함한 P. baumii는 주로 북한산 제품으로 조사되었다. 한편 P. linteus와 P. baumii는 변이가 높은 ITS1 , ITS2 부위에서도 다른 종에 비하여 높은 상동성을 나타내어 종 특이적인 유전자 탐침이 유효하지 않은 반면에 P. igniarius, P. pini, P. gilvus종에 대한 detection primer로 각각 IF1-IR3, PF1-PF3, GF2-FR4 primer는 유전자 탐침이 유효함을 확인하였다. 유연관계가 아주 가까운 P. linteus와 P. baumii에 대한 정확한 분석을 위해서는 변이가 심한 ITS 부위와 보존성이 높은 18S rDNA 부위의 염기서열을 함께 분석, 비교하여야 가능하리라 사료된다.