• Title/Summary/Keyword: ITS regions of rDNA

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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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    • v.2 no.1
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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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PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp.

  • Min, Byung-Re;Lee, Young-Mi;Choi, Yong-Keel
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.66-73
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    • 2000
  • To investigate the genetic relationship among 12 species belonging to the Fusarium section Martiella, Dlaminia, Gibbosum, Arthrosporiella, Liseola and Elegans, the internal transcribed spacer(ITS) regions of ribosomal DNA (rDNA) were amplified with primer pITS1 and pITS4 using the polymerase chain reaction(PCR). After the amplified products were digested with 7 restriction enzymes, restriction fragment length polymorphism (RFLP) patterns were analyzed. The partial nucleotide sequences of the ITS region were determined and compared. Little variation was observed in the size of the amplified product having sizes of 550bp or 570bp. Based on the RFLP analysis, the 12 species studied were divided into 5 RFLP types. In particular, strains belonging to the section Martiella were separated into three RFLP types. Interestingly, the RFLP type of F. solani f. sp. piperis was identical with that of isolates belonging to the section Elegans. In the dendrogram derived from RFLP analysis of the ITS region, the Fusarium spp. examined were divided into two major groups. In general, section Martiella excluding F. solani f. sp. piperis showed relatively low similarity with the other section. The dendrogram based on the sequencing analysis of the ITS2 region also gave the same results as that of the RFLP analysis. As expected, 5.8S, a coding region, was highly conserved, whereas the ITS2 region was more variable and informative. The difference in the ITS2 region between the length of F. solani and its formae speciales excluding F. solani f. sp. piperis and that of other species was caused by the insertion/deletion of nucleotides in positions 143-148 and 179-192.

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Chemosystematics and Molecular Phylogeny of a New Bioflocculant-Producing Aspergillus Strain Isolated from Korean Soil

  • Kim, Gi-Young;Ha, Myoung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.870-872
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    • 1999
  • The ubiquinone and G+C contents of the bioflocculant-producing fungus, a new Aspergillus strain, were detennined using high-perfonnance liquid chromatography. The internal transcribed spacers 1 and 2 (ITS1 and ITS2), and the 5.8S ribosomal DNA (rDNA) of the strain were amplified and sequenced. The strain contained ubiquinone-l0($H_2$)as a major quinone and the G+C content was 49 mol%. A phylogenetic analysis of the ITS regions indicated that the strain belonged to the genus Aspergillus according to its previously classified morphological characteristics. Based on a sequence homology search, the strain was most closely related to Petromyces muricatus (anamorph, A. muricatus; accession number, AJ005674). The sequence of a new Aspergillus strain in ITS1 and ITS2, and 5.8S rDNA showed 97% homology to P. muricatus. Therefore, the strain is believed to be a new bioflocculant-producing Aspergillus strain.

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Two Unreported Glomeromycota Fungi Isolated from Rhizospheres of Smilax china (청미래덩굴의 근권에서 분리된 2종의 Glomeromycota 미기록종)

  • Park, Hyeok;Ka, Kang-Hyeon;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.47 no.3
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    • pp.275-280
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    • 2019
  • We isolated fungal spores belonging to the phylum Glomeromycota from the rhizospheres of Smilax china, cultured in a greenhouse. We identified the isolated spores using sequence analysis of 18S partial rDNA region, internal transcribed spacer and 28S rDNA regions. We confirmed 2 unreported spores of Glomeromycota fungal species, Diversispora eburnea and Paraglomus laccatum. Here, we described the morphological characteristics and results of phylogenetic analysis of the confirmed species.

Restriction Fragment Length Ploymorphism of PCR Amplified Ribosomal DNA Among Korean Isolates of Phytophthora

  • Hong, Seung-Beom;Jee, Hyeong-Jin;Lee, Seung-Im;Go, Seung-Joo
    • The Plant Pathology Journal
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    • v.15 no.4
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    • pp.228-235
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    • 1999
  • Genetic diversity of ninety-five Korean isolates of Phytophthora was investigated on the basis of PCR-RFLP of ribosomal DNA. The isolates were previously identified as following fifteen species by mycological and cultural characteristics; P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamoni, P. citricola, P. citrophthora, P. cryptogea, P. drechsleri, P. erythroseptica, P. infestans, P. megasperma, P. nicotianae, P. palmivora and P. sojae. The regions of small subunit (SSU) and internal transcribed spacer (ITS) of rDNA were amplified with primer pair, NS1 and ITS4, by polymerase chain reaction (PCR) and digested with nine restriction enzymes. P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamomi, P. citricola, P. citrphthora, P. infestans, P. nicotianae and P. palmivora showed specific band patterns for each species. However, P. sojae and P. erythroseptica presented identical band patterns and P. cryptogea, P. drechsleri and P. megasperma were divided into six groups, which were not compatible with delineation of the species. A group originated from cucurbits showed distinct band patterns from other groups, but the other five groups were closely related within 96.0% similarity, forming one complex group. Consequently, Korean isolates of Phytophthora were divided into thirteen genetic groups and each group was readily differentiated by comparing digestion patterns of AvaII, HaeIII, MboI, HhaI and MspI. Therefore, PCR-RFLP of rDNA using the five enzymes can be used to differentiate or identify the Phytophthora species reported in Korea so far.

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Genetic Diversity and Phylogenetic Relationship in Korean Strains of Lentinus lepideus Based on PCR Polymorphism (PCR 다형성 분석에 의한 한국산 잣버섯의 유전적 다양성 및 유연관계)

  • Lee, Jae-Seong;Cho, Hae-Jin;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Lee, Yun-Hae;Jang, Myoung-Jun;Ju, Young-Chul;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.105-111
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    • 2010
  • Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

Phylogenetic Study of Korean Chrysosplenium Based on nrDNA ITS Sequences (ITS 염기서열에 의한 한국산 괭이눈속(Chrysosplenium)의 계통학적 연구)

  • Han, Jong-Won;Yang, Sun-Gyu;Kim, Hyun-Jun;Jang, Chang-Gee;Park, Jeong-Mi;Kang, Shin-Ho
    • Korean Journal of Plant Resources
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    • v.24 no.4
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    • pp.358-369
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    • 2011
  • The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA from genus Chrysosplenium were sequenced to address phylogenetic relationship. ITS including 5.8S sequence varied in length from 647 bp to 653 bp. Among them, 219 sites were variable sites with parsimony-informative. The aligned sequences were analyzed by maximum parsimony (MP) and neighbor-joining (NJ) methods. In the strict consensus trees of parsimony analysis, the monophyly of Chrysosplenium was supported by 100% bootstrap value. The first clade, C. pseudofauriei was at the basal position of the genus, and others formed two clades with high bootstrap support. The second clade included Ser. Pilosa and Ser. Oppositifolia and third clade included Ser. Alternifolia and Ser. Flagellifera. The NJ trees showed essentially the same topology. Finally, DNA sequences of ITS regions were useful phylogenetic marker in this genus. Based on the ITS and ridge seed morphological results, C. sphaerospermum Maxim. and C. valdepilosum (Ohwi) S.H. Kang & J.W. Han were discussed their scientific names and taxonomic positions.

Comparison between DNA- and cDNA-based gut microbial community analyses using 16S rRNA gene sequences (16S rRNA 유전자 서열 분석을 이용한 DNA 및 cDNA 기반 장내 미생물 군집 분석의 비교)

  • Jo, Hyejun;Hong, Jiwan;Unno, Tatsuya
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.220-225
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    • 2019
  • Studies based on microbial community analyses have increased in the recent decade since the development of next generation sequencing technology. Associations of gut microbiota with host's health are one of the major outcomes of microbial ecology filed. The major approach for microbial community analysis includes the sequencing of variable regions of 16S rRNA genes, which does not provide the information of bacterial activities. Here, we conducted RNA-based microbial community analysis and compared results obtained from DNA- and its cDNA-based microbial community analyses. Our results indicated that these two approaches differed in the ratio of Firmicutes and Bacteroidetes, known as an obesity indicator, as well as abundance of some key bacteria in gut metabolisms such as butyrate producers and probiotics strains. Therefore, cDNA-based microbial community may provide different insights regarding roles of gut microbiota compared to the previous studies where DNA-based microbial community analyses were performed.

Nucleotide Sequences of an Aphid ribosomal RNA Unit (진딧물의 전 ribosomal RNA 염기배열)

  • Kwon, Tae-Young;An, Seung-Lak;Song, Cheol;Park, Jong-Kyun;Kim, Young-Sub;Hwang, Jae-Sam;Kwon, O-Yu
    • Journal of Life Science
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    • v.8 no.1
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    • pp.32-39
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    • 1998
  • The length and G/C concent of regions of an aphid rDNA unit that spans 13,061bo with 59% G/C content. flolowing belowing below are the those results, 5’ETS is 843bp in length with 69% G/C content, 18S is 2,469bp in length with 59% G/C content, ITS I is 229bp in length with 70% G/C content, 5.8S is 160bp in length with 63% G/C content, ITS II is 325bp in length with 70% G/C content, 28S is 4, 147bp in length with 60% G/C content, IGS is 4,888bp in length with 55% G/C content.

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Phylogenetic relationships in different strains of Pholiota species based on PCR polymorphism (PCR 다형성 분석에 의한 비늘버섯 속 계통의 유연관계 분석)

  • Kwon, Woon-Hyuk;Park, Hyuk;Baek, Min-Jae;Cho, Woo-Jin;Choi, Woo-Jeong;Ahn, Chi-Beom;Shin, Do-Bin;Lee, Tae-Soo
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.69-76
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    • 2013
  • Pholiota species were collected from different geographical regions of the world. Genetic diversity and phylogenetic relationships were analyzed by rDNA-ITS sequences and RAPD polymorphism. The sizes of rDNA-ITS PCR amplicons of Pholiota spp. varied from 233~271, 158~223 and 174~219 bp, respectively. A phylogenetic tree was constructed on the ITS region sequences and Pholiota strains were classified into 8 clusters. Twenty strains in seven Pholiota spp. were classified into seven clusters by RAPD polymorphism using 15 arbitrary primers. Our experimental results suggested that rDN-ITS and RAPD analysis are useful tool for classifying Pholiota spp. and strains.