• Title/Summary/Keyword: ITS 1/5.8S rDNA/ITS2

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Secondary Structure of the Ribosomal Internal Transcribed Spacer (ITS) Region of Hypsizygus marmoreus (느티만가닥버섯의 ITS (internal transcribed spacer) 영역의 2차구조 분석)

  • Woo, Ju-Ri;Yoon, Hyeokjun;You, Young-Hyun;Lee, Chang-Yun;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1260-1266
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    • 2013
  • The ribosomal DNA (rDNA) clusters of Hypsizygus marmoreus 3-10 and H. marmoreus 1-1 were analyzed in this study. The small subunit (SSU) and intergenic spacer 2 (IGS 2) was partially sequenced. The internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), and 5S were completely sequenced. The rDNA clusters of H. marmoreus 3-10 and H. marmoreus 1-1 were 7,049 bp in length. The sequence of SSU rDNA, which corresponded to 18S rDNA, was 1,796 bp in length, and the sequence of LSU rDNA, which corresponded to 28S rDNA, was 3,348 bp in length. The ITS region that variable region and IGS region that non-transcribed spacer was 462 bp and 1,290 bp in length. The sequence of 5.8S rDNA and 5S rDNA was 153 bp and 43 bp in length, respectively. The 17 bp of the rDNA cluster in the H. marmoreus 3-10 strain was different to that in the H. marmoreus 1-1 strain, with 2 bp in the SSU, 3 bp in the ITS, 9 bp in the LSU, and 3 bp in the IGS. The analysis of the secondary structure revealed that the ITS regions of H. marmoreus 3-10 and H. marmoreus 1-1 have five stem-loop structures. Interestingly, among these structures, one different nucleotide sequence resulted in a different secondary structure in stem-loop V.

Phylogenetic and Chemical Analyses of Cirsium pendulum and Cirsium setidens Inhabiting Korea (국내에 자생하는 큰엉겅퀴와 고려엉겅퀴의 분자유전학적 및 화학적 분석)

  • Yoo, Sun-Kyun;Bae, Young-Min
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1120-1125
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    • 2012
  • Cirsium pendulum plants were collected from Hongcheon, Pyeongchang, Wonju, Yangyang in Kangwondo, Gapyeong in Gyeongkido, and Choongju in Choongcheongbukdo. Cirsium setidens plants were collected from Taebaek in Kangwondo and Bonghwa in Kyeongsangbukdo. Genomic DNA was prepared from those plants and used for the amplification of 18S rDNA, ITS1, 5.8S rDNA, ITS2, and part of 28S rDNA. The PCR products were sequenced, and the sequence was deposited in the GenBank. The comparison of those sequences has revealed that the rDNA sequences are identical for all six C. pendulum plants, but that the ITS1 and ITS2 sequences contain variable nucleotides. The two C. setidens plants had different nucleotides in 18S rDNA, ITS1, and ITS2. The comparison of the DNA sequences of C. pendulum and C. setidens collected in this study with C. pendulum of Hokkaido in Japan and C. japonicum of Anhui in China indicated that the plants of those three species are clearly divided into three distinct groups. The silymarin content of the collected plants was analyzed and turned out to be quite high. Therefore, it has been found that both C. pendulum and C. setidens plants are producing large amounts of silymarin, which has been reported to have various medicinal effects.

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

  • Bae, Young-Min
    • Journal of Life Science
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    • v.20 no.4
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    • pp.578-583
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    • 2010
  • It has been reported that extracts of globe thistle (Echinops spp.) and thistle (Circium spp., Carduus spp. and Onopordum spp.) have anti-bacterial and anti-fungal activities. Methanol extracts of Echinops setifer and Carduus spp. were used to test and see if the extracts of these plants could suppress growth of Mycobacterium smegmatis and Mycobacterium fortuitum. Although extract of Echinops setifer showed no anti-mycobacterial activities, extract of Carduus spp. showed inhibition zones when tested with filter discs. Genomic DNA was isolated from Carduus spp. and PCR was performed to clone a DNA fragment containing ITS1, 5.8S rRNA gene and ITS2. A 733-bp PCR product was obtained and its DNA sequence was reported to the GenBank (accession number GU188570). BLAST search of the obtained DNA sequence did not show a match with any DNA sequences in the Genbank. Carduus crispus and Carduus defloratus had the closest phylogenetic relationships to this plant.

Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

  • Park, Joung-Eon;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Cha, Jae-Ho;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.29 no.3
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    • pp.121-131
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    • 2001
  • This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

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A Phylogenetic Relationship between Foreign and Korean Strains of Flammulina velutipes Identified by rDNA-ITS Sequence Analysis (Flammulina velutipe의 국내 균주와 외래 균주 간의 ITS region을 이용한 계통학적 유연관계 분석)

  • Hwang, Gwang-Rip;Woo, Ju-Ri;Yoon, Hyeok-Jun;Lee, Chang-Yun;Lee, Sang-Han;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.1
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    • pp.62-73
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    • 2012
  • This study was carried out to investigate the genetic relationship of Flammulina velutipes with other species. The ribosomal DNA cluster containing 4 rRNA genes from F. velutipes 4154 were sequenced. The length of the rDNA cluster sequence was estimated at 7,403 bp long and consisted of 1,806 bp of SSU rDNA, 245 bp of ITS 1 region, 159 bp of 5.8S rDNA, 308 bp of ITS 2 region, 3,402 bp of LSU rDNA, 1,400 bp of IGS 1 region, and 83 bp of 5S rDNA. The F. velutipes 4154 genes were contained in the rDNA cluster of F. velutipes in the order of SSU rDNA - ITS 1 - 5.8S rDNA - ITS 2 - LSU rDNA - IGS 1 - 5S rDNA. The phylogenetic relationships of 20 strains of Tricholomataceae and Physalacriaceae were analyzed by conducting distance analysis using the Neighbor-joining (NJ) method. The 20 strains used in this study were divided into three groups and the strains of the genus Flammulina were related very closely to strains of Physalacria bambusae.

Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

  • Kim, Gi-Young;Lee, Goang-Jae;Ha, Myung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.28 no.1
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    • pp.11-16
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    • 2000
  • The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

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Phylogenetic Analysis of the Genus Phellinus by Comparing the Sequences of Internal Transcribed Spacers and 5.8S Ribosomal DNA (Ribosomal DNA의 Internal Transcribed Spacer(ITS) 부위의 염기서열분석에 의한 Phellinus속의 계통분석에 관한 연구)

  • Chung, Ji-Won;Kim, Gi-Young;Ha, Myung-Gui;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.124-131
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    • 1999
  • This study was carried out to identify the phylogenetic relationship among Phellinus species by comparing the DNA sequences of the 5.8S ribosomal DNA (rDNA) and the internal transcribed spacers (ITSs), ITS1 and ITS2 regions. Two primers from the 3' end of 18S rDNA and the 5' end of 28S rDNA sequences were chosen to amplify the specific ITS regions of Phellinus spp. Phellinus strains used in the study were divided into four clusters by the phylogenetic tree based on the amplified regions of ITS and 5.8S rDNA sequences. The first cluster consist of Phellinus hartigii IMSNU 32041 and Phellinus robustus IMSNU 32068, and the second cluster consists of Phellinus linteus strains and Phellinus weirianus IMSNU 32021. Phellinus laevigatus KCTC 6229, KCTC 6230 and Phellinus igniarius KCTC 6227, KCTC 6228 belong to the third cluster. Finally, Phellinus chrysoloma KCTC 6225 and Phellinus chrysoloma KCTC 6226 are the fourth cluster. In the second cluster the differentiation between Phellinus linteus strains and Phellinus weirianus species were not possible by the comparison of the ITS sequences. These results revealed that Phellinus linteus and Phellinus weirianus cannot be established the concept of species level only by the ITS sequences. Therefore, both physiological and molecular biological methods as well as the sequences of type strains are necessary to classify the strains of these two species accurately. The comparison of the ITS sequences of four Phellinus species indicated that the sequences of the ITS1 generally are more divergent than those of the ITS2. Although the ITS sequences are varied in some species, the conserved regions in both ITS1 and ITS2 are useful tool to differentiate the species. Phellinus linteus and related species have their specific sequences in the ITS1 compared to the other species.

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Phylogenetic Analysis of Dendropanax morbifera Using Nuclear Ribosomal DNA Internal Transcribed Spacer (ITS) Region Sequences (Internal transcribed spacer (ITS) region의 염기서열 분석에 의한 보길도산 황칠나무의 분자 계통학적 연구)

  • Shin, Yong Kook
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1341-1344
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    • 2016
  • Dendropanax morbifera is an endemic tree species of Korea, it is restricted to the southern parts of Korea. The internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) for Dendropanax morbifera grown at Bogil-do, Korea was determined. We investigated the sequence-based phylogenetic relationships of plants related and clarified its taxonomical position. The determined sequences consisted of 689 residues. ITS1 was 222 bp long while ITS2 was 233 bp long. The 5.8S rDNA was 160 bp long. The ITS region sequences of the Dendropanax species included in this study were obtained from GenBank. Oreopanax polycephalus was used as the outgroup. A pairwise alignment was calculated using the Clustal X program. A phylogenetic tree was constructed by the neighbor-joining method using the Tree view program. Sequence similarities among species including D. morbifera Bogil-do isolate showed the range 92.6 to 99.7% in sequence-based phylogenetic analysis using total 615 base pairs of ITS1, 5.8S rDNA and ITS2. D. morbifera Bogil-do isolate exhibited the highest degree of relatedness to D. chevalieri, sharing 99.7% ITS region similarity. D. morbifera Bogil-do isolate also showed to D. trifidus, sharing 99.4% ITS region similarity.

Comparison of ITS(Internal Transcribed Spacer) and 5.8S rDNA Sequences among varieties and Cultivars in Panax ginseng

  • Yang, Deok-Chun;Yang, Key-Jin;Yoon, Eui-Soo
    • Journal of Photoscience
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    • v.8 no.2
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    • pp.55-60
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    • 2001
  • Ginseng (Panax genus) is one of the most medicinally important genera and consists of highly regarded medicines. Among the species of Panax, the ginseng species is widely known to have most medicinal quality. P. ginseng has 3 varieties, Jakyung, Chunggyung and Hwangsook, discovered in nature with different colors of stem and fruit, Jakyung has two cultivars, Yunpoong and Chunpoong. Rigorous phylogenetic analysis of these varieties and cultivars has been conducted with sequencing of rDNA region. The sequences of ITS1, ITS2 of every varieties and cultivars within P. ginseng were identical. The sequence of 5.8S rDNAs of Hwangsook variety were different from the sequences of 5.8S rDNAs of others by only one base pair at nucleotide position 14. In phylogenetic analysis and predicted RNA secondary structure study, it is assumed that evolution has proceeded from Hwangsook to other varieties. recently.

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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.