• Title/Summary/Keyword: ITS (internal transcribed spacer)

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Isolation of a Pestalotiopsis Species Degrading Mucilage from Fruit of Opuntia ficus-indica var. Saboten

  • Huh, Yoon-Hee;Ko, Young-Hwan
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.221-226
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    • 2007
  • The high molecular-weight mucilage extracted and purified from cactus fruit of Opuntia ficus-indica var. Saboten was degraded by the cell-free culture filtrate of a fungus isolated from soil. TLC analysis of the polymeric mucilage after incubation with the fungal culture filtrate confirmed its degradation. When the degradation products were tested for their qualitative reactions with ninhydrin and phenol-sulfuric acid, only phenol-sulfuric acid gave positive development, and ninhydrin did not show any observable color reaction. This coloring reaction suggested the presence of a carbohydrate without an amino group within the mucilage. Analyses by HPLC and liquid gel permeation chromatography on sephadex G-100 also provided additional information on degradation of the mucilage by the fungal culture filtrate. The sequences of ITS-5.8S rDNA from the fungal isolate that was cultivated for the preparation of mucilage-degrading enzyme showed 99% similarity to those of Pestalotiopsis aquatica.

Characterization of a Zygomycete Fungus, Mortierella minutissima from Freshwater of Yeongsan River in Korea

  • Nguyen, Thi Thuong Thuong;Lee, Hyang Burm
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.346-349
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    • 2016
  • During a survey of fungal diversity of the order Mortierellales, a zygomycete strain, EML-YS717-1, was isolated from a freshwater sample collected from the Yeongsan River in Gwangju, Korea. Based on its morphological characteristics and a phylogenetic analysis of the internal transcribed spacer (ITS1 and ITS2) and 5.8S rDNA sequences, the strain was identified as Mortierella minutissima. To the best of our knowledge, M. minutissima, has not previously been authentically reported in Korea.

Phylogeny and Morphology of Sarcopodium terrigenum sp. nov., a Novel Fungal Species Isolated from Soil in Korea

  • Elderiny, Nabil Salah;Das, Kallol;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.49 no.2
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    • pp.175-181
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    • 2021
  • A fungal strain was isolated from a soil sample collected in Korea and designated as YW23-8. Based on a sequence analysis of the internal transcribed spacer (ITS) regions, the isolate was assigned to the genus Sarcopodium. Moreover, a phylogenetic analysis based on the concatenated nucleotide sequences of the ITS regions and the large subunit of the nuclear ribosomal RNA (LSU) gene showed that the strain YW23-8 occupies a distinct phylogenetic position within Sarcopodium. The isolate had significant differences from its closest neighbors, S. circinosetiferum, S. circinatum, S. macalpinei, and S. vanillae. Morphological features such as different conidial structures, the absence of septation in conidia, and the presence of milky white watery droplets along with the results of the phylogenetic analysis clearly distinguish YW23-8 from the closest Sarcopodium species. We therefore conclude that strain YW23-8 represents a novel species of the genus Sarcopodium for which we propose the name Sarcopodium terrigenum.

ITS sequence variations in common ragweed and giant ragweed (돼지풀 및 단풍잎돼지풀의 ITS 염기서열 변이)

  • Kim, Young-Dong;Park, Chong-Wook;Sun, Byung-Yun;Kim, Ki-Joong;Lee, Eun-Ju;Kim, Sung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.35 no.4
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    • pp.273-285
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    • 2005
  • Genetic variation of notorious invasive plants, common ragweed (Ambrosia artemisiifolia L.) and giant ragweed (Ambrosia trifida L.) were examined using the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA. A total of 18 different ITS types were detected from 156 individuals of common ragweed sampled mainly from the southern part of Korean peninsula whereas four types were identified from 46 individuals of giant ragweed. High sequence diversity observed from common ragweed in Korean populations was interpreted as multiple introduction. Genetic recombination was suggested as possible method for the production of some of the ITS types while point mutation was mainly responsible for the origin of the sequence diversity. This study provided some of basic genetic information needed for understanding of the evolutionary process in ragweed during invasion.

Molecular Authentication and Phylogenetic Relationship of Bupleurum Species by the rDNA-ITS Sequences (rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석)

  • Moon, Byeong-Cheol;Choo, Byeong-Kil;Ji, Yun-I;Yoon, Tae-Sook;Lee, A-Young;Cheon, Myeong-Sook;Kim, Bo-Bae;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.3
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    • pp.59-68
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    • 2009
  • Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. 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 adulterant Bupleurum species.

Cloning and Organization of the Ribosomal RNA Genes of the Mushroom Trichloma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.194-199
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    • 1995
  • A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

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Phylogenetic Study of Genus Haliotis In Korea by Internal Transcribed Spacer Sequence (ITS) (ITS에 의한 한국내 전복 속 분류군의 유전적 계통분류학적 연구)

  • Huh, Man-Kyu;Kim, Jung-Ho;Moon, Du-Ho
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1003-1008
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    • 2009
  • Abalone (genus Haliotis) is a woody species with a long life span that is primarily distributed throughout the world, including Asia. This species is regarded as a very important marine gastropod mollusk in Korea and China, and also in food industries around the world. We evaluated a representative sample of the five species with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within the genus. Aligned nucleotide sequences of the length of the 5.8S subunit of all taxa of Haliotis were found to constant of 160 bp nucleotides. However, aligned nucleotide sequences of the length of ITS1 were varied within genus Haliotis, varying from 272 in H. diversicolor aquatilis to 292 in H. discus hannai. Aligned nucleotide sequences of the length of ITS2, especially, vary from 722 in H. diversicolor aquatilis to 752 in H. sieboldii. Total alignment length is 763 positions, of which 78 are parsimony-informative, 57 variable but parsimony-uninformative, and 459 constant characters. H. discus hannai was similar to H. discus, while H. diversicolor aquatilis was more distinct. ITS analysis may be useful in germ-plasm classification several taxa of genus Haliotis.

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.

Phylogenetic Analysis of Phellinus linteus and Related Species Comparing the Sequences of rDNA Internal Transcribed Spacers

  • Lee, Jae-Dong;Kim, Gi-Young;Park, Joung-Eon;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Lee, Tae-Ho
    • Journal of Life Science
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    • v.11 no.2
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    • pp.126-134
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    • 2001
  • The phylogenetic tree displayed the presence of five groups in the Phellinus genus, which were distinguished based on their morphology. Most of the p. linteus appeared a cluster which was highly significant with the exception of P. linteus KACC 500122 and KACC 500411. They formed the sister taxa of P 1inteus where P. baumii, Phellinus sp. MPNU 7003, MPNU 7007, and MPNU 7010 had similar morphological characteristics. Also, P. nigricans IMSNU 32024 and P. pini var, carniformans IMSNU 32031 were grouped in the same cluster with P. igniarius KCTC 6227, KCTC 6228, and P. chrysoloma KCTC 6225 extracted from the Gen-Bank database. P. torulosus IMSNU 32028 and Phellinus sp. MPNU 7011 formed a closed group, however, these species had a distant taxa when compared with the other Phellinus species. The nucleotide sequences of the internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) including the 5.85 rDNA were determined from 24 strains of the Phellinus genus in order to analyze their phylogenetic relationship. These fungi were divided into two basic groups based on their ITS length, however, this grouping was different from that based on their morphological characteristics. Although various ITS sequences were ambiguously aligned, conserved sites were also identified. Accordingly, a neighbor-joining tree was constructed using the nucleotide sequence data of the conserved sites of the ITS regions and the 5.8S rDNA.

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Morphological and molecular evidence of the hybrid origin of Crepidiastrum ×muratagenii in Korea (홍도고들빼기의 형태 다양성 및 잡종 기원의 분자 증거)

  • JANG, Young-Jong;PARK, Boem Kyun;SON, Dong Chan;CHOI, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.52 no.2
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    • pp.85-96
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    • 2022
  • The plant "Hong-do-go-deul-ppae-gi" has been considered as Crepidiastrum × muratagenii, a hybrid between C. denticulatum and C. lanceolatum, based on its morphological traits and geographical distribution. To reveal the hybrid origin of Hong-do-go-deul-ppae-gi, we examined additional morphological traits of this plant and its putative parents (C. denticulatum, C. lanceolatum, C. platyphyllum) and analyzed one nuclear ribosomal internal transcribed spacer (ITS) region and four chloroplast regions (trnT-L, trnL-F, rpl16 intron, and rps16 intron). As a result of examining the morphological traits, putative hybrid individuals were classified into three types based on the habit, cauline leaf, outer phyllary, and achene beak traits. A molecular analysis found that the ITS sequences of Type 1 and Type 2 individuals showed additive species-specific sites of C. denticulatum and C. lanceolatum. Plastid sequences of Type 1 and Type 2 individuals showed C. denticulatum and C. lanceolatum sequences, respectively. However, Type 3 individuals had ITS and plastid sequences corresponding to C. denticulatum. Accordingly, Type 1 and Type 2 individuals not only share morphological traits with C. denticulatum and C. lanceolatum but also show additive species-specific sites for C. denticulatum and C. lanceolatum, and not C. platyphyllum, supporting its origin as a hybrid between C. denticulatum and C. lanceolatum. Type 3 had morphological traits similar to other hybrid types but was distinguished with respect to outer phyllaries and demonstrated some resemblance to C. denticulatum. In a molecular analysis, Type 3 was found to be identical with regard to the sequence of C. denticulatum and was judged to be an ecological variation of C. denticulatum.