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

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Identification of Entomopathogenic Fungus, Beauveria spp. F-101 Isolated from Thecodiplosis japonensis Using Internal Transcribed Spacer Sequence

  • Shin, Sang-Chul;Roh, Jong-Yul;Kim, Chul-Su;Park, Il-Kwon;Jeon, Mun-Jang;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.77-80
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    • 2004
  • For the development of the alternative control system against the major forest pests, Beauveria spp. F-101, isolated from a dead larva of Thecodiplosis japonensis, was selected because this isolate showed high pathogenicities against T. japonensis and Acantholyda parki. Beauveria spp. F-101 had irregular clustered conidio-phores and conidia borne on a distinctive apical zigzag extension, and it showed typical characteristic of the genus, Beauveria in morphology. For molecular based-identification, the ribosomal ITS region of Beauveria spp. F-101 was amplified with ITS1 and ITS4 primers, and cloned into pGEM- T Easy vector. The amplified PCR product was 569 bp in size and completely sequenced. The similarities of the cloned ITS sequence were 99 % and 97% to those of B. bassiana and B. brongniartii, respectively. In comparison to other species among the genus Beauveria, the ITS region of Beauveria spp. F-101 showed a similarity of 95% to B. amorpha, 95% to B. tenella, 89% to B. vermiconia and 69% to B. alba, respectively. In addition, in comparison to different genus, it had 95% similarities to Cordyceps militaris and 91% to Paecilomyces tenuipes. Accordingly, the current result suggests that Beauveria spp. F-101 was a variant of B. bassiana and it seems to be a new isolate considering sequence variation in ITS region.

Characteristics of Plantlets Redifferentiated from F1 Hybrid between Panax ginseng and Panax quinquefolius (고려인삼과 미국삼의 종간잡종으로부터 재분화된 식물체의 특성)

  • Ahn, In-Ok;Lee, Sung-Sik;Lee, Jang-Ho;Lee, Bum-Soo;In, Jun-Gyo;Yang, Deok-Chun
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.45-48
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    • 2006
  • The characteristics of plantlets redifferentiated from calli of F1 hybrid between Panax ginseng and Panax quinquefolius were investigated. Growth of plantlets redifferentiated from F1 hybrid was superior to the plants redifferentiated from Korean ginseng. Stem color of plantlets redifferentiated from F1 hybrid was more purple than that from Korean ginseng and leaf color of the former was also greener than that of the latter. Chunpoong, Yunpoong and Seonweon which are belonged to Korean ginseng showed same PCR band(A), while American ginseng showed different PCR band (B) in Internal Transcribed Spacer (ITS) region. F1 hybrid exhibited both A and B PCR band which belonged to Korean ginseng and American ginseng, respectively. F1 hybrid calli and plantlets redifferentiated from F1 hybrid calli showed same PCR band with that of F1 hybrid plant in ITS region. Therefore it was confirmed that piantlets redifferentiated from F1 hybrid exhibited genetic stability in ITS region.

Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species (DNA 바코딩과 고해상 융해곡선분석에 기반한 인삼속 식물의 종 판별)

  • Bang, Kyong Hwan;Kim, Young Chang;Lim, Ji Young;Kim, Jang Uk;Lee, Jung Woo;Kim, Dong Hwi;Kim, Kee Hong;Jo, Ick Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.6
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    • pp.439-445
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    • 2015
  • Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

Development of specific SNP molecular marker from Thistle using DNA sequences of ITS region (엉겅퀴의 ITS 영역 염기서열 분석을 통한 특이적 SNP 분자마커의 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.102-109
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    • 2018
  • Thistle is a perennial plant that is widely used for medicinal purposes. Information on the genetic diversity of thistle populations are great important for their conservation and germ plasmic utilization. Although thistle is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish them from other similar species from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from the nuclear ribosomal DNA internal transcribed spacer (ITS) regions of genomic sequences to identify distinct Korean-specific thistle species via an amplification refractory mutation system (ARMS)-PCR and high resolution melting (HRM) curve analyses. We performed molecular authentication of four different kinds of thistle species from different regions using DNA sequences in the ITS intergenic region. We also developed a quantitative PCR assay using species-specific ITS primers, which allowed us to estimate the ratio of Korean-specific thistle species using varying ratios of mixed genomic DNA templates from the two species. The SNP markers developed in this study are useful for rapidly identifying specific thistle species from different countries.

Sequence Analysis of Cochlodinium polykrikoides Isolated from Korean Coastal Waters Using Sequences of Internal Transcribed Spacers and 5.8S rDNA

  • Kim, Hak-Gyoon;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • v.35 no.3
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    • pp.158-160
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    • 2000
  • The relativity of four isolates of C. polykrikoides was determined by comparative sequence analysis based on direct sequencing of PCR amplified ribosomal DNA (the internal transcribed spacer region and the 5.8S rDNA). Sequence comparisons indicated that four isolates had the same nucleotide sites in the ITS regions, as well as a total of 585 nucleotide length and 100% homology. The molecular data revealed that C. polykrikoides in Korean coastal waters show no genetical difference.

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Phylogenetic Classification of Antrodia and Related Genera Based on Ribosomal RNA Internal Transcribed Spacer Sequences

  • Kim, Seon-Young;Park, So-Yeon;Jung, Hack-Sung
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.475-481
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    • 2001
  • Sequences of ribosomal internal transcribed spaces (ITS) obtained from two Antrobia species and two related species were compared to investigate intrageneric and intergeneric phylogenetic relationships of Antrodia. The results showed that Antrodia species causing a brown rot in wood did not form a monophyletic clade and were separated into three distinct groups. Antrodia gossypina and A. vaillantii formed a clade having rhizomorphs as a homologous character. Antrodia serialis, A. sinuosa, and A. malicola formed a group together with Daedalea, Fomitopsis, and Postia species with brown rot habit. Antrodia xantha with a trimitic hyphal system and amyloid skeletal hyphae formed another distinct clade form other Antrodia species. The Antrodia species were separated from white rot genera such as Antrodiella, Diplomitoporus, Junghuhnia, and Steccherinum, indicating the phylogenetic importance of the rot type in the classification of the Polyporaceae.

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Genetic characterization of Phellinus baumii PMO-P4 by analyzing restriction fragment length polymorphisms of nuclear ribosomal DNA internal transcribed spacers (ITS) (Ribosomal DNA의 ITS부위에 대한 RFLP 분석에 의한 Phellinus baumii PMO-P4의 유전학적 특성)

  • Chang, Yun-Hee;Kim, Tae-Rack;Kim, Hyun-Su;Yeo, Ik-Hyun;Lee, Sang-Youn;Ha, Hyo-Cheol
    • Journal of Mushroom
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    • v.4 no.2
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    • pp.43-47
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    • 2006
  • PMO-P4, being cultivated as "Sanghwang" in Korea, was proved to be P. baumii based on ITS (internal transcribed spacer) sequencing and RFLP (Restriction Fragment Length Polymorphism) patterns along with some Phellinus species including P. linteus. The similaraty of ITS sequencing between PMO-P4 and other Phellinus species was given the range of 48.6%~72.2%, showing the highest homology from P. linteus and the lowest from P. gilvus.

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Analysis of the Phylogenetic Relationships in the Genus Spiraea Based on the Nuclear Ribosomal DNA ITS Region (핵 리보솜 DNA ITS 부위에 의한 조팝나무속 식물종의 계통 관계 분석)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.22 no.3
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    • pp.285-292
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    • 2012
  • Genus Spiraea is composed of many long-lived woody species that are primarily distributed throughout Asia and Europe. In this study, we evaluated a representative sample of the 38 taxa in the world, including 14 in Korea, with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within the genus. The molecular data allowed us to resolve well-supported clades in the taxa. In 47 world accessions (38 taxa: 14 Korean taxa, 33 world taxa, and 9 overlapping taxa), total alignment length was 689 positions, of which 452 were parsimony informative, 527 variable, 75 singleton, and 159 constant characters. Although the phylogenic tree showed that many taxa of genus Spiraea were well separated from each other, many branches were not congruent with the morphological characteristics and geographical distributions of the genus. There were 430 segregating sites and the nucleotide diversity (${\pi}$) value was 0.281. Under the neutral mutation hypothesis, the probability that the Tajima test statistic (D) is positive (2.325) is more than 0.5. Therefore, there may be a site at which natural selection, which increases genetic variation, is operating.

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

  • Kamaruddin, Mudyawati;Tokoro, Masaharu;Rahman, Md. Moshiur;Arayama, Shunsuke;Hidayati, Anggi P.N.;Syafruddin, Din;Asih, Puji B.S.;Yoshikawa, Hisao;Kawahara, Ei
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.471-478
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    • 2014
  • Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

Delimitation of Russula Subgenus Amoenula in Korea Using Three Molecular Markers

  • Park, Myung Soo;Fong, Jonathan J.;Lee, Hyun;Oh, Seung-Yoon;Jung, Paul Eunil;Min, Young Ju;Seok, Soon Ja;Lim, Young Woon
    • Mycobiology
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    • v.41 no.4
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    • pp.191-201
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    • 2013
  • Distinguishing individual Russula species has been difficult due to extensive phenotypic plasticity and obscure morphological and anatomical discontinuities. Due to highly similar macroscopic features, such as the presence of a red-cap, species identification within the Russula subgenus Amoenula is particularly difficult. Three species of the subgenus Amoneula have been reported in Korea. We used a combination of morphology and three molecular markers, the internal transcribed spacer (ITS), 28S nuclear ribosomal large subunit (LSU), and RNA polymerase II gene (RPB2), for identification and study of the genetic diversity of Russula subgenus Amoenula in Korea. We identified only two species in Korea (R. mariae and R. violeipes); these two species were indistinguishable according to morphology and LSU, but were found to be reciprocally monophyletic species using ITS and RPB2. The markers, ITS, LSU, and RPB2, have been tested in the past for use as DNA barcoding markers, and findings of our study suggest that ITS and RPB2 had the best performance for the Russula subgenus Amoneula.