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

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Development of molecular markers for the differentiation of Angelica gigas Jiri line by using ARMS-PCR analysis (세발당귀(Angelica gigas Jiri)의 판별을 위한 ARMS-PCR용 분자표지 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Han, Eun-Hee;Shin, Yong-Wook;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.26-33
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    • 2021
  • Angelica is a widely used medicinal and perennial plant. Information on the genetic diversity of Angelica populations is essential for their conservation and germ plasmic utilization. Although Angelica is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish it from other similar species from different countries. This developed single nucleotide polymorphism (SNP) markers derived from nuclear ribosomal DNA internal transcribed spacer regions genomic sequences to identify distinct Korean-specific Angelica species via amplification refractory mutation system (ARMS)-PCR curve analyses. We performed molecular authentication of different kinds of Korean-specific Angelica species such as A. gigas Nakai and A. gigas Jiri using DNA sequences in the ITS intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific Angelica species from different countr.

First Report of Root Rot of Dendropanax trifidus Caused by Fusarium oxysporum in Korea (Fusarium oxysporum에 의한 황칠나무 묘목 뿌리썩음병 발생 보고)

  • Hyerin Ma;Sungyu Choi;Hyunkyu Sang;Hyun-Jun Kim
    • The Korean Journal of Mycology
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    • v.51 no.1
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    • pp.51-56
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    • 2023
  • Dendropanax trifidus belonging to the family Araliaceae, is a warm-temperate evergreen tree distributed in Jeju Island, Bogil Island, Geomun Island, Geoje Island, Wando, and Haenam in Korea. In June 2021, a root rot disease in which branches of Dendropanax trifidus seedlings turned brown and shrunk was discovered at the seedling cultivation facility in Naju-si, Republic of Korea. To identify the root rot fungus, three strains were isolated from the diseased tissues of seedlings and their mycological characteristics were investigated on potato dextrose agar. In addition, a molecular phylogenetic analysis was performed using sequences of the internal transcribed spacer (ITS) region and translation elongation factor 1-α (EF1-α) gene. The fungus was identified as Fusarium oxysporum. For pathogenicity test, the roots of seedlings were immersed in the conidia suspension of the strains and planted. After 20 days inoculation, root rot and browning symptoms were confirmed in the inoculated plants. This is the first report of F. oxysporum on D. trifidus in Korea.

Diversity of Endophytic Fungi Associated with Roots of Calanthe discolor and Cephalanthera longibracteata in Korea (새우난초와 은대난초 뿌리에서 분리한 내생균의 다양성)

  • Lee, Bong-Hyung;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.427-435
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    • 2018
  • In this study, endophytic fungi were isolated from the surface-sterilized roots of Calanthe discolor and Cephalanthera longibracteata collected from the Chungnam, Jeju, Kyungnam and Chungbuk provinces in Korea. The morphological characteristics of the obtained isolates were examined and their sequences of the internal transcribed spacer (ITS) rDNA region were analyzed using the ITS1F and ITS4 primers for species identification. Leptodontidium orchidcola showed the highest species abundance and frequency among the isolated endophytic fungi. Additionally, the community analysis revealed a high specificity between the host plants and the endophytic fungal species.

Yeasts in Internal Roots of the Rare Plant Dendropanax morbifera

  • Kim, Jong-Shik;Kim, Dae-Shin;Ko, Suk-Hyung
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.1
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    • pp.33-40
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    • 2017
  • To isolate and identify the yeast strains associated with D. morbifera, homogenized D. morbifera root samples were spread onto GPY, DG18, SCG and DOB agar media containing antibiotics, Triton X-100, and l-sorbose. Total 81 yeast isolates were analyzed by sequencing of internal transcribed spacer (ITS) region of the ribosomal DNA. The results showed that the root-associated yeast species were composed of the genera Vanderwaltozyma (40 isolates), Cryptococcus (40 isolates), and Kluyveromyces (one isolate). Moreover, the Kluyveromyces isolate exhibited high bioethanol productivity. In addition, the Vanderwaltozyma and Cryptococcus were dominant in D. morbifera roots. The specific yeast community associated with D. morbifera roots was identified by phylogenetic sequence analyses. These yeast isolates may have industrial applications as biosurfactant and bioethanol.

Detection of Laminariaceae Species Based on PCR by Family-specific ITS Primers

  • Choi, Chang-Geun;Kim, Jong-Myoung
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.157-162
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    • 2012
  • To analyze nucleotide sequence encoding internal transcribed spacer (ITS) regions specific to the Laminariaceae family, genomic DNA was isolated from six brown algae species distributed along the east coast of Korea. These included three species from the Laminariaceae family (Agarum clathratum Dumortier, Costaria costata [C. Agardh] Saunders, and Saccharina japonica Areschoug) and two species from the Alariaceae family (Undaria pinnatifida [Harvey] Suringer and Ecklonia cava Kjellman), both in the order Laminariales, and one species from the family Sargassaceae in the order Fucales (Sargassum serratifolium). Based on a sequence analysis of ITS-1 and ITS-2 for A. clathratum, C. costata, and E. cava, oligonucleotides were designed from the regions that showed sequence conservation in Laminariaceae. Following polymerase chain reaction using three sets of primers, amplification of ITS-1 and ITS-2 was detected in reactions using genomic DNA isolated from the species belonging to Laminariaceae, but not from the species belonging to the other families. The results indicate that this method can be used for the detection and identification of Laminariaceae species.

Analysis of Cultural Characteristics and Phylogenic Relationships of Collected Strains of Pholiota species

  • Cho, Yong-Hyun;Kong, Won-Sik;Kim, Gyu-Hyun;Jhune, Chang-Sung;You, Chang-Hyun;Yoo, Young-Bok;Kim, Kwang-Ho
    • Mycobiology
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    • v.31 no.4
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    • pp.200-204
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    • 2003
  • Cultural characteristics and phylogenic relationships were investigated and classified among collected strains in Pholiota spp. which contain P. adiposa, P. squarrosa, P. nameko etc. They were tested on the four different media(PDA, MCM, YM, MEA) and sawdust(Alder, Oak, Pine, Popular) substrates. There was a little variation according to the media and sawdust substrates, although PDA and popular sawdust substrate seemed to be better. Most strains showed white colonies, but some strains were brown. Mycelial growth length differed according to the strains. To classify species, the internal transcribed spacer regions(ITS) of the ribosomal DNA(rDNA) repeats from Pholiota spp. were amplified using polymerase chain reaction(PCR) and then sequenced. According to the analysis of ITS sequences, they were classified into five clusters. Their spacer regions were $644{\sim}700$ nucleotides in length. The reciprocal homologies of each ITS region among these strains were ranged from $49.6{\sim}99.9%$. The phylogenic analysis might give a criterion to classify species in the collected strains.

Variation of nuclear and mitochondrial DNAs in Korean and Chinese isolates of Clonorchis sinensis

  • Lee, Soo-Ung;Huh, Sun
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.145-148
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    • 2004
  • We compared the DNA sequence difference of isolates of Clonorchis sinensis from one Korean (Kimhae) and two Chinese areas (Guangxi and Shenyang), The sequences of nuclear rDNA (18S, internal transcribed spacer 1 and 2: ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1: cox1) were compared. A very few intraspecific nucleotide substitution of the 18S, ITS1, ITS2 and cox1 was found among three isolates of C. sinensis and a few nucleotide insertion and deletion of ITS1 were detected. The 18S, ITS1, ITS2 and cox1 sequences were highly conserved among three isolates. These findings indicated that the Korean and two Chinese isolates are similar at the DNA sequence level.

Phylogeographic Messages Encoded in the rDNA of the Commercial Mushroom Zhenghonggu@ From Fujian, China

  • Chen, Yu H.;Chen, Peng D.;Chen, Liu Y.;Ma, Li Z.
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.45-45
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    • 2014
  • Individualities of precious health mushroom called Zhenghonggu@ from respective protections scattered among all main mountains of Fujian China were collected and recognized locally, then compared with Russula griseocarnosa. Their internal transcribed spacer (ITS) region (ITS1, ITS2 and 5.8S rDNA) of the nuclear rDNA were amplified, AMOVA analyzed, nested clade analyzed and then compared with the ITS sequences of relative Russula species from other regions of China to confirm the taxonomic status of Zhenghonggu$^@$ and its population structure. Total 23 haplotypes from different protections of Fujian can be clustered into three clades similar to the three lineages of Dahongjun$^@$ from southeastern China reported by Li et al. The geographic distribution characteristic of these three phylogeny clades may be closely coupled with the vegetation regionalization and/or the differences of coenosium construction of Fagaceae that is the host of Russula griseocarnosa. The correlation of taxonomy, phylogeny and geographical distribution of Russula are discussed.

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Rapid and Accurate Species-Specific Detection of Phytophthora infestans Through Analysis of ITS Regions in Its rDNA

  • Kim, Kyoung-Su;Lee, Youn-Su
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.651-655
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    • 2000
  • Polymerase chain reaction (PCR) was used to specifically detect Phytophthora infestans by analyzing the sequences of the ribosomal internal transcribed spacer regions (ITS) in the rDNA of the Phytophthora species. Based on the sequence data, PISP-1 together with the ITS3 primer were used to detect p. infestans. A single ca. 450 bp segment was observed in P. infestans, but not in the other fungal or bacterial isolates. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using these species-specific primers, a unique band was obtained within annealing temperatures of $55^{\circ}C$-$61^{\circ}C$ and template DNA levels of 10 pg-100 ng.

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Phylogenetic Analysis of Phyllospadix iwatensis Based on Nucleotide Sequences Encoding 18S rRNA and ITS-1

  • Kim, Jong-Myoung;Choi, Chang-Geun
    • Fisheries and Aquatic Sciences
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    • v.13 no.4
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    • pp.272-277
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    • 2010
  • Seagrasses are marine angiosperms of ecological importance in providing shelter and food to aquatic species as well as maintaining the carbon cycle on earth. Phyllospadix iwatensis is a seagrass of the family Zosteraceae and is distributed along the eastern coast of Korea. The nucleotide sequences of P. iwatensis nuclear genes encoding 18S ribosomal RNA (rRNA) and internal transcribed spacer-1 (ITS-1) were determined for molecular phylogenetic analysis. Genomic DNA was isolated from P. iwatensis and used for PCR amplification of 18S rRNA and ITS-1. Examination of the 18S rRNA sequence of P. iwatensis showed a close (99% similarity) relationship to Zostera noltii, another genus of Zosteraceae, but a distant (84% similarity) evolutionary relationship to other macroalgal Laminariales species. Further discrepancies found in ITS-1 nucleotide sequences between closely related species indicate that the sequence information could be used for species identification.