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

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Development of specific single nucleotide polymorphism molecular markers for Angelica gigas Nakai (ITS 영역의 HRM 분석을 통한 참당귀(Angelica gigas Nakai)의 특이적 SNP 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.2
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    • pp.71-76
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    • 2021
  • Angelica is a perennial plant used widely for medicinal purposes. Information on the genetic diversity of Angelica populations is important for their conservation and germplasm utilization. Although Angelica is an important medicinal plant genus registered in South Korea, no molecular markers are currently available to distinguish individual species from other similar species in different countries, in particular, China and Japan. In this study, we developed single nucleotide polymorphism (SNP) markers derived from internal transcribed spacer regions of the nuclear ribosomal DNA to identify a distinct domestic species, Angelica gigas Nakai, via a high-resolution melting (HRM) curve analyses. We also performed HRM curve analysis of intentionally mixed genomic DNA samples from five Angelica species. Finally, we investigated A. gigas Nakai and A. sinensis using varying ratios of mixed genomic DNA templates. The SNP markers developed in this study are useful for rapidly identifying A. gigas species from different countries.

Causal Agents of Blossom Blight of Kiwifruit in Korea

  • Lee, Young-Sun;Han, Hyo-Shim;Kim, Gyoung-Hee;Koh, Young-Jin;Hur, Jae-Seoun;Jung, Jae-Sung
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.220-224
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    • 2009
  • The causal agents of bacterial blossom blight in kiwifruit were isolated from flowers displaying symptoms in Korea. The pathogens were characterized by biochemical and physiological tests, and identified on the basis of 16S rDNA and 16S-23S internal transcribed spacer (ITS) sequences. Pathogenicity tests demonstrated that the blossom blight of kiwifruit in Korea is caused by two pathogens, Pseudomonas syringae pv. syringae and P. fluorescens. Carbon source utilization and DNA-DNA hybridization experiments confirmed P. fluorescens as one of the causal agents of blossom blight of kiwifruit. P. syringae pv. syringae and P. fluorescens can be distinguished from each other by the symptoms they produce in flowers. P. syringae pv. syringae primarily affected the stamen, while P. fluorescens caused rotting of all internal tissues of buds or flowers.

Morphology and phylogenetic relationships of Micractinium (Chlorellaceae, Trebouxiophyceae) taxa, including three new species from Antarctica

  • Chae, Hyunsik;Lim, Sooyeon;Kim, Han Soon;Choi, Han-Gu;Kim, Ji Hee
    • ALGAE
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    • v.34 no.4
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    • pp.267-275
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    • 2019
  • Three new species of the genus Micractinium were collected from five localities on the South Shetland Islands in maritime Antarctica, and their morphological and molecular characteristics were investigated. The vegetative cells are spherical to ellipsoidal and a single chloroplast is parietal with a pyrenoid. Because of their simple morphology, no conspicuous morphological characters of new species were recognized under light microscopy. However, molecular phylogenetic relationships were inferred from the concatenated small subunit rDNA, and internal transcribed spacer (ITS) sequence data indicated that the Antarctic microalgal strains are strongly allied to the well-supported genus Micractinium, including M. pusillum, the type species of the genus, and three other species in the genus. The secondary structure of ITS2 and compensatory base changes were used to identify and describe six Antarctic Micractinium strains. Based on their morphological and molecular characteristics, we characterized three new species of Micractinium: M. simplicissimum sp. nov., M. singularis sp. nov., and M. variabile sp. nov.

Identification of Botrytis cinerea, the Cause of Post-Harvest Gray Mold on Broccoli in Korea

  • Aktaruzzaman, Md.;Afroz, Tania;Hong, Sae-Jin;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.23 no.4
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    • pp.372-378
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    • 2017
  • In this study, we identified the causative agent of post-harvest gray mold on broccoli that was stored on a farmers' cooperative in Pyeongchang, Gangwon Province, South Korea, in September 2016. The incidence of gray mold on broccoli was 10-30% after 3-5 weeks of storage at $3^{\circ}C$. Symptoms included brownish curd and gray-to-dark mycelia with abundant conidia on the infected broccoli curds. The fungus was isolated from infected fruit and cultured on potato dextrose agar. To identify the fungus, we examined the morphological characteristics and sequenced the rDNA of the fungus and confirmed its pathogenicity according to Koch's postulates. The results of the morphological examination, pathogenicity test, and sequencing of the 5.8S rDNA of the internal transcribed spacer regions (ITS1 and ITS4) and three nuclear protein-coding genes, G3PDH, HSP60, and RPB2, revealed that the causal agent of the post-harvest gray mold on broccoli was Botrytis cinerea. To our knowledge, this is the first report of post-harvest gray mold on broccoli in Korea.

Molecular Diversity of Fungal Endophytes Isolated from Garcinia mangostana and Garcinia parvifolia

  • Sim, Jiun-Horng;Khoo, Chai-Hoon;Lee, Learn-Han;Cheah, Yoke-Kqueen
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.651-658
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    • 2010
  • Garcinia is commonly found in Malaysia, but limited information is available regarding endophytic fungi associated with this plant. In this study, 24 endophytic fungi were successfully recovered from different parts of two Garcinia species. Characterization of endophytic fungi was performed based on the conserved internal transcribed spacer (ITS) region sequence analysis and the antimicrobial properties. Results revealed that fruits of the plant appeared to be the highest inhabitation site (38%) as compared with others. Glomerella sp., Guignardia sp., and Phomopsis sp. appeared to be the predominant endophytic fungi group in Garcinia mangostana and Garcinia parvifolia. Phylogenetic relationships of the isolated endophytic fungi were estimated from the sequences of the ITS region. On the other hand, antibacterial screening showed 11 of the isolates possessed positive response towards pathogenic and nonpathogenic bacteria. However, there was no direct association between certain antibacterial properties with the specific genus observed.

A New Record of Nannizziopsis mirabilis from Field Soil in Korea

  • Adhikari, Mahesh;Gurung, Sun Kumar;Kim, Sang Woo;Lee, Hyun Goo;Ju, Han Jun;Gwon, Byeong Heon;Kosol, San;Bazie, Setu;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.249-254
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    • 2018
  • Nannizziopsis mirabilis (KNU17-67) was isolated from a field soil sample collected from Jejudo in 2017. This fungal isolate was identified and described by its morphological characters and the internal transcribed spacer rDNA gene sequencing. Potato dextrose agar medium was used to study its macro and micromorphology. In addition, a molecular phylogenetic tree was constructed for its precise confirmation as N. mirabilis. This fungal isolate has not officially been reported previously in Korea.

Identification of Alternaria alternata as a Causal Agent for Leaf Blight in Syringa Species

  • Mmbaga, Margaret T.;Shi, Ainong;Kim, Mee-Sook
    • The Plant Pathology Journal
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    • v.27 no.2
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    • pp.120-127
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    • 2011
  • While many isolates of Alternaria alternata are common saprophytes on trees and shrubs, this study clearly demonstrated that A. alternata is a primary pathogen in lilac (Syringa sp.), causing a leaf-blight that affects different Syringa species. Isolates of Alternaria sp. were collected from leaf blight samples of lilacs in the field. The internal transcribed spacer (ITS) region and morphological characterization were used to identify lilac blight pathogen. Based on 100% ITS nucleotide sequence identities to the Alternaria genus in the GenBank and morphological features, these isolates were identified as A. alternata. Disease symptoms were reproduced in lilac plants inoculated with A. alternata mycelial plugs and sprayed with a fungus-free culture filtrate, indicating that pathogenesis in lilac involves secondary metabolites or toxins. Diagnostic primers were developed to detect Alternaria sp. and A. alternata in lilac leaf blight based on ITS region and four known genes associated with pathogenesis in A. alternata: mixed-linked glucanase precursor, endopolygalacturonase, hsp70, and histone genes. The results from our study indicated A. alternata is a primary pathogen in lilac leaf blight, and these diagnostic primers can be used as a tool for the fast detection of A. alternata associated with lilac leaf blight.

First Report of Gray Mold Disease of Sweet Basil (Ocimum basilicum) Caused by Botrytis cinerea in Korea

  • Aktaruzzaman, Md.;Kim, Joon-Young;Afroz, Tania;Hong, Sae-Jin;Kim, Byung-Sup
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.277-280
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    • 2015
  • In August 2015, we collected samples of gray mold from sweet basil growing in Sachunmeon, Gangneung, Gangwon Province, Korea. Symptoms included extensive growth of mycelia with gray conidia on young leaves, stems, and blossoms. The pathogen was isolated from infected leaves and blossoms and the fungus was cultured on potato dextrose agar. For identification of the fungus, morphology and rDNA sequencing analysis of the fungus were performed, which confirmed its pathogenicity according to Koch's postulates. The results of morphological examinations, pathogenicity tests, and the rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4) and the three nuclear protein-coding genes G3PDH, HSP60, and RPB2 showed that the causal agent was Botrytis cinerea. This is the first report of gray mold caused by Botrytis cinerea on sweet basil in Korea.

Phylogenetic Relationships in Korean Elaeagnus L. Based on nrDNA ITS Sequences

  • Son, OGyeong;Yoon, Chang Young;Park, SeonJoo
    • Korean Journal of Plant Resources
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    • v.27 no.6
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    • pp.671-679
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    • 2014
  • Molecular phylogenetic analyses of Korean Elaeagnus L. were conducted using seven species, one variety, one forma and four outgroups to evaluate their relationships and phylogeny. The sequences of internal transcribed spacer regions in nuclear ribosomal DNA were employed to construct phylogenetic relationships using maximum parsimony (MP) and Bayesian analysis. Molecular phylogenetic analysis revealed that Korean Elaeagnus was a polyphyly. E. umbellata var. coreana formed a subclade with E. umbellata. Additionally, the genetic difference between E. submacrophylla and E. macrophylla was very low. Moreover, E. submacrophylla formed a branch from E. macrophylla, indicating that E. submacrophylla can be regarded as a variety. However, several populations of this species were not clustered as a single clade; therefore, further study should be conducted using other molecular markers. Although E. glabra f. oxyphylla was distinct in morphological characters of leaf shape with E. glabra. But E. glabra f. oxyphylla was formed one clade by molecular phylogenetic with E. glabra. Additionally, this study clearly demonstrated that E. pungens occurs in Korea, although it was previously reported near South Korea in Japan and China. According to the results of ITS regions analyses, it showed a resolution and to verify the relationship between interspecies of Korean Elaeagnus.

Phylogenetic analysis of Viburnum (Adoxaceae) in Korea using DNA sequences

  • CHOI, Yun Gyeong;YOUM, Jung Won;LIM, Chae Eun;OH, Sang-Hun
    • Korean Journal of Plant Taxonomy
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    • v.48 no.3
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    • pp.206-217
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    • 2018
  • The nucleotide sequences of the chloroplast rbcL, matK, and psbA-trnH and nuclear internal transcribed spacer (ITS) regions were determined from all species of Viburnum in Korea with multiple accessions to reconstruct the phylogeny and to evaluate the utility of the DNA sequences as DNA barcodes. The results of phylogenetic analyses of the cpDNA and ITS data are consistent with the findings of previous studies of Viburnum. Four morphologically closely related species, V. dilatatum, V. erosum, V. japonicum, and V. wrightii, were included in a strongly supported sister clade of V. koreanum and V. opulus. Viburnum odoratissimum is suggested to be sister to the V. dilatatum/V. koreanum clade in the cpDNA data, while V. odoratissimum is a sister to V. furcatum in the ITS data. Viburnum burejaeticum and V. carlesii are strongly supported as monophyletic. Our analyses of DNA barcode regions from multiple accessions of the species of Viburnum in Korea confirm that six out of ten species in Korea can be discriminated at the species level. The V. dilatatum complex can be separated from the remaining species according to molecular data, but the resolution power to differentiate a species within the complex is weak. This study suggests that regional DNA barcodes are useful for molecular species identification in the case of Viburnum when flowering or fruiting materials are not available.