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

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Molecular Phylogeny and Morphology of Mycosphaerella nawae, the Causal Agent of Circular Leaf Spot on Persimmon

  • Lee, Seung-Yeol;Lim, Yang-Sook;Jung, Hee-Young
    • Mycobiology
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    • v.44 no.4
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    • pp.191-201
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    • 2016
  • In this study, the phylogeny and morphology of Mycosphaerella nawae (Dothideomycetes, Ascomycota) were examined using Korean and Japanese isolates, to establish the phylogenetic relationship between M. nawae and its allied species. Korean and Japanese isolates of M. nawae were collected from circular leaf spot-diseased leaves and were confirmed based on internal transcribed spacer (ITS) sequence data. Phylogenetic analysis was conducted using multiple genes, including the ITS region, 28S rDNA, ${\beta}-tubulin$, translation elongation $factor-1{\alpha}$, and actin genes. Our results revealed that M. nawae is closely related to members of the genus Phaeophleospora but are distant from the Ramularia spp. In addition, microscopic analysis revealed pseudothecia on the adaxial and abaxial surface of overwintered diseased leaves (ODL) and only on the abaxial surface of diseased leaves. Ascospores are oval to fusiform, one-septate, tapered at both ends, $1.7{\sim}3.1{\times}8.1{\sim}14.1{\mu}m$, and were observed in ODL. Conidia are oval, guttulate, one-septate, $3.5{\sim}4.9{\times}12.8{\sim}19.8{\mu}m$, and barely discernable on 30-day cultures. To our knowledge, this is the first report on the phylogeny of M. nawae, which is closely related to the genus Phaeophleospora, especially P. scytalidii.

Taxonomy and phylogeny of the genus Cryptomonas (Cryptophyceae, Cryptophyta) from Korea

  • Choi, Bomi;Son, Misun;Kim, Jong Im;Shin, Woongghi
    • ALGAE
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    • v.28 no.4
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    • pp.307-330
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    • 2013
  • The genus Cryptomonas is easily recognized by having two flagella, green brownish color, and a swaying behavior. They have relatively simple morphology, and limited diagnostic characters, which present a major difficulty in differentiating between species of the genus. To understand species delineation and phylogenetic relationships among Cryptomonas species, the nuclear-encoded internal transcribed spacer 2 (ITS2), partial large subunit (LSU) and small subunit ribosomal DNA (rDNA), and chloroplast-encoded psbA and LSU rDNA sequences were determined and used for phylogenetic analyses, using Bayesian and maximum likelihood methods. In addition, nuclear-encoded ITS2 sequences were predicted to secondary structures, and were used to determine nine species and four unidentified species from 47 strains. Sequences of helix I, II, and IIIb in ITS2 secondary structure were very useful for the identification of Cryptomonas species. However, the helix IV was the most variable region across species in alignment. The phylogenetic tree showed that fourteen species were monophyletic. However, some strains of C. obovata had chloroplasts with pyrenoid while others were without pyrenoid, which used as a key character in few species. Therefore, classification systems depending solely on morphological characters are inadequate, and require the use of molecular data.

Taxonomic study of three new Antarctic Asterochloris (Trebouxiophyceae) based on morphological and molecular data

  • Kim, Jong Im;Kim, Yong Jun;Nam, Seung Won;So, Jae Eun;Hong, Soon Gyu;Choi, Han-Gu;Shin, Woongghi
    • ALGAE
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    • v.35 no.1
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    • pp.17-32
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    • 2020
  • Asterochloris is one of the most common genera of lichen phycobionts in Trebouxiophyceae. Asterochloris phycobionts associated with the lichenized fungi Cladonia and Stereocaulon in King George Island (Antarctica) and Morro Chico (Chile), were isolated and then used to establish clonal cultures. To understand the phylogenetic relationships and species diversity of Antarctic Asterochloris species, molecular and morphological data were analyzed by using three microscopy techniques (light, confocal laser and transmission electron) and a multi-locus phylogeny with data from the nuclear-encoded internal transcribed spacer (ITS) rDNA and the actin and plastid-encoded ribulose bisphosphate carboxylase large chain (rbcL) coding genes. Morphological data of three Antarctic strains showed significant species-specific features in chloroplast while molecular data segregated the taxa into distinct three clades as well. Each species had unique molecular signatures that could be found in secondary structures of the ITS1 and ITS2. The species diversity of Antarctic Asterochloris was represented by six taxa, namely, A. glomerata, A. italiana, A. sejongensis, and three new species (A. antarctica, A. pseudoirregularis, A. stereocaulonicola).

Actual distribution and present status of a threatened aquatic plant, Sagittaria aginashi (Alismataceae), in Korea

  • Shiga, Takashi;Tsubota, Kazumasa;Choi, Hyeok Jae
    • Korean Journal of Plant Taxonomy
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    • v.47 no.3
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    • pp.180-188
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    • 2017
  • We evaluated the actual distribution, present status, and number of remnant individuals of Korean Sagittaria aginashi (Alismataceae) based on herbarium specimens and field surveys. We also reidentified Korean S. aginashi by analyzing sequences of the internal transcribed spacer (ITS) region. We found nine specimens of S. aginashi in herbaria from four localities housed at the Osaka Museum of Natural History (OSA) and at the Korea National Herbarium (KH). During our field research, we could not confirm the current statuses of two collection localities (Taereung and Ansan-si), though this was not the case for Ulsan-si. In addition, we found two new localities in Ulsan-si and Yangsan-si. These three remnant populations are distributed in wetlands in mountainous areas (elev. 480-740 m). With regard to the number of flowering and immature individuals in the three localities, there were <50 and <2,000, respectively, in total. ITS sequences demonstrated that the sequences of Korean and Japanese S. aginashi are uniform and distinct from other Sagittaria species. These results indicate that S. aginashi is a rare and threatened species in Korea. It should be listed as an endangered species on the Korean Red List and requires urgent protection by conservation programs, including the extensive surveys of other possible natural habitats.

Diversity and Plant Growth Promotion of Fungal Endophytes in Five Halophytes from the Buan Salt Marsh

  • Khalmuratova, Irina;Choi, Doo-Ho;Yoon, Hyeok-Jun;Yoon, Tae-Myung;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.31 no.3
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    • pp.408-418
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    • 2021
  • The diversity and plant growth-promoting ability of fungal endophytes that are associated with five halophytic plant species (Phragmites australis, Suaeda australis, Limonium tetragonum, Suaeda glauca Bunge, and Suaeda maritima) growing in the Buan salt marsh on the west coast of South Korea have been explored. About 188 fungal strains were isolated from these plant samples' roots and were then studied with the use of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2). The endophytic fungal strains belonged to 33 genera. Alternaria (18%) and Fusarium (12.8%), of the classes Dothideomycetes and Sordariomycetes, were most rampant in the coastal salt marsh plants. There was a higher diversity in fungal endophytes that are isolated from S. glauca Bunge than in isolates from other coastal salt marsh plants. Plant growth-promoting experiments with the use of Waito-C rice seedlings show that some of the fungal strains could encourage a more efficient growth than others. Furthermore, gibberellins (GAs) GA1, GA3, and GA9 were seen in the Sa-1-4-3 isolate (Acrostalagmus luteoalbus) culture filtrate with a gas chromatography/mass spectrometry.

Phylogenetic relationships of Coreanomecon (Papaveraceae: Papaveroideae), an endemic genus in Korea, using DNA sequences

  • YUN, Narae;OH, Sang-Hun
    • Korean Journal of Plant Taxonomy
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    • v.48 no.4
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    • pp.289-300
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    • 2018
  • Coreanomecon is a monotypic and endemic genus in Korea, distributed mainly in the southern regions. Coreanomecon is morphologically similar to Hylomecon by producing red latex, easily distinguished from Chelidonium, which produces yellow latex. Coreanomecon were merged into Hylomecon or Chelidonium depending on the authors. To understand the phylogenetic relationship of Coreanomecon, DNA sequences of chloroplast rbcL and matK and nuclear Internal Transcribed Spacer (ITS) regions were determined from the species of Papaveroideae (Papaveraceae) in Korea and analyzed with the Maximum Parsimony and Bayesian methods. Phylogenetic analyses of Papaveroideae suggest that Coreanomecon is sister to the clade of Chelidonium and Stylophorum in the ITS data and that it is sister to Hylomecon in the chloroplast (cpDNA) data. A constraining analysis using the Shimodaira-Hasegawa test (S-H test) suggested that the ITS data do not reject the sister relationship of Coreanomecon and Hylomecon. The S-H test also suggested that the cpDNA data is compatible with the placement of Coreanomecon as a sister to the clade of Chelidonium and Stylophorum. Although the conflicting phylogenetic results may stem from insufficient phylogenetic signals, they may also be associated with hybridization between Hylomecon and an ancestor of Stylophorum and Chelidonium. The results of this study suggest that Coreanomecon is a distinct lineage as an endemic genus, supporting the morphological data.

Identification and Characterization of Ceratocystis fimbriata Causing Lethal Wilt on the Lansium Tree in Indonesia

  • Suwandi, Suwandi;Irsan, Chandra;Hamidson, Harman;Umayah, Abu;Asriyani, Khoirotun Dwi
    • The Plant Pathology Journal
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    • v.37 no.2
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    • pp.124-136
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    • 2021
  • Bark canker, wood discoloration, and wilting of the duku tree (Lansium domesticum) along the watershed of Komering River, South Sumatra Province, Indonesia first appeared in 2013. The incidence of tree mortality was 100% within 3 years in badly infected orchards. A Ceratocystis species was consistently isolated from the diseased tissue and identified by morphological and sequence analyses of the internal transcribed spacer (ITS) and β-tubulin regions. Pathogenicity tests were conducted and Koch's postulates were confirmed. The fungus was also pathogenic on Acacia mangium, but was less pathogenic on mango. Partial flooding was unfavourable for disease development. Two described isolates (WRC and WBC) had minor variation in morphology and DNA sequences, but the former exhibited a more pathogenic on both duku and acacia. The ITS phylogenies grouped the most pathogenic isolate (WRC) causing wilting of the duku tree within the aggressive and widely distributed ITS5 haplotype of C. fimbriata.

Molecular and Morphological Characteristics of a New Species Collected from an Insect (Cicindela transbaicalica) in Korea

  • Lee, Ju-Heon;Ten, Leonid N.;Lim, Seong-Keun;Ryu, Jung-Joo;Avalos-Ruiz, Diane;Lee, Seung-Yeol;Jung, Hee-Young
    • Mycobiology
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    • v.50 no.3
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    • pp.181-187
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    • 2022
  • To exploit insect-derived fungi, insects were collected from seven different regions in Korea, including Gyeongbuk, Goryeong, and several fungi were isolated from them. A fungal strain designated 21-64-D was isolated from riparian tiger beetle (Cicindela transbaicalica) and morphologically identified as a species belonging to the genus Oidiodendron. Phylogenetic analysis using the nucleotide sequences of internal transcribed spacer (ITS) regions and the partial sequence of the large subunit of the nuclear ribosomal RNA (LSU) gene revealed the distinct phylogenetic position of the isolate among recognized Oidiodendron species including its closest neighbors O. chlamydosporicum, O. citrinum, O. maius, and O. pilicola. The hyphal and conidial morphology of the strain, particularly club-shaped hyphae, clearly differentiated it from its close relatives. Results indicated that 21-64-D is a novel species in the genus Oidiodendron, for which the name Oidiodendron clavatum sp. nov. is proposed.

Molecular Marker to Identify and Origin of Cnidii Rhizoma from Korea and China (천궁의 기원과 식별을 위한 분자마커)

  • Song, Im-Geun;An, Bo-Ram;Seo, Bu-Il;Park, Seon-Joo
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.1-8
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    • 2009
  • Objectives : This study was carried out to discriminate origin and molecular marker of oriental medicine "Cnidii Rhizoma" be circulated between Korea and China, which is difficult to discriminate from morphological distinction because of a fragmental materials of roots. Methods : Materials were collected randomly from a medicinal herb markets in Korea and China and be analyzed with ITS (internal transcribed spacer) regions of nuclear ribosomal DNA (nrDNA). Results : As a results, ITS regions of nrDNA was shown to be identify as three molecular markers. "Cnidii Rhizoma" was made up syster group of the genus Ligusticum L. and divided into three groups with "Tou-chun-gung", "IL-chun-gung" and "China-chun-gung". Conclusions : From the analysis of ITS regions of nrDNA, we presumed that it is the same origin of "Cnidii Rhizoma" from Korea and China because of phylogenetic tree consisted of sister groups with the genus Ligusticum than the genus Cnidium.

Rapid Identification of Diaporthe citri by Gene Sequence Analysis

  • Zar Zar Soe;Yong Ho Shin;Hyun Su Kang;Mi Jin Kim;Yong Chull Jeun
    • Research in Plant Disease
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    • v.29 no.2
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    • pp.130-136
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    • 2023
  • Citrus melanoses caused by Diaporthe citri, has been one of the serious diseases in many citrus orchards of Jeju Island. To protect melanose in citrus farms, a fast and exact diagnosis method is necessary. In this study, diseased leaves and dieback twigs were collected from a total of 49 farms within March to April in 2022. A total of 465 fungal isolates were obtained from a total of 358 isolated plant samples. Among these fungal isolates, 40 representatives of D. citri isolates which were isolated from 22 twigs and 18 leaves on 23 farms were found based on cultural characteristics on potato dextrose agar and conidial morphology. Additionally, the molecular assay was carried out and compared with those by morphological diagnosis. All isolates were identified as D. citri by analyzing the sequences at the internal transcribed spacer (ITS) rDNA region using primers of ITS1/ITS4 or at β-tubulin using primer Btdcitri-F/R. Therefore, based on the present study, where the results of morphological identification of conidial type were consistent with DNA sequence analysis of certain gene, choosing a suitable method for a fast diagnosis of citrus melanose was suggested.