• Title/Summary/Keyword: rDNA internal transcribed spacer sequence

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Identification and Pathogenicity of Neophysopella vitis Causing Rust Disease on Meliosma myriantha in Korea

  • Dong Hwan Na;Jae Sung Lee;Young-Joon Choi;Ji-Hyun Park;Hyeon-Dong Shin
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.299-303
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    • 2023
  • Rust symptoms on Meliosma myriantha trees have been noticed during disease surveys in Korea since 2010, with a high disease incidence frequently surpassing 90%. The causal fungus of the rust disease was identified as Neophysopella vitis based on the morphological investigation and molecular sequence analysis of the internal transcribed spacer (ITS) and large subunit (LSU) rDNA regions. This is the first report of rust disease caused by N. vitis on M. myriantha in Korea. A pathogenicity assay proved that M. myriantha serves as the aecial host of N. vitis as spermogonia and aeciospores were produced, which can infect the two uredinial hosts, Boston ivy (Parthenocissus tricuspidata) and Virginia creeper (Parthenocissus quinquefolia).

Occurrence of Sporendocladia bactrospora on Quercus variabilis in Korea

  • Lee, Dong-Hyeon;Seo, Sang-Tae;Lee, Sang-Hyun;Lee, Seung Kyu;Lee, Sun Keun
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.394-398
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    • 2017
  • A survey to assess the diversity of wound-associated Ophiostomatales and Microascales, the so-called ophiostomatoid fungi, on Korean native trees, was undertaken in 2017. Wounds were artificially created, and a fungus resembling a species of Sporendocladia was consistently isolated from the exposed cambium and inner bark of artificially induced wounds on Quercus variabilis. Morphological examination and DNA sequence comparisons based on the internal transcribed spacer (ITS) and 5.8S regions of the rDNA confirmed the identity of the fungus as Sporendocladia bactrospora. This is the first report on S. bactrospora occurring on Q. variabilis in Korea.

Molecular Identification of Zoysia japonica and Zoysia sinica (Zoysia Species) Based on ITS Sequence Analyses and CAPS (ITS 염기서열 분석 및 CAPS를 이용한 조이시아 속(Zoysia) 들잔디와 갯잔디의 구별)

  • Hong, Min-Ji;Yang, Dae-Hwa;Jeong, Ok-Cheol;Kim, Yang-Ji;Park, Mi-Young;Kang, Hong-Gyu;Sun, Hyeon-Jin;Kwon, Yong-Ik;Park, Shin-Young;Yang, Paul;Song, Pill-Soon;Ko, Suk-Min;Lee, Hyo-Yeon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.344-360
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    • 2017
  • Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

First Report of Sclerotinia White Rot Caused by Sclerotinia nivalis on Panax ginseng in Korea

  • Cho, Hye Sun;Shin, Jeong-Sup;Kim, Jae-Hyun;Hong, Tae-Kyun;Cho, Dae-Hui;Kang, Je Yong
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.49-54
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    • 2013
  • Sclerotinia white rot disease was observed on 5 and 6-year-old ginseng (Panax ginseng) roots in Hongchun, Cheorwon, and Yanggu, Gangwon Province, Korea from 2006 to 2010. Symptoms included a brownish watery soft rot of the roots, and black sclerotia were often found on the rotten roots. The causal agent of the disease was identified as Sclerotinia nivalis based on cultural characteristics and sequence analyses of the internal transcribed spacer region of rDNA and ${\beta}$-tubulin gene with 100% sequence similarity. Pathogenicity tests were performed on 2-year-old ginseng roots with mycelium plugs without wounds. A watery soft rot of the roots and black sclerotia were observed 10 days after inoculation. These symptoms were identical to those observed on naturally infected roots. The same fungus was re-isolated from the lesions induced by artificial inoculation. This is the first report of sclerotinia white rot caused by S. nivalis on P. ginseng in Korea.

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.

First Report of Peach Fruit Rot Caused by Fusarium avenaceum in Korea (Fusarium avenaceum에 의한 복숭아 신규 과실 썩음병 발생 보고)

  • Heo, A Yeong;Koo, Young Mo;Choi, Young-Joon;Kim, Sang Hee;Chung, Gyu Young;Choi, Hyong Woo
    • Research in Plant Disease
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    • v.26 no.1
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    • pp.48-52
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    • 2020
  • In July 2019, typical rot symptom was observed on peach fruits harvested from the fields at Andong, Korea. As the disease progressed, white and purple colored mycelial mat developed on the surface of the infected fruits. A causal pathogen was isolated from the infected fruit and cultured on potato dextrose agar media for identification. Fungal colonies on potato dextrose agar produced 3 pigments, including purple, yellow, and white colors. The isolate incited fruit rot symptoms on artificially inoculated peach fruits, from which the same fungus was isolated, fulfilling Koch's postulates. Based on the morphological characteristics and sequence analysis of rDNA internal transcribed spacer, translation elongation factor 1-alpha, and β-tubulin, the causal agent of the disease was identified as Fusarium avenaceum. This study is the first report of fruit rot of peach fruits caused by Fusarium avenaceum in Korea.

Identification and Characterization of the Causal Organism of Gummy Stem Blight in the Muskmelon (Cucumis melo L.)

  • Choi, In-Young;Choi, Jang-Nam;Choi, Dong-Chil;Sharma, Praveen Kumar;Lee, Wang-Hyu
    • Mycobiology
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    • v.38 no.3
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    • pp.166-170
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    • 2010
  • Gummy stem blight is a major foliar disease of muskmelon (Cucumis melo L.). In this study, morphological characteristics and rDNA internal transcribed spacer (ITS) sequences were analyzed to identify the causal organism of this disease. Morphological examination of the Jeonbuk isolate revealed that the percentage of monoseptal conidia ranged from 0% to 10%, and the average length $\times$ width of the conidia was 70 ($\pm$ 0.96) $\times$ 32.0 ($\pm$ 0.15) ${\mu}m$ on potato dextrose agar. The BLAST analysis showed nucleotide gaps of 1/494, 2/492, and 1/478 with identities of 485/492 (98%), 492/494 (99%), 491/494 (99%), and 476/478 (99%). The similarity in sequence identity between the rDNA ITS region of the Jeonbuk isolate and other Didymella bryoniae from BLAST searches of GenBank was 100% and was 95.0% within the group. Nucleotide sequences of the rDNA ITS region from pure culture ranged from 98.2% to 99.8%. Phylogenetic analysis with related species of D. bryoniae revealed that D. bryoniae is a monophyletic group distinguishable from other Didymella spp., including Ascochyta pinodes, Mycosphaerella pinodes, M. zeae-maydis, D. pinodes, D. applanata, D. exigua, D. rabiei, D. lentis, D. fabae, and D. vitalbina. Phylogenetic analysis, based on rDNA ITS sequence, clearly distinguished D. bryoniae and Didymella spp. from the 10 other species studied. This study identified the Jeonbuk isolate to be D. bryoniae.

Identification of Metarhizium sp. Isolated from Protaetia brevitarsis seulensis (Kolbe) Using Ribosomal DNA Sequence (흰점박이꽃무지로부터 Metarhizium속 사상균의 분리 및 ribosomal DNA 염기서열에 의한 동정)

  • 최지영;김철학;제연호;최영철;김종길;박규택;김근영
    • Korean journal of applied entomology
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    • v.42 no.1
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    • pp.65-70
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    • 2003
  • For the purpose of the protection of beneficial insects from pathogens and the development of control agent against pests, a strain of Metarhizium sp. was isolated from the infected Protaetia brevitarsis seulensis larvae in Korea. Under the scanning electron microscope, the isolate, Metarhizium sp. KMA-1, showed distinct formation of conidia on the palisade-like masse which were comprised of elongate chains and this shape is a typical feature of Metarhizium species. PCR techniques were used to identify the isolate and the primers used were designed on the basis of two kinds of rRNAs sequences, 28S rRNA and internal transcribed spacer(ITS). The specific PCR products from each primer set were amplified and the DNA sequences were determined for the similarity comparison. Sequence alignment of these fragments using GenBank database resulted in the highest homology similarity between the isolate Metarhizium sp. KMA-1 and M. anisopliae. From these results, the isolate Metarhizium sp. KMA-1 in this study was identified as M. anisopliae.

Molecular Identification of a Sea Anemone (Cnidaria: Anthozoa: Actiniaria) Obtained in Gijang, Busan (부산 기장에서 채집된 말미잘의 분자생물학적 방법을 이용한 동정)

  • Yoo, Sang Joon;Kim, Do-Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.4
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    • pp.447-452
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    • 2017
  • In this study, we tried to identify a sea anemone collected from the coast of Gijang, Busan. The anemone was morphologically similar to species belonging to the genus Anthopleura, but its morphological characteristics did not allow for confirmed identification to species level. Multiple genes from mitochondrial cytochrome oxidase III, 12S and 16S rRNA, and nuclear 18S and 28S rRNA, were amplified for multilocus sequence typing (MLST) analysis using genomic DNA extracted from the sampled anemone and a different primer set. Based on the MLST analysis, the anemone obtained in this study was identified as Anthopleura artemisia. Also, the sequence of internal transcribed spacer-2 was most closely related to A. artemisia, indicating that this single region might be useful for anemone identification. This study shows significance of molecular identification for sea anemones, and will be helpful in studies of sea anemone identification using genotyping-by-sequencing.

New Records of Creeping Ctenophores, Genus Coeloplana (Tentaculata: Platyctenida: Coeloplanidae), from Korea

  • Song, Jun-Im;Hwang, Sung-Jin;Lee, Sang-Hwa;Park, Joong-Ki
    • Animal Systematics, Evolution and Diversity
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    • v.27 no.1
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    • pp.47-52
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    • 2011
  • Creeping ctenophores, Coeloplana species, were collected by SCUBA divers throughout the year (November 2006 to June 2010) from the branches and polyp masses of encrusting dendronephthyas at a depth of 20-32m off Munseom Island (Seogwipo-si, Jeju-do, Korea). A single individual of a newly recorded species in Korea, Coeloplana bocki Komai, 1920, was collected together with C. anthostella from the same location on 16 August 2009. A large number of individuals of each species were subsequently collected from the host Dendronephthya aff. dendritica on 20 June 2010. C. bocki can be distinguished from C. anthostella Song and Hwang, 2010 and C. komaii Utinomi, 1963 by its unique blue and orange colored stripes, and/or the branching and anastomosing milky-white stripes encircling the aboral sense organ towards the margin. The detailed morphology and molecular sequence information (nuclear 18S rDNA, internal transcribed spacer 1, and mitochondrial cox1 gene sequences) for C. bocki is provided, and C. bocki and C. anthostella are compared.