• Title/Summary/Keyword: ITS rDNA sequences

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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.

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.

DNA Barcoding of Rocinela niponia (Isopoda, Cymothooidea, Aegidae) from South Korea

  • Kim, Sung Hoon;Choi, Hyun Ki;Kim, Jong Guk
    • Animal Systematics, Evolution and Diversity
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    • v.38 no.2
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    • pp.108-112
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    • 2022
  • An aegid species, Rocinela niponia Richardson, 1909, is a Far Eastern species known from Korean and Japanese waters. In this study, mitochondrial cytochrome c oxidase subunit I (COI) sequences of R. niponia were determined based on four specimens collected from the subtidal zone of Chujado Island, South Korea. We compared DNA barcoding data of this species with its congeners. As a result, there was no intra-specific genetic distance between the four COI sequences of R. niponia. Inter-specific distances between R. niponia and other five aegid species ranged from 23.8% to 35.6%. Morphological diagnosis and images of R. niponia are also provided as a valuable contribution toward the identification of Rocinela species in further taxonomic and ecological studies.

Leaf variants of Pinus and their ITS DNA sequences (소나무속 잎 변이와 그의 ITS DNA 염기서열)

  • Koo, JaChoon;Whang, Sung Soo
    • Korean Journal of Plant Taxonomy
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    • v.43 no.1
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    • pp.63-68
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    • 2013
  • ITS DNA sequences of two variants of Pinus spp. having single fasciculate leaf or two to three fasciculate leaves within an individual were analysed in order to determine their origin. Also, the same DNA locus of P. densiflora, P. rigida and P. koraiensis, distributed at the same region together with the OTUs having leaf variations, were analysed to compare with each other. Aligned sequences including ITS1, 5.8S and ITS2 were 580~584 bp in length. The 5.8S region was well conserved among all the OTUs we tested except for P. koraiensis, which has two nucleotide substitutions. The partial ITS1 region upstream of the 5.8S region showed relatively high sequence diversity compare to the ITS2 and has 181~185 bp in length. In this region, the sequences of the two variants were identical to that of P. densiflora. The ITS2 has identical for all OTUs in length and the two variants also have same sequences compare to that of P. densiflora. These two variants and samples of P. densiflora were grouped together in the UPGMA tree with 100% similarity level. The result strongly suggest that these two variants were originated from P. densiflora.

Characterization of Trichoderma spp. Associated with Green Mold of Oyster Mushroom by PCR-RFLP and Sequence Analysis of ITS Regions of rDNA

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.229-236
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    • 2005
  • Molecular profIles of PCR-RFLP and sequence analysis of internal transcribed spacer (ITS) regions of rDNA were compared between morphologically distinguishable species of Trichoderma isolated from substrates of oyster mushroom in Korea, T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, T. virens, and two unidentified species, Trichoderma sp. 1 and 2. PCR­RFLP analysis divided the Trichoderma spp. into six RFLP groups, A, B, C, D, E, and F. The RFLP groups were generally agreed with described morphological species, except that the RFLP group A containing the two unidentified species. A neighbor-joining tree based on ITS sequences well supported RFLP groups observed by RFLP analysis of ITS regions of rDNA. Additionally, the two unidentified species, Trichoderma sp. 1 and 2, which could not be distinguished by PCR­RFLP analysis, were separated in sequence analysis of ITS regions of rDNA.

New record of three hypotrich soil ciliates(Ciliophora: Hypotricha) from South Korea: Oxytricha multilineata, Mixophrya pantanalensis pantanalensis and Caudiurostyla sinensis

  • Kyu-Seok Chae;Gi-Sik Min
    • Journal of Species Research
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    • v.12 no.4
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    • pp.307-312
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    • 2023
  • Oxytricha multilineata, Mixophrya pantanalensis pantanalensis, and Caudiurostyla sinensis were isolated from soil samples collected from Cheongju-si and Yeoju-si, confirmed as new to South Korea. Oxytricha multilineata was distinguished from other congeners by seven dorsal kineties and dorsal bristles about 15 ㎛ long. Mixophrya pantanalensis pantanalensis was characterized by five to seven lithosomes and six dorsal kineties. Caudiurostyla sinensis was characterized by colorless cortical granules present, 10-14 midventral pairs, 7-9 left and 6-9 right marginal rows and four or five dorsal kineties. We determined the ribosomal DNA sequences (including 18S rDNA, ITS1, 5.8S rDNA, ITS2, and partial 28S rDNA) from above three species. And the genetic distances were compared with their congeners.

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.

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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Ansanella granifera gen. et sp. nov. (Dinophyceae), a new dinoflagellate from the coastal waters of Korea

  • Jeong, Hae Jin;Jang, Se Hyeon;Moestrup, Ojvind;Kang, Nam Seon;Lee, Sung Yeon;Potvin, Eric;Noh, Jae Hoon
    • ALGAE
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    • v.29 no.2
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    • pp.75-99
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    • 2014
  • A small dinoflagellate, Ansanella granifera gen. et sp. nov., was isolated from estuarine and marine waters, and examined by light microscopy, scanning electron microscopy, and transmission electron microscopy. In addition, the identity of the sequences (3,663-bp product) of the small subunit (SSU), internal transcribed spacer (ITS) region (ITS1, 5.8S, ITS2), and D1-D3 large subunit (LSU) rDNA were determined. This newly isolated, thin-walled dinoflagellate has a type E eyespot and a single elongated apical vesicle, and it is closely related to species belonging to the family Suessiaceae. A. granifera has 10-14 horizontal rows of amphiesmal vesicles, comparable to Biecheleria spp. and Biecheleriopsis adriatica, but greater in number than in other species of the family Suessiaceae. Unlike Biecheleria spp. and B. adriatica, A. granifera has grana-like thylakoids. Further, A. granifera lacks a nuclear fibrous connective, which is present in B. adriatica. B. adriatica and A. granifera also show a morphological difference in the shape of the margin of the cingulum. In A. granifera, the cingular margin formed a zigzag line, and in B. adriatica a straight line, especially on the dorsal side of the cell. The episome is conical with a round apex, whereas the hyposome is trapezoidal. Cells growing photosynthetically are $10.0-15.0{\mu}m$ long and $8.5-12.4{\mu}m$ wide. The cingulum is descending, the two ends displaced about its own width. Cells of A. granifera contain 5-8 peripheral chloroplasts, stalked pyrenoids, and a pusule system, but lack nuclear envelope chambers, a nuclear fibrous connective, lamellar body, rhizocysts, and a peduncle. The main accessory pigment is peridinin. The SSU, ITS regions, and D1-D3 LSU rDNA sequences differ by 1.2-7.4%, >8.8%, and >2.5%, respectively, from those of the other known genera in the order Suessiales. Moreover, the SSU rDNA sequence differed by 1-2% from that of the three most closely related species, Polarella glacialis, Pelagodinium bei, and Protodinium simplex. In addition, the ITS1-5.8S-ITS2 rDNA sequence differed by 16-19% from that of the three most closely related species, Gymnodinium corii, Pr. simplex, and Pel. bei, and the LSU rDNA sequence differed by 3-4% from that of the three most closely related species, Protodinium sp. CCMP419, B. adriatica, and Gymnodinium sp. CCMP425. A. granifera had a 51-base pair fragment in domain D2 of the large subunit of ribosomal DNA, which is absent in the genus Biecheleria. In the phylogenetic tree based on the SSU and LSU sequences, A. granifera is located in the large clade of the family Suessiaceae, but it forms an independent clade.