• Title/Summary/Keyword: ITS rDNA sequences

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First Reports of Unrecorded Mortierellomycetes and Umbelopsidomycetes Fungi from Freshwater Ecosystems in Korea

  • Jaeduk Goh;Yoosun Oh;Hye Yeon Mun
    • The Korean Journal of Mycology
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    • v.51 no.3
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    • pp.155-165
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    • 2023
  • In this study, we isolated several fungal strains from filtered water and sediment collected from rivers and streams. The strains were identified by molecular phylogenetic analyses of rDNA sequences (internal transcribed spacer [ITS], large subunit of ribosomal DNA [LSU]). The morphological characteristics of the fungi were investigated using microscopy, and the culture characteristics of fungi grown on several media were examined. We identified four species previously unknown in South Korea, namely, Dissophora globulifera, Linnemannia exigua, Mortierella rishikesha and Umbelopsis autotrophica.

PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp.

  • Min, Byung-Re;Lee, Young-Mi;Choi, Yong-Keel
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.66-73
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    • 2000
  • To investigate the genetic relationship among 12 species belonging to the Fusarium section Martiella, Dlaminia, Gibbosum, Arthrosporiella, Liseola and Elegans, the internal transcribed spacer(ITS) regions of ribosomal DNA (rDNA) were amplified with primer pITS1 and pITS4 using the polymerase chain reaction(PCR). After the amplified products were digested with 7 restriction enzymes, restriction fragment length polymorphism (RFLP) patterns were analyzed. The partial nucleotide sequences of the ITS region were determined and compared. Little variation was observed in the size of the amplified product having sizes of 550bp or 570bp. Based on the RFLP analysis, the 12 species studied were divided into 5 RFLP types. In particular, strains belonging to the section Martiella were separated into three RFLP types. Interestingly, the RFLP type of F. solani f. sp. piperis was identical with that of isolates belonging to the section Elegans. In the dendrogram derived from RFLP analysis of the ITS region, the Fusarium spp. examined were divided into two major groups. In general, section Martiella excluding F. solani f. sp. piperis showed relatively low similarity with the other section. The dendrogram based on the sequencing analysis of the ITS2 region also gave the same results as that of the RFLP analysis. As expected, 5.8S, a coding region, was highly conserved, whereas the ITS2 region was more variable and informative. The difference in the ITS2 region between the length of F. solani and its formae speciales excluding F. solani f. sp. piperis and that of other species was caused by the insertion/deletion of nucleotides in positions 143-148 and 179-192.

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Taxonomy of the Golovinomyces cynoglossi Complex (Erysiphales, Ascomycota) Disentangled by Phylogenetic Analyses and Reassessments of Morphological Traits

  • Braun, Uwe;Bradshaw, Michael;Zhao, Ting-Ting;Cho, Sung-Eun;Shin, Hyeon-Dong
    • Mycobiology
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    • v.46 no.3
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    • pp.192-204
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    • 2018
  • The name Golovinomyces cynoglossi s. lat. is traditionally applied to a complex of morphologically similar powdery mildews on hosts of the plant family Boraginaceae. The current species-level taxonomy within this complex is ambiguous due to the lack of phylogenetic examinations. The present study applied phylogenetic methods to clarify the taxonomy of G. cynoglossi s. lat. Phylogenetic analysis of rDNA ITS sequences retrieved from Asian, European and North American specimens revealed that G. cynoglossi s. lat. collections from different hosts involved several species in five clearly separated lineages. Clade I consists primarily of Golovinomyces cynoglossi s. str. on Cynoglossum. Clade III consists of Golovinomyces sequences retrieved from the host genera Symphytum and Pulmonaria. The taxa within clade III are now assigned to G. asperifoliorum comb. nov. Clade V encompasses G. cynoglossi s. lat. on the host genera Bothriospermum, Buglossoides, Echium, Myosotis, and Trigonotis. The taxa within clade V are now assigned to G. asperifolii comb. nov. The species concerned in this study were lecto- and epitypified to stabilize their nomenclature.

Four Endophytic Ascomycetes New to Korea: Cladosporium anthropophilum, C. pseudocladosporioides, Daldinia eschscholtzii, and Nigrospora chinensis

  • Lee, Dong Jae;Lee, Jae Sung;Lee, Hyang Burm;Choi, Young-Joon
    • The Korean Journal of Mycology
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    • v.47 no.3
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    • pp.187-197
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    • 2019
  • Ascomycota is the largest phylum of the Fungi, including approximately 6,600 genera. They are often isolated from soils, indoor air, and freshwater environments, but also from plants as pathogens or endophytes. In this study, four species of Ascomycota (two of Cladosporium and one of each Daldinia and Nigrospora) were collected from the leaves of four woody plants (Camellia japonica, Ginkgo biloba, Quercus sp., Vitis vinifera). Their cultural characteristics were investigated on five different media (PDA, V8A, CMA, MEA, CZA) at 3 days after incubation at $25^{\circ}C$ in darkness. BLASTn search and phylogenetic analysis were performed using the internal transcribed spacer (ITS) rDNA sequences, in addition to tef1 gene sequences for Cladosporium species. Based on the cultural, morphological, and phylogenetic data, the isolates were identified as Cladosporium anthropophilum, Cladosporium pseudocladosporioides, Daldinia eschscholtzii, and Nigrospora chinensis. Previously, some members of Cladosporium and Nigrospora have been recorded as endophytes inhabiting the leaves and stems of various plants, whereas Daldinia eschscholtzii is a wood-inhabiting endophyte or wood-decaying fungus. To our knowledge, this is the first report of these four ascomycetes in Korea.

Monitoring of Gentic Variability in Dicofol-susceptible, Dicofol-resistant, and its Reverse-selected Strains of Tetranychus urticae by RAPD-PCR

  • Song, Cheol;Park, Jin-Hee;Kim, Gil-Hah;Kwon, O-Yu;Cho, Kwang-Yun
    • Journal of Life Science
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    • v.9 no.1
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    • pp.14-16
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    • 1999
  • Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

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Phylogenetic Relationships of the Mutualistic Fungi Associated with Macrotermes subhyalinus in Oman

  • Hilal S. AlShamakhi;Abdullah M. Al-Sadi;Lyn G. Cook
    • Mycobiology
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    • v.51 no.5
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    • pp.281-287
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    • 2023
  • The symbiotic association between fungus-gardening termites Macrotermes and its fungal symbiont has a moderate degree of specificity-although the symbiotic fungi (Termitomyces) form a monophyletic clade, there is not a one-to-one association between termite species and their fungus-garden associates. Here, we aim to determine the origin and phylogenetic relationships of Termitomyces in Oman. We used sequences of the internal transcribed spacer region (ITS) and the nuclear large subunit ribosomal RNA (LSU rRNA, 25S) gene and analyzed these with sequences of Termitomyces from other geographic areas. We find no evidence for more than a single colonization of Oman by Termitomyces. Unexpectedly, we find Termitomyces in Oman is most closely related to the symbiont of M. subhyalinus in West Africa rather than to those of geographically closer populations in East Africa.

The complete plastid genome and nuclear ribosomal transcription unit sequences of Spiraea prunifolia f. simpliciflora (Rosaceae)

  • Jeongjin CHOI;Wonhee KIM;Jee Young PARK;Jong-Soo KANG;Tae-Jin YANG
    • Korean Journal of Plant Taxonomy
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    • v.53 no.1
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    • pp.32-37
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    • 2023
  • Spiraea prunifolia f. simpliciflora Nakai is a perennial shrub widely used for horticultural and medicinal purposes. We simultaneously obtained the complete plastid genome (plastome) and nuclear ribosomal gene transcription units, 45S nuclear ribosomal DNA (nrDNA) and 5S nrDNA of S. prunifolia f. simpliciflora, using Illumina short-read data. The plastome is 155,984 bp in length with a canonical quadripartite structure consisting of 84,417 bp of a large single-copy region, 18,887 bp of a short single-copy region, and 26,340 bp of two inverted repeat regions. Overall, a total of 113 genes (79 protein-coding genes, 30 tRNAs, and four rRNAs) were annotated in the plastome. The 45S nrDNA transcription unit is 5,848 bp in length: 1,809 bp, 161 bp, and 3,397 bp for 18S, 5.8S, and 26S, respectively, and 261 bp and 220 bp for internal transcribed spacer (ITS) 1 and ITS 2 regions, respectively. The 5S nrDNA unit is 512 bp, including 121 bp of 5S rRNA and 391 bp of intergenic spacer regions. Phylogenetic analyses showed that the genus Spiraea was monophyletic and sister to the clade of Sibiraea angustata, Petrophytum caespitosum and Kelseya uniflora. Within the genus Spiraea, the sections Calospira and Spiraea were monophyletic, but the sect. Glomerati was nested within the sect. Chamaedryon. In the sect. Glomerati, S. prunifolia f. simpliciflora formed a subclade with S. media, and the subclade was sister to S. thunbergii and S. mongolica. The close relationship between S. prunifolia f. simpliciflora and S. media was also supported by the nrDNA phylogeny, indicating that the plastome and nrDNA sequences assembled in this study belong to the genus Spiraea. The newly reported complete plastome and nrDNA transcription unit sequences of S. prunifolia f. simpliciflora provide useful information for further phylogenetic and evolutionary studies of the genus Spiraea, as well as the family Rosaceae.

Monitoring of Fasciola Species Contamination in Water Dropwort by COX1 Mitochondrial and ITS-2 rDNA Sequencing Analysis

  • Choi, In-Wook;Kim, Hwang-Yong;Quan, Juan-Hua;Ryu, Jae-Gee;Sun, Rubing;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.53 no.5
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    • pp.641-645
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    • 2015
  • Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.

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.

Nucleotide Sequences of an Aphid ribosomal RNA Unit (진딧물의 전 ribosomal RNA 염기배열)

  • Kwon, Tae-Young;An, Seung-Lak;Song, Cheol;Park, Jong-Kyun;Kim, Young-Sub;Hwang, Jae-Sam;Kwon, O-Yu
    • Journal of Life Science
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    • v.8 no.1
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    • pp.32-39
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    • 1998
  • The length and G/C concent of regions of an aphid rDNA unit that spans 13,061bo with 59% G/C content. flolowing belowing below are the those results, 5’ETS is 843bp in length with 69% G/C content, 18S is 2,469bp in length with 59% G/C content, ITS I is 229bp in length with 70% G/C content, 5.8S is 160bp in length with 63% G/C content, ITS II is 325bp in length with 70% G/C content, 28S is 4, 147bp in length with 60% G/C content, IGS is 4,888bp in length with 55% G/C content.

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