• Title/Summary/Keyword: Internal transcribed spacer rDNA

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Multi-loci Molecular Characterisation of Endophytic Fungi Isolated from Five Medicinal Plants of Meghalaya, India

  • Bhagobaty, Ranjan Kumar;Joshi, S.R.
    • Mycobiology
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    • v.39 no.2
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    • pp.71-78
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    • 2011
  • The phylogenetic relationships of the most dominant and morphologically cryptic endophytic fungal isolates from each of five selected medicinal plants, namely Potentilla fulgens, Osbeckia stellata, Osbeckia chinensis, Camellia caduca, and Schima khasiana of the biodiversity rich state of Meghalaya, were assessed with random amplification of polymorphic DNA and PCR-restriction fragment length polymorphism profiles. Sequencing of the internal transcribed spacer 1, small subunit rRNA and partial ${\beta}$-tubulin gene fragments was also conducted to determine the phylogenetic relationships of these isolates with fungal sequences available in Genbank, NCBI. The identity of the fungal isolates is suggested based on the molecular phylogenetic data.

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.

A Report of Eighteen Unrecorded Fungal Species in Korea (국내에서 분리된 미기록 진균 18종 보고)

  • Ahn, Geum Ran;Choi, Min Ah;Kim, Ji Eun;Seo, Eun Ji;Kim, Jun Young;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.292-303
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    • 2017
  • During a survey of culturable fungi in Korea, 18 unrecorded fungal species were isolated and identified from the indoor air of mushroom cultivation houses, the materials used for preparation of mushroom cultivation media, wild plants, and funitures. This study reports the descriptions of the 18 unrecorded fungal species: Aspergillus creber, Ceratocystis paradoxa, Colletotrichum spaethianum, Coniochaeta velutina, Coprinellus xanthothrix, Epicoccum sorghinum, Leptosphaeria rubefaciens, Myrothecium gramineum, Paraconiothyrium fuckelii, Penicillium erubescens, Penicillium melinii, Penicillium pulvillorum, Penicillium sabulosum, Penicillium turbatum, Pestalotiopsis portugalica, Pilidiella castaneicola, Rachicladosporium pini, and Umbelopsis nana. For all the identified species, the morphological characteristics including the features of colony formed on media, images of light microscopy, and molecular phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer ribosomal DNA (ITS rDNA), 18S rDNA, 28S rDNA, ${\beta}-tubulin$ gene, calmodulin gene, and translation elongation factor gene were described.

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.

Diversity of Fungi Isolated from Soil of Jeollabuk-do and Chungcheongbuk-do, Korea

  • Yadav, Dil Raj;Adhikari, Mahesh;Kim, Sang Woo;Kim, Hyun Seung;Um, Yong Hyun;Lee, Youn Su
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.30-30
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    • 2015
  • This study was conducted aiming with the assessment of fungal diversity in soil samples collected from different locations of Jeollabuk-do and Chungcheongbuk-do, Korea. Forty soil samples were collected in 2015 and fungi were isolated through serial dilution technique. Isolated fungi were purified and differentiated according to their morphological and microscopic characteristics. In total, 150 different representative isolates were recovered and the genomic DNA of each isolate was extracted by using QIAGEN$^{(R)}$ Plasmid Mini Kit (QIAGEN Sciences, USA) and the identification of fungi was carried out by sequence analysis of internal transcribed spacer (ITS) region of the 18S ribosomal DNA (18S rDNA). Recovered isolates belonged to 37 family, 67 genera and 108 species. Aspergillus spp., Penicillium spp., Trichoderma spp., Chaetomium spp. And Fusarium spp. were the most dominant taxa in this study. Out of total species, 20 species were identified as new records for Korea.

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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.

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.

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.

An Easy, Rapid, and Cost-Effective Method for DNA Extraction from Various Lichen Taxa and Specimens Suitable for Analysis of Fungal and Algal Strains

  • Park, Sook-Young;Jang, Seol-Hwa;Oh, Soon-Ok;Kim, Jung A;Hur, Jae-Seoun
    • Mycobiology
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    • v.42 no.4
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    • pp.311-316
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    • 2014
  • Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive species identification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacy in discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventional methods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cell walls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. This method involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, and precipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within 20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from the fungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa were tested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearly demonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.

Effect of Lime Sulfur on Changes of Fungal Diversity in Pear Fallen Leaves (석회유황합제가 배나무 낙엽의 진균 다양성 변화에 미치는 영향)

  • Min, Kwang-Hyun;Song, Jang Hoon;Cho, Baik Ho;Yang, Kwang-Yeol
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.281-285
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    • 2015
  • This study was conducted to examine changes in the fungal community on fallen leaves of pear by treatment with lime sulfur. Although the lime sulfur could reduce the primary inoculum of several pathogens on spring season, the effect of lime sulfur has not been well determined scientifically. Fallen leaves infected by pear diseases in pear orchards in Naju were collected and treated with lime sulfur or water as a control. To determine the fungal diversity from each treatment, rDNA internal transcribed spacer (ITS) regions were analyzed after extraction of fungal genomic DNA from lime sulfur-treated or water-treated fallen leaves, respectively. The most common fungal species were Ascomycota and Basidiomycota in both treated leaves. However, the population dynamics of several fungal species including Alternari sp., Cladosporium sp., and Phomopsis sp., which are known as pear pathogens for skin sooty dapple disease, were quite different from each treated leaves. These results indicated that lime sulfur treatment led to changes of fungal communities on pear fallen leaves and could be applicable as a dormant spray.