• Title/Summary/Keyword: Internal transcribed spacer 2 (ITS2)

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First Report of Leaf Spot of Datura metel Caused by Alternaria tenuissima in Korea

  • Aktaruzzaman, Md.;Kim, Joon-Young;Afroz, Tania;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.330-333
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    • 2015
  • In June 2013, we collected leaf spot disease samples of Datura metel from Gangneung, Gangwon Province, Korea. The symptoms observed were small circular to oval dark brown spots with irregular in shape or remained circular with concentric rings. We isolated the pathogen from infected leaves and cultured the fungus on potato dextrose agar. We examined the fungus morphologically and confirmed its pathogenicity according to Koch's postulates. The results of morphological examinations, pathogenicity tests, and the rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4), glycerol-3-phosphate dehydrogenase (G3PDH) and the RNA polymerase II second largest subunit (RPB2) gene sequence revealed that the causal agent was Alternaria tenuissima. To the best of our knowledge, this is the first report of leaf spot of D. metel caused by A. tenuissima in Korea as well as worldwide.

Characterization of a Zygomycete Fungus, Mortierella minutissima from Freshwater of Yeongsan River in Korea

  • Nguyen, Thi Thuong Thuong;Lee, Hyang Burm
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.346-349
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    • 2016
  • During a survey of fungal diversity of the order Mortierellales, a zygomycete strain, EML-YS717-1, was isolated from a freshwater sample collected from the Yeongsan River in Gwangju, Korea. Based on its morphological characteristics and a phylogenetic analysis of the internal transcribed spacer (ITS1 and ITS2) and 5.8S rDNA sequences, the strain was identified as Mortierella minutissima. To the best of our knowledge, M. minutissima, has not previously been authentically reported in Korea.

Phylogeny and Morphology of Sarcopodium terrigenum sp. nov., a Novel Fungal Species Isolated from Soil in Korea

  • Elderiny, Nabil Salah;Das, Kallol;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.49 no.2
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    • pp.175-181
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    • 2021
  • A fungal strain was isolated from a soil sample collected in Korea and designated as YW23-8. Based on a sequence analysis of the internal transcribed spacer (ITS) regions, the isolate was assigned to the genus Sarcopodium. Moreover, a phylogenetic analysis based on the concatenated nucleotide sequences of the ITS regions and the large subunit of the nuclear ribosomal RNA (LSU) gene showed that the strain YW23-8 occupies a distinct phylogenetic position within Sarcopodium. The isolate had significant differences from its closest neighbors, S. circinosetiferum, S. circinatum, S. macalpinei, and S. vanillae. Morphological features such as different conidial structures, the absence of septation in conidia, and the presence of milky white watery droplets along with the results of the phylogenetic analysis clearly distinguish YW23-8 from the closest Sarcopodium species. We therefore conclude that strain YW23-8 represents a novel species of the genus Sarcopodium for which we propose the name Sarcopodium terrigenum.

A Rapid and Universal Direct PCR Method for Macrofungi

  • Park, Mi-Jeong;Lee, Hyorim;Ryoo, Rhim;Jang, Yeongseon;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.455-467
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    • 2021
  • Macrofungi are valuable resources as novel drug candidates, new biomaterials, and edible materials. Recently, genetic approaches pertaining to macrofungi have been continuously growing for their identification, molecular breeding, and genetic engineering. However, purification and amplification of fungal DNA is challenging because of the rigid cell wall and presence of PCR inhibitory metabolites. Here, we established a direct PCR method to provide a rapid and efficient method for PCR-grade macrofungal DNA preparation applicable to both conventional PCR and real-time PCR. We first optimized the procedure of lysis and PCR using the mycelia of Lentinula edodes, one of the most widely consumed macrofungal species. Lysates prepared by neutralizing with (NH4)2SO4 after heating the mycelia in a mixture of TE buffer and KOH at 65℃ for 10 min showed successful amplification in both conventional and real-time PCR. Moreover, the addition of bovine serum albumin to the PCR mixture enhanced the amplification in conventional PCR. Using this method, we successfully amplified not only internal transcribed spacer fragments but also low-copy genes ranging in length from 500 to 3,000 bp. Next, we applied this method to 62 different species (54 genera) of macrofungi, including edible mushrooms, such as Pleurotus ostreatus, and medicinal mushrooms such as Cordyceps militaris. It was found that our method is widely applicable to both ascomycetes and basidiomycetes. We expect that our method will contribute to accelerating PCR-based approaches, such as molecular identification, DNA marker typing, gene cloning, and transformant screening, in macrofungal studies.

Phylogenetic Study of Genus Haliotis In Korea by Internal Transcribed Spacer Sequence (ITS) (ITS에 의한 한국내 전복 속 분류군의 유전적 계통분류학적 연구)

  • Huh, Man-Kyu;Kim, Jung-Ho;Moon, Du-Ho
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1003-1008
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    • 2009
  • Abalone (genus Haliotis) is a woody species with a long life span that is primarily distributed throughout the world, including Asia. This species is regarded as a very important marine gastropod mollusk in Korea and China, and also in food industries around the world. We evaluated a representative sample of the five species with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within the genus. Aligned nucleotide sequences of the length of the 5.8S subunit of all taxa of Haliotis were found to constant of 160 bp nucleotides. However, aligned nucleotide sequences of the length of ITS1 were varied within genus Haliotis, varying from 272 in H. diversicolor aquatilis to 292 in H. discus hannai. Aligned nucleotide sequences of the length of ITS2, especially, vary from 722 in H. diversicolor aquatilis to 752 in H. sieboldii. Total alignment length is 763 positions, of which 78 are parsimony-informative, 57 variable but parsimony-uninformative, and 459 constant characters. H. discus hannai was similar to H. discus, while H. diversicolor aquatilis was more distinct. ITS analysis may be useful in germ-plasm classification several taxa of genus Haliotis.

Genetic Variation in Flammulina velutipes (팽이버섯의 유전적 변이)

  • Kim, Jong-Bong;Jeong, Ja-In
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1434-1442
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    • 2011
  • A genetic variation within 29 strains of F. velutipes was analyzed by internal transcribed spacer (ITS) sequence analysis and random amplified polymorphic DNA (RAPD). Seven hundred and twenty base pairs were sequenced during the analysis of the ITS region, but no significant variation was observed among the 29 strains of F. velutipes. Sixteen out of 40 random primers amplified polymorphic RAPD fragment patterns. The polymorphic levels of RAPD bands by some primers (OPA-2,4,3,9,10,20) were very high in all 29 strains, with 3,030 fragments ranging between 200 and 2,000 bp. Intraspecific genetic dissimilarity of the 29 strains was calculated to range from 3.3% to 45% by Nei-Li's method using these 3,030 RAPD bands. The genetic variation among Korean strains was relatively high, with dissimilarities ranging between 17% and 38.6%. In the Neighbor-Joining analysis using the genetic dissimilarities based on RAPD, all 29 strains were classified into 5 clusters. Strains in each cluster showed specific characteristics according to their origin and strains. These results suggested that OPA and OPB primers could be used for developing molecular genetic markers and screening of unidentified (F. velutipes) strains.

Diaporthe eleutharrhenae sp. nov. Associated with a Critically Endangered Liana in China

  • Song, Shijie;Landrein, Sven
    • Mycobiology
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    • v.50 no.2
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    • pp.99-103
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    • 2022
  • A new fungus isolated from the leaves of Eleutharrhena macrocarpa in southern Yunnan, China is described using morphological and molecular evidence. Phylogenetic trees based on the combined nuclear ribosomal DNA internal transcribed spacer (ITS), translation elongation factor-1α (TEF1), and β-tubulin gene (TUB2) sequences showed that Diaporthe eleutharrhenae sp. nov. is sister to Diaporthe chinensis N.I. de Silva, Lumyong & K.D. Hyde and morphologically differs in shorter alpha conidia (5-8.5× 1.5-2 ㎛) and the presence of beta conidia. This study also resolves a nomenclatural problem, as two taxa were published using the same name. To avoid confusion, the unrelated D. chinensis H. Dong, J. W. Xia & X. G. Zhang is here renamed as D. dongii (H. Dong, J. W. Xia & X. G. Zhang) S. J. Song & Landrein, sp. nov. in honor of the author that described this species. Study and description of fungi associated with threatened tropical species could help to understand their ecology as well as the potential spread of fungi onto cultivated crop species.

Control Efficacy of Controlled Atmosphere and Temperature Treatment System Against the Hawthorn Spider Mite, Tetranychus viennensis (환경조절열처리 기술을 이용한 벚나무응애(Tetranychus viennensis) 살비 효과)

  • Son, Ye-Rim;Lee, Jong-Ho;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.51 no.2
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    • pp.131-140
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    • 2012
  • The hawthorn spider mite, Tetranychus viennensis, is a pest of apples and a quarantine pest from some countries that import apples from Korea. A controlled atmosphere and temperature treatment system (CATTS) was developed as an alternative disinfestation method to methyl bromide fumigation treatment, and has been applied to control various insects and other arthropod pests on fruits. We applied CATTS to disinfect T. viennensis under conditions that were previously developed to control the peach fruit moth, Carposina sasakii. First, T. viennensis was sampled from Japanese apricot, Prunus mume, and identified by its morphological characters. In addition, both cytochrome oxidase I (COI) and internal transcribed spacer (ITS) sequences supported the morphological identification. Second, the heat-tolerant developmental stage was determined in T. viennensis. When a $46^{\circ}C$ heat treatment was applied to egg, nymph, and adult stages of T. viennensis, adults were the most tolerant stage. Third, when heat temperature was used along with 1% $O_2$ and 15% $CO_2$, the mites showed a significant increase in susceptibility to the heat treatment. Finally, CATTS at $46^{\circ}C$ with 15% $CO_2$ and 1% $O_2$ for 30 min resulted in 100% mortality of all T. viennensis development stages. These results indicated that CATTS isapplicable to disinfest T. viennensis in post-harvest apples.

Phylogenetic Relationships of Coprinoid Taxa and an Agaric-like Gastroid Taxon Based on the Sequences of Internal Transcribed Spacer (ITS) Regions (ITS 영역의 염기서열을 이용한 먹물버섯류 및 주름버섯 유사 복균류와의 계통학적 유연관계)

  • Park, Dong-Suk;Go, Seung-Joo;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.27 no.6 s.93
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    • pp.406-411
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    • 1999
  • The phylogenetic relationships of black-spored basidioid taxa (mainly coprinoid taxa) and an agaric-like gastroid taxon were studied. The sequences of internal transcribed spacer regions (ITS) partially including 17S, 25S and 5.8S from 14 species (Coprinus comatus, C. atramentarius, C. micaceus, C. cinereus, C. disseminatus, C. rhizophorus, C. radians, C. echinosporus, Psathyrella candolleana, Podaxis pistillaris, Conocybe lactea, Bolbitius demangei, Agaricus balzei, and Stropharia rugosoannulata) were compared. The reciprocal homologies of ITS sequences among these species were in the range of $38.7{\sim}77.2%$. Black-spored taxa were classified into four clusters. Cluster I comprised C. micaceus, C. radians, and C. disseminata. Cluster II is consisted of C. cinereus, C. echinosporus, C. rhizophorus, and C. atramentarius. On the other hand, C. comatus is in cluster III with Agaricus balzei and Podaxis pistillaris even though this species is belonging to the section Coprinus in morphological aspect. Psathyrelloid taxon is included in cluster II. The question of the origin of secotioid (agaric-like) fungi has been taken, though largely on theoritical suggestions as to whether gastroid taxa give rise to agarics with secotioid taxa as intermediates or whether secotioid taxa are evolutionary novelities arising from many distinct groups of agarics. In this relationship, it was shown that secotioid taxon evolved from within agaric species.

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Identification of Orchid Mycorrhizal Fungi Isolated from Terrestrial Orchids in Mt. Hambaek, Korea (함백산의 난초과 식물의 뿌리에서 난균근균의 분리 및 동정)

  • Lee, Bong-Hyung;Han, Han-Kyeol;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.43 no.2
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    • pp.129-132
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    • 2015
  • In this study, orchid mycorrhizal fungi (OMF) were isolated from four terrestrial orchids on Mt. Hambeak, Platnathera chlorantha, Platnathera mandarinorum, Cephalanthera falcate, and Cephalanthera longibracteata. OMF were identified using morphological and sequences analysis of fungal internal transcribed spacer (ITS) regions by specific primer of basidiomycetous orchid mycorrhizas; ITS1-OF and ITS4-OF. Four species of orchid mycorrhizal fungi were identified as Ceratobasidium sp, Epulorhiza anaticula, Tulasnella calospora and Tulasnella sp.