• Title/Summary/Keyword: ITS rDNA

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Genetic Diversity of Penicillium isolates Isolated from Pears with Postharvest Decay in Storage (수확 후 저장 배에서 분리한 Penicillium속 균의 유전적 다양성)

  • Han, Do-Suk;Hong, Sung-Kee;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.11-18
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    • 2012
  • This study was carried out to identify the genetic diversity of Penicillium isolates that were isolated from pears with postharvest decay in storage. URP-PCR was used to detect DNA diversity of 84 Penicillium isolates. Based on URP-PCR profiles, 18 Penicillium isolates were selected and their PCR polymorphic bands were produced by additional primers URP1F, URP2R, URP2F, and URP4R. UPGMA cluster analysis using the polymorphic bands showed four clustered groups and futhermore cultural and morphological features characterized the 18 Penicillium isolates. Group 1 was dominant, which occupies 70% in the four clustered groups and identified as P. expansum based on ITS sequence and morphological features.

Phylogenetic Analysis of the Genus Gliocladium and its Related Taxa by Comparing the Sequences of Internal Transcribed Spacers and 5.8S r-DNA (Ribosomal DNA의 Internal Transcribed Spacer(ITS) 부위의 염기서열 분석에 의한 Gliocladium 속과 근연속에 관한 계통 분류학적 연구)

  • Park, Ju-Young;Kim, Gi-Young;Ha, Myoung-Gyu;Shin, Young-Kook;Park, Yong-Ha;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.27 no.3 s.90
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    • pp.191-197
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    • 1999
  • The phylogenetic position of Gliocladium and its related taxa were investigated, using the neighbor-joining method of the sequences from internal transcribed spacers (ITS1 and ITS2) and 5.8S ribosomal DNA (rDNA). It was focused especially on the generic concept by comparing with the related genera such as Trichoderma, Hypocrea, Verticillium, Penicillium and Talaromyces. Gliocladium species and its related genus were divided into three groups by the phylogenetic analysis using the neighbor-joining method. The first group includes Penicillium-like strains such as Penicillium, Tararomyces, Verticillium and one species of Gliocladium (G. cibotii JCM 9203 and JCM 9206). Especially, Gliocladium cibotii JCM 9203 is thought to be the similar species with Verticillium bulbillosum JCM 9214. Between these two species, Gliocladium cibotii and Verticillium bulbillosum, the intraspecies concept needs to examined with culture condition. and morphological properties. The second group includes two species Verticillium, Verticillium tricorpus and Verticillium albo-atrum which extracted from the GenBank database in NCBI (National Center for Biotechnology Information). Trichoderma-like strains, such as Trichoderma, Hypocrea and several species of Gliocladium are included in the third group. Also, Gliocladium penicillioides IFO 5869 and Gliocladium catenulatum ATCC 10523 formed the subgroup of Trichoderma-like strains. The species of Gliocladium were dispersed in Trichoderma-like and Penicillinum-like group, and only one species of Gliocladium cihotii used in our study was located in Penicillium-like genus group. The species of Verticillium appeared in all three groups and the species of Trichoderma formed the monophylogeny with Hypocrea (telemorph). Also, Gliocladium virens was grouped with Trichoderma harzianum with a high bootstrap value, supporting that Gliocladium virens is to be placed in Trichoderma. The results suggest that Gliocladium is polyphyletic, and is more Trichoderma-like than Penicillium-like.

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Fungi Isolated from Pine Wood Nematode, Its Vector Japanese Pine Sawyer, and the Nematode-Infected Japanese Black Pine Wood in Korea

  • Hyun, Min-Woo;Kim, Ji-Hye;Suh, Dong-Yeon;Lee, Seung-Kyu;Kim, Seong-Hwan
    • Mycobiology
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    • v.35 no.3
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    • pp.159-161
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    • 2007
  • Pine tree death caused by pine wood nematode(PWN) involves phoretic relationships between PWN and its vector Japanese pine sawyer beetle(JPS). In an effort to understand the diversity of fungi involved in PWN life cycle, a total of 176 fungal isolates were collected from PWNs, adults and larvae of JPS, PWN-diseased Japanese black pine that was cut down in 2005 at Jinju, Korea. Based on microscopic observation and colony morphology, and sequence analysis of the ITS rDNA, the fungal isolates were identified at the level of genus. Three genera including Mucor, Ophiostoma, and Penicillium were identified from PWN. Two genera of Ophiostoma and Penicillium were discovered from JPS larvae. Frpm JPS adult beetles, nine genera of Aspergillus, Gibberalla, Hypocrea, Irpex, Leptosphaeria, Ophiostoma, Penicillium, and Plectosphaerella and unknown basidio-mycetes were found. Ten genera from PWN-infected weed were confirmed as Bionectria, Botrytis, Camarops, Fusarium, Hypocrea, Nectria, Mucor, Ophiostoma, Penicillium, and Trichoderma. Penicillium and Ophiostoma were commonly distributed on PWN and its vector and host. This is first report of the fungi associated with PWN and its vector and host in Korea.

Diversity of Endophytic Fungi Associated with Roots of Calanthe discolor and Cephalanthera longibracteata in Korea (새우난초와 은대난초 뿌리에서 분리한 내생균의 다양성)

  • Lee, Bong-Hyung;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.427-435
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    • 2018
  • In this study, endophytic fungi were isolated from the surface-sterilized roots of Calanthe discolor and Cephalanthera longibracteata collected from the Chungnam, Jeju, Kyungnam and Chungbuk provinces in Korea. The morphological characteristics of the obtained isolates were examined and their sequences of the internal transcribed spacer (ITS) rDNA region were analyzed using the ITS1F and ITS4 primers for species identification. Leptodontidium orchidcola showed the highest species abundance and frequency among the isolated endophytic fungi. Additionally, the community analysis revealed a high specificity between the host plants and the endophytic fungal species.

Vegetative Growth and Phylogenetic Relationship of Commercially Cultivated Strains of Pleurotus eryngii based on ITS sequence and RAPD

  • Alam, Nuhu;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • v.37 no.4
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    • pp.258-266
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    • 2009
  • Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

Identification and Phylogenetic Relationship of Dermatophytes Based on RFLP Analysis and Nucleotide Sequence of Internal Transcribed Spacer (ITS)1 in Nuclear Ribosome DNA (ITS-RFLP와 ITS1 염기서열 분석에 의한 피부사상균의 동정과 계통적 유연관계)

  • Choi, Yeon-Hwa;Lee, Yeong-Seon;Yoo, Jae-Il;Kim, Bong-Su
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.49-60
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    • 2000
  • ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

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Morphological and Phylogenetic Characteristics of a Nematophagous Fungus, Drechslerella brochopaga Kan-23 (국내 미기록종 선충포식성 곰팡이 Drechslerella brochopaga Kan-23의 형태 및 계통분류)

  • Cho, Chun-Hwi;Kang, Doo-Sun;Kim, Yoon-Ji;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.63-68
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    • 2008
  • Strain Kan-23 was extracted from nematophagous fungi, which were isolated from the soil sample of oriental melon field. The strain exhibited the slow-growing characteristic forming conidia after prolonged incubation for 30 days. Morphological features of strain Kan-23 were observed under scanning electron microscope (SEM). It possesses erect conidiophores which contain $2{\sim}3$ side branches, with each branch producing $5{\sim}10$ conidia. The size of conidiophores were between $160{\sim}450\;{\mu}m$. Conidia were ellipsoidal with three septa[septum] in each conidium. Strain Kan-23 captured nematodes by means of giant constricting rings, which were observed in the glucose peptone agar medium. ITS region of rDNA sequence was analyzed. On the basis of the high sequence similarity of ITS region (99%), the Kan-23 strain was closely related to Drechslerella brochopaga (U51950). This is the first report on Drechslerella brochopaga as a nematophagous fungus in Korea.

Morphological and molecular characteristics of Paralecithodendrium longiforme (Digenea: Lecithodendriidae) adults and cercariae from Chinese pipistrelle bats and viviparid snails in Thailand

  • Thitichai Arttra;Pheravut Wongsawad;Chalobol Wongsawad;Nattawadee Nantarat;Preeyaporn Butboonchoo;Jong-Yil Chai
    • Parasites, Hosts and Diseases
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    • v.62 no.1
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    • pp.85-97
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    • 2024
  • This study aimed to describe the morphological and molecular characteristics of Paralecithodendrium longiforme (Digenea: Lecithodendriidae) adults and cercariae isolated in Thailand. Adult flukes were isolated from the Chinese pipistrelle bat (Hypsugo sp.), and cercariae were detected in the viviparid snail (Filopaludina martensi martensi) from Chiang Mai province. The morphological characteristics were observed and described using conventional methods, and the molecular characteristics with internal transcribed spacer 2 (ITS2) and 28S rDNA gene sequences. The adult flukes were fusiform, 0.84-0.98 mm in length, and 0.37-0.49 mm in width, and were distinguishable from other species by the presence of longitudinal uterine coils. The cercariae were nonvirgulate xiphidiocercariae, with the oral sucker bigger than the acetabulum, the tail without fin fold, a body size of 117.5-138.3×48.3-52.2 ㎛, and a tail size of 100.7-103.7×15.0-18.9 ㎛. Molecular studies revealed that the adults and cercariae shared 99.3% (ITS2) and 99.6% (28S rDNA) homology with each other. They were phylogenetically close to P. longiforme with an identity of 94.5% for ITS2 and 98.7% for 28S rDNA. This study provides new information on the natural definitive host and first intermediate host of P. longiforme in Thailand. The discovery of its cercarial stage in Filopaludina snails highlights the importance of monitoring the associated second intermediate host and prevention and control of this potentially zoonotic trematode.

Ligand and Dimerization Dependent Transactivation Capability of Aromatic Hydrocarbon Receptor

  • Park, Hyun-Sung
    • BMB Reports
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    • v.32 no.3
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    • pp.279-287
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    • 1999
  • The aromatic hydrocarbon receptor (AhR) is a cytosolic protein that binds the environmental pollutant, dioxin. The liganded AhR translocates into the nucleus where it heterimerizes with a constitutive nuclear protein, AhR nuclear translocator (Arnt). The N-terminal regions of both AhR and Arnt contain basic helix-loop-helix (bHLH) and Per-AhR-Arnt-Sim (PAS) motifs that are required for DNA binding, dimerization, and ligand binding whereas the C-terminal regions of both AhR and Arnt contain transactivation domains. Here, results from the mammalian two-hybrid system indicate that Arnt can make a homodimer but AhR cannot. In the presence of dioxin, the interaction between AhR and Arnt is stronger than that of the Arnt homodimer, suggesting that Arnt prefers to make a heterodimer with the liganded AhR rather than a homodimer. Transfection analyses using the GAL4-driven reporter system suggest that AhR's N-terminal region represses its own transactivation domain, as well as exogenous transactivation domains such as Sp 1 and VP16. Interestingly, the repressed transactivation domains of AhR are activated by ligand-dependent heterodimerization with Arnt. These observations suggest that heterodimerzation with Arnt is necessary not only for DNA binding but also for activation of the repressed transactivation capability of AhR.

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Study on Molecular Phylogenetics of Korean Arisaema Species Based on Universal DNA Barcodes (범용성 DNA 바코드 분석 기반 한국산 천남성속(Arisaema) 식물의 분자계통학적 연구)

  • Noh, Pureum;Han, Kyeongsuk;Kim, Wook Jin;Yang, Sungyu;Choi, Goya;Ko, Sung Chul;Moon, Byeong Cheol
    • Korean Journal of Plant Resources
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    • v.31 no.1
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    • pp.37-51
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    • 2018
  • Molecular phylogenetic analysis was conducted to evaluate the taxonomic relationships of genus Arisaema L. distributed in Korea and the molecular phylogenetic characteristics of three authentic Arisaema species for the herbal medicine Arisaematis Rhizoma (the rhizomes of A. amurense, A. heterophyllum, and A. erubescens). The sequences of three DNA barcodes (rDNA-ITS, matK, and rbcL) were analyzed using 50 samples of nine taxa consisted of eight Korean and one Chinese Arisaema with one outgroup (Dracunculus vulgaris). Both individual and combined phylogenetic analyses of three DNA barcode sequences revealed that the treated nine taxa are independently classified into six distinct clades (Clade I, A. amurense f. amurense and A. amurense f. serratum; Clade II, A. serratum and A. takesimense; Clade III, A. ringens; Clade IV, A. erubescens; Clade V, A. heterophyllum; Clade VI, A. thunbergii subsp. thunbergii and A. thunbergii subsp. geomundoense). These six clades were reasonably divided into three individual sections, Pedatisecta, Sinarisaema, and Tortuosa. Futhermore, the results of comparative DNA barcode sequences analyses provided a significant information for the taxonomic reconsideration of Arisaema L. at the specific and intraspecific level. However, we could not confirm the taxonomic characteristics or identity among the three authentic medicinal species through the molecular phylogenetic analyses of genus Arisaema L. for Arisaematis Rhizoma.