• Title/Summary/Keyword: ITs rDNA

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Isolation and Identification of Rice Root Endophytic Antagonistic Serratia marcescens (벼 뿌리 내생 항균성 Serratia marcescens의 분리 및 동정)

  • Lee, Sook-Kyung;Song, Wan-Yeob;Kim, Hyung-Moo
    • Research in Plant Disease
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    • v.10 no.1
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    • pp.63-68
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    • 2004
  • Twenty-three strains of Serratia sp., isolated from surface-sterilized rice roots collected in Chonbuk and Chungnam province, were identified and characterized. They were Gram-negative, rod shaped and red pigmented typically and their endophytism was confirmed by inoculation and reisolation of the strains in planta. Their antifungal activity against 4 rice pathogenic fungi was compared and ranged from 62.4 to 85.2% against Rhizoctonia solani and 68.0 to 88.5% against Pyricularia grisea. Among the 23 strains tested, strain Rsm220 showed the strongest inhibition activity against 4 pathogenic fungi. The strain was, therefore, selected as a biocontrol candidate for both the pathogens and its bacteriological characteristics and 165 rDNA sequences were analyzed. Phenotypic and biochemical characteristics of the selected Rsm220 were highly related to the type strain of S. marcescens and 165 rDNA sequencing of Rsm220 showed a homology of 98.2% to the type strain of S. marcescens. The strain Rsm220 was identified as S. marcescens and the inhibition result of this endophytic strain indicates that it is a potential biocontrol agent for R. solani and R grisea.

Identification of Unrecorded Endophytic Fungi Isolated from Leaves of Woody Plants in Jejudo, Korea (제주도에 서식하는 목본 식물의 잎에서 분리한 미기록 내생균)

  • Lee, Bong-Hyung;Kim, Dong-Yeo;Park, Hyeok;Eo, Ju-Kyeong;Lee, Hyang Burm;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.252-258
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    • 2016
  • In this study, endophytic fungi were isolated from the leaves of five species of woody plants in Jeju, Korea, namely Chamaecyparis obtusa, Cryptomeria japonica, Torreya nucifera, Ilex crenata, and Camellia japonica. The isolated fungal endophytes were identified based on their morphological and molecular characteristics including a sequence analysis of the internal transcribed spacer and 26S regions of rDNA and ${\beta}$-tubulin genes. Ten species of fungal endophytes have not been previously reported in Korea, namely Mycosphaerella aleuritidis, Neofusicoccum eucalyptorum, Neofusicoccum parvum, Phyllosticta citrichinensis, Phyllosticta cryptomeriae, Phomopsis cotoneastri, Sphaerulina rhododendricola, Guignardia mangiferae, Lophodermium jiangnanense, and Lophodermium minus.

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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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.

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.

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.

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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