• Title/Summary/Keyword: Internal Transcribed Spacer

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Genetic Relationships of Internal Transcribed Spacer (ITS) Regions on Entomopathogenic Fungi by RFLP (Entomopathogenic Fungi의 ITS 영역에 대한 RFLP 분석)

  • Choi, In-Young;You, Young-Jin;Choi, Joung-Sik;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.28 no.2
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    • pp.112-117
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    • 2000
  • A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

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Analysis of ITS DNA Sequences of the Viola albida Complex (태백제비꽃군 ITS DNA 염기서열 분석)

  • Whang, Sung-Soo
    • Korean Journal of Plant Resources
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    • v.19 no.5
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    • pp.628-633
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    • 2006
  • ITS DNA sequences from five individuals, representative of five groups designated according to the degree of leaf teeth and lobes from simple to palmate compound leaf in the Viola albida complex, established and further analysed in order to solve the taxonomic difficulty. A total 702 bp was sequenced at the 5.8S ribosomal DNA and internal transcribed spacer 1 and 2. The 5.8S coding region is 163 bp, and has no sequence variations. The ITS1 and ITS2 noncoding regions have a little bit sequence variations, and those were further analysed by the methods of the analysis of variance (ANOVA), the analysis of sequence divergence and the phylogenetic analysis. The result of ANOVA showed no significant differences among individuals investigated. The analysis of sequence divergence with Kimura 2-parameter distance revealed that in-groups showed much less than 0.05 in absolute value among individuals, while two out groups more than 0.05, V. grypoceras and V. orientalis. This result appeared that the sequence divergence among in-groups was not yet occurred in the species level but situated at somewhere below the species level. In the phylogenetic analysis, two outgroups formed the basal clades in order. Five individuals in-groups formed a clade. The clade was, however, not very robust as around 50% in bootstrap value, suggesting that this result was not meaningful in the phylogenetic point of views.

Pulmonary Aspergillosis Secondary to Open Fracture of Pneumatic Bone in a Brown Hawk Owl (Ninox scutulata) (솔부엉이의 함기골 개방골절에 의한 속발성 폐 아스퍼질러스 감염 증례)

  • Han, Jae-Ik;Lee, Young-Sun;Lee, Sook-Jin;Kang, Hyo-Min;Jang, Hye-Jin;Chang, Dong-Woo;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.30 no.3
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    • pp.218-221
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    • 2013
  • A flightless brown hawk owl (Ninox scutulata) weighing 180 g was rescued and referred to the teaching veterinary hospital with humerus open fracture. On physical examination and radiography, open fracture of the left humerus was found. After 3 days, orthopedic surgery was operated with intramedullary pins and wires. The bird died 2 days after surgery with anorexia. On necropsy, multiple green nodules with 2-3 mm in diameter were observed at the surface and inside of the left lung. Numerous conidial heads and spores were seen in the center of foci in the histopathological examination. The mycelia penetrated the surrounding pulmonary parenchyma, showing inflammation and necrosis. The fungus was isolated from the lung and cultured on Sabouraud Dextrose Agar at $30^{\circ}C$ for 7 days. The colony was blue-green color with a powdery surface. The fungus was identified as Aspergillus fumigatus by DNA analysis, including the internal transcribed spacer region, partial ${\beta}$-tubulin, and the calmodulin gene. This case was diagnosed as pulmonary aspergillosis secondary to open fracture of pneumatic bone in a brown hawk owl.

Gibberellin A7 production by Aspergillus tubingensis YH103 and cultural characteristics of endophytic fungi isolated from Tetragonia tetragonoides in Dokdo islands (독도 번행초에서 분리된 내생균류의 배양적 특성과 Aspergillus tubingensis YH103의 gibberellin A7의 생산)

  • You, Young-Hyun;Park, Jong Myong;Lim, Sung Hwan;Kang, Sang-Mo;Park, Jong-Han;Lee, In-Jung;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.32-39
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    • 2016
  • Coastal plant species Tetragonia tetragonoides (Pall.) Kuntze native to the Dokdo islands was sampled and then 17 endophytic fungi were purely isolated based on morphological differences. The fungal isolates were characterized by their growth properties under NaCl concentration or pH gradient. Culture filtrates of the 17 fungal isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity. As the results, YH103 strain showed the highest plant growth-promoting activity among them. Phylogenetic analysis of the isolates was done by the maximum likelihood method based on partial internal transcribed spacer region (ITS region: contaning ITS1, 5.8S, and ITS2), beta-tubulin (BenA), and calmodulin (CaM) gene sequences. Chromatographic analysis of the strain YH103 culture filtrate showed the existence of gibberellins ($GA_4$, $GA_7$, $GA_8$, and $GA_{19}$). Finally, the strain YH103 was identified as Aspergillus tubingensis by microscopic observation and molecular analysis and, to our knowledge, this is the first report of GAs producing A. tubingensis.

Screening and Characteristics of Useful Fungi for Brewing from Commercial Nuruk in Chungcheong Provinces (충청지역 누룩에서 양조용 우수 곰팡이의 탐색 및 특성)

  • Baek, Seong-Yeol;Yun, Hye-Ju;Choi, Hye-Sun;Hong, Seung-Beom;Koo, Bon-Sung;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.373-378
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    • 2010
  • Studies on standardization and quality upgrade of nuruk which is a basic component in brewing are required to increase the quality level of Korean traditional rice wines and to develop the technology for practical use of it. It is important to isolate best strains, to improve the properties and effectively preserve them for brewing industry. In this study, 16 commercial nuruk samples were obtained from the commercial markets located in Chungcheong areas in Korea. 174 fungal strains were isolated from the samples on DG18 medium using a dilution plating method and then screened for enzyme activity and acid production. The active strains were identified based on the morphological characteristics and ITS sequence analysis. Out of 174 strains, 12 strains showed high amylase activity. Especially, Rhizopus sp. CN084, CN174, Aspergillus sp. CN161 and Mycocladus sp. CN042 showed high saccharogenic power and dextrinogenic enzyme activity on cooked wheat bran medium. On the other hand, Aspergillus sp. CN010, CN161, Rhizopus sp. CN105, CN168 and Rhizomucor sp. CN088 produced high acid production on the same medium. Our results showed that the active strains may be used as microbial sources for nuruk starter with good quality in brewing.

New Species of the Genus Metschnikowia Isolated from Flowers in Indonesia, Metschnikowia cibodasensis sp. nov.

  • Sjamsuridzal, Wellyzar;Oetari, Ariyanti;Nakashima, Chiharu;Kanti, Atit;Saraswati, Rasti;Widyastuti, Yantyati;Ando, Katsuhiko
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.905-912
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    • 2013
  • A novel species, Metschnikowia cibodasensis, is proposed to accommodate eight strains (ID03-$0093^T$, ID03-0094, ID03-0095, ID03-0096, ID03-0097, ID03-0098, ID03-0099, and ID03-0109) isolated from flowers of Saurauia pendula, Berberis nepalensis, and Brunfelsia americana in Cibodas Botanical Garden, West Java, Indonesia. The type strain of M. cibodasensis is ID03-$0093^T$ (= NBRC $101693^T$ =UICC $Y-335^T$ = BTCC-$Y25^T$). The common features of M. cibodasensis are a spherical to ellipsoidopedunculate shaped ascus, which contains one or two needle-shaped ascospores, and lyse at maturity. Asci generally develop directly from vegetative cells but sometimes from chlamydospores. The neighbor-joining tree based on the D1/D2 domain of nuclear large subunit (nLSU) ribosomal DNA sequences strongly supports that M. cibodasensis (eight strains) and its closest teleomorphic species, M. reukaufii, are different species by a 100% bootstrap value. The type strain of M. cibodasensis, ID03-$0093^T$, differed from M. reukaufii NBRC $1679^T$ by six nt (five substitutions and one deletion) in their D1/D2 region of nLSU rDNA, and by 18 nt (five deletions, four insertions, and nine substitutions) in their internal transcribed spacer regions of rDNA, respectively. Four strains representative of M. cibodasensis (ID03-$0093^T$, ID03-0095, ID03-0096, and ID03-0099) showed a mol% G+C content of $44.05{\pm}0.25%$, whereas that of M. reukaufii NBRC $1679^T$ was 41.3%. The low value of DNA-DNA homology (5-16%) in four strains of M. cibodasensis and M. reukaufii NBRC $1679^T$ strongly supported that these strains represent a distinct species.

Phylogeny of Scopolia Jacq. s. str. based on ITS sequences (ITS 염기서열에 의한 미치광이풀속의 계통)

  • Kim, Young-Dong;Paik, Jin-Hyub;Kim, Sung-Hee;Hong, Suk-Pyo
    • Korean Journal of Plant Taxonomy
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    • v.33 no.4
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    • pp.373-386
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    • 2003
  • Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA were determined for 14 individuals representing eight taxa from Scopolia s. str. and related genera, Anisodus and Atropanthe. We found that the ITS sequences of Korean endemic species, S. parviflora, are significantly different from its allied species, S. japonica. This is contradictory to traditional taxonomic treatments in which those species are regarded as conspecific. S. parviflora exhibited closer relationship to S. carniolica, which is disjunctly distributed in Europe. In spite of substantially high sequence divergence between S. japonica and S. parvlflora/S. carniolica clade, morphological resemblance is evident among the species. Morphological stasis concept (retardation of morphological differentiation or evolution of similar characters among the disjuncts in a similar ecological habitat) was referred to understand this rather unusual evolutionary feature. S. lutescens, another Korean endemic species, shared almost identical ITS sequences with S. parviflora. Lack of diagnostic character distinguishing the taxa suggests that they are conspecific. Anisodus carniolicoides, which was originally described in Scopolia, was grouped with A. luridus and A. tanguticus. The monophyletic Anisodus formed a sister group relationship with a monotypic genus Atropanthe.

Bacterial and fungal community composition across the soil depth profiles in a fallow field

  • Ko, Daegeun;Yoo, Gayoung;Yun, Seong-Taek;Jun, Seong-Chun;Chung, Haegeun
    • Journal of Ecology and Environment
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    • v.41 no.9
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    • pp.271-280
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    • 2017
  • Background: Soil microorganisms play key roles in nutrient cycling and are distributed throughout the soil profile. Currently, there is little information about the characteristics of the microbial communities along the soil depth because most studies focus on microorganisms inhabiting the soil surface. To better understand the functions and composition of microbial communities and the biogeochemical factors that shape them at different soil depths, we analyzed microbial activities and bacterial and fungal community composition in soils up to a 120 cm depth at a fallow field located in central Korea. To examine the vertical difference of microbial activities and community composition, ${\beta}$-1,4-glucosidase, cellobiohydrolase, ${\beta}$-1,4-xylosidase, ${\beta}$-1,4-N-acetylglucosaminidase, and acid phosphatase activities were analyzed and barcoded pyrosequencing of 16S rRNA genes (bacteria) and internal transcribed spacer region (fungi) was conducted. Results: The activity of all the soil enzymes analyzed, along with soil C concentration, declined with soil depth. For example, acid phosphatase activity was $125.9({\pm}5.7({\pm}1SE))$, $30.9({\pm}0.9)$, $15.7({\pm}0.6)$, $6.7({\pm}0.9)$, and $3.3({\pm}0.3)nmol\;g^{-1}\;h^{-1}$ at 0-15, 15-30, 30-60, 60-90, and 90-120 cm soil depths, respectively. Among the bacterial groups, the abundance of Proteobacteria (38.5, 23.2, 23.3, 26.1, and 17.5% at 0-15, 15-30, 30-60, 60-90, and 90-120 cm soil depths, respectively) and Firmicutes (12.8, 11.3, 8.6, 4.3, and 0.4% at 0-15, 15-30, 30-60, 60-90, and 90-120 cm soil depths, respectively) decreased with soil depth. On the other hand, the abundance of Ascomycota (51.2, 48.6, 65.7, 46.1, and 45.7% at 15, 30, 60, 90, and 120 cm depths, respectively), a dominant fungal group at this site, showed no clear trend along the soil profile. Conclusions: Our results show that soil C availability can determine soil enzyme activity at different soil depths and that bacterial communities have a clear trend along the soil depth at this study site. These metagenomics studies, along with other studies on microbial functions, are expected to enhance our understanding on the complexity of soil microbial communities and their relationship with biogeochemical factors.

Taxonomical Classification and Species-specific Detection of Genus Some Phellinus using Phylotype (Phylotype에 의한 수종의 Phellinus속의 분류체계 확립 및 종간구별을 위한 신속동정법 개발)

  • Kim, Cheng-Yun;Lee, Jae-Yun;Kim, Gi-Young;Lee, Ki-Won;Park, Jae-Min;Kim, Mun-Ok;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.121-128
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    • 2003
  • This study was carried out to identify the phylogenetic relationship of Phellinus species and to know its distribution by comparing the DNA sequences of internal transcribed spacer regions(ITS1 and IST2) and 5.8S ribosomal DNA (rDNA) repeat unit. The Phellinus species had their specific sequences in IST1 and 2 regions depending on suedes. The comparison of the ITS sequences of standard strains indicated that the sequences of ITS1 were more variable than those of ITS2. Nine strains of the commercial products of Phellinus species used in this study were identified as P. lintues, P. baumii, P. igniarius, and P. pini. Most of commercial species were P. pini and P. baumii, and P. gilvus was not found. Also, P. linteus was only found in form of mycelial culture rather than fruiting body. Moreover, the species-specific primers were designed based on ITS sequence data. Each species-specific primers were bound in P. lintues(ITSF-PL2R), P. baumii(PB1F-ITS4R), P. igniarius(IF1-IR3), P. pini(PF1-PR3), and P. gilvus(GF2-GR4), respectively. These primer sets would be useful fer the detection of specific-species among unidentified Phellinus species rapidly.

Diversity and Plant Growth Promoting Capacity of Endophytic Fungi Associated with Halophytic Plants from the West Coast of Korea

  • Khalmuratova, Irina;Kim, Hyun;Nam, Yoon-Jong;Oh, Yoosun;Jeong, Min-Ji;Choi, Hye-Rim;You, Young-Hyun;Choo, Yeon-Sik;Lee, In-Jung;Shin, Jae-Ho;Yoon, Hyeokjun;Kim, Jong-Guk
    • Mycobiology
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    • v.43 no.4
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    • pp.373-383
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    • 2015
  • Five halophytic plant species, Suaeda maritima, Limonium tetragonum, Suaeda australis, Phragmites australis, and Suaeda glauca Bunge, which are native to the Muan salt marsh of South Korea, were examined for fungal endophytes by sequencing the internal transcribed spacer (ITS) region containing ITS1, 5.8S rRNA, and ITS2. In total, 160 endophytic fungal strains were isolated and identified from the roots of the 5 plant species. Taxonomically, all 160 strains belonged to the phyla Ascomycota, Basidiomycota, and Zygomycota. The most dominant genus was Fusarium, followed by the genera Penicillium and Alternaria. Subsequently, using 5 statistical methods, the diversity indices of the endophytes were determined at genus level. Among these halophytic plants, P. australis was found to host the greatest diversity of endophytic fungi. Culture filtrates of endophytic fungi were treated to Waito-C rice seedlings for plant growth-promoting effects. The fungal strain Su-3-4-3 isolated from S. glauca Bunge provide the maximum plant length (20.1 cm) in comparison with wild-type Gibberella fujikuroi (19.6 cm). Consequently, chromatographic analysis of the culture filtrate of Su-3-4-3 showed the presence of physiologically active gibberellins, $GA_1$ (0.465 ng/mL), $GA_3$ (1.808 ng/mL) along with other physiologically inactive $GA_9$ (0.054 ng/mL) and $GA_{24}$ (0.044 ng/mL). The fungal isolate Su-3-4-3 was identified as Talaromyces pinophilus.