• Title/Summary/Keyword: soil DNA

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Direct Detection of Cylindrocarpon destructans, Root Rot Pathogen of Ginseng by Nested PCR from Soil Samples

  • Jang, Chang-Soon;Lim, Jin-Ha;Seo, Mun-Won;Song, Jeong-Young;Kim, Hong-Gi
    • Mycobiology
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    • v.38 no.1
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    • pp.33-38
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    • 2010
  • We have successfully applied the nested PCR to detect Cylindrocarpon destructans, a major pathogen causing root rot disease from ginseng seedlings in our former study. The PCR assay, in this study, was used to detect the pathogen from soils. The nested PCR using internal transcribed spacer (ITS) 1, 4 primer set and Dest 1, 4 primer set maintained the specificity in soils containing various microorganisms. For a soil DNA extraction method targeting chlamydospores, when several cell wall disrupting methods were tested, the combination of lyophilization and grinding with glass beads, which broke almost all the chlamydospores, was the strongest. The DNA extraction method which was completed based on the above was simple and time-saving because of exclusion of unnecessary stages, and efficient to apply in soils. As three ginseng fields whose histories were known were analyzed, the PCR assay resulted as our expectation derived from the field information. The direct PCR method will be utilized as a reliable and rapid tool for detecting and monitoring C. destructans in ginseng fields.

Isolation and Characterization of Diesel Oil Degrading Bacterium, Pseudomonas sp. GENECO 1 Isolated from Oil Contaminated Soil (유류 오염 토양으로부터 분리한 디젤 분해 세균 Pseudomonas sp. GENECO 1의 분리 및 특성 규명)

  • 이종광;김무훈;박형수
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.102-107
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    • 2003
  • With the enrichment culture technique, bacterial strains which degrade diesel oil were isolated from soil contaminated with diesel oil. One of the isolates named GENECO 1 showed the highest activity for emulsification of diesel oil as well as the highest growth rate. This strain, GENECO 1, was identified as a Pseudomonas sp. based on its biochemical, physiological characteristics and 16S rDNA sequences. The optimal cultural conditions for cell growth and oil emulsifying activity of its culture were as follow; $30^{\circ}C$ for temperature, 7.0 for pH. Diesel oil degradation was analysed by the gas chromatography. More than 95% of 1% treated diesel oil were converted into a form no longer extractable by mixed organic solvents after 96 hours incubation.

Bacillus subtilis as a Tool for Screening Soil Metagenomic Libraries for Antimicrobial Activities

  • Biver, Sophie;Steels, Sebastien;Portetelle, Daniel;Vandenbol, Micheline
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.850-855
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    • 2013
  • Finding new antimicrobial activities by functional metagenomics has been shown to depend on the heterologous host used to express the foreign DNA. Therefore, efforts are devoted to developing new tools for constructing metagenomic libraries in shuttle vectors replicatable in phylogenetically distinct hosts. Here we evaluated the use of the Escherichia coli-Bacillus subtilis shuttle vector pHT01 to construct a forest-soil metagenomic library. This library was screened in both hosts for antimicrobial activities against four opportunistic bacteria: Proteus vulgaris, Bacillus cereus, Staphylococcus epidermidis, and Micrococcus luteus. A new antibacterial activity against B. cereus was found upon screening in B. subtilis. The new antimicrobial agent, sensitive to proteinase K, was not active when the corresponding DNA fragment was expressed in E. coli. Our results validate the use of pHT01 as a shuttle vector and B. subtilis as a host to isolate new activities by functional metagenomics.

Sixteen New Records of Ascomycetes from Crop Field Soil in Korea

  • Adhikari, Mahesh;Kim, Sangwoo;Kim, Hyun Seung;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.271-288
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    • 2016
  • The present study reports 16 species of Ascomycota that were previously unknown in Korea, namely Acremonium cellulolyticus (KNU14-25), Acremonium zonatum (KNU14-15), Chaetomium madrasense (KNU14-9), Cladosporium silenes (KNU 14-18-1), Humicolopsis cephalosporioides (KNU15-3), Leptosphaerulina chartarum (KNU14-16), Paecilomyces marquandii (KNU14-8), Paecilomyces tenuis (KNU14-18-2), Paraphaeosphaeria sporulosa (KNU15-2), Penicillium rubidurum (KNU14-12), Pochonia suchlasporia (KNU15-6), Sporothrix inflata (KNU15-8), Thielavia hyrcaniae (KNU15-1), Thielavia terricola (KNU14-23-1), Xylogone sphaerospora (KNU15-7), and Zopfiella longicaudata (KNU15-5). These fungal species were isolated from soil samples collected from various regions of Korea and identified based on their morphological characteristics and rDNA internal transcribed spacer sequence data. Full descriptions and illustrations for each species are provided.

First Report of Xenoroussoella triseptata Isolated from Soil in Korea

  • Jung-Joo Ryu;Seung-Yeol Lee;In-Kyu Kang;Leonid N. Ten;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.50 no.3
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    • pp.195-204
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    • 2022
  • A fungal strain, designated KNUF-20-NI009, was isolated from soil collected from Gunsan-si, Jeollabuk-do, Korea. The isolate showed cultural features typical of the genus Xenoroussoella. Colonies cultivated on malt extract agar were olivaceous-brown to pale olivaceous-white at the margins, with undersides of dark olivaceous to olivaceous-brown and a white margin. The conidia, with a size range of 2.7-5.1×1.6-3.3 ㎛ ($\bar{x}=3.6\times2.6{\mu}m$, n=50), were globoid to ellipsoid in shape, hyaline when immature, becoming light brown to golden-brown when mature, and characterized by 1 or 2 guttules. Multi-locus sequence analysis based on a combined dataset of internal transcribed spacer regions (ITS), large subunit rDNA (LSU), small subunit rDNA (SSU), translation elongation factor 1-alpha (TEF1α), and RNA polymerase II largest subunit (RPB2) sequences revealed KNUF-20-NI009 to be a strain of Xenoroussoella triseptata. This is the first report of this species in Korea.

Restriction Analyses of PCR Amplified Partial SSU Ribosomal DNA to Distinguish Arbuscular Mycorrhizal Fungi from Other Fungi Colonizing Plant Roots

  • Lee, Jae-Koo;Tae, Moon-Sung;Eom, Ahn-Heum;Lee, Sang-Sun
    • Mycobiology
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    • v.31 no.2
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    • pp.68-73
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    • 2003
  • Roots of Glycine max and Miscanthus sinensis and soil samples were collected from various field sites at Goesan, Chungbuk in Korea. Microscopic observations of the roots indicated high colonization rates of both arbuscular mycorrhizal fungi(AMF) and other fungi. The partial small subunit of ribosomal DNA genes were amplified with the genomic DNA extracted from their roots by nested polymerase chain reaction(PCR) with universal primer NS1 and fungal specific primers AML Restriction fragment length polymorphism(RFLP) was analyzed using the combinations of three restriction enzymes, HinfI, AluI and AsuC21. Nucleotides sequence analysis revealed that ten sequences from Miscanthus sinensis and one sequence from Glycine max were close to those of arbuscular mycorrhizal fungi. Also, 33% of total clones amplified with NS31-AM1 primers from M. sinensis and 97% from G. max were close to Fusarium oxysporum or other pathogenic fungi, and they were successfully distinguished from AME Results suggested that these techniques could help to distinguish arbuscular mycorrhizal fungi from root pathogenic fungi in the plant roots. Especially, DNA amplified by these primers showed distinct polymorphisms between AMF and plant pathogenic species of Fusarium when digested with AsuC21.

Cloning of \beta-mananase gene from Aeromonas sp. in E. coli (토양에서 분리한 Aeromonas sp 로 부터 \beta-mannanase 유전자의 클로닝)

  • 박봉환;강대경;김하근
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.201-205
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    • 2001
  • A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Aeromonas sp. A genomic DNA library constructed from Aeromonas, sp that secrets a $\beta$-mannanase was screened for mannan hydrolytic acticity. Recombinant $\beta$-mannanase activity was detercted on the basis of the clear zones around Escherichia coli colonies grown on a LB medium supplemented locust bean gum, EcoRI restriction analysis of plasmid prepared from recombinant E. coli which showed a $\beta$-mannanase activity revealed 10 kb DNA insert, The optimum pH and temperature for the activity of reconmbinant $\beta$-mannanase were 6.0 and $50^{\circ}C$ respectively and were identical to those of the native enzyme.

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Phylogenetic Inter- and Intrarelationships of the Genus Microbispora of the Family Streptosporangiaceae Based on 16S Ribosomal DNA Sequences (16S Ribosomal DNA 염기서열 분석에 근거한 Streptosporangiaceae과 Microbispora 속의 계통 관계)

  • Lee, Soon-Dong
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.429-434
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    • 2003
  • The 16S rDNA sequences of nine strains, two type strains of validated Microbispora species and a strain of invalidated Microbispora species, and six soil isolates, were determined and compared with those of representatives of the family Streptosporangiaceae. The phylogenetic analysis indicated that all of the validated species of the genus Microbispora consistently formed a monophyletic unit and were well separated from the other genera of the family Streptosporangiaceae. All the isolates were placed to the genus Microbispora, whereas an invalidated Microbispora species, Microbispora griseoalba IMSNU $22049^{T}$ (= KCTC $9314^{T}$), was closely related to members of the genus Nocardia.

DNA Toposiomerase I Inhibitor by Streptomyces sp. 7489 (방선균주 7489가 생산하는 DNA Topoisomerase I 저해제에 관한 연구)

  • Lee, Dong-Sun;Ha, Sang-Chul;Lee, Sang-Yong;Kim, Jong-Guk;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.101-104
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    • 1996
  • During the screening of inhibitor of DNA topoisomerase I from microbial secondary metabolites, Streptomyces melanosporofaciens 7489 which was capable of producing high level of inhibitor was selected from soil. The active compound (7489-1) was purified from the culture broth by solvent extraction, silica gel column chromatography and HPLC. The inhibitor was identified as dibutyl phthalate by spectroscopic methods of UV, $^{1}H$-NMR, $^{13}C$-NMR, DEPT and EI-MS. 7489-1 showed a strong inhibitory activity against topoisomerase I with 10 ${\mu}$M of $IC_{50}$ value.

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Characterization of Soil Microorganism from Humus and Indigenous Microorganism Amendments

  • Jan, Umair;Feiwen, Rui;Masood, Jan;Chun, Se Chul
    • Mycobiology
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    • v.48 no.5
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    • pp.392-398
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    • 2020
  • This study was conducted to understand the dynamics of microbial communities of soil microorganisms, and their distribution and abundance in the indigenous microorganisms (IMOs) manipulated from humus collected from the forest near the crop field. The soil microorganisms originated from humus and artificially cultured microbial-based soil amendments were characterized by molecular and biochemical analyses. The bacterial population (2 × 106~13 × 106 CFU/g sample) was approximately 100-fold abundant than the fungal population (2 × 104~8 × 104 CFU/g sample). The 16S rDNA and ITS sequence analyses showed that the bacterial and fungal communities in humus and IMOs were mainly composed of Bacillus and Pseudomonas, and Trichoderma and Aspergillus species, respectively. Some of the bacterial isolates from the humus and IMOs showed strong inhibitory activity against soil-borne pathogenic fungi Fusarium oxysporum and Sclerotinia sclerotiorum. These bacteria also showed the siderophore production activity as well as phosphate solubilizing activity, which are requisite traits for biological control of plant pathogenic fungi. These results suggest that humus and IMOs could be a useful resource for sustainable agriculture.