• 제목/요약/키워드: 16S-rRNA

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Isolation and Identification of Endophytic Bacteria with Plant Growth Promoting Activity and Biocontrol Potential from Wild Pistachio Trees

  • Etminani, Faegheh;Harighi, Behrouz
    • The Plant Pathology Journal
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    • 제34권3호
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    • pp.208-217
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    • 2018
  • In this study, samples were collected from the leaves and stems of healthy wild Pistachio trees (Pistacia atlantica L.) from various locations of Baneh and Marivan regions, Iran. In total, 61 endophytic bacteria were isolated and grouped according to phenotypic properties. Ten selected isolates from each group were further identified by partial sequencing of the 16S rRNA gene. Based on the results, isolates were identified as bacteria belonging to Pseudomonas, Stenotrophomonas, Bacillus, Pantoea and Serratia genus. The ability of these isolates was evaluated to phytohormone production such as auxin and gibberellin, siderophore production, phosphate solubilization, atmospheric nitrogen fixation, protease and hydrogen cyanide production. All strains were able to produce the plant growth hormone auxin and gibberellin in different amounts. The majority of strains were able to solubilize phosphate. The results of atmospheric nitrogen fixation ability, protease and siderophore production were varied among strains. Only Ba66 could produce a low amount of hydrogen cyanide. The results of biocontrol assay showed that Pb78 and Sp15 strains had the highest and lowest inhibition effects on bacterial plant pathogens, Pseudomonas syringae pv. syringae Pss20 and Pseudomonas tolaasii Pt18 under in vitro condition. Pb3, Pb24 and Pb71 strains significantly promote root formation on carrot slices. To our knowledge this is the first report of the isolation of endophytic bacterial strains belonging to Pantoea, Bacillus, Pseudomonas, Serratia and Stenotrophomonas genus from wild pistachio trees with plant growth promoting potential and biocontrol activity.

젓갈에서 분리한 Bacillus subtilis SW-1의 항암활성 (Antitumor activity of Bacillus subtilis SW-1 isolated from Jeotgal)

  • 박종기;조용운;최영우;정영기;갈상완
    • 생명과학회지
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    • 제14권5호
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    • pp.815-820
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    • 2004
  • 본 연구에서 미생물유래의 종양 예방 및 억제제 개발을 목적으로 우리나라 전통 발효식품인 젓갈에서 항암 활성이 우수한 미생물을 분리, 동정 하였다. 젓갈로부터 항암물질을 생산하는 균주 SW-1을 분리하여 형태학적, 생리학적 특성을 검토한 결과, 본 균주는 Bacillus subtillis SW-1으로 동정되었다. 본 균주로부터 항암물질은 실험에 사용한 두가지의 암세포에 강한 활성을 나타내었다. 그 중에서 폐암 세포인 A549에 가장 강한 활성을 나타내었다. 항암물질 생산을 위한 최적 배양조건을 검토한 결과, 3.0% soluble starch, 1.0% yeast extract 였다. 무기염의 경우는 활성에는 그다지 영향을 미치지 않았다. 배지의 초기 pH 및 배양온도에 따른 항암물질의 생산성을 검토한 결과 초기 pH 5.0배양온도 3$0^{\circ}C$에서 25시 간배양 했을 때 최대 활성을 보였다. Chromosomal DNA단편화 실험에서 Bacillus subtilis SW-1의 배양상등액을 처리했을 때 DNA가 사닥다리모양으로 분해되는 Apoptosis(계획된 세포죽임)를 일으키는 것으로 확인되었다. 이 미생물이 생성하는 물질을 현재 HPLC, GC등을 이용하여 분리 중에 있으며 현재까지의 결과로는 당 중합체 인 것으로 분석된다.

Enzymatic Transformation of Ginsenoside Rb1 by Lactobacillus pentosus Strain 6105 from Kimchi

  • Kim, Se-Hwa;Min, Jin-Woo;Quan, Lin-Hu;Lee, Sung-Young;Yang, Dong-Uk;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • 제36권3호
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    • pp.291-297
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    • 2012
  • Ginsenoside (ginseng saponin), the principal component of ginseng, is responsible for the pharmacological and biological activities of ginseng. We isolated lactic acid bacteria from Kimchi using esculin agar, to produce ${\beta}$-glucosidase. We focused on the bio-transformation of ginsenoside. Phylogenetic analysis was performed by comparing the 16S rRNA sequences. We identified the strain as Lactobacillus (strain 6105). In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside Rb1 to catalyse the reaction. A carbon substrate, such as cellobiose, lactose, and sucrose, resulted in the highest yields of ${\beta}$-glucosidase activity. Biotransformations of ginsenoside Rb1 were analyzed using TLC and HPLC. Our results confirmed that the microbial enzyme of strain 6105 significantly transformed ginsenoside as follows: Rb1${\rightarrow}$gypenoside XVII, Rd${\rightarrow}$F2 into compound K. Our results indicate that this is the best possible way to obtain specific ginsenosides using microbial enzymes from 6105 culture.

Isolation and Characterization of Oligotrophic Bacteria Possessing Induced Systemic Disease Resistance against Plant Pathogens

  • Han, Song-Hee;Kang, Beom-Ryong;Lee, Jang-Hoon;Kim, Hyun-Jung;Park, Ju-Yeon;Kim, Jeong-Jun;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • 제28권1호
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    • pp.68-74
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    • 2012
  • Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

Diversity and Characterization of Endophytic Bacteria Associated with Tidal Flat Plants and their Antagonistic Effects on Oomycetous Plant Pathogens

  • Bibi, Fehmida;Yasir, Muhammad;Song, Geun-Cheol;Lee, Sang-Yeol;Chung, Young-Ryun
    • The Plant Pathology Journal
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    • 제28권1호
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    • pp.20-31
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    • 2012
  • Endophytic bacterial communities of tidal flat plants antagonistic to oomycete plant pathogens were studied by the isolation of 256 root colonizing endophytic bacteria from surface-disinfected root tissues of six plants ($Rosa$ $rugosa$, $Suaeda$ $maritima$, $Vitex$ $rotundifolia$, $Carex$ $scabrifolia$, $Glehnia$ $littoralis$ and $Elymus$ $mollis$) growing in a tidal flat area of Namhae Island, Korea. To understand the antagonistic potential, an $in$ $vitro$ antagonistic assay was performed to characterize and identify strains that were antagonistic to the oomycete plant pathogens $Phytophthora$ $capsici$ and $Pythium$ $ultimum$ from the total population. Nine percent of the total number of isolated bacteria exhibited in vitro inhibitory activity against target plant pathogenic oomycetes. Taxonomic and phylogenetic placement of the antagonistic bacteria was investigated by analysis of the 16S rRNA gene sequences. The sequence analysis classified the antagonistic strains into four major classes of the domain bacteria ($Firmicutes$, ${\alpha}-Proteobacteria$, ${\gamma}-Proteobacteria$ and $Actinomycetes$) and 10 different genera. Further production of secondary metabolites, hydrolytic enzymes and plant growth promoting traits were determined for the putative new species of antagonistic endophytic bacteria. These new strains could not be identified as known species of ${\alpha}-Proteobacteria$, and so may represent novel bacterial taxa. The unexpected high antagonistic bacterial diversity associated with the tidal flat plants may be indicative of their importance in tidal flat plants as a promising source of novel antimicrobial compounds and biocontrol agents.

디젤로 오염된 군부대 토양에 대하여 토착미생물 4종을 이용한 생분해법의 TPH 제거 효율 규명 (TPH Removal of the Biodegradation Process Using 4 Indigenous Microorganisms for the Diesel Contaminated Soil in a Military Camp)

  • 박민호;이민희
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제17권3호
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    • pp.49-58
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    • 2012
  • Batch experiments using indigenous and commercialized adventive microorganisms were performed to investigate the feasibility of the biodegradation process for the diesel contaminated soil, which was taken in US Military Camp 'Hialeah', Korea. TPH concentration of the soil was determined as 3,819 mg/kg. Four indigenous microorganisms having high TPH degradation activity were isolated from the soil and by 16S rRNA gene sequence analysis, they were identified as Arthrobacter sp., Burkholderia sp., Cupriavidus sp. and Bacillus sp.. Two kinds of commercialized solutions cultured with adventive microorganisms were also used for the experiments. Various biodegradation conditions such as the amount of microorganism, water content and the temperature were applied to decide the optimal bioavailability condition in the experiments. In the case of soils without additional microorganisms (on the natural attenuation condition), 35% of initial TPH was removed from the soil by inhabitant microorganisms in soil for 30 days. When the commercialized microorganism cultured solutions were added into the soil, their average TPH removal efficiencies were 64%, and 54%, respectively, which were higher than that without additional microorganisms. When indigenous microorganisms isolated from the contaminated soil were added into the soil, TPH removal efficiency increased up to 95% (for Bacillus sp.). According to the calculation of the average biodegradation rates for Bacillus sp., the remediation goal (87% of the removal efficiency: 500 mg/kg) for the soil would reach within 24 days. Results suggested that TPH removal efficiency of biodegradation by injecting indigenous microorganisms is better than those by injecting commercialized adventive microorganisms and only by using the natural attenuation.

멕시코 B 광산 슬러지에 존재하는 생물학적 광물화 미생물의 특성에 관한 연구 (Isolation and Identification of Bacteria Involved with Biomineralization at B Mine Sludge in Mexico)

  • 김준하;윤성열;박윤수;이재영
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제22권2호
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    • pp.41-51
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    • 2017
  • Microbial processes that bind heavy metals and form minerals are widespread, and they represent a basic aspect of biogeochemistry. Some microorganisms can crystallize minerals by secreting a specific enzyme. In particular, calcite ($CaCO_3$) precipitation is an important part of biomineralization, and has been studied extensively because of its wide application in civil engineering technology. This process provides an effective way to stabilize heavy metals within a relatively stable crystal phase. In this study, biomineralization of calcite by three urea-hydrolyzing indigenous bacterial strains was investigated by microbiological analyses. Three bacterial strains were isolated from the sludge of B mine in Mexico and each bacterial strain was identified by the cellular fatty acid composition and 16S rRNA partial sequencing analysis. The results of the identification analysis showed that these strains were closest to Sporosarcina pasteurii, Kurthia gibsonii, and Paenibacillus polymyxa. We found that the optimum conditions for growth of these indigenous bacteria were $30-40^{\circ}C$ and pH range of 7-8. Microbiological analyses showed the possibility that the bioaccumulated heavy metals ions were deposited around the cell as crystalline carbonate minerals under the optimum conditions. The findings of our study suggest that the indigenous bacterial strains play an important role in heavy metal immobilization.

교차오염방지를 위한 공기정화제품에서 미생물 오염도 분석 (Monitoring of Microbial Contamination in Air Purifier for Preventing Cross-contamination)

  • 염승목;김영목;이명숙
    • 한국식품위생안전성학회지
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    • 제31권3호
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    • pp.201-209
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    • 2016
  • 실내 공기질 관리의 중요성이 대두되면서 쾌적한 실내 환경에 도움을 주는 공기청정 기능과 습도 조절 기능을 동시에 갖춘 제습기와 공기청정기가 각광받고 있다. 하지만 오랜 기간 동안 공기정화제품을 사용하게 될 시에는 필터가 오염되어 본연의 기능을 상실하게 되는 것으로 알려져 있지만 이에 대한 구체적인 연구나 보고는 드문 편이다. 이에 본 연구에서는 가정과 사무실 등에서 사용한 공기정화제품을 수거하여 주요 부위별 미생물 오염도 및 주요 오염 미생물들을 분석하였다. 그 결과, 4 종류의 공기정화제품에서 오염도가 높은 부위는 필터부위, 물이 직접 닿는 부위 및 공기가 외부로부터 직접적으로 들어오는 입구부위 등에서 미생물학적 오염도가 가장 높았다. 하지만 공기정화제품은 사용하는 환경과 조건에 따라서 미생물학적 오염도 및 오염 미생물의 성상은 각각 다르게 나타났다. 하지만 이들 공기정화제품들에는 공통적으로 Staphylococcus sp., Micrococcus sp. 그리고 Bacillus sp.의 세균과 Cladosporium sp. 및 Penicillium sp.의 진균이 공통적으로 오염되어 있는 우점종인 것으로 분석되었다.

Amplified Ribosomal DNA Restriction Analysis (ARDRA) 방법을 이용한 국내 분리 Acinetobacter calcoaceticus - Acinetobacter baumannii Complex 균주의 유전자종 동정 (Genomic Species Identification of Acinetobacter calcoaceticus - Acinetobacter baumannii Complex Strains by Amplified Ribosomal DNA Restriction Analysis (ARDRA))

  • 오재영;조재위;박종천;이제철
    • 대한미생물학회지
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    • 제35권1호
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    • pp.69-76
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    • 2000
  • Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.

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2016년 한국 경기도의 3월 황사기간 동안 부유세균 군집과 다양성에 대한 메타지노믹 분석 (Metagenomic Analysis of Airborne Bacteria Community and Diversity in Gyeonggi-do, Korea, during March 2016, Asian Dust Event)

  • 장준형;김지혜;배경선;김정명;이원석;정현미;박상정;서태근
    • 한국환경보건학회지
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    • 제43권6호
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    • pp.491-498
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    • 2017
  • Objective: Bacterial abundance and community compositions have been examined in Asian dust events, clarifying their impacts on public health. This study aims to determine the bacterial community compositions and viable bacteria in Asian dust particles in the Asian dust or non-Asian dust event of March 2016. Methods: The dust samples were collected using the high volume air sampler or high volume cascade impactor, and bacterial 16S rRNA genes were amplified using PCR, followed by pyrosequencing. Bacterial diversity index, richness estimate and community composition in the particles were analyzed from the sequencing data using Mothur software. Results: The results showed that the diversity and richness during Asian dust events were higher than them in non-Asian dust events. The total bacterial community analysis showed that at the phylum Proteobacteria, Actinobacteria and Firmicutes were the most dominant of Asian dust events and non-Asian dust events. In addition, the bacterial colony counts were higher during Asian dust event, comparing with non-Asian dust event. Conclusions: This study showed that bacterial community and richness of Asian dust samples was more complex and higher than non-Asian dust samples in Gyeonggi-do, Korea, which could affect public health and environment. Thus, the continuous monitoring of Asian dust could be an alternative for managing airborne bacteria.