• 제목/요약/키워드: dilution plating

검색결과 28건 처리시간 0.02초

Flavimonas oryzihabitans KU21의 원형질체 생성, 재생 및 융합 (Spheroplast Formation, Regeneration and Fusion of Flavimonas oryzihabitans KU21)

  • 이수연;임영복;박용근;이영록
    • 미생물학회지
    • /
    • 제31권4호
    • /
    • pp.318-325
    • /
    • 1993
  • 아닐린 분해균주 Flavimonas oryzihabitans KU21 의 원형질체 생성, 재생, 그리고 융합 등의 최적 조건을 조사하였다. 세포를 37.deg.C 로 prewarming 시킨 0.2 M Tris-Hcl(pH8.0) 완총액으로 현탁시킹후 0.5% 이상 원형질체로 전환되었으며, 이때 효소처리는 37.deg.C 에서 진탕하지 않고 처리했을 때가 가장 효과적이었다. MgCl/sub 2/ 와 CaCl/sub 2/ 수용액은 원형질체의 안정성을 높였고 완층액에 0.8% BSA 를 첨가함으로써 상온에서 4시간 까지 원형질체의 생존력을 80% 까지 유지할 수 있었다. 원형질체의 재생은 overlaying 방법이 가장 효과적이어서 3.8% 의 재생을 보였다. Rich regeneration medium 에 20 mM CaCl/sub 2/ 를 첨가하였을 때 재생율이 약 3.5배 증가하였고 완충액에 0.8% BSA 를 첨가했을 때는 4.5배의 증가율을 나타내었다. 삼투안정제로 0.5 M sucrose 를 첨가한 rich regeneration medium 에서 F. oryzihabitans 의 원형질체는 top plating 하여 6시나 이후부터 재생되기 시작하여 11 시간까지 80% 의 재생이 이루어졌다. 융합원으로 40.deg. PEC6000 과 CaCl/sub 2/ 를 사용하여 F, oryzihabitans 의 종내 원형질체 융합을 유도하였을때 recombinants 의 생성율은 2.0 * 10/sup -5/-3.6 * oryzihabitans 의 종내 원형질체 융합을 유도하였을때 recombinants 의 셍성율은 2.0 * 10/sup -5/-3.6 * 10/sup -5/ 이었으며 recombinants 들은 여러 세대 후에도 분리되지 않고 안정하였다.

  • PDF

Sufflavibacter maritimus gen. nov., sp. nov., Novel Flavobacteriaceae Bacteria Isolated from Marine Environments

  • Kwon, Kae-Kyoung;Yang, Seung-Jo;Lee, Hee-Soon;Cho, Jang-Cheon;Kim, Sang-Jin
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권8호
    • /
    • pp.1379-1384
    • /
    • 2007
  • Four Gram-negative, chemoheterotrophic, non-motile, yellow-colored strains were isolated from the East Sea or from deep-sea sediments of Nankai Trough by standard dilution plating. Characterization by polyphasic approaches indicated that the four strains are members of the same species. Phylogenetic analyses based on 16S rRNA gene sequences revealed that the strains formed a coherent and novel genus-level lineage within the family Flavobacteriaceae. The dominant cellular fatty acids were i-C15:0, 3-OH i-C17:0, and 2-OH i-C15:0 and/or C16:1 ${\omega}7c$. Predominance of 2-OH i-C15:0 and/or C16:1 ${\omega}7c$ clearly differentiated the strains from closely related members. The DNA G+C contents ranged 35.1-36.2 mol%. It is proposed, from the polyphasic evidence, that the strains should be placed into a novel genus and species named Sufflavibacter maritimus gen. nov., sp. nov., with strain $IMCC1001^T(=KCCM\;42359^T=NBRC\;102039^T)$ as the type strain.

구리 함유 활성 탄소 섬유의 항균 특성 (Antibacterial Activity of Activated Carbon Fibers Containing Copper Metal)

  • 박수진;김병주;이종문
    • 폴리머
    • /
    • 제27권3호
    • /
    • pp.235-241
    • /
    • 2003
  • 항균 활성이 없는 폴리아크릴로나이트릴계 활성 탄소 섬유에 항균 활성을 부여하기 위해 반응성이 큰 전이금속인 구리를 전해도금 방식으로 도입시켜, 활성 탄소 섬유의 항균 활성 및 기공 특성에 미치는 영향을 고찰해 보았다. 항균 활성 시험은 병원성 제균으로서 그람 양성균인 황색 포도상 구균 (Staphylococcus aureus)과 비병원성의 그람 음성균인 대장균 (Klebsiella pnemoniae)을 대상으로 그 효과를 측정하였으며, 활성 탄소 섬유의 기공 특성은 BET식, Boer의 t-plot, 그리고 H-K식을 이용하여 확인하였다. 도입되는 구리의 양이 많아질수록 활성 탄소 섬유의 비표면적, 총 기공 부피, 미세기공 부피 등이 감소되는 것이 관찰되었으며, 반면 항균 활성은 S. aureus 및 K. pnemoniae에서 증가되었다.

'Bring to Lab' of 19 Novel Species Among 60 Isolates Retrieved from a Freshwater Pond

  • Song, Jae-Ho;Yang, Seung-Jo;Cho, Jang-Cheon
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권1호
    • /
    • pp.168-175
    • /
    • 2007
  • We report here on the cultivation of numerous novel bacterial species from a eutrophic freshwater pond. A total of 60 strains, 15 strains per each culture medium, were obtained from the surface of a eutrophic freshwater pond by employing a conventional dilution-plating method with four different kinds of culture media, including R2A, 1/10R2A, PCA, and 1/10PCA. Among the 60 strains isolated, 27 strains showed less than 97% 16S rRNA gene sequence similarities to validly published species, and thus they are considered to comprise 19 novel species. Of the 27 strains assigned to the novel species, the majority of the strains (20 strains) were affiliated with the Alphaproteobacteria and Betaproteobacteria. The remaining 7 strains were affiliated with the Gammaproteobacteria, Firmicutes, Actinobacteria, and Deinococci. Because we have isolated 19 novel species from a usual freshwater pond using a conventional culturing technique, our results suggest that an unexplored ecosystem, even if it looks like a common ecosystem found elsewhere, harbors diverse unidentified microbes, which will be definitely further characterized.

Pseudomonas putida Strain 17 Isolated from Replant Soil Promotes Tomato Growth and Inhibits Conidial Germination of Soilborne Plant Pathogens

  • Lee, Sang-Woo;Ahn, Il-Pyung;Lim, Jae-Wook;Lee, Yong-Hwan
    • The Plant Pathology Journal
    • /
    • 제21권3호
    • /
    • pp.244-251
    • /
    • 2005
  • The induction of growth promotion on numerous crops by rhizobacteria is a well documented phenomenon. In case of tomato (Lycopersicon esculentum), fruit yield is higher in replant soil than that in fresh soil. To investigate what kind of rhizobacterium is involved, microbial community in rhizosphere and on rhizoplane of tomato plants from each soil was analyzed by dilution plating on selective media. Many Gram-negative bacteria and actinomycetes were isolated from tomato in replant soil. One Gram-negative rhizobacterium isolated was identified as Pseudomonas putida based on its biochemical characteristics, fatty acid methyl ester analysis and 16S rDNA sequence. This bacterium designated strain 17 inhibited the growth of Pseudomonas corrugata, and increased growth of tomato seedlings. In addition, its culture filtrate inhibited conidial germination of plant-pathogenic fungi such as Fusarium oxysporum f. sp. radicis-lycopersici, F. oxysporum f. sp. cucumerinum, and Nectria radicicola. Scanning electron microscopy revealed strain 17 colonized and persisted on the epidermal surfaces of tomato radicles and roots. These results suggest that P. putida strain 17 may serve as a biological control agent to suppress multiple soil-borne diseases for tomato plants. Increased microbial populations that suppress deleterious microorganisms including pathogens could be one of the major factors in increased tomato yield in replant soil.

A Super-Absorbent Polymer Combination Promotes Bacterial Aggressiveness Uncoupled from the Epiphytic Population

  • Lee, Bo-Young;Kim, Dal-Soo;Ryu, Choong-Min
    • The Plant Pathology Journal
    • /
    • 제24권3호
    • /
    • pp.283-288
    • /
    • 2008
  • Plant leaf surface is an important niche for diverse epiphytic microbes, including bacteria and fungi. Plant leaf surface plays a critical frontline defense against pathogen infections. The objective of our study was to evaluate the effectiveness of a starch-based super-absorbent polymer(SAP) combination, which enhances water potential and nutrient availability to plant leaves. We evaluated the effect of SAP on the maintenance of bacterial populations. In order to monitor bacterial populations in situ, a SAP mixture containing Pseudomonas syringae pv. tabaci that expressed recombinant green fluorescent protein(GFPuv) was spray-challenged onto whole leaves of Nicotiana benthamiana. The SAP combination treatment enhanced bacterial robustness, as indicated by disease severity and incidence. Unexpectedly, bacterial numbers were not significantly different between leaves treated with the SAP combination and those treated with water alone. Furthermore, young leaves treated with the SAP combination had more severe symptoms and a greater number of bacterial spots caused by primary and secondary infections compared to young leaves treated with the water control. In contrast, bacterial cell numbers did not statistically differ between the two groups, which indicated that measurement of viable GFP-based bacterial spots may provide a more sensitive methodology for assessing virulence of bacterial pathogens than methods that require dilution plating following maceration of bacterial-inoculated leaf tissue. Our study suggests that the SAP combination successfully increased bacterial aggressiveness, which could either be used to promote the ability of biological agents to control weedy plants or increase the robustness of saprophytic epiphytes against competition from potentially harmful microbes.

Rapid Quantification of Salmonella in Seafood Using Real-Time PCR Assay

  • Kumar, Rakesh;Surendran, P.K.;Thampuran, Nirmala
    • Journal of Microbiology and Biotechnology
    • /
    • 제20권3호
    • /
    • pp.569-573
    • /
    • 2010
  • A quantitative detection method for Salmonella in seafood was developed using a SYBR Green-based real-time PCR assay. The assay was developed using pure Salmonella DNA at different dilution levels [i.e., 1,000 to 2 genome equivalents (GE)]. The sensitivity of the real-time assay for Salmonella in seeded seafood samples was determined, and the minimum detection level was 20 CFU/g, whereas a detection level of 2 CFU/ml was obtained for pure culture in water with an efficiency of ${\geq}85%$. The real-time assay was evaluated in repeated experiments with seeded seafood samples and the regression coefficient ($R^2$) values were calculated. The performance of the real-time assay was further assessed with naturally contaminated seafood samples, where 4 out of 9 seafood samples tested positive for Salmonella and harbored cells <100 GE/g, which were not detected by direct plating on Salmonella Chromagar media. Thus, the method developed here will be useful for the rapid quantification of Salmonella in seafood, as the assay can be completed within 2-3 h. In addition, with the ability to detect a low number of Salmonella cells in seafood, this proposed method can be used to generate quantitative data on Salmonella in seafood, facilitating the implementation of control measures for Salmonella contamination in seafood at harvest and post-harvest levels.

S. mutans에 대한 대나무 숯의 항균력에 관한 연구 (Antibacterial effect of bamboo charcoal on Streptococcus mutans)

  • 최미숙;안권숙
    • 한국치위생학회지
    • /
    • 제14권1호
    • /
    • pp.95-100
    • /
    • 2014
  • Objectives : The purpose of the present study was to investigate the effect of bamboo charcoal on Streptococcus mutans which is one of the most important causative agents of dental caries. Methods : S. mutans was incubated with or without bamboo charcoal and then changes were observed in its cell viability and antibacterial effect. Oral epithelial cells viabillity(human gingival fibroblast, HGF) was performed using MTT assay. Antibacterial effect was analyzed using a dilution plating method and agar diffusion method. Results : Oral epithelial cells, human gingival fibroblast (HGF) showed a tendency to increase in bamboo charcoal treatment solution concentrations(0.5, 1, 2, 3, 5, 10%). The bamboo charcoal had an antibacterial effect on S. mutans. Antibacterial effect of bamboo charcoal for the bacterium was 58%. Charcoal concentration of 2% and 5% in the inhibition zone showed a minimal growth, but the concentration of 10% bamboo charcoal in inhibition zone revealed a conspicuous antibacterial activity. Conclusions : Overall results suggested that the bamboo charcoal proved to be bactericidal effect on S. mutans.

Confirmation of Two Undescribed Fungal Species from Dokdo of Korea Based on Current Classification System Using Multi Loci

  • Lee, Hye Won;Nguyen, Thi Thuong Thuong;Yeon, Hye;Lee, Haengsub;Kim, Changmu;Lee, Hyang Burm
    • Mycobiology
    • /
    • 제43권4호
    • /
    • pp.392-401
    • /
    • 2015
  • Using dilution plating method, 47 fungal isolates were obtained from a soil sample collected from Dokdo in the East Sea of Korea in 2013. In this study, two fungal isolates, EML-MFS30-1 and EML-DDSF4, were confirmed as undescribed species, Metarhizium guizhouense and Mortierella oligospora in Korea based on current classification system using multi loci including rDNA internal transcribed spacer, large subunit, small subunit, and ${\beta}$-tubulin (BTUB) genes. Herein, detailed morphological descriptions on characters of the undescribed fungal species as well as their molecular phylogenetic status are provided with comparisons to related species.

사람 평활면 치아우식에서 분리한 Neisseria sp. KEM232 균주의 유전체 서열 분석 (Complete genome sequence of Neisseria sp. KEM232 isolated from a human smooth surface caries)

  • 김은미;성치남
    • 미생물학회지
    • /
    • 제54권1호
    • /
    • pp.81-83
    • /
    • 2018
  • Neisseria 속 균주 KEM232는 사람 평활면 치아우식 부위로부터 분리하였다. 균주 KEM232의 유전체는 G + C 비율이 58.5%, 2,369개의 유전자와 2,210개의 단백질 코딩 유전자, 108개의 위유전자, 51개의 RNA 유전자 그리고 한 개의 CRISPR array를 포함한 단일 원형 염색체로 구성되었으며 그 크기는 2,371,912 bp였다. 균주 KEM232의 최 근연종은 Neisseria baciliformis 로서 두 균주 사이의 16S rRNA 유전자 염기서열의 유사도는 96.8% 그리고 유전체의 평균 염기 동일성은 84%였다.