• Title/Summary/Keyword: Methylobacterium

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Biochemical and Molecular Characterization of High Population Density Bacteria Isolated from Sunflower

  • Goes, Kelly Campos Guerra Pinheiro De;Fisher, Maria Luisa De Castro;Cattelan, Alexandre Jose;Nogueira, Marco Antonio;Carvalho, Claudio Guilherme Portela De;Oliveira, Andre Luiz Martinez De
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.437-447
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    • 2012
  • Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plant-growth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 ${\mu}M$ auxin per ${\mu}g$ of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.

Isolation and Characterization of a Pink-Pigmented Facultative Methylotrophic Bacterium (분홍색 통성 메탄올 자화세균의 분리 및 특성)

  • 양석훈;김영민
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.63-69
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    • 1989
  • A pink-pigmented facultative methylotrophic bacterium, Methylobacterium sp. strain SY1, was isolated from soil through methanol-enrichment culture technique. The isolate was gram-negative, slightly curved rod, and motile by a single polarly inserted flagellum. The colony was smooth, bright pink, and slimy. The guanine plus cytosine content of the KNA was 66%. The cell was obigately aerobic and exhibited both catalase and oxidase activities. Carotenoid pigment and poly-$\beta$-hydroxybutyrate were present. It was found to have three kinds of plasmid with molecular weights 45,000, 38,500 and 23,000. Growth with methanol(0.5%) was fast ($t_{d}$=6.5h) and was optimal at $30^{\circ}C$ and at pH 7.0. The isolate could grow on several sugars, organic acids, amino acids, amines, and alcohols in addition to the methanol. Methanol was found to be assimilated through the serine pathway.

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Study on the Chlorine-Resistant Bacteria Isolated from Water Pipe Network (상수도관망에서 분리한 잔류염소 내성균에 관한 연구)

  • Hyun, Jae-Yeoul;Yoon, Jong-Ho
    • Journal of Korean Society on Water Environment
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    • v.27 no.3
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    • pp.334-341
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    • 2011
  • The free residual chlorine of tap water samples, collected from 266 faucets on the water pipe network in Daegu City, was between 0.1 and 0.79 mg/L. On microorganic tests, general bacteria and the coliform goup were not detected and thus the tap water was turned out to be fit to drink. In particular, samples of which free residual chlorine was 0.1 mg/L and over were cultured in R2A agar media at $25^{\circ}C$ for 7 days, and as a result heterotrophic bacteria were detected in 65.9% of samples; (1). The closer tap water got to the faucet from the stilling basin, the lower residual chlorine concentration became but the more the bacterial count became. And, more bacteria were detected in the R2A agar medium than in the PCA medium. (2). In the case of separated strains, most colonies were reddish or yellowish. 16S rRNA sequence was identified as Methylobacterium sp. and Williamsia sp., and yellow strain was identified as Sphingomonas sp., Sphingobium sp., Novosphingobium sp., Blastomonas sp., Rhodococcus sp. and Microbacterium sp. White strain was identified as Staphylococcus sp. (3). Sterilized tap water in polyethylene bottles was inoculated with separated strain and was left as it was for 2 months. As a result, bio-film was observed in tap water inoculated with Methylobacterium sp. and Sphingomonas sp. It was found that heterotrophic bacteria increased when free residual chlorine was removed from tap water in the water pipe network. Thus, there is a need to determine a base value for heterotrophic bacteria in order to check the cleanliness of tap water in the water pipe network.

Nodulation Experiment by Cross-Inoculation of Nitrogen-Fixing Bacteria Isolated from Root Nodules of Several Leguminous Plants

  • Ahyeon Cho;Alpana Joshi;Hor-Gil Hur;Ji-Hoon Lee
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.570-579
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    • 2024
  • Root-nodule nitrogen-fixing bacteria are known for being specific to particular legumes. This study isolated the endophytic root-nodule bacteria from the nodules of legumes and examined them to determine whether they could be used to promote the formation of nodules in other legumes. Forty-six isolates were collected from five leguminous plants and screened for housekeeping (16S rRNA), nitrogen fixation (nifH), and nodulation (nodC) genes. Based on the 16S rRNA gene sequencing and phylogenetic analysis, the bacterial isolates WC15, WC16, WC24, and GM5 were identified as Rhizobium, Sphingomonas, Methylobacterium, and Bradyrhizobium, respectively. The four isolates were found to have the nifH gene, and the study confirmed that one isolate (GM5) had both the nifH and nodC genes. The Salkowski method was used to measure the isolated bacteria for their capacity to produce phytohormone indole acetic acid (IAA). Additional experiments were performed to examine the effect of the isolated bacteria on root morphology and nodulation. Among the four tested isolates, both WC24 and GM5 induced nodulation in Glycine max. The gene expression studies revealed that GM5 had a higher expression of the nifH gene. The existence and expression of the nitrogen-fixing genes implied that the tested strain had the ability to fix the atmospheric nitrogen. These findings demonstrated that a nitrogen-fixing bacterium, Methylobacterium (WC24), isolated from a Trifolium repens, induced the formation of root nodules in non-host leguminous plants (Glycine max). This suggested the potential application of these rhizobia as biofertilizer. Further studies are required to verify the N2-fixing efficiency of the isolates.

Optimization of Growth Medium and Poly-$\beta$-hydroxybutyric Acid Production from Methanol in Methylobacterium organophilum (메탄올로부터 Methylobacterium organophilum에 의한 Poly-$\beta$-hydroxybutyric Acid의 생산과 배지성분의 최적화)

  • Choi, Joon-H;Kim, Jung H.;M. Daniel;J.M. Lebeault
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.392-396
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    • 1989
  • Methylobacterium organophilum, a facultative methylotroph was cultivated on a methanol as a sole carbon and energy source. The cell growth was affected by the various components of minimal synthetic medium and the medium composition was optimized with 0.5% (v/v) methanol at pH 6.8 and at 3$0^{\circ}C$. The maximum specific growth rate of M. organophilum was achieved to 0.26 hr$^{-1}$ in the optimized medium which has following composition: Methanol, 0.5% (v/v):(NH$_4$)$_2$SO$_4$, 1.0g/l:KH$_2$PO$_4$, 2.13g/l:KH$_2$PO$_4$, 1.305g/ι:MgSO$_4$.7$H_2O$. 45g/l and trace elements (CaCl$_2$.2$H_2O$, 3.3mg:FeSO$_4$.7$H_2O$, 1.3mg:MnSO$_4$.4$H_2O$, 130$\mu\textrm{g}$:ZnSO$_4$.5$H_2O$, 40$\mu\textrm{g}$:Na$_2$MoO$_4$.2$H_2O$, 40$\mu\textrm{g}$:CoCl$_2$.6$H_2O$, 40$\mu\textrm{g}$:H$_3$BO$_3$, 30$\mu\textrm{g}$ per liter). By the limitation of nitrogen and deficiency of Mn$^{+2}$ or Fe$^{+2}$, the cell growth was significantly repressed. Methanol greatly repressed the cell growth and the complete inhibition was observed at concentration above 4% (v/v). In order to overcome the methanol inhibition and to prevent the methanol limitation, intermittent feeding of methanol was conducted by a D.O.-stat technique. PHB production by M. organophilum was stimulated by deficiency of nutrients such as NH$_{4}^{+}$, SO$_{4}^{-2}$, $Mg^{+2}$, $K^{+}$, or PO$_{4}^{-3}$ in the medium. The maximum PHB content was obtained as 58% of dry cell weight under deficiency of potassium ion in the optimized synthetic medium.

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Correlation between Phylogeny and Zn-Resistance in Methylobacterium Species Isolated from Non-Polluted Soil Environments

  • Kim, Hong-Ik;Kazunori Nakamura;Hiroshi Oyaizu
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.10a
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    • pp.118-124
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    • 2000
  • Zn-resistant Methylosobacterium strains were isolated from several non-polluted soil samples collected in all over Japan. Zn-resistant Methylosobacterium strains were predominantly detected in all soil samples and they were also characterized as a UV-tolerant bacteria. The MIC test revealed that the isolates have high zinc resistance in comparison with that of reference Methylosobacterium strains obtained from culture collections. The 16S rDNA-based phylogenetic analysis showed that all strains were divided into two clusters designated as cluster A and cluster B in the present study. All isolates were distributed only in the cluster A. The phylogenetic clustering also well coincided with the differences in the pattern of carbon source utilization.

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The first record of nine bacterial species belonging to the phylum Proteobacteria in Korea

  • Kim, Dong-Uk;Kang, Myung-Suk;Kim, Ju-Young;Kim, Myung Kyum
    • Journal of Species Research
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    • v.6 no.3
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    • pp.214-223
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    • 2017
  • As part of a larger study with the aim to discover indigenous prokaryotic species in Korea, nine bacterial strains were isolated and assigned to the phylum Proteobacteria in 2016. High 16S rRNA gene sequence similarity (>98.5%) and formation of a robust phylogenetic clades with known species indicated that each strain belongs to an independent and predefined bacterial species. This is the first report of these nine species in Korea: two strains of the Methylobacterium, two strains of the Microvirga, one strain of the Pantoea, and four strains of the Psychrobacter, all within the Proteobacteria. Gram reaction, colony and cell morphology, basic biochemical characteristics, and isolation sources are also described in the species description section.

Growth kinetics and chlorine resistance of heterotrophic bacteria isolated from young biofilms formed on a model drinking water distribution system (모델 상수관망에 형성된 초기 생물막에서 분리한 종속영양세균의 생장 동역학 및 염소 내성)

  • Park, Se-Keun;Kim, Yeong-Kwan;Oh, Young-Sook;Choi, Sung-Chan
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.355-363
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    • 2015
  • The present work quantified the growth of young biofilm in a model distribution system that was fed with chlorinated drinking water at a hydraulic retention time of 2 h. Bacterial biofilms grew on the surface of polyvinyl chloride (PVC) slides at a specific growth rate of $0.14{\pm}0.09day^{-1}$ for total bacteria and $0.16{\pm}0.08day^{-1}$ for heterotrophic bacteria, reaching $3.1{\times}10^4cells/cm^2$ and $6.6{\times}10^3CFU/cm^2$ after 10 days, respectively. The specific growth rates of biofilm-forming bacteria were found to be much higher than those of bulk-phase bacteria, suggesting that biofilm bacteria account for a major part of the bacterial production in this model system. Biofilm isolates exhibited characteristic kinetic properties, as determined by ${\mu}_{max}$ and $K_S$ values using the Monod model, in a defined growth medium containing various amounts of acetate. The lowest ${\mu}_{max}$ value was observed in bacterial species belonging to the genus Methylobacterium, and their slow growth seemed to confer high resistance to chlorine treatment (0.5 mg/L for 10 min). $K_S$ values (inversely related to substrate affinity) of Sphingomonas were two orders of magnitude lower for acetate carbon than those of other isolates. The Sphingomonas isolates may have obligate-oligotrophic characteristics, since the lower $K_S$ values allow them to thrive under nutrient-deficient conditions. These results provide a better understanding and control of multi-species bacterial biofilms that develop within days in a drinking water distribution system.

A Study Regarding Bacterial Contamination of Surfaces in Dental Offices (치과진료실 내 표면 세균 오염에 관한 연구)

  • Yun, Kyoung-Ok;Kim, Hye-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.279-285
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    • 2015
  • This study studied samples taken off surfaces at three sites (Unit chairs, light handles, cuspidors) of 19 dental hospitals and 28 clinics located in Gyeonggi-do and Incheon, South Korea. The bacterial contamination levels of surfaces were $44.82{\times}10^3CFU/mL$ in cuspidors, higher than in unit chairs ($5.47{\times}10^3CFU/mL$) and light handles ($16.28{\times}10^3CFU/mL$). The values were statistically higher at dental hospitals than at dental clinics, the greater number of patients being associated with the higher bacterial cell count in the cuspidors. The results of identifying the strains isolated purely from surfaces at dental clinic showed Gram positive 47.3% and Gram negative 52.7%. Among Gram positive, the most numberous bacteria were Micrococcus luteus (10.9%), Bacillus pumilus (3.6%), and Staphylococcus aureus (3.6%). Among Gram negative, the most numberous bacteria were Acinetobacter ursingii (5.5%), Brevundimonas diminuta (4.5%), Chryseobacterium (Flavo.) indologenes (CDC IIb) (4.5%), and Methylobacterium sp. (4.5%). This study measures the level of bacterial contamination and identifies the strains isolated in dental clinics. It recognizes the importance of infection control, and the results of the study may be considered as the basis for establishing specific plans for prevention of infection.

Microbiological Water Quality of Water Purifiers at Elementary Schools in Gunsan Area (군산지역 초등학교 정수기 물의 미생물학적 수질)

  • Seo, Lan-Ju;Park, Suhk-Hwan;Lee, Geon-Hyoung
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.74-81
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    • 2009
  • In this research, we investigated the actual conditions of water purification systems at ten elementary schools located in Gunsan, Korea from July to December, 2007. The results were as follows; The population densities of heterotrophic bacteria in water purifiers ranged from 0 to $1.2{\pm}0.2{\times}10^4$ CFU/ml and those of tap water were in the range from 0 to $1.9{\pm}0.3{\times}10^4$ CFU/ml during investigation periods. Ninety percentage of purified water samples in July and September, 87.2% in October and November, and 93.7% in December turned out not to be suitable for drinking. The seasonal variation of the population densities of heterotrophic bacteria from purified waters was not notable. The total coliform, Salmonella and Shigella were not detected in purified water and tap water during investigation periods. Forty-five species of bacteria were isolated from water purifiers. The identified bacterial genera were Sphingomonas, Methylobacterium, Caulobacter, Novosphingobium, Bosea, Brevundimonas, Aminobacter, Ralstonia, Mitsuaria, Variovorax, Acidovorax, Massilia, Pseudomonas, Acinetobacter, Aeromonas, Bacillus, Staphylococcus, Brevibacillus, Microbacterium, Lapillicoccus, Micrococcus, Arthrobacter, Janibacter, Flavobacterium, Chryseobacterium, and Hymenobacter: Among the isolates, opportunistic pathogens such as Pseudomonas fluorescens, Staphylococcus epidermidis, Flavobacterium johnsoniae, and Acinetobacter johnsonii were also found.