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Sample records for nascent pseudomonas fluorescens

  1. 40 CFR 180.1114 - Pseudomonas fluorescens A506, Pseudomonas fluorescens 1629RS, and Pseudomonas syringae 742RS...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Pseudomonas fluorescens A506, Pseudomonas fluorescens 1629RS, and Pseudomonas syringae 742RS; exemptions from the requirement of a tolerance... Tolerances § 180.1114 Pseudomonas fluorescens A506, Pseudomonas fluorescens 1629RS, and Pseudomonas syringae...

  2. Growth of Pseudomonas fluorescens on Cassava Starch ...

    African Journals Online (AJOL)

    The potential of local strains of microorganism (Pseudomonas fluorescens) in polyhydroxbutyrate production was investigated in this study. This was with a view to establishing the capabilities of local strains of microorganisms on utilizing renewable and locally available substrates in polyhydroxybutyrate production.

  3. Characterization and identification of Pseudomonas fluorescens ...

    African Journals Online (AJOL)

    Crystal violet (CV), a recalcitrant molecule, used commonly as biological stain and commercial dye, was subjected to biodegradation in this study by Pseudomonas fluorescens NCIM 2100 and its biodegraded metabolic products were identified by UV, 1HNMR and IR Spectrophotometry. During degradation process CV dye ...

  4. Biosynthesis and regulation of cyclic lipopeptides in Pseudomonas fluorescens

    OpenAIRE

    Bruijn, de, I.

    2009-01-01

    Cyclic lipopeptides (CLPs) are surfactant and antibiotic metabolites produced by a variety of bacterial genera. For the genus Pseudomonas, many structurally different CLPs have been identified. CLPs play an important role in surface motility of Pseudomonas strains, but also in virulence and attachment/detachment to and from surfaces. In this Ph.D. thesis project, two new CLP biosynthesis clusters were identified in Pseudomonas fluorescens and fully sequenced. In P. fluorescens strain SBW2...

  5. Pseudomonas fluorescens' view of the periodic table.

    Science.gov (United States)

    Workentine, Matthew L; Harrison, Joe J; Stenroos, Pernilla U; Ceri, Howard; Turner, Raymond J

    2008-01-01

    Growth in a biofilm modulates microbial metal susceptibility, sometimes increasing the ability of microorganisms to withstand toxic metal species by several orders of magnitude. In this study, a high-throughput metal toxicity screen was initiated with the aim of correlating biological toxicity data in planktonic and biofilm cells to the physiochemical properties of metal ions. To this end, Pseudomonas fluorescens ATCC 13525 was grown in the Calgary Biofilm Device (CBD) and biofilms and planktonic cells of this microorganism were exposed to gradient arrays of different metal ions. These arrays included 44 different metals with representative compounds that spanned every group of the periodic table (except for the halogens and noble gases). The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum biofilm eradication concentration (MBEC) values were obtained after exposing the biofilms to metal ions for 4 h. Using these values, metal ion toxicity was correlated to the following ion-specific physicochemical parameters: standard reduction-oxidation potential, electronegativity, the solubility product of the corresponding metal-sulfide complex, the Pearson softness index, electron density and the covalent index. When the ions were grouped according to outer shell electron structure, we found that heavy metal ions gave the strongest correlations to these parameters and were more toxic on average than the other classes of the ions. Correlations were different for biofilms than for planktonic cells, indicating that chemical mechanisms of metal ion toxicity differ between the two modes of growth. We suggest that biofilms can specifically counter the toxic effects of certain physicochemical parameters, which may contribute to the increased ability of biofilms to withstand metal toxicity.

  6. Detection of Pseudomonas fluorescens from broth, water and ...

    African Journals Online (AJOL)

    Loop mediated isothermal amplification is rapid, highly sensitive and specifically developed method for detection of bacterial infections. AprX gene for alkaline metalloprotease of Pseudomonas fluorescens was used to design four primers and loop mediated isothermal amplification (LAMP) conditions were standardized for ...

  7. Detection of Pseudomonas fluorescens from broth, water and ...

    African Journals Online (AJOL)

    sonal

    2015-04-08

    Apr 8, 2015 ... Full Length Research Paper. Detection of Pseudomonas fluorescens from broth, water and infected tissues by loop-mediated isothermal amplification (LAMP) method. Pooja Saharan, Joginder S. Duhan and Suresh K. Gahlawat*. Department of Biotechnology, Chaudhary Devi Lal University, Sirsa-125 055, ...

  8. Adaptive synonymous mutations in an experimentally evolved Pseudomonas fluorescens population

    DEFF Research Database (Denmark)

    Bailey, Susan; Hinz, Aaron; Kassen, Rees

    2014-01-01

    in an experimentally evolved population of Pseudomonas fluorescens. We show experimentally that these mutations increase fitness by an amount comparable to non-synonymous mutations and that the fitness increases stem from increased gene expression. These results provide unequivocal evidence that synonymous mutations...

  9. Genome Sequence of the Biocontrol Strain Pseudomonas fluorescens F113

    Science.gov (United States)

    Redondo-Nieto, Miguel; Barret, Matthieu; Morrisey, John P.; Germaine, Kieran; Martínez-Granero, Francisco; Barahona, Emma; Navazo, Ana; Sánchez-Contreras, María; Moynihan, Jennifer A.; Giddens, Stephen R.; Coppoolse, Eric R.; Muriel, Candela; Stiekema, Willem J.; Rainey, Paul B.; Dowling, David; O'Gara, Fergal; Martín, Marta

    2012-01-01

    Pseudomonas fluorescens F113 is a plant growth-promoting rhizobacterium (PGPR) that has biocontrol activity against fungal plant pathogens and is a model for rhizosphere colonization. Here, we present its complete genome sequence, which shows that besides a core genome very similar to those of other strains sequenced within this species, F113 possesses a wide array of genes encoding specialized functions for thriving in the rhizosphere and interacting with eukaryotic organisms. PMID:22328765

  10. Lethality and Developmental Delay of Drosophila melanogaster Following Ingestion of Selected Pseudomonas fluorescens Strains

    Science.gov (United States)

    Pseudomonas fluorescens secretes antimicrobial compounds that promote plant health and provide protection from pathogens. We used a non-invasive feeding assay to study the toxicity of P. fluorescens strains Pf0-1, SBW25, and Pf-5 to Drosophila melanogaster. The three strains of P. fluorescens varie...

  11. 40 CFR 180.1200 - Pseudomonas fluorescens strain PRA-25; temporary exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Pseudomonas fluorescens strain PRA-25... RESIDUES IN FOOD Exemptions From Tolerances § 180.1200 Pseudomonas fluorescens strain PRA-25; temporary... established for residues of the microbial pesticide, pseudomonas fluorescens strain PRA-25 when used on peas...

  12. Combined inoculation of Pseudomonas fluorescens and Trichoderma harzianum for enhancing plant growth of vanilla (Vanilla planifolia).

    Science.gov (United States)

    Sandheep, A R; Asok, A K; Jisha, M S

    2013-06-15

    This study was conducted to evaluate the plant growth promoting efficiency of combined inoculation of rhizobacteria on Vanilla plants. Based on the in vitro performance of indigenous Trichoderma spp. and Pseudomonas spp., four effective antagonists were selected and screened under greenhouse experiment for their growth enhancement potential. The maximum percentage of growth enhancement were observed in the combination of Trichoderma harzianum with Pseudomonas fluorescens treatment followed by Pseudomonas fluorescens, Trichoderma harzianum, Pseudomonas putida and Trichoderma virens, respectively in decreasing order. Combined inoculation of Trichoderma harzianum and Pseudomonas fluorescens registered the maximum length of vine (82.88 cm), highest number of leaves (26.67/plant), recorded the highest fresh weight of shoots (61.54 g plant(-1)), fresh weight of roots (4.46 g plant(-1)) and dry weight of shoot (4.56 g plant(-1)) where as the highest dry weight of roots (2.0806 g plant(-1)) were achieved with treatments of Pseudomonas fluorescens. Among the inoculated strains, combined inoculation of Trichoderma harzianum and Pseudomonas fluorescens recorded the maximum nitrogen uptake (61.28 mg plant(-1)) followed by the combined inoculation of Trichoderma harzianum (std) and Pseudomonas fluorescens (std) (55.03 mg plant(-1)) and the highest phosphorus uptake (38.80 mg plant(-1)) was recorded in dual inoculation of Trichoderma harzianum and Pseudomonas fluorescens.

  13. Uji Lapangan Formula Cair Pseudomonas fluorescens P60 terhadap Layu Fusarium pada Tanaman Tomat

    Directory of Open Access Journals (Sweden)

    Loekas Soesanto

    2011-12-01

    Full Text Available A research aimed at knowing 1 the effect of Pseudomonas fluorescens P60 in liquid formula on Fusarium wilt of tomato, 2 the effect of P. fluorescens P60 in the formula on tomato growth and yield, and 3 P. fluorescens P60 mechanisms on tomata was carried out at tomato field of Selomoyo Village, Kaliangkrik Subdistrict, Magelang Regency at altitude of 826 m above sea level. Randomized block design was used with seven treatments and four replicates. The treatments were control, with P. fluorescens P60 soaked for 15 min and without fungicide, pathogen without P. fluorescens P60 with fungicide (PBG1, pathogen with P. fluorescens P60 without fungicide, pathogen with pouring P. fluorescens P60 1, 3, and 5 times. Result indicated that application of formulated P. fluorescens P60 for 5 times decreased the disease intensity as high as 26.77%, and late population of the pathogen but increased P. fluorescens P60 as high as 4.54×1010 cfu ml-1. P. fluorescens P60 affected growth and yield of tomato. P. Fluorescens P60 induced tomato resistance by increasing qualitatively its phenolic compound content (saponin, tannin, glycoside.   Penelitian dengan tujuan untuk mengetahui: 1 pengaruh Pseudomonas fluorescens P60 dalam formula cair terhadap penyakit layu fusarium pada tanaman tomat, 2 pengaruh P. fluorescens P60 dalam formula cair terhadap pertumbuhan dan produksi tanaman tomat, dan 3 mekanisme P. fluorescens P60 pada tanaman tomat dilakukan di lahan Desa Selomoyo, Kecamatan Kaliangkrik, Kabupaten Magelang dengan ketinggian 826 m di atas permukaan laut. Penelitian menggunakan Rancangan Acak Kelompok (RAK, dengan 7 perlakuan dan jumlah ulangan 4 kali, dan setiap unit terdiri atas 8 tanaman. Perlakuan tersebut meliputi kontrol; dengan P. fluorescens P60 rendam 15 menit dan tanpa fungisida; dengan patogen; tanpa P. fluorescens P60; dengan fungisida (PBG1; patogen, tanpa P. fluorescens P60, tanpa fungisida; patogen, dengan penyiraman P. fluorescens P60 1 kali

  14. Factors Affecting Zebra Mussel Kill by the Bacterium Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Daniel P. Molloy

    2004-02-24

    The specific purpose of this research project was to identify factors that affect zebra mussel kill by the bacterium Pseudomonas fluorescens. Test results obtained during this three-year project identified the following key variables as affecting mussel kill: treatment concentration, treatment duration, mussel siphoning activity, dissolved oxygen concentration, water temperature, and naturally suspended particle load. Using this latter information, the project culminated in a series of pipe tests which achieved high mussel kill inside power plants under once-through conditions using service water in artificial pipes.

  15. Effect of Pseudomonas fluorescens on Buried Steel Pipeline Corrosion.

    Science.gov (United States)

    Spark, Amy J; Law, David W; Ward, Liam P; Cole, Ivan S; Best, Adam S

    2017-08-01

    Buried steel infrastructure can be a source of iron ions for bacterial species, leading to microbiologically influenced corrosion (MIC). Localized corrosion of pipelines due to MIC is one of the key failure mechanisms of buried steel pipelines. In order to better understand the mechanisms of localized corrosion in soil, semisolid agar has been developed as an analogue for soil. Here, Pseudomonas fluorescens has been introduced to the system to understand how bacteria interact with steel. Through electrochemical testing including open circuit potentials, potentiodynamic scans, anodic potential holds, and electrochemical impedance spectroscopy it has been shown that P. fluorescens increases the rate of corrosion. Time for oxide and biofilms to develop was shown to not impact on the rate of corrosion but did alter the consistency of biofilm present and the viability of P. fluorescens following electrochemical testing. The proposed mechanism for increased corrosion rates of carbon steel involves the interactions of pyoverdine with the steel, preventing the formation of a cohesive passive layer, after initial cell attachment, followed by the formation of a metal concentration gradient on the steel surface.

  16. Genomic and Genetic Diversity within the Pseudomonas fluorescens Complex.

    Directory of Open Access Journals (Sweden)

    Daniel Garrido-Sanz

    Full Text Available The Pseudomonas fluorescens complex includes Pseudomonas strains that have been taxonomically assigned to more than fifty different species, many of which have been described as plant growth-promoting rhizobacteria (PGPR with potential applications in biocontrol and biofertilization. So far the phylogeny of this complex has been analyzed according to phenotypic traits, 16S rDNA, MLSA and inferred by whole-genome analysis. However, since most of the type strains have not been fully sequenced and new species are frequently described, correlation between taxonomy and phylogenomic analysis is missing. In recent years, the genomes of a large number of strains have been sequenced, showing important genomic heterogeneity and providing information suitable for genomic studies that are important to understand the genomic and genetic diversity shown by strains of this complex. Based on MLSA and several whole-genome sequence-based analyses of 93 sequenced strains, we have divided the P. fluorescens complex into eight phylogenomic groups that agree with previous works based on type strains. Digital DDH (dDDH identified 69 species and 75 subspecies within the 93 genomes. The eight groups corresponded to clustering with a threshold of 31.8% dDDH, in full agreement with our MLSA. The Average Nucleotide Identity (ANI approach showed inconsistencies regarding the assignment to species and to the eight groups. The small core genome of 1,334 CDSs and the large pan-genome of 30,848 CDSs, show the large diversity and genetic heterogeneity of the P. fluorescens complex. However, a low number of strains were enough to explain most of the CDSs diversity at core and strain-specific genomic fractions. Finally, the identification and analysis of group-specific genome and the screening for distinctive characters revealed a phylogenomic distribution of traits among the groups that provided insights into biocontrol and bioremediation applications as well as their role as

  17. ANOMALOUS BLUE COLOURING OF MOZZARELLA CHEESE INTENTIONALLY CONTAMINATED WITH PIGMENT PRODUCING STRAINS OF PSEUDOMONAS FLUORESCENS

    Directory of Open Access Journals (Sweden)

    P. Sechi

    2011-04-01

    Full Text Available In summer 2010 a large outbreak of anomalous blue coloration of mozzarella cheese was recorded in Italy and some northern European countries. Official laboratory analysis and health authorities linked the outbreak to the contamination of processing water with strains of Pseudomonas fluorescens, although several expert raised the question of how to unequivocally link the blue coloring to the presence of the micro-organism. In an attempt to set-up a method to determine whether a given Pseudomonas spp. strain is responsible of the defect, an in vitro system for the evaluation of blue colouring of mozzarella cheese intentionally contaminated with strains of Pseudomonas fluorescens. was developed The system is aimed to ascertain whether P. fluorescens strains, isolated from mozzarella cheese with anomalous blue coloration, are able to reproduce the blue coloration under controlled experimental condition. 96 trials of experimental inoculation of mozzarella cheese in different preservation liquids, were conducted using various suspension of Pseudomonas spp. (P. fluorescens ATCC 13525, P. fluorescens CFBP 3150, one P. fluorescens field strain isolated from blue-colored mozzarella cheese and P. aeruginosa ATCC 10145 as positive control at different concentrations and incubated at different temperatures. Growth curve of all Pseudomonas spp. strains tested demonstrated that after three days of incubation the concentration was generally higher than 106 CFU/g of mozzarella cheese incubated in Tryptic Soy Broth (TSB, and higher than 105 CFU/g of mozzarella cheese incubated in preservation liquid. All mozzarella cheeses inoculated with the field strain of Pseudomonas fluorescens showed the characteristic anomalous blue coloration, which is often associated with Pseudomonas fluorescens contamination of water used during mozzarella cheesemaking. With the proposed system, which enabled a considerable amount of samples to be analysed under controlled experimental

  18. Effects of cysteine on growth, protease production, and catalase activity of Pseudomonas fluorescens.

    OpenAIRE

    Himelbloom, B H; Hassan, H M

    1986-01-01

    Cysteine inhibits growth of and protease production by Pseudomonas fluorescens NC3. Catalase activity in P. fluorescens NC3 was increased by cysteine. The addition of exogenous hydrogen peroxide did not increase catalase activity, thus suggesting a role for the endogenous generation of hydrogen peroxide via the autoxidation of cysteine.

  19. Three Strains of Pseudomonas fluorescens Exhibit Differential Toxicity Against Drosophila melanogaster

    Science.gov (United States)

    Three strains of Pseudomonas fluorescens were tested for toxicity to Drosophila melanogaster in an insect feeding assay. Insect eggs were placed on the surface of a non-nutritive agar plate supplemented with a food source that was non-inoculated or inoculated with P. fluorescens Pf0-1, SBW25, or Pf-...

  20. High pressure inactivation of Pseudomonas in black truffle - comparison with Pseudomonas fluorescens in tryptone soya broth

    Science.gov (United States)

    Ballestra, Patricia; Verret, Catherine; Cruz, Christian; Largeteau, Alain; Demazeau, Gerard; El Moueffak, Abdelhamid

    2010-03-01

    Pseudomonas is one of the most common genera in black Perigord truffle. Its inactivation by high pressure (100-500 MPa/10 min) applied on truffles at sub-zero or low temperatures was studied and compared with those of Pseudomonas fluorescens in tryptone soya broth. Pressurization of truffles at 300 MPa/4 °C reduced the bacterial count of Pseudomonas by 5.3 log cycles. Higher pressures of 400 or 500 MPa, at 4 °C or 20 °C, allowed us to slightly increase the level of destruction to the value of ca. 6.5 log cycles but did not permit us to completely inactivate Pseudomonas. The results showed a residual charge of about 10 CFU/g. Pressure-shift freezing of truffles, which consists in applying a pressure of 200 MPa/-18 °C for 10 min and then quickly releasing this pressure to induce freezing, reduced the population of Pseudomonas by 3.3 log cycles. The level of inactivation was higher than those obtained with conventional freezing. Endogenous Pseudomonas in truffle was shown to be more resistant to high pressure treatments than P. fluorescens used for inoculation of broths.

  1. Boolean models of biosurfactants production in Pseudomonas fluorescens.

    Directory of Open Access Journals (Sweden)

    Adrien Richard

    Full Text Available Cyclolipopeptides (CLPs are biosurfactants produced by numerous Pseudomonas fluorescens strains. CLP production is known to be regulated at least by the GacA/GacS two-component pathway, but the full regulatory network is yet largely unknown. In the clinical strain MFN1032, CLP production is abolished by a mutation in the phospholipase C gene (plcC and not restored by plcC complementation. Their production is also subject to phenotypic variation. We used a modelling approach with Boolean networks, which takes into account all these observations concerning CLP production without any assumption on the topology of the considered network. Intensive computation yielded numerous models that satisfy these properties. All models minimizing the number of components point to a bistability in CLP production, which requires the presence of a yet unknown key self-inducible regulator. Furthermore, all suggest that a set of yet unexplained phenotypic variants might also be due to this epigenetic switch. The simplest of these Boolean networks was used to propose a biological regulatory network for CLP production. This modelling approach has allowed a possible regulation to be unravelled and an unusual behaviour of CLP production in P. fluorescens to be explained.

  2. Bioreporter pseudomonas fluorescens HK44 immobilized in a silica matrix

    Directory of Open Access Journals (Sweden)

    Trogl J.

    2003-01-01

    Full Text Available The bioluminescent bioreporter Pseudomonas fluorescens HK44, the whole cell bacterial biosensor that responds to naphthalene and its metabolites via the production of visible light, was immobilized into a silica matrix by the sol-gel technique. The bioluminescence intensities were measured in the maximum of the bioluminescence band at X = 500 nm. The immobilized cells (>105 cells per g silica matrix produced light after induction by salicylate (cone. > 10 g/l, naphthalene and aminobenzoic acid. The bioluminescence intensities induced by 2,3-dihydroxynaphthalene 3-hydroxybenzoic acid and 4-hydroxybenzoic acid were comparable to a negative control. The cells in the silica layers on glass slides produced light in response to the presence of an inductor at least 8 months after immobilization, and >50 induction cycles. The results showed that these test slides could be used as assays for the multiple determination of water pollution.

  3. Promotion of plant growth by Pseudomonas fluorescens strain SS101 via novel volatile organic compounds

    NARCIS (Netherlands)

    Park, Yong-Soon; Dutta, Swarnalee; Ann, Mina; Raaijmakers, Jos M.; Park, Kyungseok

    2015-01-01

    Abstract Volatile organic compounds (VOCs) from plant growth-promoting rhizobacteria (PGPR) play key roles in modulating plant growth and induced systemic resistance (ISR) to pathogens. Despite their significance, the physiological functions of the specific VOCs produced by Pseudomonas fluorescens

  4. Metabolic and transcriptomic changes induced in Arabidopsis by the rhizobacterium Pseudomonas fluorescens SS101

    NARCIS (Netherlands)

    Mortel, van de J.E.; Vos, de R.C.H.; Dekkers, E.; Pineda, A.; Guillod, L.; Bouwmeester, K.; Loon, van J.J.A.; Dicke, M.; Raaijmakers, J.M.

    2012-01-01

    Systemic resistance induced in plants by nonpathogenic rhizobacteria is typically effective against multiple pathogens. Here, we show that root-colonizing Pseudomonas fluorescens strain SS101 (Pf.SS101) enhanced resistance in Arabidopsis (Arabidopsis thaliana) against several bacterial pathogens,

  5. Discovery of new regulatory genes of lipopeptide biosynthesis in Pseudomonas fluorescens

    NARCIS (Netherlands)

    Song, C.; Aundy, K.; Mortel, van de J.E.; Raaijmakers, J.M.

    2014-01-01

    Pseudomonas fluorescens SS101 produces the cyclic lipopeptide massetolide with diverse functions in antimicrobial activity, motility, and biofilm formation. To understand how massetolide biosynthesis is genetically regulated in SS101, c. 8000 random plasposon mutants were screened for reduced or

  6. Pseudomonas putida and Pseudomonas fluorescens Species Group Recovery from Human Homes Varies Seasonally and by Environment.

    Directory of Open Access Journals (Sweden)

    Susanna K Remold

    Full Text Available By shedding light on variation in time as well as in space, long-term biogeographic studies can help us define organisms' distribution patterns and understand their underlying drivers. Here we examine distributions of Pseudomonas in and around 15 human homes, focusing on the P. putida and P. fluorescens species groups. We describe recovery from 10,941 samples collected during up to 8 visits per home, occurring on average 2.6 times per year. We collected a mean of 141 samples per visit, from sites in most rooms of the house, from the surrounding yards, and from human and pet occupants. We recovered Pseudomonas in 9.7% of samples, with the majority of isolates being from the P. putida and P. fluorescens species groups (approximately 62% and 23% of Pseudomonas samples recovered respectively. Although representatives of both groups were recovered from every season, every house, and every type of environment sampled, recovery was highly variable across houses and samplings. Whereas recovery of P. putida group was higher in summer and fall than in winter and spring, P. fluorescens group isolates were most often recovered in spring. P. putida group recovery from soils was substantially higher than its recovery from all other environment types, while higher P. fluorescens group recovery from soils than from other sites was much less pronounced. Both species groups were recovered from skin and upper respiratory tract samples from healthy humans and pets, although this occurred infrequently. This study indicates that even species that are able to survive under a broad range of conditions can be rare and variable in their distributions in space and in time. For such groups, determining patterns and causes of stochastic and seasonal variability may be more important for understanding the processes driving their biogeography than the identity of the types of environments in which they can be found.

  7. Pseudomonas putida and Pseudomonas fluorescens Species Group Recovery from Human Homes Varies Seasonally and by Environment.

    Science.gov (United States)

    Remold, Susanna K; Purdy-Gibson, Megan E; France, Michael T; Hundley, Thomas C

    2015-01-01

    By shedding light on variation in time as well as in space, long-term biogeographic studies can help us define organisms' distribution patterns and understand their underlying drivers. Here we examine distributions of Pseudomonas in and around 15 human homes, focusing on the P. putida and P. fluorescens species groups. We describe recovery from 10,941 samples collected during up to 8 visits per home, occurring on average 2.6 times per year. We collected a mean of 141 samples per visit, from sites in most rooms of the house, from the surrounding yards, and from human and pet occupants. We recovered Pseudomonas in 9.7% of samples, with the majority of isolates being from the P. putida and P. fluorescens species groups (approximately 62% and 23% of Pseudomonas samples recovered respectively). Although representatives of both groups were recovered from every season, every house, and every type of environment sampled, recovery was highly variable across houses and samplings. Whereas recovery of P. putida group was higher in summer and fall than in winter and spring, P. fluorescens group isolates were most often recovered in spring. P. putida group recovery from soils was substantially higher than its recovery from all other environment types, while higher P. fluorescens group recovery from soils than from other sites was much less pronounced. Both species groups were recovered from skin and upper respiratory tract samples from healthy humans and pets, although this occurred infrequently. This study indicates that even species that are able to survive under a broad range of conditions can be rare and variable in their distributions in space and in time. For such groups, determining patterns and causes of stochastic and seasonal variability may be more important for understanding the processes driving their biogeography than the identity of the types of environments in which they can be found.

  8. Expression analysis of the gacS mutant of Pseudomonas fluorescens SBW25

    NARCIS (Netherlands)

    Cheng, Xu; Bruijn, de Irene; Voort, van der M.; Raaijmakers, Jos

    2013-01-01

    Pseudomonas species are ubiquitous in plant-associated environments and produce an array of volatiles, enzymes and antimicrobials. The biosynthesis of many metabolites is regulated by the GacS/GacA two-component regulatory system. Transcriptome analysis of Pseudomonas fluorescens SBW25 revealed that

  9. The effect of phylogenetically different bacteria on the fitness of Pseudomonas fluorescens in sand microcosms.

    Directory of Open Access Journals (Sweden)

    Olaf Tyc

    Full Text Available In most environments many microorganisms live in close vicinity and can interact in various ways. Recent studies suggest that bacteria are able to sense and respond to the presence of neighbouring bacteria in the environment and alter their response accordingly. This ability might be an important strategy in complex habitats such as soils, with great implications for shaping the microbial community structure. Here, we used a sand microcosm approach to investigate how Pseudomonas fluorescens Pf0-1 responds to the presence of monocultures or mixtures of two phylogenetically different bacteria, a Gram-negative (Pedobacter sp. V48 and a Gram-positive (Bacillus sp. V102 under two nutrient conditions. Results revealed that under both nutrient poor and nutrient rich conditions confrontation with the Gram-positive Bacillus sp. V102 strain led to significant lower cell numbers of Pseudomonas fluorescens Pf0-1, whereas confrontation with the Gram-negative Pedobacter sp. V48 strain did not affect the growth of Pseudomonas fluorescens Pf0-1. However, when Pseudomonas fluorescens Pf0-1 was confronted with the mixture of both strains, no significant effect on the growth of Pseudomonas fluorescens Pf0-1 was observed. Quantitative real-time PCR data showed up-regulation of genes involved in the production of a broad-spectrum antibiotic in Pseudomonas fluorescens Pf0-1 when confronted with Pedobacter sp. V48, but not in the presence of Bacillus sp. V102. The results provide evidence that the performance of bacteria in soil depends strongly on the identity of neighbouring bacteria and that inter-specific interactions are an important factor in determining microbial community structure.

  10. Control of Fusarium verticillioides, cause of ear rot of maize, by Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Nayaka, Siddaiah Chandra; Shankar, Akarere C. Udaya; Reddy, Munagala S.

    2009-01-01

    and the formulations, in comparison with the control, increased plant growth and vigour as measured by seed germination, seedling vigour, plant height, 1000 seed weight and yield. P. fluorescens pure culture used as seed treatment and as spray treatment enhanced the growth parameters and reduced the incidence of F...... elicited considerable attention over the past decade owing to their association with animal disease syndromes. Hence, the present study was conducted to evaluate ecofriendly approaches by using a maize rhizosphere isolate of Pseudomonas fluorescens (Trev.) Mig. and its formulation to control ear rot....... The field trials included both seed treatment and foliar spray. For all the experiments, P. fluorescens was formulated using corn starch, wheat bran and talc powder. In each case there were three different treatments of P. fluorescens, a non-treated control and chemical control. RESULTS: Pure culture...

  11. Switching Catalysis from Hydrolysis to Perhydrolysis in Pseudomonas fluorescens Esterase

    Energy Technology Data Exchange (ETDEWEB)

    Yin, D.; Bernhardt, P; Morley, K; Jiang, Y; Cheeseman, J; Purpero, V; Schrag, J; Kazlauskas, R

    2010-01-01

    Many serine hydrolases catalyze perhydrolysis, the reversible formation of peracids from carboxylic acids and hydrogen peroxide. Recently, we showed that a single amino acid substitution in the alcohol binding pocket, L29P, in Pseudomonas fluorescens (SIK WI) aryl esterase (PFE) increased the specificity constant of PFE for peracetic acid formation >100-fold [Bernhardt et al. (2005) Angew. Chem., Int. Ed. 44, 2742]. In this paper, we extend this work to address the three following questions. First, what is the molecular basis of the increase in perhydrolysis activity? We previously proposed that the L29P substitution creates a hydrogen bond between the enzyme and hydrogen peroxide in the transition state. Here we report two X-ray structures of L29P PFE that support this proposal. Both structures show a main chain carbonyl oxygen closer to the active site serine as expected. One structure further shows acetate in the active site in an orientation consistent with reaction by an acyl-enzyme mechanism. We also detected an acyl-enzyme intermediate in the hydrolysis of {var_epsilon}-caprolactone by mass spectrometry. Second, can we further increase perhydrolysis activity? We discovered that the reverse reaction, hydrolysis of peracetic acid to acetic acid and hydrogen peroxide, occurs at nearly the diffusion limited rate. Since the reverse reaction cannot increase further, neither can the forward reaction. Consistent with this prediction, two variants with additional amino acid substitutions showed 2-fold higher k{sub cat}, but K{sub m} also increased so the specificity constant, k{sub cat}/K{sub m}, remained similar. Third, how does the L29P substitution change the esterase activity? Ester hydrolysis decreased for most esters (75-fold for ethyl acetate) but not for methyl esters. In contrast, L29P PFE catalyzed hydrolysis of {var_epsilon}-caprolactone five times more efficiently than wild-type PFE. Molecular modeling suggests that moving the carbonyl group closer to the

  12. Transport of a genetically modified Pseudomonas fluorescens and its parent strain through undisturbed tropical soil cores

    NARCIS (Netherlands)

    Guimaraes, V.F.; Cruz, I.V.; Hagler, A.N.; Mendonca-Hagler, L.C.; Elsas, van J.D.

    1997-01-01

    The transport of a genetically modified strain of Pseudomonas fluorescens, BR12, and its parent, BR5, was studied after irrigation of undisturbed clayey and sandy soil cores, simulating heavy tropical rainfall (56.6 mm/h). Vertical transport of both inoculant strains was detected in all soil cores.

  13. Effects of gamma irradiation on the survival of Pseudomonas fluorescens inoculated on romaine lettuce

    Science.gov (United States)

    Irradiation of fresh fruits and vegetables has been utilized as a post-harvest intervention measure for inactivation of microbial pathogens. Pseudomonas fluorescens 2-79, Q8R1, Q287 are non-plant pathogenic, non-pectolytic strains of a potential biocontrol agent. We tested the sensitivity of these s...

  14. Sensitivity of Pseudomonas fluorescens to gamma irradiation following surface inoculations on romaine lettuce and baby spinach

    Science.gov (United States)

    Irradiation of fresh fruits and vegetables is a post-harvest intervention measure often used to inactivate pathogenic food-borne microbes. We evaluated the sensitivity of Pseudomonas fluorescens strains (2-79, Q8R1, Q287) to gamma irradiation following surface inoculations on romaine lettuce and spi...

  15. Evolutionary history of the phl gene cluster in the plant-associated bacterium Pseudomonas fluorescens

    NARCIS (Netherlands)

    Moynihan, J.A.; Morrissey, J.P.; Coppoolse, E.; Stiekema, W.J.; O'Gara, F.; Boyd, E.F.

    2009-01-01

    Pseudomonas fluorescens is of agricultural and economic importance as a biological control agent largely because of its plant-association and production of secondary metabolites, in particular 2, 4-diacetylphloroglucinol (2, 4-DAPG). This polyketide, which is encoded by the eight gene phl cluster,

  16. Improved Performance of Pseudomonas fluorescens lipase by covalent immobilization onto Amberzyme

    NARCIS (Netherlands)

    Aslan, Yakup; Handayani, Nurrahmi; Stavila, Erythrina; Loos, Katja

    2013-01-01

    Objective: In this study, the conditions of covalent immobilization of Pseudomonas fluorescens lipase onto an oxirane-activated support (Amberzyme) were optimized to obtain a high activity yield. Furthermore, the operational and storage stabilities of immobilized lipase were tested. Methods: Optimum

  17. Water flow induced transport of Pseudomonas fluorescens cells through soil columns as affected by inoculant treatment

    NARCIS (Netherlands)

    Hekman, W.E.; Heijnen, C.E.; Trevors, J.T.; Elsas, van J.D.

    1994-01-01

    Water flow induced transport of Pseudomonas fluorescens cells through soil columns was measured as affected by the inoculant treatment. Bacterial cells were introduced into the topsoil of columns, either encapsulated in alginate beads of different types or mixed with bentonite clay in concentrations

  18. Contribution of the Pseudomonas fluorescens MFE01 Type VI Secretion System to Biofilm Formation.

    Directory of Open Access Journals (Sweden)

    Mathias Gallique

    Full Text Available Type VI secretion systems (T6SSs are widespread in Gram-negative bacteria, including Pseudomonas. These macromolecular machineries inject toxins directly into prokaryotic or eukaryotic prey cells. Hcp proteins are structural components of the extracellular part of this machinery. We recently reported that MFE01, an avirulent strain of Pseudomonas fluorescens, possesses at least two hcp genes, hcp1 and hcp2, encoding proteins playing important roles in interbacterial interactions. Indeed, P. fluorescens MFE01 can immobilise and kill diverse bacteria of various origins through the action of the Hcp1 or Hcp2 proteins of the T6SS. We show here that another Hcp protein, Hcp3, is involved in killing prey cells during co-culture on solid medium. Even after the mutation of hcp1, hcp2, or hcp3, MFE01 impaired biofilm formation by MFP05, a P. fluorescens strain isolated from human skin. These mutations did not reduce P. fluorescens MFE01 biofilm formation, but the three Hcp proteins were required for the completion of biofilm maturation. Moreover, a mutant with a disruption of one of the unique core component genes, MFE01ΔtssC, was unable to produce its own biofilm or inhibit MFP05 biofilm formation. Finally, MFE01 did not produce detectable N-acyl-homoserine lactones for quorum sensing, a phenomenon reported for many other P. fluorescens strains. Our results suggest a role for the T6SS in communication between bacterial cells, in this strain, under biofilm conditions.

  19. Pseudomonas fluorescens alters epithelial permeability and translocates across Caco-2/TC7 intestinal cells

    Directory of Open Access Journals (Sweden)

    Madi Amar

    2010-11-01

    Full Text Available Abstract Background Pseudomonas fluorescens has long been considered as a psychrotrophic microorganism. Recently, we have shown that clinical strains of P. fluorescens (biovar 1 are able to adapt at a growth temperature of 37°C or above and induce a specific inflammatory response. Interestingly, a highly specific antigen of P. fluorescens, I2, is detected in the serum of patients with Crohn's disease but the possible role of this bacterium in the disease has not yet been explored. In the present study, we examined the ability of a psychrotrophic and a clinical strain of P. fluorescens to modulate the permeability of a Caco-2/TC7 intestinal epithelial model, reorganize the actin cytoskeleton, invade the target cells and translocate across the epithelium. The behaviour of these two strains was compared to that of the well known opportunistic pathogen P. aeruginosa PAO1. Results Both strains of P. fluorescens were found to decrease the transepithelial resistance (TER of Caco-2/TC7 differentiated monolayers. This was associated with an increase in paracellular permeability and F-actin microfilaments rearrangements. Moreover, the invasion and translocation tests demonstrated that the two strains used in this study can invade and translocate across the differentiated Caco-2/TC7 cell monolayers. Conclusions The present work shows for the first time, that P. fluorescens is able to alter the intestinal epithelial barrier function by disorganizing the F-actin microfilament network. Moreover, we reveal that independently of their origins, the two P. fluorescens strains can translocate across differentiated Caco-2/TC7 cell monolayers by using the transcellular pathway. These findings could, at least in part, explain the presence of the P. fluorescens specific I2 antigen in the serum of patients with Crohn's disease.

  20. Iron-regulated metabolites of plant growth-promoting Pseudomonas fluorescens WCS374 : Their role in induced systemic resistance

    NARCIS (Netherlands)

    Djavaheri, M.

    2007-01-01

    The plant growth-promoting rhizobacterium Pseudomonas fluorescens WCS374r effectively suppresses fusarium wilt in radish by induced systemic resistance (ISR). In radish, WCS374r-mediated ISR depends partly on iron-regulated metabolites. Under iron-limiting conditions, P. fluorescens WCS374r produces

  1. HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS

    Directory of Open Access Journals (Sweden)

    ANTONIUS SUWANTO

    2003-01-01

    Full Text Available A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR. The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli carrying the recombinant plasmids with Pseudomotws fluorescens 5100, a phyllosphere bacterium, was performed. Pseudomonas fluorescens 5100 exconjugants were examined for constitutive expression of chitinase employing a spectrophotometric assay; they showed stronger chitin degradation activity than Escherichia coli transformants. Using a fungal antagonism plate assay, this chitinolytic P. fluorescens, however, could not inhibit selected phytopathogenic fungi.

  2. Pseudomine, an isoxazolidone with siderophoric activity from Pseudomonas fluorescens AH2 isolated from Lake Victorian Nile Perch

    DEFF Research Database (Denmark)

    Anthoni, U.; Christophersen, C.; Nielsen, P.H.

    1995-01-01

    A siderophore, pseudomonine, and sodium salicylate were isolated from the culture broth of iron-deficient cultures of Pseudomonas fluorescens AH2 isolated from the surface of spoiled Nile Perch from Lake Victoria......A siderophore, pseudomonine, and sodium salicylate were isolated from the culture broth of iron-deficient cultures of Pseudomonas fluorescens AH2 isolated from the surface of spoiled Nile Perch from Lake Victoria...

  3. Mutation Rate, Spectrum, Topology, and Context-Dependency in the DNA Mismatch Repair-Deficient Pseudomonas fluorescens ATCC948

    OpenAIRE

    Long, Hongan; Sung, Way; Miller, Samuel F.; Ackerman, Matthew S.; Doak, Thomas G.; Lynch, Michael

    2014-01-01

    High levels of genetic diversity exist among natural isolates of the bacterium Pseudomonas fluorescens, and are especially elevated around the replication terminus of the genome, where strain-specific genes are found. In an effort to understand the role of genetic variation in the evolution of Pseudomonas, we analyzed 31,106 base substitutions from 45 mutation accumulation lines of P. fluorescens ATCC948, naturally deficient for mismatch repair, yielding a base-substitution mutation rate of 2...

  4. Systematic investigations on the biodegradation and viscosity reduction of long chain hydrocarbons using Pseudomonas aeruginosa and Pseudomonas fluorescens.

    Science.gov (United States)

    Sakthipriya, N; Doble, Mukesh; Sangwai, Jitendra S

    2016-03-01

    The use of microorganisms has been researched extensively for possible applications related to hydrocarbon degradation in the petroleum industry. However, attempts to improve the effect of microorganisms on the viscosity of hydrocarbons, which find potential use in the development of robust models for biodegradation, have been rarely documented. This study investigates the degradation of long chain hydrocarbons, such as hexadecane and eicosane using Pseudomonas fluorescens PMMD3 (P. fluorescens) and Pseudomonas aeruginosa CPCL (P. aeruginosa). P. aeruginosa used here is isolated from petroleum contaminated sediments and the P. fluorescens is from the coastal area, and both have hydrocarbon degrading genes. The degradation of hydrocarbons is studied using carbon profiling and reduction in viscosity pre- and post-degradation of hydrocarbons. The carbon profiling has been obtained using gas chromatography-mass spectroscopy (GC-MS), and Fourier transform infrared spectrometer (FTIR) results. GC-MS results have indicated an improved biodegradation of hydrocarbons by 77-93% in one day. The yield coefficients of biomass (YX/S) for P. aeruginosa and P. fluorescens using hexadecane as a carbon source are 1.35 and 0.81 g g(-1), and the corresponding values with eicosane are 0.84 and 0.88 g g(-1). The viscosity of hexadecane is reduced by the order of 53 and 47%, while that of eicosane was reduced by 53 and 65%, using P. aeruginosa and P. fluorescens, respectively. This study also presents information on the activity of enzymes responsible for the hydrocarbon degradation. Pseudomonas species have shown their use in potential applications for bioremediation, oil-spill treatment, and flow assurance. We believe that this study will also provide stringent tests for possible model development for the bioremediation of long chain paraffins suitable for oilfield applications.

  5. Genomic and genetic analyses of diversity and plant interactions of Pseudomonas fluorescens.

    Science.gov (United States)

    Silby, Mark W; Cerdeño-Tárraga, Ana M; Vernikos, Georgios S; Giddens, Stephen R; Jackson, Robert W; Preston, Gail M; Zhang, Xue-Xian; Moon, Christina D; Gehrig, Stefanie M; Godfrey, Scott A C; Knight, Christopher G; Malone, Jacob G; Robinson, Zena; Spiers, Andrew J; Harris, Simon; Challis, Gregory L; Yaxley, Alice M; Harris, David; Seeger, Kathy; Murphy, Lee; Rutter, Simon; Squares, Rob; Quail, Michael A; Saunders, Elizabeth; Mavromatis, Konstantinos; Brettin, Thomas S; Bentley, Stephen D; Hothersall, Joanne; Stephens, Elton; Thomas, Christopher M; Parkhill, Julian; Levy, Stuart B; Rainey, Paul B; Thomson, Nicholas R

    2009-01-01

    Pseudomonas fluorescens are common soil bacteria that can improve plant health through nutrient cycling, pathogen antagonism and induction of plant defenses. The genome sequences of strains SBW25 and Pf0-1 were determined and compared to each other and with P. fluorescens Pf-5. A functional genomic in vivo expression technology (IVET) screen provided insight into genes used by P. fluorescens in its natural environment and an improved understanding of the ecological significance of diversity within this species. Comparisons of three P. fluorescens genomes (SBW25, Pf0-1, Pf-5) revealed considerable divergence: 61% of genes are shared, the majority located near the replication origin. Phylogenetic and average amino acid identity analyses showed a low overall relationship. A functional screen of SBW25 defined 125 plant-induced genes including a range of functions specific to the plant environment. Orthologues of 83 of these exist in Pf0-1 and Pf-5, with 73 shared by both strains. The P. fluorescens genomes carry numerous complex repetitive DNA sequences, some resembling Miniature Inverted-repeat Transposable Elements (MITEs). In SBW25, repeat density and distribution revealed 'repeat deserts' lacking repeats, covering approximately 40% of the genome. P. fluorescens genomes are highly diverse. Strain-specific regions around the replication terminus suggest genome compartmentalization. The genomic heterogeneity among the three strains is reminiscent of a species complex rather than a single species. That 42% of plant-inducible genes were not shared by all strains reinforces this conclusion and shows that ecological success requires specialized and core functions. The diversity also indicates the significant size of genetic information within the Pseudomonas pan genome.

  6. Getting the ecology into interactions between plants and the plant growth-promoting bacterium Pseudomonas fluorescens.

    Science.gov (United States)

    Hol, W H Gera; Bezemer, T Martijn; Biere, Arjen

    2013-01-01

    Plant growth-promoting rhizobacteria (PGPR) are increasingly appreciated for their contributions to primary productivity through promotion of growth and triggering of induced systemic resistance in plants. Here we focus on the beneficial effects of one particular species of PGPR (Pseudomonas fluorescens) on plants through induced plant defense. This model organism has provided much understanding of the underlying molecular mechanisms of PGPR-induced plant defense. However, this knowledge can only be appreciated at full value once we know to what extent these mechanisms also occur under more realistic, species-diverse conditions as are occurring in the plant rhizosphere. To provide the necessary ecological context, we review the literature to compare the effect of P. fluorescens on induced plant defense when it is present as a single species or in combination with other soil dwelling species. Specifically, we discuss combinations with other plant mutualists (bacterial or fungal), plant pathogens (bacterial or fungal), bacterivores (nematode or protozoa), and decomposers. Synergistic interactions between P. fluorescens and other plant mutualists are much more commonly reported than antagonistic interactions. Recent developments have enabled screenings of P. fluorescens genomes for defense traits and this could help with selection of strains with likely positive interactions on biocontrol. However, studies that examine the effects of multiple herbivores, pathogens, or herbivores and pathogens together on the effectiveness of PGPR to induce plant defenses are underrepresented and we are not aware of any study that has examined interactions between P. fluorescens and bacterivores or decomposers. As co-occurring soil organisms can enhance but also reduce the effectiveness of PGPR, a better understanding of the biotic factors modulating P. fluorescens-plant interactions will improve the effectiveness of introducing P. fluorescens to enhance plant production and defense.

  7. Inhibition of quorum sensing-controlled virulence factors and biofilm formation in Pseudomonas fluorescens by cinnamaldehyde.

    Science.gov (United States)

    Li, Tingting; Wang, Dangfeng; Liu, Nan; Ma, Yan; Ding, Ting; Mei, Yongchao; Li, Jianrong

    2018-03-23

    Pseudomonas fluorescens, an important food spoiling bacteria, uses quorum sensing to control biofilm formation and motility. To date, only a few compounds targeting the LuxR-based quorum sensing system of P. fluorescens have been identified. In the present study, the quorum sensing inhibitory effect of cinnamaldehyde at sublethal concentrations was investigated in terms of inhibition of the extracellular protease, biofilm formation, and swimming and swarming motility. The total volatile basic nitrogen value was also measured to evaluate the effect of cinnamaldehyde on quality preservation of turbot fillets stored at 4 ± 1 °C for 15 days. The results showed that cinnamaldehyde significantly inhibited quorum sensing-dependent factors in P. fluorescens and extended the storage life of turbot. Unexpectedly, cinnamaldehyde did not interfere with production of AHLs (N-acylhomoserine lactones) by P. fluorescens, as shown by measurement of AHL production using GC-MS. Molecular docking analysis revealed that cinnamaldehyde can interact with the LuxR-type protein of P. fluorescens, which could constitute the molecular basis of the quorum sensing inhibition observed. These findings strongly suggest that cinnamaldehyde is a quorum sensing inhibitor with great potential for the preservation of aquatic products to guarantee food safety. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Investigation of Pseudomonas fluorescens strain 3JW1 on preventing and reducing aflatoxin contaminations in peanuts.

    Directory of Open Access Journals (Sweden)

    Xiaona Yang

    Full Text Available Pseudomonas fluorescens strain 3JW1, which has a broad-spectrum antimicrobial activity, was studied to investigate whether it affects the amounts of aflatoxin B1 (AFB1 produced by Aspergillus flavus. It was found that the bacterium reduced the amounts of AFB1 in potato dextrose broth (PDB and peanut medium by 97.8% and 99.4%, respectively. It also reduced AFB1 by ~183 μg/kg (55.8% when applied onto peanut kernels. This strain reduced AFB1 via three mechanisms. First, it significantly inhibited A. flavus growth; second, our data showed that strain 3JW1 inhibits aflatoxin biosynthesis by A. flavus; and third, P. fluorescens strain 3JW1 is capable of degrading AFB1 at a rate as high as 88.3% in 96 hours. This is the first report demonstrating that Pseudomonas fluorescens can reduce toxin contamination caused by A. flavus on peanut kernels. Our findings indicate that P. fluorescens strain 3JW1 had multiple effects including reducing A. flavus infection and aflatoxin contamination. And the results also highlight the potential applications of the strain 3JW1 for the biological control of aflatoxin contamination in peanuts and other susceptible crops.

  9. Biosynthesis and regulation of cyclic lipopeptides in Pseudomonas fluorescens

    NARCIS (Netherlands)

    Bruijn, de I.

    2009-01-01

    Cyclic lipopeptides (CLPs) are surfactant and antibiotic metabolites produced by a variety of bacterial
    genera. For the genus Pseudomonas, many structurally different CLPs have been identified. CLPs play an
    important role in surface motility of Pseudomonas strains, but also in virulence

  10. Effect of Naphthalene and Salicylate Analogues on the Bioluminescence of Bioreporter Pseudomonas Fluorescens HK44.

    Czech Academy of Sciences Publication Activity Database

    Trögl, Josef; Kuncová, Gabriela; Kubicová, L.; Pařík, P.; Hálová, Jaroslava; Demnerová, K.; Ripp, S.; Sayler, G. S.

    2007-01-01

    Roč. 52, 1 (2007) , s. 3-14 ISSN 0015-5632 R&D Projects: GA ČR(CZ) GA104/05/2637; GA ČR(CZ) GA203/06/1244 Institutional research plan: CEZ:AV0Z40720504; CEZ:AV0Z40320502 Keywords : pseudomonas fluorescens HK44 * bioluminescence * bioluminescence Subject RIV: CE - Biochemistry Impact factor: 0.989, year: 2007

  11. Selectivity of Whole Cell Optical Biosensor with Immobilized Bioreporter Pseudomonas Fluorescens HK44

    Czech Academy of Sciences Publication Activity Database

    Trögl, Josef; Ripp, S.; Kuncová, Gabriela; Sayler, G. S.; Churavá, Alena; Pařík, P.; Demnerová, K.; Hálová, Jaroslava; Kubicová, L.

    2005-01-01

    Roč. 107, č. 1 (2005), s. 98-103 ISSN 0925-4005. [European Conference on Optical Chemical Sensors and Biosensors EUROPT(R)ODE /7./. Madrid, 04.04.2004-07.04.2004] R&D Projects: GA ČR(CZ) GA104/01/0461 Institutional research plan: CEZ:AV0Z40720504 Keywords : biosensor * Pseudomonas fluorescens HK44 * naphthalene Subject RIV: CE - Biochemistry Impact factor: 2.646, year: 2005

  12. Controlled-release fertilizers combined with Pseudomonas fluorescens rhizobacteria inoculum improve growth in Pinus halepensis seedlings

    OpenAIRE

    Dominguez-Nuñez JA; Delgado-Alvez D; Berrocal-Lobo M; Anriquez A; Albanesi A

    2015-01-01

    Pinus halepensis seedlings are currently used to regenerate arid Mediterranean regions. Optimized methods for seedling fertilization in nurseries improve plant growth and are essential for successful reforestation. Previously, we showed that inoculation of P. halepensis seedlings with Pseudomonas fluorescens CECT 844 rhizobacteria improved plant growth and N uptake. The aim of this study was to determine the physiological and morphological response of P. halepensis seedlings to a combined tre...

  13. Pseudomonas fluorescens in dairy products: a new case of blue mozzarella in Sardinia region

    Directory of Open Access Journals (Sweden)

    Chiara Nogarol

    2013-02-01

    Full Text Available Following a case of blue mozzarella occurred in Sardinia region, 14 isolates of Pseudomonas fluorescens have been isolated. Data analysis of pulsed-field-gel-electrophoresis profiles allowed to divide the isolates in two different clades, genetically unrelated. The presence of these two strains, deriving from nearby productions, confirmed the high diffusion of this microorganism. Multiples contamination sources (raw materials, processing surfaces and water supply made this specie one of the most relevant of the dairy productions chain.

  14. Isolation and characterization of a T7-like lytic phage for Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2008-10-01

    Full Text Available Abstract Background Despite the proven relevance of Pseudomonas fluorescens as a spoilage microorganism in milk, fresh meats and refrigerated food products and the recognized potential of bacteriophages as sanitation agents, so far no phages specific for P. fluorescens isolates from dairy industry have been closely characterized in view of their lytic efficiency. Here we describe the isolation and characterization of a lytic phage capable to infect a variety of P. fluorescens strains isolated from Portuguese and United States dairy industries. Results Several phages were isolated which showed a different host spectrum and efficiency of lysis. One of the phages, phage ϕIBB-PF7A, was studied in detail due to its efficient lysis of a wide spectrum of P. fluorescens strains and ribotypes. Phage ϕIBB-PF7A with a head diameter of about 63 nm and a tail size of about 13 × 8 nm belongs morphologically to the Podoviridae family and resembles a typical T7-like phage, as analyzed by transmission electron microscopy (TEM. The phage growth cycle with a detected latent period of 15 min, an eclipse period of 10 min, a burst size of 153 plaque forming units per infected cell, its genome size of approximately 42 kbp, and the size and N-terminal sequence of one of the protein bands, which gave similarity to the major capsid protein 10A, are consistent with this classification. Conclusion The isolated T7-like phage, phage ϕIBB-PF7A, is fast and efficient in lysing different P. fluorescens strains and may be a good candidate to be used as a sanitation agent to control the prevalence of spoilage causing P. fluorescens strains in dairy and food related environments.

  15. Genome-wide analysis of bacterial determinants of plant growth promotion and induced systemic resistance by Pseudomonas fluorescens

    NARCIS (Netherlands)

    Cheng, Xu; Etalo, Desalegn W.; van de Mortel, Judith E.; Dekkers, Ester; Nguyen, Linh; Medema, Marnix H; Raaijmakers, Jos M.

    2017-01-01

    Pseudomonas fluorescens strain SS101 (Pf.SS101) promotes growth of Arabidopsis thaliana, enhances greening and lateral root formation, and induces systemic resistance (ISR) against the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). Here, targeted and untargeted approaches were adopted to

  16. Inhibition of Vibrio anguillarum by Pseudomonas fluorescens AH2, a possible probiotic treatment of fish

    DEFF Research Database (Denmark)

    Gram, Lone; Melchiorsen, Jette; Spanggaard, Bettina

    1999-01-01

    To study the possible use of probiotics in fish farming, we evaluated the in vitro and in vivo antagonism of antibacterial strain Pseudomonas fluorescens strain AH2 against the fish- pathogenic bacterium Vibrio anguillarum. As iron is important in virulence and bacterial interactions, the effect....... fluorescens AH2 inhibited the growth of V. anguillarum during coculture, independently of the iron concentration, when the initial count of the antagonist was 100 to 1,000 times greater that of the fish pathogen. These in vitro results were successfully repeated in vivo. A probiotic effect in vivo was tested...... by exposing rainbow trout (Oncorynchus mykiss Walbaum) to P. fluorescens AH2 at a density of 10(5) CFU/ml for 5 days before a challenge with V. anguillarum at 10(4) to 10(5) CFU/ml for 1 h. Some fish were also exposed to P. fluorescens AH2 at 10(7) CFU/ml during the 1-h infection. The combined probiotic...

  17. Inactivation of Pseudomonas fluorescens in skim milk by combinations of pulsed electric fields and organic acids.

    Science.gov (United States)

    Fernández-Molina, Juan J; Altunakar, Bilge; Bermúdez-Aguirre, Daniela; Swanson, Barry G; Barbosa-Cánovas, Gustavo V

    2005-06-01

    Pseudomonas fluorescens suspended in skim milk was inactivated by application of pulsed electric fields (PEF) either alone or in combination with acetic or propionic acid. The initial concentration of microorganisms ranged from 10(5) to 10(6) CFU/ml. Addition of acetic acid and propionic acid to skim milk inactivated 0.24 and 0.48 log CFU/ml P. fluorescens, respectively. Sets of 10, 20, and 30 pulses were applied to the skim milk using exponentially decaying pulses with pulse lengths of 2 micros and pulse frequencies of 3 Hz. Treatment temperature was maintained between 16 and 20 degrees C. In the absence of organic acids, PEF treatment of skim milk at field intensities of 31 and 38 kV/cm reduced P. fluorescens populations by 1.0 to 1.8 and by 1.2 to 1.9 log CFU/ml, respectively. Additions of acetic and propionic acid to the skim milk in a pH range of 5.0 to 5.3 and PEF treatment at 31, 33, and 34 kV/cm, and 36, 37, and 38 kV/cm reduced the population of P. fluorescens by 1.4 and 1.8 log CFU/ml, respectively. No synergistic effect resulted from the combination of PEF with acetic or propionic acid.

  18. Interactions of trivalent and tetravalent heavy metal-siderophore complexes with pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Yoshida, T.; Ozaki, T.; Ohnuki, T.; Francis, A.J.

    2004-01-01

    We investigated the interactions of the Fe(III)-, Eu(III)-, and Hf(IV)-desferrioxamine B (DFO) complexes with the Gram-negative aerobic bacterium Pseudomonas fluorescens. Potentiometric titration of 1:1 Fe(III)-, Eu(III)-, and Hf(IV)-DFO complexes showed that Hf(IV) formed a strong complex with DFO whose stability was comparable to that of the Fe(III)-DFO complex, while Eu(III) formed a weaker one. DFO in a growth medium was not degraded by P. fluorescens. Contact of P. fluorescens cells with the Fe(III)-, Eu(III)-, and Hf(IV)-DFO complexes at pH 4-9 revealed that there was negligible adsorption of Hf(IV) and Fe(III), whereas Eu(III) was dissociated from DFO and was readily adsorbed by the cells. These results suggest that Fe(III) and Hf(IV) form stable complexes with DFO and are not adsorbed by P. fluorescens cells. Europium(III) forms a weaker complex with DFO than Fe(III) and Hf(IV) do and its DFO complex is readily dissociated in the presence of the cells. (orig.)

  19. Growth of Pseudomonas fluorescens in modified atmosphere packaged tofu.

    Science.gov (United States)

    Stoops, J; Maes, P; Claes, J; Van Campenhout, L

    2012-03-01

    The objective was to study the growth of Pseudomonas in a food product (tofu) where it typically occurs as a spoilage organism, and when this product is stored under modified atmosphere. A Pseudomonas strain was isolated from the endogenous microflora of tofu. Tofu was inoculated with the strain, packaged in different gas conditions (air, 100% N(2), 30% CO(2) /70% N(2) or 100% CO(2)) and stored under refrigerated conditions. Microbial loads and the headspace gas composition were monitored during storage. The strain was capable of growing in atmospheres containing no or limited amounts of oxygen and increased amounts of carbon dioxide. Even when 100% CO(2) was used, growth could not be inhibited completely. in contrast to the general characteristics of the genus Pseudomonas (strictly aerobic, highly sensitive to CO(2)), it should not be expected in the food industry that removing oxygen from the food package and increasing the carbon dioxide content, combined with cold storage, will easily avoid spoilage by Pseudomonas species. Guarantee of hygienic standards and combination of strategies with other microbial growth inhibiting measures should be implemented. © Kempen University College. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  20. Production and properties of biosurfactants from a newly isolated Pseudomonas fluorescens HW-6 growing on hexandecane

    Energy Technology Data Exchange (ETDEWEB)

    Vasileva-Tonkova, E.; Galabova, D. [Bulgarian Academy of Sciences, Dept. of Microbial Biochemistry, Sofia (Bulgaria); Stoimenova, E.; Lalchev, Z. [Dept. of Biochemistry, Sofia Univ. ' ' St. Kliment Ohridski' ' , Sofia (Bulgaria)

    2006-07-15

    The newly isolated from industrial wastewater Pseudomonas fluorescens strain HW-6 produced glycolipid biosurfactants at high concentrations (1.4-2.0 g 1{sup -1}) when grown on hexadecane as a sole carbon source. Biosurfactants decreased the surface tension of the air/water interface by 35 mN m{sup -1} and possessed a low critical micelle concentration value of 20 mg 1{sup -1}, which indicated high surface activity. They efficiently emulsified aromatic hydrocarbons, kerosene, n-paraffins and mineral oils. Biosurfactant production contributed to a significant increase in cell hydrophobicity correlated with an increased growth of the strain on hexadecane. The results suggested that the newly isolated strain of Ps. fluorescens and produced glycolipid biosurfactants with effective surface and emulsifying properties are very promising and could find application for bioremediation of hydrocarbon-polluted sites. (orig.)

  1. Interaction of the psychrotroph Pseudomonas fluorescens In5 with phytopathogens in cold soils

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Olsson, Stefan; Stougaard, Peter

    Aim: Potato cultivation in southwest Greenland, at Inneruulalik, omits the use of pesticides while relying on limited crop rotations and despite the presence of plant pathogens in the soil does not suffer from major disease outbreaks. Previously, we have shown that the soil at Inneruulalik...... is disease suppressive owing to the presence of diverse antimicrobial microorganisms. Using culture-based approaches, the psychrotroph Pseudomonas fluorescens In5 was previously isolated from the soil microbiome. The aim of this study is to unravel key biocontrol traits underpinning the antagonistic activity...... of this cold-active bacterium against phytopathogens. Method: A combination of different technologies including genomics, transcriptomics and metabolomics are being used to explore the interaction of the psychrotroph P. fluorescens In5 in dual-culture with diverse plant pathogens. To date, molecular genetics...

  2. Pseudomonas fluorescens biofilms subjected to phage phiIBB-PF7A

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2008-10-01

    Full Text Available Abstract Background Pseudomonas fluorescens is an important food spoilage organism, usually found in the form of biofilms. Bacterial biofilms are inherently resistant to a variety of antimicrobial agents, therefore alternative methods to biofilm control, such as bacteriophages (phages have been suggested. Phage behavior on biofilms is still poorly investigated and needs further understanding. Here we describe the application of phage ϕIBB-PF7, a newly isolated phage, to control P. fluorescens biofilms. The biofilms were formed under static or dynamic conditions and with or without renewal of medium. Results Conditions for biofilm formation influenced the feature of the biofilm and the morphology of P. fluorescens. Biomass removal due to phage activity varied between 63 and 91% depending on the biofilm age and the conditions under which the biofilm had been formed and phages applied. Removal of the biofilm by phage treatment was faster in younger biofilms, but the same number of surviving cells was detected in all tested biofilms, after only 4 h of treatment, even in older biofilms. Under static conditions, a 3 log higher number of phage progeny remained either inside the biofilm matrix or attached to the substratum surface than under dynamic conditions, pointing to the importance of experimental conditions for the efficacy of phage entrapment into the biofilm. Conclusion Phage ϕIBB-PF7A is highly efficient in removing P. fluorescens biofilms within a short time interval. The conditions of biofilm formation and applied during phage infection are critical for the efficacy of the sanitation process. The integration of phages into the biofilm matrix and their entrapment to the surface may be further beneficial factors when phage treatment is considered alone or in addition to chemical biocides in industrial environments where P. fluorescens causes serious spoilage.

  3. Genetic Control of Plant Root Colonization by the Biocontrol agent, Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Cole, Benjamin J.; Fletcher, Meghan; Waters, Jordan; Wetmore, Kelly; Blow, Matthew J.; Deutschbauer, Adam M.; Dangl, Jeffry L.; Visel, Axel

    2015-03-19

    Plant growth promoting rhizobacteria (PGPR) are a critical component of plant root ecosystems. PGPR promote plant growth by solubilizing inaccessible minerals, suppressing pathogenic microorganisms in the soil, and directly stimulating growth through hormone synthesis. Pseudomonas fluorescens is a well-established PGPR isolated from wheat roots that can also colonize the root system of the model plant, Arabidopsis thaliana. We have created barcoded transposon insertion mutant libraries suitable for genome-wide transposon-mediated mutagenesis followed by sequencing (TnSeq). These libraries consist of over 105 independent insertions, collectively providing loss-of-function mutants for nearly all genes in the P.fluorescens genome. Each insertion mutant can be unambiguously identified by a randomized 20 nucleotide sequence (barcode) engineered into the transposon sequence. We used these libraries in a gnotobiotic assay to examine the colonization ability of P.fluorescens on A.thaliana roots. Taking advantage of the ability to distinguish individual colonization events using barcode sequences, we assessed the timing and microbial concentration dependence of colonization of the rhizoplane niche. These data provide direct insight into the dynamics of plant root colonization in an in vivo system and define baseline parameters for the systematic identification of the bacterial genes and molecular pathways using TnSeq assays. Having determined parameters that facilitate potential colonization of roots by thousands of independent insertion mutants in a single assay, we are currently establishing a genome-wide functional map of genes required for root colonization in P.fluorescens. Importantly, the approach developed and optimized here for P.fluorescens>A.thaliana colonization will be applicable to a wide range of plant-microbe interactions, including biofuel feedstock plants and microbes known or hypothesized to impact on biofuel-relevant traits including biomass productivity

  4. Analysis of Bacillus thuringiensis Population Dynamics and Its Interaction With Pseudomonas fluorescens in Soil

    Science.gov (United States)

    Rojas-Ruiz, Norma Elena; Sansinenea-Royano, Estibaliz; Cedillo-Ramirez, Maria Lilia; Marsch-Moreno, Rodolfo; Sanchez-Alonso, Patricia; Vazquez-Cruz, Candelario

    2015-01-01

    Background: Bacillus thuringiensis is the most successful biological control agent, however, studies so far have shown that B. thuringiensis is very sensitive to environmental factors such as soil moisture and pH. Ultraviolet light from the sun had been considered as the main limiting factor for its persistence in soil and it has recently been shown that the antagonism exerted by other native soil organisms, such as Pseudomonas fluorescens, is a determining factor in the persistence of this bacterium under in vitro culture conditions. Objectives: The aim of the present investigation was to analyze the population dynamics of B. thuringiensis and its interaction with P. fluorescens using microbiological and molecular methods in soil, under different conditions, and to determinate the effect of nutrients and moisture on its interaction. Materials and Methods: The monitoring was performed by microbiological methods, such as viable count of bacteria, and molecular methods such as Polymerase Chain Reaction (PCR) and hybridization, using the direct extraction of DNA from populations of inoculated soil. Results: The analysis of the interaction between B. thuringiensis and P. fluorescens in soil indicated that the disappearance of B. thuringiensis IPS82 is not dependent on the moisture but the composition of nutrients that may be affecting the secretion of toxic compounds in the environment of P. fluorescens. The results showed that the recovered cells were mostly spores and not vegetative cells in all proved treatments. The molecular methods were effective for monitoring bacterial population inoculated in soil. Conclusions: Bacillus thuringiensis is very sensitive to the interaction of P. fluorescens, however is capable to survive in soil due to its capacity of sporulate. Some of the cells in the form of spores germinated and folded slightly and remained in a constant cycle of sporulation and germination. This confirms that B. thuringiensis IPS82 can germinate, grow and

  5. Transcriptomic analysis of the response of Pseudomonas fluorescens to epigallocatechin gallate by RNA-seq.

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    Xiaoxiang Liu

    Full Text Available Epigallocatechin gallate (EGCG is a main constituent of green tea polyphenols that are widely used as food preservatives and are considered to be safe for consumption. However, the underlying antimicrobial mechanism of EGCG and the bacterial response to EGCG are not clearly understood. In the present study, a genome-wide transcriptional analysis of a typical spoilage bacterium, Pseudomonas fluorescens that responded to EGCG was performed using RNA-seq technology. A total of 26,365,414 and 23,287,092 clean reads were generated from P. fluorescens treated with or without 1 mM EGCG and the clean reads were aligned to the reference genome. Differential expression analysis revealed 291 upregulated genes and 134 downregulated genes and the differentially expressed genes (DEGs were verified using RT-qPCR. Most of the DGEs involved in iron uptake, antioxidation, DNA repair, efflux system, cell envelope and cell-surface component synthesis were significantly upregulated by EGCG treatment, while most genes associated with energy production were downregulated. These transcriptomic changes are likely to be adaptive responses of P. fluorescens to iron limitation and oxidative stress, as well as DNA and envelope damage caused by EGCG. The expression of specific genes encoding the extra-cytoplasmic function sigma factor (PvdS, RpoE and AlgU and the two-component sensor histidine kinase (BaeS and RpfG were markedly changed by EGCG treatment, which may play important roles in regulating the stress responses of P. fluorescens to EGCG. The present data provides important insights into the molecular action of EGCG and the possible cross-resistance mediated by EGCG on P. fluorescens, which may ultimately contribute to the optimal application of green tea polyphenols in food preservation.

  6. Supervivencia de Pseudomonas fluorescens en suelos con diferente contenido de materia orgánica Pseudomonas fluorescens survival in soils with different contents of organic matter

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    E.B.R. Perotti

    2005-06-01

    Full Text Available Pseudomonas fluorescens es una bacteria PGPR (plant growth promoting rhizobacteria, heterótrofa, capaz de combatir fitopatógenos edáficos. Su supervivencia podría estar favorecida por el elevado contenido de materia orgánica del suelo (MOS. Para probarlo, se inocularon, en condiciones de laboratorio, tres cepas de P. fluorescens: UP61, C7R12, y P190 (nativa de Balcarce, Buenos Aires en suelos rizosféricos de tomate representativos de diferentes zonas de Argentina: suelo Argiudol (Balcarce, y Zavalla, Santa Fe y suelo Torrifluvens (Cipolletti, Río Negro (MOS %: 7,2; 4,3 y 2,6 respectivamente. Los resultados indicaron que la supervivencia de P. fluorescens en los suelos Argiudoles fue similar; aunque las pendientes de las curvas de supervivencia en el suelo de Zavalla fueron menores que las observadas en el suelo de Balcarce. La producción de CO2 fue superior en el suelo de Balcarce que en el suelo de Zavalla (4,3 y 2,8 mmol.g-1suelo, esta diferencia podría ser explicada por la existencia de una mayor presión competitiva por parte de la microflora nativa. La supervivencia en el suelo Torrifluvens resultó mínima, lo que sería atribuible a su elevada conductividad eléctrica más que al menor contenido de MOS. La cepa UP61 presentó en todos los casos la mejor supervivencia.Pseudomonas fluorescens are plant growth promoting rhizobacteria (PGPR. The survival of this inoculated heterotrophic bacterium may be affected by soil organic matter content (SOM. To confirm this hypothesis, three strains of P. fluorescens: UP61, C7R12 y P190 (native of Balcarce, Buenos Aires were inoculated, in laboratory conditions, into three argentine rhizospheric soils: two Argiudolls (Balcarce, and Zavalla, Santa Fe and a Torrifluvens (Cipolletti, Río Negro with different SOM: 7,2; 4,3; and 2,6%, respectibily. The results indicated that the all three isolates survival in general was not different. The slopes of the regression curves in Zavalla soil were very

  7. The Influence of Pseudomonas fluorescens on Corrosion Products of Archaeological Tin-Bronze Analogues

    Science.gov (United States)

    Ghiara, G.; Grande, C.; Ferrando, S.; Piccardo, P.

    2018-01-01

    In this study, tin-bronze analogues of archaeological objects were investigated in the presence of an aerobic Pseudomonas fluorescens strain in a solution, containing chlorides, sulfates, carbonates and nitrates according to a previous archaeological characterization. Classical fixation protocols were employed in order to verify the attachment capacity of such bacteria. In addition, classical metallurgical analytical techniques were used to detect the effect of bacteria on the formation of uncommon corrosion products in such an environment. Results indicate quite a good attachment capacity of the bacteria to the metallic surface and the formation of the uncommon corrosion products sulfates and sulfides is probably connected to the bacterial metabolism.

  8. Exposure-related effects of Pseudomonas fluorescens (Pf-CL145A) on juvenile unionid mussels

    Science.gov (United States)

    Weber, Kerry L.; Luoma, James A.; Mayer, Denise A.; Aloisi, Douglas B.; Eckert, Nathan L.

    2015-01-01

    The exposure-related effects of a commercially prepared spray-dried powder (SDP) or freeze-dried powder (FDP) formulation of Pseudomonas fluorescens (strain CL145A) on the survival of seven species of newly metamorphosed (<72 hours old) freshwater unionid mussels was evaluated. Forty unionid mussels of each species were randomly distributed to test chambers and each species independently exposed for 24 hours to a static dose of either SDP (four species: Lampsilis cardium, Lampsilis siliquoidea, Lampsilis higginsii, andLigumia recta) or FDP (three species: Obovaria olivaria, Actinonaias ligamentina, andMegalonaias nervosa).

  9. Biodegradation of didecyldimethylammonium chloride by Pseudomonas fluorescens TN4 isolated from activated sludge.

    Science.gov (United States)

    Nishihara, T; Okamoto, T; Nishiyama, N

    2000-04-01

    Bacteria that degrade didecyldimethylammonium chloride (DDAC) were isolated from activated sludge from a municipal sewage treatment plant by enrichment culture with DDAC as a sole carbon source. One of the isolates, Pseudomonas fluorescens TN4, degraded DDAC to produce decyldimethylamine and subsequently, dimethylamine, as the intermediates. The TN4 strain also assimilated the other quaternary ammonium compounds (QACs), alkyltrimethyl- and alkylbenzyldimethyl-ammonium salts, but not alkylpyridinium salts. TN4 was highly resistant to these QACs and degraded them by an N-dealkylation process. These data mean that there are some QAC-resistant and QAC-degrading bacteria such as TN4 in the environment.

  10. Sorption of Eu(III) on Pseudomonas fluorescens in the presence of citric acid

    International Nuclear Information System (INIS)

    Suzuki, Yoshinori; Tsushima, Satoru; Yamamoto, Ichiro; Nankawa, Takuya; Yoshida, Takahiro; Ozaki, Takuo; Ohnuki, Toshihiko; Francis, Arokiasamy J.; Enokida, Youichi

    2005-01-01

    We studied the sorption of Eu(III) on Pseudomonas fluorescens in the absence and presence of citric acid by a batch method. The cells were placed in a solution containing 2 μM of Eu(III) and 0, 100, or 1000 μM of citric acid at pH 3 9 for 5 hours. In the absence of citric acid, almost 100% of Eu(III) was sorbed on P. fluorescens at pHs below 7; above 7, sorption decreased with an increase in pH. The time course of Eu(III) sorption on P. fluorescens showed that a fraction of it was desorbed into the solution at alkaline pHs, suggesting that the bacterium may release some exudates. With citric acid present, we found that at higher concentrations there was lower sorption of Eu(III), reflecting the formation of Eu(III)-citrate complexes with the Eu(III)-cell-surface complexes. This decrease in Eu(III) sorption was significant in alkaline pHs. These findings suggest that citric acid which is ubiquitously found in the environment enhances migration of trivalent actinides in the alkaline environment. (author)

  11. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere

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    Hyun Gi Kong

    2016-04-01

    Full Text Available Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.

  12. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere.

    Science.gov (United States)

    Kong, Hyun Gi; Kim, Nam Hee; Lee, Seung Yeup; Lee, Seon-Woo

    2016-04-01

    Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.

  13. Rhizobacteria Pseudomonas fluorescens and Azospirillum sp. association enhances growth of Lactuca sativa L. under tropical conditions

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    Amael APONTE

    2017-06-01

    Full Text Available The selection of microorganisms that enhance plant growth and confer biotic and abiotic tolerance to crops constitutes a biotechnology currently gaining importance on a global scale. The aim of this investigation was to evaluate the effects of inoculating rhizobacteria to lettuce (Lactuca sativa L. on seed germination and vegetative development in order to use isolates as potential biofertilizers under tropical conditions. Five isolates of Pseudomonas fluorescens (Pf and one of Azospirillum sp. (Az were inoculated to seeds using a bacterial suspension of 1.5*108 CFU*mL-1. In vitro, none of the isolates promoted germination. In vivo, isolates promoted growth and acted as stress alleviators by conferring tolerance to high temperatures (≥ 30 °C. The highest seedling emergence percentages were induced by the association of P.fluorescens with Azospirillum. This association also promoted the highest leaf-area in 25 d seedlings and exhibited a significantly higher dry-weight in 40 d plants compared to the control (P≤0.05 supporting the advantages of bio-consortiums over individual strains. The strains were able to produce dependent L-tryptophan indole-3-acetic acid (IAA, to solubilize phosphorous in vitro and tolerated at least 5%-salt stress. The results indicate that isolate Pf (26 and Az possess plant growth promoting rhizobacteria (PGPR traits and should be further assessed. This study suggests that P. fluorescens and Azospirillum act synergically and are able to trigger an induced-tolerance mechanism in lettuce under abiotic stress.

  14. Nonribosomal peptides, key biocontrol components for Pseudomonas fluorescens In5, isolated from a Greenlandic suppressive soil.

    Science.gov (United States)

    Michelsen, Charlotte F; Watrous, Jeramie; Glaring, Mikkel A; Kersten, Roland; Koyama, Nobuhiro; Dorrestein, Pieter C; Stougaard, Peter

    2015-03-17

    Potatoes are cultivated in southwest Greenland without the use of pesticides and with limited crop rotation. Despite the fact that plant-pathogenic fungi are present, no severe-disease outbreaks have yet been observed. In this report, we document that a potato soil at Inneruulalik in southern Greenland is suppressive against Rhizoctonia solani Ag3 and uncover the suppressive antifungal mechanism of a highly potent biocontrol bacterium, Pseudomonas fluorescens In5, isolated from the suppressive potato soil. A combination of molecular genetics, genomics, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) imaging mass spectrometry (IMS) revealed an antifungal genomic island in P. fluorescens In5 encoding two nonribosomal peptides, nunamycin and nunapeptin, which are key components for the biocontrol activity by strain In5 in vitro and in soil microcosm experiments. Furthermore, complex microbial behaviors were highlighted. Whereas nunamycin was demonstrated to inhibit the mycelial growth of R. solani Ag3, but not that of Pythium aphanidermatum, nunapeptin instead inhibited P. aphanidermatum but not R. solani Ag3. Moreover, the synthesis of nunamycin by P. fluorescens In5 was inhibited in the presence of P. aphanidermatum. Further characterization of the two peptides revealed nunamycin to be a monochlorinated 9-amino-acid cyclic lipopeptide with similarity to members of the syringomycin group, whereas nunapeptin was a 22-amino-acid cyclic lipopeptide with similarity to corpeptin and syringopeptin. Crop rotation and systematic pest management are used to only a limited extent in Greenlandic potato farming. Nonetheless, although plant-pathogenic fungi are present in the soil, the farmers do not experience major plant disease outbreaks. Here, we show that a Greenlandic potato soil is suppressive against Rhizoctonia solani, and we unravel the key biocontrol components for Pseudomonas fluorescens In5, one of the potent biocontrol bacteria

  15. Identification and characterization of a novel nitrilase from Pseudomonas fluorescens Pf-5.

    Science.gov (United States)

    Kim, Jung-Soo; Tiwari, Manish Kumar; Moon, Hee-Jung; Jeya, Marimuthu; Ramu, Thangadurai; Oh, Deok-Kun; Kim, In-Won; Lee, Jung-Kul

    2009-05-01

    Nitrile groups are catabolized to the corresponding acid and ammonia through one-step reaction involving a nitrilase. Here, we report the use of bioinformatic and biochemical tools to identify and characterize the nitrilase (NitPf5) from Pseudomonas fluorescens Pf-5. The nitPf5 gene was identified via sequence analysis of the whole genome of P. fluorescens Pf-5 and subsequently cloned and overexpressed in Escherichia coli. DNA sequence analysis revealed an open-reading frame of 921 bp, capable of encoding a polypeptide of 307 amino acids residues with a calculated isoelectric point of pH 5.4. The enzyme had an optimal pH and temperature of 7.0 degrees C and 45 degrees C, respectively, with a specific activity of 1.7 and 1.9 micromol min(-1) mg protein(-1) for succinonitrile and fumaronitrile, respectively. The molecular weight of the nitrilase as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography was 33,000 and 138,000 Da, respectively, suggesting that the enzyme is homotetrameric. Among various nitriles, dinitriles were the preferred substrate of NitPf5 with a K (m) = 17.9 mM and k (cat)/K (m) = 0.5 mM(-1) s(-1) for succinonitrile. Homology modeling and docking studies of dinitrile and mononitrile substrate into the active site of NitPf5 shed light on the substrate specificity of NitPf5. Although nitrilases have been characterized from several other sources, P. fluorescens Pf-5 nitrilase NitPf5 is distinguished from other nitrilases by its high specific activity toward dinitriles, which make P. fluorescens NitPf5 useful for industrial applications, including enzymatic synthesis of various cyanocarboxylic acids.

  16. Study of the Population Dynamics of Listeria Monocytogenes and Pseudomonas Fluorescens in Buffalo Mozzarella by Means of Challenge Testing

    Science.gov (United States)

    Nava, Donatella; Capo, Salvatore; Caligiuri, Vincenzo; Giaccone, Valerio; Biondi, Loredana; Vaccaro, Gerardo Francesco; Guarino, Achille; Capuano, Federico

    2016-01-01

    Campania’s buffalo mozzarella is a greatly appreciated cheese in Italy and worldwide. From a microbiological standpoint, it is a highly perishable food and potentially at risk of contamination by pathogens such as Listeria monocytogenes (L. monocytogenes). The present paper reports the results of a challenge test carried out to assess the population dynamics of L. monocytogenes, alone and in presence of Pseudomonas fluorescens (P. fluorescens), in buffalo mozzarella. For this purpose buffalo mozzarella samples were contaminated with L. monocytogenes alone or combined with P. fluorescens. In samples where L. monocytogenes was inoculated alone, the bacterial load remained unchanged. On the contrary, in samples contaminated with L. monocytogenes and P. fluorescens, the growth of L. monocytogenes increased. PMID:27853707

  17. Study of the population dynamics of Listeria monocytogenes and Pseudomonas fluorescens in buffalo mozzarella by means of challenge testing

    Directory of Open Access Journals (Sweden)

    Donatella Nava

    2016-06-01

    Full Text Available Campania’s buffalo mozzarella is a greatly appreciated cheese in Italy and worldwide. From a microbiological standpoint, it is a highly perishable food and potentially at risk of contamination by pathogens such as Listeria monocytogenes (L. monocytogenes. The present paper reports the results of a challenge test carried out to assess the population dynamics of L. monocytogenes, alone and in presence of Pseudomonas fluorescens (P. fluorescens, in buffalo mozzarella. For this purpose buffalo mozzarella samples were contaminated with L. monocytogenes alone or combined with P. fluorescens. In samples where L. monocytogenes was inoculated alone, the bacterial load remained unchanged. On the contrary, in samples contaminated with L. monocytogenes and P. fluorescens, the growth of L. monocytogenes increased.

  18. Characterization of the biocontrol activity of pseudomonas fluorescens strain X reveals novel genes regulated by glucose.

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    Gerasimos F Kremmydas

    Full Text Available Pseudomonas fluorescens strain X, a bacterial isolate from the rhizosphere of bean seedlings, has the ability to suppress damping-off caused by the oomycete Pythium ultimum. To determine the genes controlling the biocontrol activity of strain X, transposon mutagenesis, sequencing and complementation was performed. Results indicate that, biocontrol ability of this isolate is attributed to gcd gene encoding glucose dehydrogenase, genes encoding its co-enzyme pyrroloquinoline quinone (PQQ, and two genes (sup5 and sup6 which seem to be organized in a putative operon. This operon (named supX consists of five genes, one of which encodes a non-ribosomal peptide synthase. A unique binding site for a GntR-type transcriptional factor is localized upstream of the supX putative operon. Synteny comparison of the genes in supX revealed that they are common in the genus Pseudomonas, but with a low degree of similarity. supX shows high similarity only to the mangotoxin operon of Ps. syringae pv. syringae UMAF0158. Quantitative real-time PCR analysis indicated that transcription of supX is strongly reduced in the gcd and PQQ-minus mutants of Ps. fluorescens strain X. On the contrary, transcription of supX in the wild type is enhanced by glucose and transcription levels that appear to be higher during the stationary phase. Gcd, which uses PQQ as a cofactor, catalyses the oxidation of glucose to gluconic acid, which controls the activity of the GntR family of transcriptional factors. The genes in the supX putative operon have not been implicated before in the biocontrol of plant pathogens by pseudomonads. They are involved in the biosynthesis of an antimicrobial compound by Ps. fluorescens strain X and their transcription is controlled by glucose, possibly through the activity of a GntR-type transcriptional factor binding upstream of this putative operon.

  19. Survival of a Rifampicin-Resistant Pseudomonas fluorescens Strain in Nine Mollisols

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    Tami L. Stubbs

    2014-01-01

    Full Text Available Pseudomonas fluorescens strain D7 (P.f. D7 is a naturally occurring soil bacterium that shows promise as a biological herbicide to inhibit growth of annual grass weeds, including downy brome (Bromus tectorum L., in crop- and rangelands. Pseudomonas fluorescens strain D7rif (P.f. D7rif is a rifampicin-resistant strain of P.f. D7. One of the greatest obstacles to successful biological weed control is survival of the organism under field conditions. Nine soils in the taxonomic order of Mollisols, collected from downy brome-infested areas of the Western and Central United States, were inoculated with P.f. D7rif and incubated in the laboratory to determine the effects of soil type, soil properties, incubation temperature, and soil water potential on survival of P.f. D7rif over 63 days. Silt loam soils from Lind, Washington, and Moro, Oregon, sustained the highest P.f. D7rif populations, and recovery was the lowest from Pendleton, Oregon soil. Survival and recovery of P.f. D7rif varied with soil type and temperature but not with the two soil water potentials tested. After 63 days, P.f. D7rif was recovered at levels greater than log 5.5 colony forming units (CFU g−1 soil from five of the nine test soils, a level adequate to suppress downy brome under field or range conditions.

  20. Kinetic modelling of enzyme inactivation : kinetics of heat inactivation of the extracellular proteinase from Pseudomonas fluorescens 22F

    NARCIS (Netherlands)

    Schokker, E.P.

    1997-01-01

    The kinetics of heat inactivation of the extracellular proteinase from Pseudomonas fluorescens 22F was studied. It was established, by making use of kinetic modelling, that heat inactivation in the temperature range 35 - 70 °C was most likely caused

  1. Gac-mediated changes in pyrroloquinoline quinone biosynthesis enhance the antimicrobial activity of Pseudomonas fluorescens SBW25

    NARCIS (Netherlands)

    Cheng, X.; Voort, van der M.; Raaijmakers, J.M.

    2015-01-01

    In Pseudomonas species, production of secondary metabolites and exoenzymes is regulated by the GacS/GacA two-component regulatory system. In P. fluorescens SBW25, mutations in the Gac-system cause major transcriptional changes and abolished production of the lipopeptide viscosin and of an

  2. Roles of Rhizoxin and 2,4-diacetylphloroglucinol in Suppression of Fusarium spp. by the Rhizobacterium Pseudomonas fluorescens Pf-5

    Science.gov (United States)

    Pseudomonas fluorescens strain Pf-5 is a rhizosphere bacterium that acts as a biocontrol agent of soilborne plant diseases and produces at least 10 different secondary metabolites, including several with antifungal properties. We derived site-directed mutants of Pf-5 with single and multiple mutatio...

  3. Characterization of toxin complex gene clusters and insect toxicity of bacteria representing four subgroups of Pseudomonas fluorescens

    Science.gov (United States)

    Ten strains representing four lineages of Pseudomonas (P. chlororaphis, P. corrugata, P. koreensis, and P. fluorescens subgroups) were evaluated for toxicity to the tobacco hornworm Manduca sexta and the fruit fly Drosophila melanogaster. The three strains within the P. chlororaphis subgroup exhibi...

  4. Assessment of DAPG-producing Pseudomonas fluorescens for management of Meloidogyne incognita and Fusarium oxysporum on watermelon

    Science.gov (United States)

    Pseudomonas fluorescens isolates Clinto 1R, Wayne 1R and Wood 1R, which produce the antibiotic 2,4-diacetylphloroglucinol (DAPG), can suppress soilborne diseases and promote plant growth. Consequently, these beneficial bacterial isolates were tested on watermelon plants for suppression of Meloidogy...

  5. LETHALITY OF PSEUDOMONAS FLUORESCENS STRAIN CLO145A TO THE 2 ZEBRA MUSSEL SPECIES PRESENT IN NORTH AMERICA

    Energy Technology Data Exchange (ETDEWEB)

    Daniel P. Molloy

    2001-10-28

    These experiments indicated that bacterial strain CL0145A of Pseudomonas fluorescens is equally lethal to the 2 zebra mussel species present in North America, Dreissena polymorpha and Dreissena bugensis. Thus, this bacterial strain should be equally effective at killing zebra mussels in power plant pipes, irrespective of which species is present.

  6. LETHALITY OF PSEUDOMONAS FLUORESCENS STRAIN CLO145A TO THE 2 ZEBRA MUSSEL SPECIES PRESENT IN NORTH AMERICA

    International Nuclear Information System (INIS)

    Molloy, Daniel P.

    2001-01-01

    These experiments indicated that bacterial strain CL0145A of Pseudomonas fluorescens is equally lethal to the 2 zebra mussel species present in North America, Dreissena polymorpha and Dreissena bugensis. Thus, this bacterial strain should be equally effective at killing zebra mussels in power plant pipes, irrespective of which species is present

  7. Metabolic engineering of Pseudomonas fluorescens for the production of vanillin from ferulic acid.

    Science.gov (United States)

    Di Gioia, Diana; Luziatelli, Francesca; Negroni, Andrea; Ficca, Anna Grazia; Fava, Fabio; Ruzzi, Maurizio

    2011-12-20

    Vanillin is one of the most important flavors in the food industry and there is great interest in its production through biotechnological processes starting from natural substrates such as ferulic acid. Among bacteria, recombinant Escherichia coli strains are the most efficient vanillin producers, whereas Pseudomonas spp. strains, although possessing a broader metabolic versatility, rapidly metabolize various phenolic compounds including vanillin. In order to develop a robust Pseudomonas strain that can produce vanillin in high yields and at high productivity, the vanillin dehydrogenase (vdh)-encoding gene of Pseudomonas fluorescens BF13 strain was inactivated via targeted mutagenesis. The results demonstrated that engineered derivatives of strain BF13 accumulate vanillin if inactivation of vdh is associated with concurrent expression of structural genes for feruloyl-CoA synthetase (fcs) and hydratase/aldolase (ech) from a low-copy plasmid. The conversion of ferulic acid to vanillin was enhanced by optimization of growth conditions, growth phase and parameters of the bioconversion process. The developed strain produced up to 8.41 mM vanillin, which is the highest final titer of vanillin produced by a Pseudomonas strain to date and opens new perspectives in the use of bacterial biocatalysts for biotechnological production of vanillin from agro-industrial wastes which contain ferulic acid. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L.

    Directory of Open Access Journals (Sweden)

    ANDHARE A. PRASAD

    Full Text Available ABSTRACT We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU and one biocontrol agent (Pseudomonas fluorescens strain PF1 for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

  9. Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L.).

    Science.gov (United States)

    Prasad, Andhare A; Babu, Subramanian

    2017-01-01

    We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU) and one biocontrol agent (Pseudomonas fluorescens strain PF1) for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

  10. Chitinolytic activity of Pseudomonas fluorescens isolates from barley and sugar beet rhizosphere.

    Science.gov (United States)

    Neiendam Nielsen M; Sørensen

    1999-11-01

    Twelve isolates of Pseudomonas fluorescens, isolated from barley and sugar beet rhizosphere as antagonists towards the plant pathogenic microfungi Rhizoctonia solani and Pythium ultimum, showed chitinolytic activity in batch cultures when grown in media without exogenous chitin. Enzyme tests in cultures demonstrated a complete array of chitinolytic enzymes. Endochitinase and chitobiosidase activities seemed to be tightly coupled in the isolates, while there was no relationship to N-acetyl-glucosaminidase activity. Endochitinase activity, showing hydrolysis of chitin polymers, was found to be extracellular in all isolates, although the most active isolates also retained a cell-bound fraction. Isoelectric focusing gel electrophoresis of supernatants containing extracellular enzyme activity showed that all isolates produced one native endochitinase in logarithmic phase. This enzyme was subsequently modified into several isozymes by extracellular processing as the cultures aged in stationary phase. The 12 isolates could be grouped into seven isozymic patterns. Detailed studies of three selected isolates showed that extracellular release of endochitinase activity also took place in stationary phase. The results, however, indicated that in stationary phase regulation of both the overall production of native enzyme and the subsequent formation of isozymes were different among the P. fluorescens isolates.

  11. Physiological and biochemical changes in Matricaria chamomilla induced by Pseudomonas fluorescens and water deficit stress

    Directory of Open Access Journals (Sweden)

    Hamid MOHAMMADI

    2018-04-01

    Full Text Available Environmental stresses and rhizosphere microorganisms affect growth parameters and accumulation of active ingredients especially in plants with medicinal properties. The present study examined the effects of chamomile (Matricaria chamomilla L. seedling inoculation with Pseudomonas fluorescens PF-135 strain on its growth parameters, photosynthetic pigments, proline, malondialdehyde (MDA, and hydrogen peroxide (H2O2 content, and essential oil concentration at both regular watering and water deficit experiments. Based on the obtained results, water deficit stress reduced root dry mass, and flower fresh and dry mass as well. However, amount of H2O2 and MDA in root and shoot tissues were considerably lower in inoculated plants compared to non-inoculated ones under both normal watering and water deficit regimes. It indicates that lipid peroxidation and production of reactive oxygen species has been diminished in inoculated plants. Also, essential oil content in inoculated plants significantly increased compared with that of non-inoculated ones under water deficit stress condition. It can be concluded that P. fluorescens PF-135 strain has an outstanding potential to alleviate adverse effects of water deficit on plant growth, and hence can be used as an excellent PGPR in order to boost chamomile productivity especially under water deficit stress condition.

  12. Three Alginate Lyases from Marine Bacterium Pseudomonas fluorescens HZJ216: Purification and Characterization

    Energy Technology Data Exchange (ETDEWEB)

    Liyan, Li [Ocean University of China, Qingdao, PRC; Jiang, Xiaolu [Ocean University of China, Qingdao, PRC; Wang, Peng [Ocean University of China, Qingdao, PRC; Guan, Huashi [Ocean University of China, Qingdao, PRC; Guo, Hong [ORNL

    2010-01-01

    Three alginate lyases (A, B, and C) from an alginate-degrading marine bacterium strain HZJ216 isolated from brown seaweed in the Yellow Sea of China and identified preliminarily as Pseudomonas fluorescens are purified, and their biochemical properties are described. Molecular masses of the three enzymes are determined by SDS-PAGE to be 60.25, 36, and 23 kDa with isoelectric points of 4, 4.36, and 4.59, respectively. Investigations of these enzymes at different pH and temperatures show that they are most active at pH 7.0 and 35 C. Alginate lyases A and B are stable in the pH range of 5.0 9.0, while alginate lyase C is stable in the pH range of 5.0 7.0. Among the metal ions tested, additions of Na+, K+, and Mg2+ ions can enhance the enzyme activities while Fe2+, Fe3+, Ba2+, and Zn2+ ions show inhibitory effects. The substrate specificity results demonstrate that alginate lyase C has the specificity for G block while alginate lyases A and B have the activities for both M and G blocks. It is the first report about extracellular alginate lyases with high alginate-degrading activity from P. fluorescens.

  13. When cheese gets the blues: Pseudomonas fluorescens as the causative agent of cheese spoilage.

    Science.gov (United States)

    Martin, N H; Murphy, S C; Ralyea, R D; Wiedmann, M; Boor, K J

    2011-06-01

    A bacterial contamination of fresh, low-acid cheese that resulted in production of a blue fluorescent pigment on the surface of the cheese was determined to be caused by Pseudomonas fluorescens biovar IV, a gram-negative bacteria that produces a blue, nondiffusible pigment as well as the soluble pigment pyoverdin, which fluoresces under UV light. Ten isolates collected from contaminated cheese and environmental samples were initially identified as P. fluorescens using 16S rDNA sequencing, but only 8 of the isolates produced blue pigment and fluoresced under UV light when re-inoculated onto fresh, low-acid cheese. The Biolog Metabolic Fingerprint system (Biolog Inc., Hayward, CA) and the Analytical Profile Index (BioMerieux Vitek Inc., Hazelwood, MO) for nonenteric gram-negative species as well as EcoRI ribotyping did not differentiate between the isolates that produced blue color and those that did not. Pulsed field gel electrophoresis with the enzyme XbaI was able to distinguish between the isolates that produced pigment and those that did not and allowed for identification of a specific environmental site (i.e., an overhead cheese vat agitator system) as the likely source of product contamination. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Antimicrobial activity of olive mill wastewater extract against Pseudomonas fluorescens isolated from mozzarella cheese

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    Rossana Roila

    2016-05-01

    Full Text Available Olive mill wastewater polyphenol extract was tested for antimicrobial activity against 64 strains of Pseudomonas fluorescens responsible for mozzarella discolouration. The extract showed a minimum inhibitory concentration (MIC50 value of 5 mg/mL and a MIC90 value of 7 mg/mL. The MBC50 and MBC90 values corresponded to 6 and 8 mg/mL, respectively. The MIC concentration (7 mg/mL was demonstrated to have a bacteriostatic effect while maintaining the bacterial concentration on the levels of the inoculum for 48 hours. The 3/2 MIC concentration was responsible for four logs CFU/mL depletion in colony count after 24 h. As the extract concentration decreased from MIC value, no inhibitory effects were recorded.

  15. Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48

    Science.gov (United States)

    Gamal, Rawia F.; Abdelhady, Hemmat M.; Khodair, Taha A.; El-Tayeb, Tarek S.; Hassan, Enas A.; Aboutaleb, Khadiga A.

    2013-01-01

    The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO) as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%). Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L) at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%). Semi-scale application (10 L working volume) increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform–hypochlorite dispersion extraction. Gas chromatography (GC) analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%). A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98–99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838. PMID:24294253

  16. Exposure-related effects of formulated Pseudomonas fluorescens strain CL145A to glochidia from seven unionid mussel species

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Severson, Todd J.; Schreier, Theresa M.; Mayer, Denise A.; Aloisi, Douglas B.; Eckert, Nathan L.

    2015-01-01

    The study was completed to evaluate the exposure-related effects of a biopesticide for dreissenid mussel (Dreissena polymorpha, zebra mussel and Dreissena rostriformis bugensis, quagga mussel) control on glochidia from unionid mussels endemic to the Great Lakes and Upper Mississippi River Basins. The commercially prepared biopesticide was either a spray-dried powder (SDP) or freeze-dried powder (FDP) formulation of Pseudomonas fluorescens, strain CL145A. Glochidia of the unionid mussel species Lampsilis cardium, Lampsilis siliquoidea,Lampsilis higginsii, Ligumia recta, Obovaria olivaria, and Actinonaias ligamentina were exposed to SDP-formulated P. fluorescens andLampsilis cardium and Megalonaias nervosa were exposed to FDP-formulated P. fluorescens.

  17. Antigen 43 from Escherichia coli induces inter- and intraspecies cell aggregation and changes in colony morphology of Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Kjærgaard, Kristian; Schembri, Mark; Hasman, Henrik

    2000-01-01

    with a distinct frizzy colony morphology in E. coli. Here we show that Ag43 can be expressed in a functional form on the surface of the environmentally important Pseudomonas fluorescens strain SBW25 with ensuing cell aggregation and frizzy colony types. Using green fluorescence protein-tagged cells, we......Antigen 43 (Ag43) is a surface-displayed autotransporter protein of Escherichia coli. By virtue of its self-association characteristics, this protein is able to mediate autoaggregation and flocculation off. coli cells in static cultures. Additionally, surface display of Ag43 is associated...... demonstrate that Ag43 can be used as a tool to provide interspecies cell aggregation between E. coli and P. fluorescens. Furthermore, Ag43 expression enhances biofilm formation in P. fluorescens to glass surfaces. The versatility of this protein was also reflected in Ag43 surface display in a variety of other...

  18. Pseudomonas fluorescens strain CL145A - a biopesticide for the control of zebra and quagga mussels (Bivalvia: Dreissenidae).

    Science.gov (United States)

    Molloy, Daniel P; Mayer, Denise A; Gaylo, Michael J; Morse, John T; Presti, Kathleen T; Sawyko, Paul M; Karatayev, Alexander Y; Burlakova, Lyubov E; Laruelle, Franck; Nishikawa, Kimi C; Griffin, Barbara H

    2013-05-01

    Zebra mussels (Dreissena polymorpha) and quagga mussels (Dreissena rostriformis bugensis) are the "poster children" of high-impact aquatic invasive species. In an effort to develop an effective and environmentally acceptable method to control their fouling of raw-water conduits, we have investigated the potential use of bacteria and their natural metabolic products as selective biological control agents. An outcome of this effort was the discovery of Pseudomonas fluorescens strain CL145A - an environmental isolate that kills these dreissenid mussels by intoxication (i.e., not infection). In the present paper, we use molecular methods to reconfirm that CL145A is a strain of the species P. fluorescens, and provide a phylogenetic analysis of the strain in relation to other Pseudomonas spp. We also provide evidence that the natural product lethal to dreissenids is associated with the cell wall of P. fluorescens CL145A, is a heat-labile secondary metabolite, and has degradable toxicity within 24 h when applied to water. CL145A appears to be an unusual strain of P. fluorescens since it was the only one among the ten strains tested to cause high mussel mortality. Pipe trials conducted under once-through conditions indicated: (1) P. fluorescens CL145A cells were efficacious against both zebra and quagga mussels, with high mortalities achieved against both species, and (2) as long as the total quantity of bacterial cells applied during the entire treatment period was the same, similar mussel mortality could be achieved in treatments lasting 1.5-12.0 h, with longer treatment durations achieving lower mortalities. The efficacy data presented herein, in combination with prior demonstration of its low risk of non-target impact, indicate that P. fluorescens CL145A cells have significant promise as an effective and environmentally safe control agent against these invasive mussels. Copyright © 2012 Elsevier Inc. All rights reserved.

  19. Effects of Pseudomonas fluorescens on the Water Parameters of Mycorrhizal and Non-Mycorrhizal Seedlings of Pinus halepensis

    Directory of Open Access Journals (Sweden)

    José A. Saiz de Omeñaca

    2013-08-01

    Full Text Available Inoculation of forest seedlings with mycorrhizal fungi and rhizobacteria can improve the morphological and physiological qualities of plants, especially those used for regeneration of arid areas. In this paper, under standard nursery conditions, Aleppo pine seedlings were inoculated with Pseudomonas fluorescens CECT 5281 rhizobacteria. Some of these seedlings were also inoculated with the ectomycorrhizal fungus Pisolithus tinctorius. Five months after the inoculations, we examined the growth, water parameters (osmotic potential at full turgor [Ψπfull], osmotic potential at zero turgor [Ψπ0], and the tissue modulus of elasticity near full turgor [Emax], mycorrhizal colonisation, and concentration of macronutrients (N, P, K, Ca and Mg in the seedlings. Subsequently, a trial was conducted to assess the root growth potential. P. fluorescens CECT 5281 decreased the cellular osmotic potential of P. halepensis seedlings but increased its elasticity. P. tinctorius + P. fluorescens caused osmotic adjustment at zero turgor and increased tissue elasticity, which improved tolerance to water stress. All inoculations improved the growth and nutrition of the seedlings but caused non-significant effects on root growth potential. The co-inoculation Pisolithus tinctorius + Pseudomonas fluorescens at the nursery may be a suitable technique for producing improved seedling material for restoration purposes.

  20. Efficacy of Pseudomonas fluorescens (Pf-CL145A) spray dried powder for controlling zebra mussels adhering to test substrates

    Science.gov (United States)

    Luoma, James A.; Severson, Todd J.; Weber, Kerry L.; Mayer, Denise A.

    2015-01-01

    A mobile bioassay trailer was used to assess the efficacy of Pseudomonas fluorescens (Pf-CL145A) spray dried powder (SDP) formulation for controlling zebra mussels (Dreissena polymorpha) from two midwestern lakes: Lake Carlos (Alexandria, Minnesota) and Shawano Lake (Shawano, Wisconsin). The effects of SDP exposure concentration and exposure duration on zebra mussel survival were evaluated along with the evaluation of a benthic injection application technique to reduce the amount of SDP required to induce zebra mortality.

  1. INFLUÊNCIA DA ATIVIDADE ENZIMÁTICA DE PSEUDOMONAS FLUORESCENS 041 EM LABNEH

    Directory of Open Access Journals (Sweden)

    Andreza Angélica Ferreira

    2012-04-01

    Full Text Available A refrigeração do leite proporciona a seleção de bactérias psicrotróficas deteriorantes produtoras de enzimas termoresistentes que podem comprometer a qualidade do leite e derivados. O objetivo deste trabalho foi avaliar as implicações causadas pela atividade enzimática de Pseudomonas fluorescens 041 na produção de Labneh. O leite pasteurizado foi inoculado intencionalmente com, aproximadamente, 106 UFC.mL-1 de P. fluorescens 041 e Labneh foi produzido imediatamente após inoculação no leite (tempo 0 e após 48, 72 e 96 horas de inoculação e armazenamento a 4 ºC. A qualidade físico-química do leite, do Labneh e do soro resultante da fabricação foi determinada. O rendimento prático, o rendimento técnico ajustado e o aproveitamento final de sólidos no Labneh em relação ao volume de leite (coeficiente GL também foram avaliados. Constatou-se alterações nas características físico-químicas do soro e do Labneh fabricado com leite armazenado a partir de 48 horas. Também, observou-se um aumento significativo em litros de leite destinado à produção de um quilo do produto ao longo do tempo de estocagem do leite inoculado com P. fluorescens 041, sendo os rendimentos prático, técnico ajustado e o coeficiente GL afetados pelo fator tempo. Portanto, a redução do tempo de estocagem do leite sob refrigeração e a prevenção da contaminação da matéria-prima por meio da adoção de boas práticas na cadeia do leite são medidas a serem adotadas para assegurar a qualidade e o rendimento do produto final.

  2. Impact du séchage sur la viabilité de Pseudomonas fluorescens (synthèse bibliographique

    Directory of Open Access Journals (Sweden)

    Mputu Kanyinda, JN.

    2014-01-01

    Full Text Available Impact of drying on Pseudomonas fluorescens viability. A review. Drying Pseudomonas fluorescens makes for more economical storage, transportation and marketing. The aim of the drying process is to stop and to stabilize all biological activity for optimal storage, compatible with the conservation of the maximum desired viability of the microorganisms. However, the viability rate of the bacteria after drying depends on the operating conditions of the drying process. One of the most important criteria to consider during the drying of biologically active products is the quality of the final dried product. Freeze-drying is the drying method most commonly used for Pseudomonas fluorescens. After their production, the bacteria are harvested by centrifugation and are freeze-dried, but the changes in temperature induced by freeze-drying are not without consequence for the cells. The freeze-drying process induces cell damage: peroxidation of fatty acids and proteins and DNA oxidation. However, use of protective compounds during freeze-drying and during storage increases significantly the rate of cell viability.

  3. Broad-range antagonistic rhizobacteria Pseudomonas fluorescens and Serratia plymuthica suppress Agrobacterium crown gall tumours on tomato plants.

    Science.gov (United States)

    Dandurishvili, N; Toklikishvili, N; Ovadis, M; Eliashvili, P; Giorgobiani, N; Keshelava, R; Tediashvili, M; Vainstein, A; Khmel, I; Szegedi, E; Chernin, L

    2011-01-01

    To examine the biocontrol activity of broad-range antagonists Serratia plymuthica IC1270, Pseudomonas fluorescens Q8r1-96 and P. fluorescens B-4117 against tumourigenic strains of Agrobacterium tumefaciens and A. vitis. Under greenhouse conditions, the antagonists, applied via root soak prior to injecting Agrobacterium strains into the wounded stems, significantly suppressed tumour development on tomato seedlings. A derivative of P. fluorescens Q8r1-96 tagged with a gfp reporter, as well as P. fluorescens B-4117 and S. plymuthica IC1270 marked with rifampicin resistance, stably persisted in tomato tissues for at least 1 month. Mutants of P. fluorescens Q8r1-96 and S. plymuthica IC1270 deficient in 2,4-diacetylphloroglucinol or pyrrolnitrin production, respectively, also proficiently suppressed the tumour development, indicating that these antibiotics are not responsible for the observed biocontrol effect on crown gall disease. The volatile organic compounds (VOCs) produced by the tested P. fluorescens and S. plymuthica strains inhibited the growth of A. tumefaciens and A. vitis strains in vitro. Solid-phase microextraction-gas chromatography-mass spectrometry analysis revealed dimethyl disulfide (DMDS) as the major headspace volatile produced by S. plymuthica IC1270; it strongly suppressed Agrobacterium growth in vitro and was emitted by tomato plants treated with S. plymuthica IC1270. 1-Undecene was the main volatile emitted by the examined P. fluorescens strains, with other volatiles, including DMDS, being detected in only relatively low quantities. S. plymuthica IC1270, P. fluorescens B-4117 and P. fluorescens Q8r1-96 can be used as novel biocontrol agents of pathogenic Agrobacterium. VOCs, and specifically DMDS, might be involved in the suppression of oncogenicity in tomato plants. However, the role of specific volatiles in the biocontrol activity remains to be elucidated. The advantage of applying these antagonists lies in their multiple activities against a

  4. A type VI secretion system is involved in Pseudomonas fluorescens bacterial competition.

    Directory of Open Access Journals (Sweden)

    Victorien Decoin

    Full Text Available Protein secretion systems are crucial mediators of bacterial interactions with other organisms. Among them, the type VI secretion system (T6SS is widespread in Gram-negative bacteria and appears to inject toxins into competitor bacteria and/or eukaryotic cells. Major human pathogens, such as Vibrio cholerae, Burkholderia and Pseudomonas aeruginosa, express T6SSs. Bacteria prevent self-intoxication by their own T6SS toxins by producing immunity proteins, which interact with the cognate toxins. We describe here an environmental P. fluorescens strain, MFE01, displaying an uncommon oversecretion of Hcp (hemolysin-coregulated protein and VgrG (valine-glycine repeat protein G into the culture medium. These proteins are characteristic components of a functional T6SS. The aim of this study was to attribute a role to this energy-consuming overexpression of the T6SS. The genome of MFE01 contains at least two hcp genes (hcp1 and hcp2, suggesting that there may be two putative T6SS clusters. Phenotypic studies have shown that MFE01 is avirulent against various eukaryotic cell models (amebas, plant or animal cell models, but has antibacterial activity against a wide range of competitor bacteria, including rhizobacteria and clinical bacteria. Depending on the prey cell, mutagenesis of the hcp2 gene in MFE01 abolishes or reduces this antibacterial killing activity. Moreover, the introduction of T6SS immunity proteins from S. marcescens, which is not killed by MFE01, protects E. coli against MFE01 killing. These findings suggest that the protein encoded by hcp2 is involved in the killing activity of MFE01 mediated by effectors of the T6SS targeting the peptidoglycan of Gram-negative bacteria. Our results indicate that MFE01 can protect potato tubers against Pectobacterium atrosepticum, which causes tuber soft rot. Pseudomonas fluorescens is often described as a major PGPR (plant growth-promoting rhizobacterium, and our results suggest that there may be a

  5. Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48

    Directory of Open Access Journals (Sweden)

    Rawia F. Gamal

    2013-01-01

    Full Text Available The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%. Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%. Semi-scale application (10 L working volume increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform-hypochlorite dispersion extraction. Gas chromatography (GC analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%. A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98-99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838.

  6. Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling

    Directory of Open Access Journals (Sweden)

    Thiruvengadam Raguchander

    2009-12-01

    Full Text Available Abstract Background Plant Growth Promoting Rhizobacteria (PGPR, Pseudomonas fluorescens strain KH-1 was found to exhibit plant growth promotional activity in rice under both in-vitro and in-vivo conditions. But the mechanism underlying such promotional activity of P. fluorescens is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to P. fluorescens treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry. Results Priming of P. fluorescens, 23 different proteins found to be differentially expressed in rice leaf sheaths and MS analysis revealed the differential expression of some important proteins namely putative p23 co-chaperone, Thioredoxin h- rice, Ribulose-bisphosphate carboxylase large chain precursor, Nucleotide diPhosphate kinase, Proteosome sub unit protein and putative glutathione S-transferase protein. Conclusion Functional analyses of the differential proteins were reported to be directly or indirectly involved in growth promotion in plants. Thus, this study confirms the primary role of PGPR strain KH-1 in rice plant growth promotion.

  7. Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling.

    Science.gov (United States)

    Kandasamy, Saveetha; Loganathan, Karthiba; Muthuraj, Raveendran; Duraisamy, Saravanakumar; Seetharaman, Suresh; Thiruvengadam, Raguchander; Ponnusamy, Balasubramanian; Ramasamy, Samiyappan

    2009-12-24

    Plant Growth Promoting Rhizobacteria (PGPR), Pseudomonas fluorescens strain KH-1 was found to exhibit plant growth promotional activity in rice under both in-vitro and in-vivo conditions. But the mechanism underlying such promotional activity of P. fluorescens is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to P. fluorescens treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry. Priming of P. fluorescens, 23 different proteins found to be differentially expressed in rice leaf sheaths and MS analysis revealed the differential expression of some important proteins namely putative p23 co-chaperone, Thioredoxin h- rice, Ribulose-bisphosphate carboxylase large chain precursor, Nucleotide diPhosphate kinase, Proteosome sub unit protein and putative glutathione S-transferase protein. Functional analyses of the differential proteins were reported to be directly or indirectly involved in growth promotion in plants. Thus, this study confirms the primary role of PGPR strain KH-1 in rice plant growth promotion.

  8. Antimicrobial activity of four essential oils against pigmenting Pseudomonas fluorescens and biofilmproducing Staphylococcus aureus of dairy origin

    Directory of Open Access Journals (Sweden)

    Francesca Pedonese

    2017-12-01

    Full Text Available Essential oils (EOs are mixtures of secondary metabolites of plant origin with many useful properties, among which the antimicrobial activity is also of interest for the food industry. EOs can exert their antimicrobial potential both directly, in food products and active packaging, and indirectly, as sanitizing and anti-biofilm agents of food facility surfaces. Aim of this research was to evaluate the antimicrobial activity of four EOs (bergamot, cinnamon, manuka and thyme against Pseudomonas fluorescens and Staphylococcus aureus isolated from milk and dairy products. The chemical composition of EOs was evaluated by Gas Chromatography-Mass Spectrometry analysis. Minimum Inhibitory Concentration values were determined by a microplate method against 9 Ps. fluorescens from marketed mozzarella with blue discoloration defect, and 3 biofilm-producing S. aureus from milk. Reference ATCC strains were included. Pigment production activity by Ps. fluorescens was assessed both in culture and in cheese. EOs of manuka (leptospermone 23% and thyme (carvacrol 30%, pcymene 20%, thymol 15% showed the highest antimicrobial activity against S. aureus, MIC values were 0.012%-0.024% and 0.024% v/v, respectively; meanwhile EOs from thyme and cinnamon (cinnamaldehyde 55% exhibited the best activity against Ps. fluorescens with MIC values of 0.098%-0.195% and 0.195%-0.391% v/v, respectively. The antimicrobial activity of these EOs is promising and they could be exploited in the dairy production chain.

  9. Detoxification of selenite and mercury by reduction and mutual protection in the assimilation of both elements by Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Belzile, Nelson [Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario, P3E 2C6 (Canada)]. E-mail: nbelzile@laurentian.ca; Wu Gaojun [Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario, P3E 2C6 (Canada); Chen, Yu-Wei [Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario, P3E 2C6 (Canada); Appanna, Vasu D. [Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario, P3E 2C6 (Canada)

    2006-08-31

    A study on the assimilation and detoxification of selenium and mercury and on the interaction between these two elements was conducted on Pseudomonas fluorescens. P. fluorescens was able to convert separately both elements to their elemental forms, which are less toxic and biologically less available. To study the converting mechanism of selenite to elemental Se, cells were grown in the presence of various selenite concentrations and several parameters such as extracellular protein concentrations, pH, carbohydrate concentrations, isocitrate dehydrogenase (ICDH) and malic enzyme were monitored. Transmission electron microscopy (TEM) and various analytical methods were applied to confirm the interaction between selenium and cell. The former appeared as a red precipitate localized predominantly in the consumed culture medium. P. fluorescens also resisted to the toxic effect of mercury by converting Hg{sup 2+} to the volatile and less toxic form Hg . Mercury reductase was likely responsible for the conversion of Hg{sup 2+} to Hg . More importantly, the interaction between mercury and selenium was also studied. The presence of selenite significantly reduced the accumulation of mercury in P. fluorescens. It was also interesting to note that mercury appeared to behave as a protecting agent against selenium intoxication as the bioaccumulation of Se was also inhibited by this metal. The formation of Se-Hg complexes could explain this mutual protective effect. No precipitate of elemental Se could be detected when Hg was present in the cultures.

  10. Detoxification of selenite and mercury by reduction and mutual protection in the assimilation of both elements by Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Belzile, Nelson; Wu Gaojun; Chen, Yu-Wei; Appanna, Vasu D.

    2006-01-01

    A study on the assimilation and detoxification of selenium and mercury and on the interaction between these two elements was conducted on Pseudomonas fluorescens. P. fluorescens was able to convert separately both elements to their elemental forms, which are less toxic and biologically less available. To study the converting mechanism of selenite to elemental Se, cells were grown in the presence of various selenite concentrations and several parameters such as extracellular protein concentrations, pH, carbohydrate concentrations, isocitrate dehydrogenase (ICDH) and malic enzyme were monitored. Transmission electron microscopy (TEM) and various analytical methods were applied to confirm the interaction between selenium and cell. The former appeared as a red precipitate localized predominantly in the consumed culture medium. P. fluorescens also resisted to the toxic effect of mercury by converting Hg 2+ to the volatile and less toxic form Hg . Mercury reductase was likely responsible for the conversion of Hg 2+ to Hg . More importantly, the interaction between mercury and selenium was also studied. The presence of selenite significantly reduced the accumulation of mercury in P. fluorescens. It was also interesting to note that mercury appeared to behave as a protecting agent against selenium intoxication as the bioaccumulation of Se was also inhibited by this metal. The formation of Se-Hg complexes could explain this mutual protective effect. No precipitate of elemental Se could be detected when Hg was present in the cultures

  11. Antimicrobial activity of four essential oils against pigmenting Pseudomonas fluorescens and biofilmproducing Staphylococcus aureus of dairy origin

    Science.gov (United States)

    Pedonese, Francesca; Fratini, Filippo; Pistelli, Luisa; Porta, Federica Maria; Ciccio, Pierluigi Di; Fischetti, Roberto; Turchi, Barbara; Nuvoloni, Roberta

    2017-01-01

    Essential oils (EOs) are mixtures of secondary metabolites of plant origin with many useful properties, among which the antimicrobial activity is also of interest for the food industry. EOs can exert their antimicrobial potential both directly, in food products and active packaging, and indirectly, as sanitizing and anti-biofilm agents of food facility surfaces. Aim of this research was to evaluate the antimicrobial activity of four EOs (bergamot, cinnamon, manuka and thyme) against Pseudomonas fluorescens and Staphylococcus aureus isolated from milk and dairy products. The chemical composition of EOs was evaluated by Gas Chromatography-Mass Spectrometry analysis. Minimum Inhibitory Concentration values were determined by a microplate method against 9 Ps. fluorescens from marketed mozzarella with blue discoloration defect, and 3 biofilm-producing S. aureus from milk. Reference ATCC strains were included. Pigment production activity by Ps. fluorescens was assessed both in culture and in cheese. EOs of manuka (leptospermone 23%) and thyme (carvacrol 30%, pcymene 20%, thymol 15%) showed the highest antimicrobial activity against S. aureus, MIC values were 0.012%-0.024% and 0.024% v/v, respectively; meanwhile EOs from thyme and cinnamon (cinnamaldehyde 55%) exhibited the best activity against Ps. fluorescens with MIC values of 0.098%-0.195% and 0.195%-0.391% v/v, respectively. The antimicrobial activity of these EOs is promising and they could be exploited in the dairy production chain. PMID:29564238

  12. Metabolic and transcriptomic changes induced in Arabidopsis by the rhizobacterium Pseudomonas fluorescens SS101.

    Science.gov (United States)

    van de Mortel, Judith E; de Vos, Ric C H; Dekkers, Ester; Pineda, Ana; Guillod, Leandre; Bouwmeester, Klaas; van Loon, Joop J A; Dicke, Marcel; Raaijmakers, Jos M

    2012-12-01

    Systemic resistance induced in plants by nonpathogenic rhizobacteria is typically effective against multiple pathogens. Here, we show that root-colonizing Pseudomonas fluorescens strain SS101 (Pf.SS101) enhanced resistance in Arabidopsis (Arabidopsis thaliana) against several bacterial pathogens, including Pseudomonas syringae pv tomato (Pst) and the insect pest Spodoptera exigua. Transcriptomic analysis and bioassays with specific Arabidopsis mutants revealed that, unlike many other rhizobacteria, the Pf.SS101-induced resistance response to Pst is dependent on salicylic acid signaling and not on jasmonic acid and ethylene signaling. Genome-wide transcriptomic and untargeted metabolomic analyses showed that in roots and leaves of Arabidopsis plants treated with Pf.SS101, approximately 1,910 genes and 50 metabolites were differentially regulated relative to untreated plants. Integration of both sets of "omics" data pointed to a prominent role of camalexin and glucosinolates in the Pf.SS101-induced resistance response. Subsequent bioassays with seven Arabidopsis mutants (myb51, cyp79B2cyp79B3, cyp81F2, pen2, cyp71A12, cyp71A13, and myb28myb29) disrupted in the biosynthesis pathways for these plant secondary metabolites showed that camalexin and glucosinolates are indeed required for the induction of Pst resistance by Pf.SS101. Also for the insect S. exigua, the indolic glucosinolates appeared to play a role in the Pf.SS101-induced resistance response. This study provides, to our knowledge for the first time, insight into the substantial biochemical and temporal transcriptional changes in Arabidopsis associated with the salicylic acid-dependent resistance response induced by specific rhizobacteria.

  13. Pseudomonas fluorescens R68 assisted enhancement in growth and fertilizer utilization of Amaranthus tricolor (L.).

    Science.gov (United States)

    Jimtha John, C; Jishma, P; Karthika, N R; Nidheesh, K S; Ray, J G; Mathew, Jyothis; Radhakrishnan, E K

    2017-08-01

    Plant probiotic potential of rhizosphere microbiome and its role in phytofertilizer mobilization are largely unexplored. In the current study, the rhizobacterium Pseudomonas fluorescens R68 (PFR68) isolated from Western Ghat was analyzed for its growth enhancement effect on the leafy vegetable Amaranthus tricolor (L.). One month of field growth of PFR68 inoculated A. tricolor has found to have enhanced growth parameters such as leaf number (1.57 fold), root number (1.76 fold), shoot length (1.28 fold) and fresh weight (2.31 fold). The treatment also improved soil fertility in terms of Nitrogen, Phosphorus and Potassium content. Most remarkably, application of PFR68 alone and 50% of recommended NPK dose along with PFR68 has resulted in enhanced growth of A. tricolor comparable to plants treated with full dose of NPK. In addition to this, application of PFR68 along with 50% NPK augmented the available Nitrogen and Phosphorus content in soil. This indicates the potential of selected organism in enrichment of soil health and enhancement of crop productivity. In conclusion, field performance of PFR68 on growth of A. tricolor confirms its promises to develop into plant probiotic formulation.

  14. Effects of high intensity pulsed electric field and thermal treatments on a lipase from Pseudomonas fluorescens.

    Science.gov (United States)

    Bendicho, S; Estela, C; Giner, J; Barbosa-Cánovas, G V; Martin, O

    2002-01-01

    Milk and dairy products may contain microorganisms capable of secreting lipases that cause sensory defects and technological problems in the dairy industry. In this study, the effects of thermal and high-intensity pulsed electric field (HIPEF) treatments on an extracellular lipase from Pseudomonas fluorescens, suspended in a simulated skim milk ultrafiltrate (SMUF) have been evaluated. Heat treatments applied were up to 30 min from 50 to 90 degrees C. HIPEF treatments were carried out using pilot plant facilities in a batch or continuous flow mode, where treatment chambers consisted of parallel and coaxial configuration, respectively. Samples were subjected to up to 80 pulses at electric field intensities ranging from 16.4 to 37.3 kV/cm. This resulted in a lipase that was quite resistant to heat and also to HIPEF. High (75 degrees C-15 s) and low pasteurization treatments (63 degrees C-30 min) led to inactivations of 5 and 20%, respectively. Using the batch-mode HIPEF equipment, a 62.1% maximum activity depletion was achieved after 80 pulses at 27.4 kV/cm. However, when HIPEF treatments were applied in the continuous flow mode, an inactivation rate of just 13% was achieved, after applying 80 pulses at 37.3 kV/cm and 3.5 Hz. The results of both heat and HIPEF treatments on enzyme inactivation were adjusted with good agreement to a first-order kinetic model (R2 > 62.3%).

  15. Modeling high-intensity pulsed electric field inactivation of a lipase from Pseudomonas fluorescens.

    Science.gov (United States)

    Soliva-Fortuny, R; Bendicho-Porta, S; Martín-Belloso, O

    2006-11-01

    The inactivation kinetics of a lipase from Pseudomonas fluorescens (EC 3.1.1.3.) were studied in a simulated skim milk ultrafiltrate treated with high-intensity pulsed electric fields. Samples were subjected to electric field intensities ranging from 16.4 to 27.4 kV/cm for up to 314.5 micros, thus achieving a maximum inactivation of 62.1%. The suitability of describing experimental data using mechanistic first-order kinetics and an empirical model based on the Weibull distribution function is discussed. In addition, different mathematical expressions relating the residual activity values to field strength and treatment time are supplied. A first-order fractional conversion model predicted residual activity with good accuracy (A(f) = 1.018). A mechanistic insight of the model kinetics was that experimental values were the consequence of different structural organizations of the enzyme, with uneven resistance to the pulsed electric field treatments. The Weibull model was also useful in predicting the energy density necessary to achieve lipase inactivation.

  16. Pit formation on stainless steel surfaces pre-treated with biosurfactants produced by Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Dagbert, Catherine; Meylheuc, Thierry; Bellon-Fontaine, Marie-Noelle

    2008-01-01

    Today, it is widely established that the surface tension of water can be reduced by some microorganisms capable of synthesizing surface-active compounds called biosurfactants (BS). BS characteristics depend on the microorganism that produces them and therefore, on the microorganism culture conditions. Some studies on chemical surfactants have shown that the adsorption of surface-active compounds plays a major role in corrosion; indeed they are used as a good corrosion inhibition tool. The purpose of this study was first, to estimate the importance and behavior of the stainless steels passive film on the adsorption of BS, produced by the Gram negative bacteria Pseudomonas fluorescens, and secondly, to study the impact of these treatments on the pitting corrosion. In this paper, the galvanostatic polarization technique, used as accelerated method for determining the characteristic pit potentials on stainless steels, is examined. Pit growth, shape and cover formation were also observed. The surface topography of the corroded specimens was investigated using field emission scanning electron microscopy (FESEM)

  17. Pit formation on stainless steel surfaces pre-treated with biosurfactants produced by Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Dagbert, Catherine [ECP-LGPM, Grande Voie des Vignes, 92295 Chatenay-Malabry (France)], E-mail: catherine.dagbert@ecp.fr; Meylheuc, Thierry; Bellon-Fontaine, Marie-Noelle [INRA, UMR 763 Bioadhesion et Hygiene des Materiaux, F-91300 Massy (France); AGROPARISTECH, UMR 763 Bioadhesion et Hygiene des Materiaux, F-91300 Massy (France)

    2008-12-01

    Today, it is widely established that the surface tension of water can be reduced by some microorganisms capable of synthesizing surface-active compounds called biosurfactants (BS). BS characteristics depend on the microorganism that produces them and therefore, on the microorganism culture conditions. Some studies on chemical surfactants have shown that the adsorption of surface-active compounds plays a major role in corrosion; indeed they are used as a good corrosion inhibition tool. The purpose of this study was first, to estimate the importance and behavior of the stainless steels passive film on the adsorption of BS, produced by the Gram negative bacteria Pseudomonas fluorescens, and secondly, to study the impact of these treatments on the pitting corrosion. In this paper, the galvanostatic polarization technique, used as accelerated method for determining the characteristic pit potentials on stainless steels, is examined. Pit growth, shape and cover formation were also observed. The surface topography of the corroded specimens was investigated using field emission scanning electron microscopy (FESEM)

  18. Expression, Purification, Crystallization and Preliminary X-ray Analysis of Pseudomonas fluorescens AlgK

    Energy Technology Data Exchange (ETDEWEB)

    Keiski,C.; Yip, P.; Robinson, H.; Burrows, L.; Howell, P.

    2007-01-01

    AlgK is an outer-membrane lipoprotein involved in the biosynthesis of alginate in Pseudomonads and Azotobacter vinelandii. A recombinant form of Pseudomonas fluorescens AlgK with a C-terminal polyhistidine affinity tag has been expressed and purified from the periplasm of Escherichia coli cells and diffraction-quality crystals of AlgK have been grown using the hanging-drop vapour-diffusion method. The crystals grow as flat plates with unit-cell parameters a = 79.09, b = 107.85, c = 119.15 {angstrom}, = 96.97{sup o}. The crystals exhibit the symmetry of space group P2{sub 1} and diffract to a minimum d-spacing of 2.5 {angstrom} at Station X29 of the National Synchrotron Light Source, Brookhaven National Laboratory. On the basis of the Matthews coefficient (V{sub M} = 2.53 {angstrom}{sup 3} Da{sup -1}), four protein molecules are estimated to be present in the asymmetric unit.

  19. Small RNA-dependent expression of secondary metabolism is controlled by Krebs cycle function in Pseudomonas fluorescens.

    Science.gov (United States)

    Takeuchi, Kasumi; Kiefer, Patrick; Reimmann, Cornelia; Keel, Christoph; Dubuis, Christophe; Rolli, Joëlle; Vorholt, Julia A; Haas, Dieter

    2009-12-11

    Pseudomonas fluorescens CHA0, an antagonist of phytopathogenic fungi in the rhizosphere of crop plants, elaborates and excretes several secondary metabolites with antibiotic properties. Their synthesis depends on three small RNAs (RsmX, RsmY, and RsmZ), whose expression is positively controlled by the GacS-GacA two-component system at high cell population densities. To find regulatory links between primary and secondary metabolism in P. fluorescens and in the related species Pseudomonas aeruginosa, we searched for null mutations that affected central carbon metabolism as well as the expression of rsmY-gfp and rsmZ-gfp reporter constructs but without slowing down the growth rate in rich media. Mutation in the pycAB genes (for pyruvate carboxylase) led to down-regulation of rsmXYZ and secondary metabolism, whereas mutation in fumA (for a fumarase isoenzyme) resulted in up-regulation of the three small RNAs and secondary metabolism in the absence of detectable nutrient limitation. These effects required the GacS sensor kinase but not the accessory sensors RetS and LadS. An analysis of intracellular metabolites in P. fluorescens revealed a strong positive correlation between small RNA expression and the pools of 2-oxoglutarate, succinate, and fumarate. We conclude that Krebs cycle intermediates (already known to control GacA-dependent virulence factors in P. aeruginosa) exert a critical trigger function in secondary metabolism via the expression of GacA-dependent small RNAs.

  20. Characterization of a Pseudomonas fluorescens strain from tomato rhizosphere and its use for integrated management of tomato damping-off

    Energy Technology Data Exchange (ETDEWEB)

    Jayaraj, J; Parthasarathi, T.; Radhakrishnan, N.V. [Annamalai University, Annamalainagar (India). Faculty of Agriculture

    2007-10-15

    A highly antagonistic Pseudomonas fluorescens strain was isolated from tomato rhizosphere and characterized for its in vitro and in vivo biocontrol potential against Pythium aphanidermatum. The identified Pseudomonas fluorescens strain (PfT-8) was capable of producing high levels of chitinase, beta-1,3-glucanase, cellulase, fungitoxic metabolites and siderophores. Seven different carrier formulations including a talc-based powder, lignite-based powder, peat-based powder, lignite + fly ash-based powder, wettable powder, bentonite-paste and polyethylene glycol (PEG) paste were prepared utilizing PfT-8. Shelf life was evaluated for up to 6 months of storage at ambient room temperature (28{sup o}C). Biocontrol efficacy of formulations was studied under greenhouse and field conditions. Among the formulations, peat, lignite, lignite+fly-ash and bentonite paste based formulations maintained higher propagule number than others and also showed greater biocontrol potential. However, propagule number gradually decreased with time. Soil incorporation of organic amendments and specifically poultry manure and FYM, significantly reduced damping-off incidence and also augmented the rhizosphere population of the marked P. fluorescens strain that was resistant to streptomycin and rifampicin.

  1. Biocidal Activity of Formaldehyde and Nonformaldehyde Biocides toward Mycobacterium immunogenum and Pseudomonas fluorescens in Pure and Mixed Suspensions in Synthetic Metalworking Fluid and Saline

    OpenAIRE

    Selvaraju, Suresh B.; Khan, Izhar U. H.; Yadav, Jagjit S.

    2005-01-01

    The microbicidal activity of four different biocides was studied in synthetic metalworking fluid (MWF) against Mycobacterium immunogenum, a suspected causative agent for hypersensitivity pneumonitis, and Pseudomonas fluorescens, a representative for the predominant gram-negative bacterial contaminants of MWF. The results indicated that M. immunogenum is more resistant than P. fluorescens to the tested formaldehyde-releasing biocides (Grotan and Bioban), isothiazolone (Kathon), and phenolic bi...

  2. Identification and Characterization of the emhABC Efflux System for Polycyclic Aromatic Hydrocarbons in Pseudomonas fluorescens cLP6a

    OpenAIRE

    Hearn, Elizabeth M.; Dennis, Jonathan J.; Gray, Murray R.; Foght, Julia M.

    2003-01-01

    The hydrocarbon-degrading environmental isolate Pseudomonas fluorescens LP6a possesses an active efflux mechanism for the polycyclic aromatic hydrocarbons phenanthrene, anthracene, and fluoranthene but not for naphthalene or toluene. PCR was used to detect efflux pump genes belonging to the resistance-nodulation-cell division (RND) superfamily in a plasmid-cured derivative, P. fluorescens cLP6a, which is unable to metabolize hydrocarbons. One RND pump, whose gene was identified in P. fluoresc...

  3. Distribution of a Take-All Suppressive Strain of Pseudomonas fluorescens on Seminal Roots of Winter Wheat.

    Science.gov (United States)

    Weller, D M

    1984-10-01

    An antibiotic-resistant strain of Pseudomonas fluorescens, that suppresses take-all of wheat, was used to study the distribution of the bacteria on seminal roots of wheat after being introduced onto seeds. Cells of P. fluorescens were isolated from the entire length of the root, and density of the introduced bacteria declined with the distance from the base of the root. Maximum populations of 10 to 10 CFU and 10 to 10 CFU per cm of root were detected on sections of roots near the seed and root tip, respectively. The introduced bacteria competed well with indigenous bacteria, comprising at least 25% of the fluorescent pseudomonads detected by plate counts for 48 days after planting.

  4. Genetic characterization of psp encoding the DING protein in Pseudomonas fluorescens SBW25

    Directory of Open Access Journals (Sweden)

    Zhang Xue-Xian

    2007-12-01

    Full Text Available Abstract Background DING proteins constitute a conserved and broadly distributed set of proteins found in bacteria, fungi, plants and animals (including humans. Characterization of DING proteins from animal and plant tissues indicated ligand-binding ability suggesting a role for DING proteins in cell signaling and biomineralization. Surprisingly, the genes encoding DING proteins in eukaryotes have not been identified in the eukaryotic genome or EST databases. Recent discovery of a DING homologue (named Psp here in the genome of Pseudomonas fluorescens SBW25 provided a unique opportunity to investigate the physiological roles of DING proteins. P. fluorescens SBW25 is a model bacterium that can efficiently colonize plant surfaces and enhance plant health. In this report we genetically characterize Psp with a focus on conditions under which psp is expressed and the protein exported. Results Psp is closely related to the periplasmic Pi binding component of the ABC-type phosphate transporter system (Pst. psp is flanked by a gene cluster predicted to function as a type II protein secretion system (Hxc. Deletion analysis combined with chromosomally integrated 'lacZ fusions showed that both psp and pstC are induced by Pi limitation and that pstC is required for competitive growth of the bacterium in Pi limited medium. hxcR is not regulated by Pi limitation. Psp was detected (using anti-DING serum in the supernatant of wild-type culture but was greatly reduced in the supernatant of an isogenic strain carrying an hxcR mutation (ΔhxcR. A promoter fusion between hxcR and a promoterless copy of a gene ('dapB essential for growth in the plant environment showed that expression of hxcR is elevated during colonization of sugar beet seedlings. A similar analysis of psp showed that it is not induced in the plant environment. Conclusion Psp gene is expressed under conditions of Pi limitation. It is an exoprotein secreted mainly via the Hxc type II secretion

  5. Genetic characterization of psp encoding the DING protein in Pseudomonas fluorescens SBW25.

    Science.gov (United States)

    Zhang, Xue-Xian; Scott, Ken; Meffin, Rebecca; Rainey, Paul B

    2007-12-18

    DING proteins constitute a conserved and broadly distributed set of proteins found in bacteria, fungi, plants and animals (including humans). Characterization of DING proteins from animal and plant tissues indicated ligand-binding ability suggesting a role for DING proteins in cell signaling and biomineralization. Surprisingly, the genes encoding DING proteins in eukaryotes have not been identified in the eukaryotic genome or EST databases. Recent discovery of a DING homologue (named Psp here) in the genome of Pseudomonas fluorescens SBW25 provided a unique opportunity to investigate the physiological roles of DING proteins. P. fluorescens SBW25 is a model bacterium that can efficiently colonize plant surfaces and enhance plant health. In this report we genetically characterize Psp with a focus on conditions under which psp is expressed and the protein exported. Psp is closely related to the periplasmic Pi binding component of the ABC-type phosphate transporter system (Pst). psp is flanked by a gene cluster predicted to function as a type II protein secretion system (Hxc). Deletion analysis combined with chromosomally integrated 'lacZ fusions showed that both psp and pstC are induced by Pi limitation and that pstC is required for competitive growth of the bacterium in Pi limited medium. hxcR is not regulated by Pi limitation. Psp was detected (using anti-DING serum) in the supernatant of wild-type culture but was greatly reduced in the supernatant of an isogenic strain carrying an hxcR mutation (DeltahxcR). A promoter fusion between hxcR and a promoterless copy of a gene ('dapB) essential for growth in the plant environment showed that expression of hxcR is elevated during colonization of sugar beet seedlings. A similar analysis of psp showed that it is not induced in the plant environment. Psp gene is expressed under conditions of Pi limitation. It is an exoprotein secreted mainly via the Hxc type II secretion system, whose expression is elevated on plant surfaces

  6. Effect of Eu(III) on the degradation of malic acid by Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Nankawa, T.; Ozaki, T.; Ohnuki, T.; Suzuki, Y.; Francis, A.J.

    2005-01-01

    Full text of publication follows: The transuranic elements, such as Am(III) and Cm(III), are highly toxic because they emit high-energy α particles and have long half-lives. To estimate their long-term environmental behavior, we need to elucidate degradation of actinide-organic complexes by microorganisms. We studied the biodegradation of Eu(III)-malic acid complexes by Pseudomonas fluorescens. Malic acid is ubiquitous in the environment and is one of the microbial metabolites that is part of the tri-carboxylic acid (TCA) cycle. Europium(III) is a good analogue for Am(III) and Cm(III). To investigate the effect of Eu(III) on the degradation of malic acid by P. fluorescens, we compared the degradation behavior of Eu(III)-malic acid complexes to that of Fe(III) and Al(III)-malic acid complexes. In the medium containing 1 mM malic acid and 0-0.5 mM Fe(III), malic acid was degraded completely. In the medium containing 1 mM malic acid and 0.05-0.5 mM Al(III), malic acid was degraded until the concentration of malic acid became equal to that of Al(III), indicating that Al(III)-malic acid complex with 1: 1 molar ratio was recalcitrant to biodegradation. In the medium containing 1 mM malic acid and 0.05-0.5 mM Eu(III), degradation of malic acid was not observed. The effect of metals on degradation of malic acid was in the order of Fe(III) < Al(III) < Eu(III). The stability constants of 1:1 Fe(III)-, Al(III)-, and Eu(III)-malic acid complexes are 7.1, 4.6, and 4.9, respectively. These results indicate that degradability of malic acid does not depend on the stability constants of metal-malic acid complexes. We found that 10 mM malic acid was degraded in the presence of 0.05 and 0.1 mM Eu(III) but 1 mM malic acid was not degraded in the presence of 0.05 and 0.1 mM Eu(III). The degradation rate of malic acid increased with a decreasing ratio of Eu(III) to malic acid. (authors)

  7. Purification and characterization of an exo-type β-N-acetylglucosaminidase from Pseudomonas fluorescens JK-0412.

    Science.gov (United States)

    Park, J K; Kim, W J; Park, Y I

    2011-01-01

    To purify and characterize an exo-acting chitinolytic enzyme produced from a Gram-negative bacterium Pseudomonas fluorescens JK-0412. A chitinolytic bacterial strain that showed confluent growth on a minimal medium containing powder chitin as the sole carbon source was isolated and identified based on a 16S ribosomal DNA sequence analysis and named Ps. fluorescens JK-0412. From the culture filtrates of this strain, a chito-oligosaccharides-degrading enzyme was purified to apparent homogeneity with a molecular mass of 50 kDa on SDS-PAGE gels. The kinetics, optimum pH and temperature, and substrate specificity of the purified enzyme (named as NagA) were determined. An extracellular chitinolytic enzyme was purified from the Ps. fluorescens JK-0412 and shown to be an exo-type β-N-acetylglucosaminidase yielding GlcNAc as the final product from the natural chito-oligosaccharides, (GlcNAc)(n) , n = 2-5. As NagA is secreted extracellularly in the presence of colloidal chitin, Ps. fluorescens JK-0412 can be recognized as a potent producer for industry-level and cost-effective production of chitinolytic enzyme. This enzyme appears to have potential applications as an efficient tool for the degradation of chitinous materials and industry-level production of GlcNAc. To the best of our knowledge, this is the first report on an exo-type chitinolytic enzyme of Pseudomonas species. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology.

  8. Comparative genomics of Pseudomonas fluorescens subclade III strains from human lungs.

    Science.gov (United States)

    Scales, Brittan S; Erb-Downward, John R; Huffnagle, Ian M; LiPuma, John J; Huffnagle, Gary B

    2015-12-07

    While the taxonomy and genomics of environmental strains from the P. fluorescens species-complex has been reported, little is known about P. fluorescens strains from clinical samples. In this report, we provide the first genomic analysis of P. fluorescens strains in which human vs. environmental isolates are compared. Seven P. fluorescens strains were isolated from respiratory samples from cystic fibrosis (CF) patients. The clinical strains could grow at a higher temperature (>34 °C) than has been reported for environmental strains. Draft genomes were generated for all of the clinical strains, and multi-locus sequence analysis placed them within subclade III of the P. fluorescens species-complex. All strains encoded type- II, -III, -IV, and -VI secretion systems, as well as the widespread colonization island (WCI). This is the first description of a WCI in P. fluorescens strains. All strains also encoded a complete I2/PfiT locus and showed evidence of horizontal gene transfer. The clinical strains were found to differ from the environmental strains in the number of genes involved in metal resistance, which may be a possible adaptation to chronic antibiotic exposure in the CF lung. This is the largest comparative genomics analysis of P. fluorescens subclade III strains to date and includes the first clinical isolates. At a global level, the clinical P. fluorescens subclade III strains were largely indistinguishable from environmental P. fluorescens subclade III strains, supporting the idea that identifying strains as 'environmental' vs 'clinical' is not a phenotypic trait. Rather, strains within P. fluorescens subclade III will colonize and persist in any niche that provides the requirements necessary for growth.

  9. The effect of rhamnolipid biosurfactant produced by Pseudomonas fluorescens on model bacterial strains and isolates from industrial wastewater.

    Science.gov (United States)

    Vasileva-Tonkova, Evgenia; Sotirova, Anna; Galabova, Danka

    2011-02-01

    In this study, the effect of rhamnolipid biosurfactant produced by Pseudomonas fluorescens on bacterial strains, laboratory strains, and isolates from industrial wastewater was investigated. It was shown that biosurfactant, depending on the concentration, has a neutral or detrimental effect on the growth and protein release of model Gram (+) strain Bacillus subtilis 168. The growth and protein release of model Gram (-) strain Pseudomonas aeruginosa 1390 was not influenced by the presence of biosurfactant in the medium. Rhamnolipid biosurfactant at the used concentrations supported the growth of some slow growing on hexadecane bacterial isolates, members of the microbial community. Changes in cell surface hydrophobicity and permeability of some Gram (+) and Gram (-) isolates in the presence of rhamnolipid biosurfactant were followed in experiments in vitro. It was found that bacterial cells treated with biosurfactant became more or less hydrophobic than untreated cells depending on individual characteristics and abilities of the strains. For all treated strains, an increase in the amount of released protein was observed with increasing the amount of biosurfactant, probably due to increased cell permeability as a result of changes in the organization of cell surface structures. The results obtained could contribute to clarify the relationships between members of the microbial community as well as suggest the efficiency of surface properties of rhamnolipid biosurfactant from Pseudomonas fluorescens making it potentially applicable in bioremediation of hydrocarbon-polluted environments.

  10. Biodegradation of phenol, salicylic acid, benzenesulfonic acid, and iomeprol by Pseudomonas fluorescens in the capillary fringe

    Science.gov (United States)

    Hack, Norman; Reinwand, Christian; Abbt-Braun, Gudrun; Horn, Harald; Frimmel, Fritz H.

    2015-12-01

    Mass transfer and biological transformation phenomena in the capillary fringe were studied using phenol, salicylic acid, benzenesulfonic acid, and the iodinated X-ray contrast agent iomeprol as model organic compounds and the microorganism strain Pseudomonas fluorescens. Three experimental approaches were used: Batch experiments (uniform water saturation and transport by diffusion), in static columns (with a gradient of water saturation and advective transport in the capillaries) and in a flow-through cell (with a gradient of water saturation and transport by horizontal and vertical flow: 2-dimension flow-through microcosm). The reactors employed for the experiments were filled with quartz sand of defined particle size distribution (dp = 200…600 μm, porosity ε = 0.42). Batch experiments showed that phenol and salicylic acid have a high, whereas benzenesulfonic acid and iomeprol have a quite low potential for biodegradation under aerobic conditions and in a matrix nearly close to water saturation. Batch experiments under anoxic conditions with nitrate as electron acceptor revealed that the biodegradation of the model compounds was lower than under aerobic conditions. Nevertheless, the experiments showed that the moisture content was also responsible for an optimized transport in the liquid phase of a porous medium. Biodegradation in the capillary fringe was found to be influenced by both the moisture content and availability of the dissolved substrate, as seen in static column experiments. The gas-liquid mass transfer of oxygen also played an important role for the biological activity. In static column experiments under aerobic conditions, the highest biodegradation was found in the capillary fringe (e.g. βt/β0 (phenol) = 0 after t = 6 d) relative to the zone below the water table and unsaturated zone. The highest biodegradation occurred in the flow-through cell experiment where the height of the capillary fringe was largest.

  11. D-glucose-6-phosphate dehydrogenase (Entner-Doudoroff enzyme) from Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Lessmann, D.; Schimz, K.L.; Kurz, G.

    1975-01-01

    The existence of two different D-glucose-6-phosphate dehydrogenases in Pseudomonas fluorescens has been demonstrated. Based on their different specificity and their different metabolic regulation one enzyme is appointed to the Entner-Doudoroff pathway and the other to the hexose monophosphate pathway. A procedure is described for the isolation of that D-glucose-6-phosphate dehydrogenase which forms part of the Entner-Doudoroff pathway (Entner-Doudoroff enzyme). A 950-fold purification was achieved with an overall yield of 44%. The final preparation, having a specific activity of about 300μmol NADH formed per min per mg protein, was shown to be homogeneous. The molecular weight of the Entner-Doudoroff enzyme has been determined to be 220,000 by gel permeation chromatography, and that of the other enzyme (Zwischenferment) has been shown to be 265,000. The pI of the Entner-Doudoroff enzyme has been shown to be 5.24 and that of the Zwischenferment 4.27. The Entner-Doudoroff enzyme is stable in the range of pH 6 to 10.5 and shows its maximal acivity at pH 8.9. The Entner-Doudoroff enzyme showed specificity for NAD + as well as for NADP + and exhibited homotropic effects for D-glucose 6-phosphate. It is inhibited by ATP which acts as a negative allosteric effector. Other nucleoside triphosphates as well as ADP are also inhibitory. The enzyme catalyzes the transfer of the axial hydrogen at carbon-1 of β-D-glucopyranose 6-phosphate to the si face of carbon-4 of the nicotinamide ring and must be classified as B-side stereospecific dehydrogenase. (orig.) [de

  12. Biodegradation of phenol, salicylic acid, benzenesulfonic acid, and iomeprol by Pseudomonas fluorescens in the capillary fringe.

    Science.gov (United States)

    Hack, Norman; Reinwand, Christian; Abbt-Braun, Gudrun; Horn, Harald; Frimmel, Fritz H

    2015-12-01

    Mass transfer and biological transformation phenomena in the capillary fringe were studied using phenol, salicylic acid, benzenesulfonic acid, and the iodinated X-ray contrast agent iomeprol as model organic compounds and the microorganism strain Pseudomonas fluorescens. Three experimental approaches were used: Batch experiments (uniform water saturation and transport by diffusion), in static columns (with a gradient of water saturation and advective transport in the capillaries) and in a flow-through cell (with a gradient of water saturation and transport by horizontal and vertical flow: 2-dimension flow-through microcosm). The reactors employed for the experiments were filled with quartz sand of defined particle size distribution (dp=200...600 μm, porosity ε=0.42). Batch experiments showed that phenol and salicylic acid have a high, whereas benzenesulfonic acid and iomeprol have a quite low potential for biodegradation under aerobic conditions and in a matrix nearly close to water saturation. Batch experiments under anoxic conditions with nitrate as electron acceptor revealed that the biodegradation of the model compounds was lower than under aerobic conditions. Nevertheless, the experiments showed that the moisture content was also responsible for an optimized transport in the liquid phase of a porous medium. Biodegradation in the capillary fringe was found to be influenced by both the moisture content and availability of the dissolved substrate, as seen in static column experiments. The gas-liquid mass transfer of oxygen also played an important role for the biological activity. In static column experiments under aerobic conditions, the highest biodegradation was found in the capillary fringe (e.g. βt/β0 (phenol)=0 after t=6 d) relative to the zone below the water table and unsaturated zone. The highest biodegradation occurred in the flow-through cell experiment where the height of the capillary fringe was largest. Copyright © 2015 Elsevier B.V. All rights

  13. Biosurfactant production by Pseudomonas fluorescens growing on molasses and its application in phenol degradation

    Science.gov (United States)

    Suryantia, Venty; Marliyana, Soerya Dewi; Wulandari, Astri

    2015-12-01

    A molasses based medium for the biosurfactant production by Pseudomonas fluorescens was developed, where the effect of pre-treated of molasses and medium composition were evaluated. Biosurfactant production was followed by measuring optical density (OD), surface tension and emulsifying index (E24) over 12 days of fermentation. The optimum condition for the biosurfactant production was obtained when a medium containing of 8 g/L nutrient broth, 5 g/L NaCl, 1 g/L NH4NO3 and 5% v/v pre-treated molasses with centrifugation was used as media with 3 days of fermentation. The biosurfactant was identified as a rhamnolipid type biosurfactant which had critical micelle concentration (CMC) value of 801 mg/L and was able to reduce the surface tension of the water from 80 mN/m to 51 mN/m. The biosurfactants had water in oil (w/o) emulsion type. Biosurfactant was able to emulsify various hydrocarbons, which were able to decrase the interfacial tension about 50-75% when benzyl chloride, anisaldehyde and palm oil were used as immiscible compounds. The biosurfactant exhibited the E24 value of about 50% and the stable emulsion was reached up to 30 days when lubricant was used as an immiscible compound. Up to 68% of phenol was degraded in the presence of biosurfactant within 15 days, whereas only 56% of phenol was degraded in the absence of biosurfactant. Overall, the results exhibited that molasses are recommended for the rhamnolipids production which possessed good surface-active properties and had potential application in the enhancement of phenol degradation.

  14. IMPACT OF SIPHONING ACTIVITY AND NATURALLY SUSPENDED PARTICLE LOAD ON MUSSEL KILL by PSEUDOMONAS FLUORESCENS

    International Nuclear Information System (INIS)

    Daniel Molloy

    2003-01-01

    Under this USDOE-NETL contract, the bacterium Pseudomonas fluorescens is being developed as a biocontrol agent for zebra mussels. The specific purpose of the contract is to identify biotic and abiotic factors that affect mussel kill. Ingestion of these bacteria by zebra mussels is required to achieve kill, and tests evaluating factors that relate to mussel feeding are contained in this report. Specifically the impact of the following two factors were investigated: (1) Mussel siphoning behavior--In nature, zebra mussels typically have their two shells spread apart and their inhalant siphon tube extended from between their shells for taking food particles into their mantle cavities (Fig. 1). Our tests indicated that there is a direct correlation between mussel siphoning activity and mussel mortality achieved by a bacterial treatment. Therefore, to encourage mussel feeding on bacteria, future pipe treatments within power plants should be carried out using procedures which minimize disturbance to mussel siphoning. 2. Naturally suspended particle loads--Since bacterial cells are lethal only if ingested by mussels, waters containing very high levels of naturally suspended particles might reduce the mortality that can be achieved by a bacterial treatment. If true, this inhibition might occur as a result of particle exclusion, i.e., there could be reduced ingestion of bacterial cells since they represent a reduced percentage of all particles ingested. Our tests indicated that a range of particle concentrations that might naturally exist in a turbid river did not inhibit mussel kill by the bacterial cells, but that an artificially high load of natural particles was capable of causing a reduction in kill. To be conservative, therefore, future pipe treatments should be timed to occur when intake waters have relatively low quantities of naturally suspended particulate matter

  15. IMPACT OF SIPHONING ACTIVITY AND NATURALLY SUSPENDED PARTICLE LOAD ON MUSSEL KILL by PSEUDOMONAS FLUORESCENS

    Energy Technology Data Exchange (ETDEWEB)

    Daniel Molloy

    2003-08-04

    Under this USDOE-NETL contract, the bacterium Pseudomonas fluorescens is being developed as a biocontrol agent for zebra mussels. The specific purpose of the contract is to identify biotic and abiotic factors that affect mussel kill. Ingestion of these bacteria by zebra mussels is required to achieve kill, and tests evaluating factors that relate to mussel feeding are contained in this report. Specifically the impact of the following two factors were investigated: (1) Mussel siphoning behavior--In nature, zebra mussels typically have their two shells spread apart and their inhalant siphon tube extended from between their shells for taking food particles into their mantle cavities (Fig. 1). Our tests indicated that there is a direct correlation between mussel siphoning activity and mussel mortality achieved by a bacterial treatment. Therefore, to encourage mussel feeding on bacteria, future pipe treatments within power plants should be carried out using procedures which minimize disturbance to mussel siphoning. 2. Naturally suspended particle loads--Since bacterial cells are lethal only if ingested by mussels, waters containing very high levels of naturally suspended particles might reduce the mortality that can be achieved by a bacterial treatment. If true, this inhibition might occur as a result of particle exclusion, i.e., there could be reduced ingestion of bacterial cells since they represent a reduced percentage of all particles ingested. Our tests indicated that a range of particle concentrations that might naturally exist in a turbid river did not inhibit mussel kill by the bacterial cells, but that an artificially high load of natural particles was capable of causing a reduction in kill. To be conservative, therefore, future pipe treatments should be timed to occur when intake waters have relatively low quantities of naturally suspended particulate matter.

  16. A phosphate-starvation-inducible outermembrane protein of Pseudomonas fluorescens Ag1 as an immunological phosphate-starvation marker

    DEFF Research Database (Denmark)

    Leopold, Kristine; Jacobsen, Susanne; Nybroe, Ole

    1997-01-01

    A phosphate-starvation-inducible outer-membrane protein of Pseudomonas fluorescens Ag1, expressed at phosphate concentrations below0.08-0.13 mM, was purified and characterized. The purification method involved separation of outer-membrane proteins by SDS-PAGE andextraction of the protein from...... nitrocellulose or PVDF membranes after electrotransfer of proteins to the membranes. The N-terminal amino acidsequence of the purified protein, called Psi1, did not show homology to any known proteins, and in contrast to the phosphate-specific porin OprP ofP. aeruginosa its mobility in SDS-PAGE was not affected...

  17. Binding sites of mosquitocidal toxins of Pseudomonas fluorescens and Bacillus subtilis on pupae and larvae of Culex quinquefasciatus.

    Science.gov (United States)

    Mary, K Athisaya; Paily, K P; Hoti, S L; Balaraman, K

    2015-01-01

    Two of the potential bacterial isolates, viz., Pseudomonas fluorescens (VCRC B-426) and Bacillus subtilis (VCRC B-471) whose toxins kill the mosquito pupae/larvae have been identified at our center. As the mode of action of these bacteria are not known, an attempt was made to find out the binding sites of the toxic proteins through immunological methods. Antibodies were raised in BALB/c mice and egg yolk system of chicken layers against the mosquitocidal proteins. The antibodies showed specific binding on to the cephalic and thoracic cuticle of the pupae as well as the paddles of the larvae, indicating the binding of the mosquitocidal proteins.

  18. Extracellular Protease of Pseudomonas fluorescens CHA0, a Biocontrol Factor with Activity against the Root-Knot Nematode Meloidogyne incognita

    OpenAIRE

    Siddiqui, Imran Ali; Haas, Dieter; Heeb, Stephan

    2005-01-01

    In Pseudomonas fluorescens CHA0, mutation of the GacA-controlled aprA gene (encoding the major extracellular protease) or the gacA regulatory gene resulted in reduced biocontrol activity against the root-knot nematode Meloidogyne incognita during tomato and soybean infection. Culture supernatants of strain CHA0 inhibited egg hatching and induced mortality of M. incognita juveniles more strongly than did supernatants of aprA and gacA mutants, suggesting that AprA protease contributes to biocon...

  19. Extracellular Protease of Pseudomonas fluorescens CHA0, a Biocontrol Factor with Activity against the Root-Knot Nematode Meloidogyne incognita

    Science.gov (United States)

    Siddiqui, Imran Ali; Haas, Dieter; Heeb, Stephan

    2005-01-01

    In Pseudomonas fluorescens CHA0, mutation of the GacA-controlled aprA gene (encoding the major extracellular protease) or the gacA regulatory gene resulted in reduced biocontrol activity against the root-knot nematode Meloidogyne incognita during tomato and soybean infection. Culture supernatants of strain CHA0 inhibited egg hatching and induced mortality of M. incognita juveniles more strongly than did supernatants of aprA and gacA mutants, suggesting that AprA protease contributes to biocontrol. PMID:16151170

  20. Expression of Fap amyloids in Pseudomonas aeruginosa, P. fluorescens, and P. putida results in aggregation and increased biofilm formation

    DEFF Research Database (Denmark)

    Dueholm, Morten Simonsen; Søndergaard, Mads; Nilsson, Martin

    2013-01-01

    fluorescens Pf-5, and P. putida F1 to express Fap fibrils, and investigated the effect of Fap expression on aggregation and biofilm formation. The fap operon in all three Pseudomonas species conferred the ability to express Fap fibrils as shown using a recombinant approach. This Fap overexpression consistently......The fap operon, encoding functional amyloids in Pseudomonas (Fap), is present in most pseudomonads, but so far the expression and importance for biofilm formation has only been investigated for P. fluorescens strain UK4. In this study, we demonstrate the capacity of P. aeruginosa PAO1, P...

  1. Phytohormone production and colonization of canola (Brassica napus L.) roots by Pseudomonas fluorescens 6-8 under gnotobiotic conditions.

    Science.gov (United States)

    Pallai, Rajash; Hynes, Russell K; Verma, Brij; Nelson, Louise M

    2012-02-01

    Pseudomonas fluorescens 6-8, a rhizosphere isolate previously shown to enhance root elongation of canola ( Brassica napus L.), was characterized for its ability to produce indole-3-acetic acid and cytokinins in pure culture and in the rhizosphere of canola under gnotobiotic conditions in comparison with the cytokinin-producing strain P. fluorescens G20-18 and its mutant CNT2. Strain 6-8 produced isopentenyl adenosine, zeatin riboside, and dihydroxyzeatin riboside at levels similar to those of G20-18, but only very low concentrations of indole-3-acetic acid. In a gnotobiotic assay canola inoculated with 6-8 and G20-18 had higher concentrations of isopentenyl adenosine and zeatin riboside in the rhizosphere and greater root length than the noninoculated control. The ability of strain 6-8 to colonize canola roots was assessed following transformation with the green fluorescent protein and inoculation onto canola seed in a gnotobiotic assay. Higher populations of strain 6-8 were observed on the proximal region of the root closest to the seed than on the mid and distal portions 9 days after seed inoculation. The ability of P. fluorescens 6-8 to produce cytokinins, colonize the roots of canola seedlings, and enhance root elongation may contribute to its ability to survive in the rhizosphere and may benefit seedling growth.

  2. Pseudomonas fluorescens filamentous hemagglutinin, an iron-regulated protein, is an important virulence factor that modulates bacterial pathogenicity

    Directory of Open Access Journals (Sweden)

    Yuan-yuan Sun

    2016-08-01

    Full Text Available Pseudomonas fluorescens is a common bacterial pathogen to a wide range of aquaculture animals including various species of fish. In this study, we employed proteomic analysis and identified filamentous hemagglutinin (FHA as an iron-responsive protein secreted by TSS, a pathogenic P. fluorescens isolate. In vitro study showed that compared to the wild type, the fha mutant TSSfha (i exhibited a largely similar vegetative growth profile but significantly retarded in the ability of biofilm growth and producing extracellular matrix, (ii displayed no apparent flagella and motility, (iii was defective in the attachment to host cells and unable to form self-aggregation, (iv displayed markedly reduced capacity of hemagglutination and surviving in host serum. In vivo infection analysis revealed that TSSfha was significantly attenuated in the ability of dissemination in fish tissues and inducing host mortality, and that antibody blocking of the natural FHA produced by the wild type TSS impaired the infectivity of the pathogen. Furthermore, when introduced into turbot as a subunit vaccine, recombinant FHA elicited a significant protection against lethal TSS challenge. Taken together, these results indicate for the first time that P. fluorescens FHA is a key virulence factor essential to multiple biological processes associated with pathogenicity.

  3. Pseudomonas fluorescens induces strain-dependent and strain-independent host plant responses in defense networks, primary metabolism and photosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Pelletier, Dale A [ORNL; Morrell-Falvey, Jennifer L [ORNL; Karve, Abhijit A [ORNL; Lu, Tse-Yuan S [ORNL; Tschaplinski, Timothy J [ORNL; Tuskan, Gerald A [ORNL; Chen, Jay [ORNL; Martin, Madhavi Z [ORNL; Jawdy, Sara [ORNL; Weston, David [ORNL; Doktycz, Mitchel John [ORNL; Schadt, Christopher Warren [ORNL

    2012-01-01

    Colonization of plants by nonpathogenic Pseudomonas fluorescens strains can confer enhanced defense capacity against a broad spectrum of pathogens. Few studies, however, have linked defense pathway regulation to primary metabolism and physiology. In this study, physiological data, metabolites, and transcript profiles are integrated to elucidate how molecular networks initiated at the root-microbe interface influence shoot metabolism and whole-plant performance. Experiments with Arabidopsis thaliana were performed using the newly identified P. fluorescens GM30 or P. fluorescens Pf-5 strains. Co-expression networks indicated that Pf-5 and GM30 induced a subnetwork specific to roots enriched for genes participating in RNA regulation, protein degradation, and hormonal metabolism. In contrast, only GM30 induced a subnetwork enriched for calcium signaling, sugar and nutrient signaling, and auxin metabolism, suggesting strain dependence in network architecture. In addition, one subnetwork present in shoots was enriched for genes in secondary metabolism, photosynthetic light reactions, and hormone metabolism. Metabolite analysis indicated that this network initiated changes in carbohydrate and amino acid metabolism. Consistent with this, we observed strain-specific responses in tryptophan and phenylalanine abundance. Both strains reduced host plant carbon gain and fitness, yet provided a clear fitness benefit when plants were challenged with the pathogen P. syringae DC3000.

  4. Isolation and characterization of antimicrobial cyclic dipeptides from Pseudomonas fluorescens and their efficacy on sorghum grain mold fungi.

    Science.gov (United States)

    Sajeli Begum, Ahil; Basha, Shaik Ameer; Raghavendra, Govardhanam; Kumar, Mallela Venkata Nagesh; Singh, Yukthi; Patil, Jagannath V; Tanemura, Yuhei; Fujimoto, Yoshinori

    2014-01-01

    This study was aimed at isolation and characterization of natural antifungal compounds for grain mold, a key parasitic fungal disease of sorghum. Pseudomonas fluorescens strain isolated from rhizosphere of groundnut crop was selected as a source. Its biocontrolling ability was assessed by testing some biochemical attributes such as phosphate-solubilization, and HCN, NH3 , indole-3-acetic acid, and siderophore production. The strain showed positive result for all except indole-3-acetic acid, revealing its suitability for a further study. The antibiotic-sensitivity pattern of the strain against 43 antibiotics was also established, which showed resistance to 15 antibiotics. The efficacy of P. fluorescens strain against grain mold was identified by dual culture technique. Hundred percent inhibition was found against Fusarium moniliforme, an important causative agent of this disease. The strain was fermented for secondary metabolites and extracted with AcOEt. Chromatographic separation of the extract yielded four known compounds, cyclo(L-Pro-L-Phe) (1), cyclo(trans-4-hydroxy-L-Pro-L-Leu) (2), cyclo(trans-4-hydroxy-L-Pro-L-Phe) (3), and cyclo(Gly-L-Pro) (4), which were characterized by spectral analysis and optical rotation. The crude extract, a mixture of 2 and 3, and isolated 1 were proved to be significantly effective against grain mold fungi. This is the first report on production of these cyclic dipeptides by P. fluorescens and their antagonistic properties. Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.

  5. Listeria monocytogenes Impact on Mature or Old Pseudomonas fluorescens Biofilms During Growth at 4 and 20°C.

    Science.gov (United States)

    Puga, Carmen H; Orgaz, Belen; SanJose, Carmen

    2016-01-01

    Changes in spatial organization, as observed by confocal laser scanning microscopy (CLSM), viable cell content, biovolume, and substratum surface coverage of the biofilms formed on glass by Pseudomonas fluorescens resulting from co-culture with Listeria monocytogenes, were examined. Two strains of L. monocytogenes, two culture temperatures and two biofilm developmental stages were investigated. Both L. monocytogenes strains, a persistently sampled isolate (collected repeatedly along 3 years from a meat factory) and Scott A, induced shrinkage in matrix volume, both at 20°C and 4°C, in mature or old biofilms, without loss of P. fluorescens cell count per surface unit. The nearly homogeneous pattern of surface coverage shown by mono-species P. fluorescens biofilms, turned into more irregular layouts in co-culture with L. monocytogenes. The upper layer of both mono and dual-species biofilms turned to predominantly consist of matrix, with plenty of viable cells underneath, in old biofilms cultured at 20°C, but not in those grown at 4°C. Between 15 and 56% of the substratum area was covered by biofilm, the extent depending on temperature, time and L. monocytogenes strain. Real biofilms in food-related surfaces may thus be very heterogeneous regarding their superficial components, i.e., those more accessible to disinfectants. It is therefore a hygienic challenge to choose an adequate agent to disrupt them.

  6. Listeria monocytogenes impact on mature or old Pseudomonas fluorescens biofilms during growth at 4 and 20ºC

    Directory of Open Access Journals (Sweden)

    Puga eCH

    2016-02-01

    Full Text Available Changes in spatial organization, as observed by confocal laser scanning microscopy (CLSM, viable cell content, biovolume and substratum surface coverage of the biofilms formed on glass by Pseudomonas fluorescens resulting from co-culture with Listeria monocytogenes, were examined. Two strains of L. monocytogenes, two culture temperatures and two biofilm developmental stages were investigated. Both L. monocytogenes strains, a persistently sampled isolate (collected repeatedly along 3 years from a meat factory and Scott A, induced shrinkage in matrix volume, both at 20ºC and 4ºC, in mature or old biofilms, without loss of P. fluorescens cell count per surface unit. The nearly homogeneous pattern of surface coverage shown by mono-species P. fluorescens biofilms, turned into more irregular layouts in co-culture with L. monocytogenes. The upper layer of both mono and dual-species biofilms turned to predominantly consist of matrix, with plenty of viable cells underneath, in old biofilms cultured at 20ºC, but not in those grown at 4ºC. Between 15 and 56% of the substratum area was covered by biofilm, the extent depending on temperature, time and L. monocytogenes strain. Real biofilms in food-related surfaces may thus be very heterogeneous regarding their superficial components, i.e. those more accessible to disinfectants. It is therefore a hygienic challenge to choose an adequate agent to disrupt them.

  7. Enlightening mineral iron sensing in Pseudomonas fluorescens by surface active maghemite nanoparticles: Involvement of the OprF porin.

    Science.gov (United States)

    Magro, Massimiliano; Fasolato, Luca; Bonaiuto, Emanuela; Andreani, Nadia Andrea; Baratella, Davide; Corraducci, Vittorino; Miotto, Giovanni; Cardazzo, Barbara; Vianello, Fabio

    2016-10-01

    Mineral iron(III) recognition by bacteria is considered a matter of debate. The peculiar surface chemistry of novel naked magnetic nanoparticles, called SAMNs (surface active maghemite nanoparticles) characterized by solvent exposed Fe(3+) sites on their surface, was exploited for studying mineral iron sensing in Pseudomonas fluorescens. SAMNs were applied for mimicking Fe(3+) ions in solution, acting as magnetically drivable probes to evaluate putative Fe(3+) recognition sites on the microorganism surface. Culture broths and nano-bio-conjugates were characterized by UV-Vis spectroscopy and mass spectrometry. The whole heritage of a membrane porin (OprF) of P. fluorescens Ps_22 cells was recognized and firmly bound by SAMNs. The binding of nanoparticles to OprF porin was correlated to a drastic inhibition of a siderophore (pyoverdine) biosynthesis and to the stimulation of the production and rate of formation of a secondary siderophore. The analysis of metabolic pathways, based on P. fluorescens Ps_22 genomic information, evidenced that this putative secondary siderophore does not belong to a selection of the most common siderophores. In the scenario of an adhesion mechanism, it is plausible to consider OprF as the biological component deputed to the mineral iron sensing in P. fluorescens Ps_22, as well as one key of siderophore regulation. The present work sheds light on mineral iron sensing in microorganisms. Peculiar colloidal naked iron oxide nanoparticles offer a useful approach for probing the adhesion of bacterial surface on mineral iron for the identification of the specific recognition site for this iron uptake regulation in microorganisms. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Multistate outbreak of Pseudomonas fluorescens bloodstream infection after exposure to contaminated heparinized saline flush prepared by a compounding pharmacy.

    Science.gov (United States)

    Gershman, Mark D; Kennedy, Donald J; Noble-Wang, Judith; Kim, Curi; Gullion, Jessica; Kacica, Marilyn; Jensen, Bette; Pascoe, Neil; Saiman, Lisa; McHale, Jean; Wilkins, Melinda; Schoonmaker-Bopp, Dianna; Clayton, Joshua; Arduino, Matthew; Srinivasan, Arjun

    2008-12-01

    Pharmaceutical compounding, the manipulation of ingredients to create a customized medication, is a widespread practice. In January 2005, the Centers for Disease Control and Prevention was notified of 4 cases of Pseudomonas fluorescens bacteremia that were traced to contaminated heparinized saline intravenous flush syringes prepared as a compounded medical product. We reviewed medical records of symptomatic patients with P. fluorescens-positive cultures of blood specimens or sections of explanted catheters, reviewed the production process of syringes, performed syringe cultures, compared isolates by pulsed-field gel electrophoresis (PFGE), and examined catheters by scanning electron microscopy. We identified 80 patients in 6 states with P. fluorescens-positive cultures during December 2004-March 2006. Sixty-four patients (80%) had received a diagnosis of cancer. Seventy-four (99%) of 75 patients for whom information about catheter type was available had long-term indwelling catheters. Thirty-three (41%) of 80 cases were diagnosed 84-421 days after the patient's last potential exposure to a contaminated flush (delayed-onset cases). Compared with patients with early infection onset, more patients with delayed infection onset had venous ports (100% versus 50%; P <.001). By PFGE, clinical isolates from 50 (98%) of 51 patients were related to isolates cultured from unopened syringes. Scanning electron microscopy of explanted catheters revealed biofilms containing organisms morphologically consistent with P. fluorescens. This outbreak underscores important challenges in ensuring the safety of compounded pharmaceuticals and demonstrates the potential for substantially delayed infections after exposures to contaminated infusates. Exposures to compounded products should be considered when investigating outbreaks. Patients exposed to contaminated infusates require careful follow-up, because infections can occur long after exposure.

  9. Cloning and expression of Pseudomonas fluorescens 26-2 lipase gene in Pichia pastoris and characterizing for transesterification.

    Science.gov (United States)

    Yang, Jiangke; Zhang, Bo; Yan, Yunjun

    2009-11-01

    Pseudomonas lipases are important biocatalysts widely used in a variety of industrial fields. An extracellular lipase gene lipA with 1,854-bp open reading frame was cloned from Pseudomonas fluorescens 26-2. The multialignment assay of the putative amino acid and the secondary structure prediction revealed this enzyme could be classified into the lipolytic subfamily I.3 and secreted via adenosine-triphosphate-binding cassette pathway. The lipA gene was integrated into Pichia pastoris GS115, and the methanol-inducible recombinants with Mut(S) and Mut(+) phenotypes were acquired. The characteristics and the transesterification capacity shown by this enzyme suggested it is a useful biocatalyst for biodiesel preparation.

  10. Pseudomonas fluorescens Pf-5 genome-wide mutant screen for resistance to the antimicrobial peptide alfalfa snakin-1.

    Science.gov (United States)

    Ayub, Nicolás D; Fox, Ana R; García, Araceli N; Mozzicafreddo, Matteo; Cuccioloni, Massimiliano; Angeletti, Mauro; Pagano, Elba; Soto, Gabriela

    2015-01-01

    Snakin-1, a peptide produced by higher plants, has broad-spectrum antibiotic activity, inhibiting organisms ranging from Bacteria to Eukaryotes. However, the mode of action against target organisms is poorly understood. As a first step to elucidate the mechanism, we screened a mutation library of Pseudomonas fluorescens Pf-5 in LB and agar medium supplemented with alfalfa snakin-1 (MsSN1). We identified three biofilm formation-related Pseudomonas mutants that showed increased resistance to MsSN1. Genetic, physiological and bioinformatics analysis validated the results of the mutant screens, indicating that bacterial adhesion protein lapA is probably the target of MsSN1. Collectively, these findings suggest that snakin-1 acts on microbial adhesion properties. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Identification of anthranilate and benzoate metabolic operons of Pseudomonas fluorescens and functional characterization of their promoter regions

    Directory of Open Access Journals (Sweden)

    Lee Vincent D

    2006-01-01

    Full Text Available Abstract Background In an effort to identify alternate recombinant gene expression systems in Pseudomonas fluorescens, we identified genes encoding two native metabolic pathways that were inducible with inexpensive compounds: the anthranilate operon (antABC and the benzoate operon (benABCD. Results The antABC and benABCD operons were identified by homology to the Acinetobacter sp. anthranilate operon and Pseudomonas putida benzoate operon, and were confirmed to be regulated by anthranilate or benzoate, respectively. Fusions of the putative promoter regions to the E. coli lacZ gene were constructed to confirm inducible gene expression. Each operon was found to be controlled by an AraC family transcriptional activator, located immediately upstream of the first structural gene in each respective operon (antR or benR. Conclusion We have found the anthranilate and benzoate promoters to be useful for tightly controlling recombinant gene expression at both small (

  12. Mutation rate, spectrum, topology, and context-dependency in the DNA mismatch repair-deficient Pseudomonas fluorescens ATCC948.

    Science.gov (United States)

    Long, Hongan; Sung, Way; Miller, Samuel F; Ackerman, Matthew S; Doak, Thomas G; Lynch, Michael

    2014-12-23

    High levels of genetic diversity exist among natural isolates of the bacterium Pseudomonas fluorescens, and are especially elevated around the replication terminus of the genome, where strain-specific genes are found. In an effort to understand the role of genetic variation in the evolution of Pseudomonas, we analyzed 31,106 base substitutions from 45 mutation accumulation lines of P. fluorescens ATCC948, naturally deficient for mismatch repair, yielding a base-substitution mutation rate of 2.34 × 10(-8) per site per generation (SE: 0.01 × 10(-8)) and a small-insertion-deletion mutation rate of 1.65 × 10(-9) per site per generation (SE: 0.03 × 10(-9)). We find that the spectrum of mutations in prophage regions, which often contain virulence factors and antibiotic resistance, is highly similar to that in the intergenic regions of the host genome. Our results show that the mutation rate varies around the chromosome, with the lowest mutation rate found near the origin of replication. Consistent with observations from other studies, we find that site-specific mutation rates are heavily influenced by the immediately flanking nucleotides, indicating that mutations are context dependent. © The Author(s) 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  13. Impact of mutations in hemA and hemH genes on pyoverdine production by Pseudomonas fluorescens ATCC17400.

    Science.gov (United States)

    Baysse, C; Matthijs, S; Pattery, T; Cornelis, P

    2001-11-27

    A Pseudomonas fluorescens Tn5 mutant, with decreased production of the siderophore pyoverdine, was obtained, with the transposon inserted in the hemA gene coding for glutamyl tRNA reductase, the enzyme that catalyzes the first step of heme biosynthesis. Since this mutant was leaky, a second round of transposition was needed to obtain a second mutant completely auxotrophic for the heme precursor delta-aminolevulinate (ALA). Pyoverdine production by this mutant is ALA-dependent at concentrations above those needed to sustain growth. A transposon mutant in the hemH gene that encodes the enzyme ferrochelatase showing a characteristic red fluorescence upon UV exposure as a result of porphyrins accumulation, was obtained by selecting transconjugants on LB medium containing hemin. The DeltahemH mutant was characterized and the corresponding hemH gene sequenced. Antibodies against P. fluorescens HemH detected the protein both in soluble and membrane fractions of the wild-type and confirmed the absence of the enzyme in the mutant. The DeltahemH mutant failed to produce pyoverdine, but the production of the siderophore was restored by introduction of the Pseudomonas aeruginosa hemH gene in trans. These results indicate that de novo heme biosynthesis is needed for a normal level of siderophore pyoverdine production.

  14. Dissolution of fluorapatite by Pseudomonas fluorescens P35 resulting in fluorine release

    Science.gov (United States)

    Zhou, Jianping; Wang, Hongmei; Cravotta, Charles A.; Dong, Qiang; Xiang, Xing

    2017-01-01

    Chemical weathering of fluorine-bearing minerals is widely accepted as the main mechanism for the release of fluorine (F) to groundwater. Here, we propose a potential mechanism of F release via microbial dissolution of fluorapatite (Ca5(PO4)3F), which has been neglected previously. Batch culture experiments were conducted at 30°C with a phosphate-solubilizing bacteria strain, Pseudomonas fluorescens P35, and rock phosphates as the sole source of phosphate for microbial growth in parallel with abiotic controls. Rock phosphates consisted of 55–91% of fluorapatite and 5–10% of dolomite before microbial dissolution as indicated by X-ray diffraction (XRD). Mineral composition and morphology changed after microbial dissolution characterized by the disappearance of dolomite and the development of etched cavities on rock phosphate surfaces. The pH of media used was approximately 7.4 at the beginning and increased gradually to 7.7 in abiotic controls; with the inoculum, the pH decreased to acidic values of 3.7–3.8 after 27 h. Phosphate, calcium, and fluoride were released from the rock phosphate to the acidified medium. At 42 h, the concentration of F reached 8.1–10.3 mg L−1. The elevated F concentration was two times higher than the F levels in groundwater in regions diagnosed with fluorosis, and was toxic to the bacteria, as demonstrated by a precipitous decrease in live cells. Geochemical modeling demonstrated that the oxidation of glucose (the carbon source for microbial growth in the medium) to gluconic acid could decrease the pH to 3.7–3.8 and result in the dissolution of fluorapatite and dolomite. Dolomite and fluorapatite remained unsaturated, while concentrations of dissolved phosphorus (P), calcium (Ca), and F increased throughout the time course Fluorite reached saturation [saturation index (SI) 0.22–0.42] after 42 h in rock phosphate–amended biotic systems. However, fluorite was not detected in XRD patterns of the final residue from

  15. Synthesis of silver nanoparticles by endosymbiont Pseudomonas fluorescens CA 417 and their bactericidal activity.

    Science.gov (United States)

    Syed, Baker; M N, Nagendra Prasad; B L, Dhananjaya; K, Mohan Kumar; S, Yallappa; S, Satish

    2016-12-01

    The present study emphasizes on biogenic synthesis of silver nanoparticles and their bactericidal activity against human and phytopathogens. Nanoparticle synthesis was performed using endosymbiont Pseudomonas fluorescens CA 417 inhabiting Coffea arabica L. Synthesized nanoparticles were characterized using hyphenated spectroscopic techniques such as UV-vis spectroscopy which revealed maximum absorption 425nm. Fourier transform infrared spectroscopy (FTIR) analysis revealed the possible functional groups mediating and stabilizing silver nanoparticles with predominant peaks occurring at 3346 corresponding to hydroxyl group, 1635 corresponding carbonyl group and 680 to aromatic group. X-ray diffraction (XRD) analysis revealed the Bragg's diffraction pattern with distinct peaks at 38° 44°, 64° and 78° revealing the face-centered cubic (fcc) metallic crystal corresponding to the (111), (200), (220) and (311) facets of the crystal planes at 2θ angle. The energy dispersive X-ray spectroscopy (EDS) analysis revealed presence of high intense absorption peak at 3keV is a typical characteristic of nano-crystalline silver which confirmed the presence of elemental silver. TEM analysis revealed the size of the nanoparticles to be in the range 5-50nm with polydisperse nature of synthesized nanoparticles bearing myriad shapes. The particle size determined by Dynamic light scattering (DLS) method revealed average size to be 20.66nm. The synthesized silver nanoparticles exhibited significant antibacterial activity against panel of test pathogens. The results showed Klebsiella pneumoniae (MTCC 7407) and Xanthomonas campestris to be more sensitive among the test human pathogen and phyto-pathogen respectively. The study also reports synergistic effect of silver nanoparticles in combination with kanamycin which displayed increased fold activity up to 58.3% against Klebsiella pneumoniae (MTCC 7407). The results of the present investigation are promising enough and attribute towards

  16. Draft Genome Sequences of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8, Soil Bacteria That Cooperate To Degrade the Poly- -D-Glutamic Acid Anthrax Capsule

    KAUST Repository

    Stabler, R. A.

    2013-01-24

    A mixed culture of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8 degraded poly-γ-d-glutamic acid; when the 2 strains were cultured separately, no hydrolytic activity was apparent. Here we report the draft genome sequences of both soil isolates.

  17. A combined microcosm and mesocosm approach to examine factors affecting survival and mortality of Pseudomonas fluorescens Ag1 in seawater

    DEFF Research Database (Denmark)

    Ahl, Thomas; Christoffersen, K.; Riemann, B.

    1995-01-01

    Abstract: The survival of Pseudomonas fluorescens Ag1 in seawater of Roskilde Fjord (Denmark) was evaluated by a series of laboratory microcosm and field-based mesocosm experiments. In sterile seawater microcosms, culturability of Ag1 was negatively influenced by high salinity (34 versus 8...

  18. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian Fog

    2017-01-01

    Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun-nup regulon. Organization of the regulon is similar to clusters found in other CLP-p...

  19. Isolation and characterization of two new lipopeptide biosurfactants produced by Pseudomonas fluorescens BD5 isolated from water from the Arctic Archipelago of Svalbard

    Czech Academy of Sciences Publication Activity Database

    Janek, T.; Lukaszewicz, M.; Řezanka, Tomáš; Krasowska, A.

    2010-01-01

    Roč. 101, č. 15 (2010), s. 6118-6123 ISSN 0960-8524 Institutional research plan: CEZ:AV0Z50200510 Keywords : Pseudomonas fluorescens BD5 * Lipopeptide * Pseudofactins Subject RIV: EE - Microbiology, Virology Impact factor: 4.365, year: 2010

  20. Structure Revision of N-Mercapto-4-formylcarbostyril Produced by Pseudomonas fluorescens G308 to 2-(2-Hydroxyphenyl)thiazole-4-carbaldehyde [aeruginaldehyde

    DEFF Research Database (Denmark)

    Guillemyn, Karel; Ballet, Steven; Ye, Lumeng

    2014-01-01

    An antibiotic substance isolated from Pseudomonas fluorescens strain G308 was earlier assigned the structure of N-mercapto-4-formylcarbostyril, but computational predictions of the H-1 and C-13 NMR magnetic shielding tensors show this structure to be incompatible with the published spectroscopic ...

  1. The Pseudomonas fluorescens Siderophore Pyoverdine Weakens Arabidopsis thaliana Defense in Favor of Growth in Iron-Deficient Conditions.

    Science.gov (United States)

    Trapet, Pauline; Avoscan, Laure; Klinguer, Agnès; Pateyron, Stéphanie; Citerne, Sylvie; Chervin, Christian; Mazurier, Sylvie; Lemanceau, Philippe; Wendehenne, David; Besson-Bard, Angélique

    2016-05-01

    Pyoverdines are siderophores synthesized by fluorescent Pseudomonas spp. Under iron-limiting conditions, these high-affinity ferric iron chelators are excreted by bacteria in the soil to acquire iron. Pyoverdines produced by beneficial Pseudomonas spp. ameliorate plant growth. Here, we investigate the physiological incidence and mode of action of pyoverdine from Pseudomonas fluorescens C7R12 on Arabidopsis (Arabidopsis thaliana) plants grown under iron-sufficient or iron-deficient conditions. Pyoverdine was provided to the medium in its iron-free structure (apo-pyoverdine), thus mimicking a situation in which it is produced by bacteria. Remarkably, apo-pyoverdine abolished the iron-deficiency phenotype and restored the growth of plants maintained in the iron-deprived medium. In contrast to a P. fluorescens C7R12 strain impaired in apo-pyoverdine production, the wild-type C7R12 reduced the accumulation of anthocyanins in plants grown in iron-deficient conditions. Under this condition, apo-pyoverdine modulated the expression of around 2,000 genes. Notably, apo-pyoverdine positively regulated the expression of genes related to development and iron acquisition/redistribution while it repressed the expression of defense-related genes. Accordingly, the growth-promoting effect of apo-pyoverdine in plants grown under iron-deficient conditions was impaired in iron-regulated transporter1 and ferric chelate reductase2 knockout mutants and was prioritized over immunity, as highlighted by an increased susceptibility to Botrytis cinerea This process was accompanied by an overexpression of the transcription factor HBI1, a key node for the cross talk between growth and immunity. This study reveals an unprecedented mode of action of pyoverdine in Arabidopsis and demonstrates that its incidence on physiological traits depends on the plant iron status. © 2016 American Society of Plant Biologists. All Rights Reserved.

  2. The Pseudomonas fluorescens Siderophore Pyoverdine Weakens Arabidopsis thaliana Defense in Favor of Growth in Iron-Deficient Conditions1

    Science.gov (United States)

    Trapet, Pauline; Avoscan, Laure; Klinguer, Agnès; Pateyron, Stéphanie; Chervin, Christian; Mazurier, Sylvie; Lemanceau, Philippe; Wendehenne, David; Besson-Bard, Angélique

    2016-01-01

    Pyoverdines are siderophores synthesized by fluorescent Pseudomonas spp. Under iron-limiting conditions, these high-affinity ferric iron chelators are excreted by bacteria in the soil to acquire iron. Pyoverdines produced by beneficial Pseudomonas spp. ameliorate plant growth. Here, we investigate the physiological incidence and mode of action of pyoverdine from Pseudomonas fluorescens C7R12 on Arabidopsis (Arabidopsis thaliana) plants grown under iron-sufficient or iron-deficient conditions. Pyoverdine was provided to the medium in its iron-free structure (apo-pyoverdine), thus mimicking a situation in which it is produced by bacteria. Remarkably, apo-pyoverdine abolished the iron-deficiency phenotype and restored the growth of plants maintained in the iron-deprived medium. In contrast to a P. fluorescens C7R12 strain impaired in apo-pyoverdine production, the wild-type C7R12 reduced the accumulation of anthocyanins in plants grown in iron-deficient conditions. Under this condition, apo-pyoverdine modulated the expression of around 2,000 genes. Notably, apo-pyoverdine positively regulated the expression of genes related to development and iron acquisition/redistribution while it repressed the expression of defense-related genes. Accordingly, the growth-promoting effect of apo-pyoverdine in plants grown under iron-deficient conditions was impaired in iron-regulated transporter1 and ferric chelate reductase2 knockout mutants and was prioritized over immunity, as highlighted by an increased susceptibility to Botrytis cinerea. This process was accompanied by an overexpression of the transcription factor HBI1, a key node for the cross talk between growth and immunity. This study reveals an unprecedented mode of action of pyoverdine in Arabidopsis and demonstrates that its incidence on physiological traits depends on the plant iron status. PMID:26956666

  3. Nitrilase from Pseudomonas fluorescens EBC191: cloning and heterologous expression of the gene and biochemical characterization of the recombinant enzyme.

    Science.gov (United States)

    Kiziak, Christoph; Conradt, Doris; Stolz, Andreas; Mattes, Ralf; Klein, Joachim

    2005-11-01

    The gene encoding an enantioselective arylacetonitrilase was identified on a 3.8 kb DNA fragment from the genomic DNA of Pseudomonas fluorescens EBC191. The gene was isolated, sequenced and cloned into the L-rhamnose-inducible expression vector pJOE2775. The nitrilase was produced in large quantities and purified as a histidine-tagged enzyme from crude extracts of L-rhamnose-induced cells of Escherichia coli JM109. The purified nitrilase was significantly stabilized during storage by the addition of 1 M ammonium sulfate. The temperature optimum (50 degrees C), pH optimum (pH 6.5), and specific activity of the recombinant nitrilase were similar to those of the native enzyme from P. fluorescens EBC191. The enzyme hydrolysed various phenylacetonitriles with different substituents in the 2-position and also heterocyclic and bicyclic arylacetonitriles to the corresponding carboxylic acids. The conversion of most arylacetonitriles was accompanied by the formation of different amounts of amides as by-products. The relative amounts of amides formed from different nitriles increased with an increasing negative inductive effect of the substituent in the 2-position. The acids and amides that were formed from chiral nitriles demonstrated in most cases opposite enantiomeric excesses. Thus mandelonitrile was converted by the nitrilase preferentially to R-mandelic acid and S-mandelic acid amide. The nitrilase gene is physically linked in the genome of P. fluorescens with genes encoding the degradative pathway for mandelic acid. This might suggest a natural function of the nitrilase in the degradation of mandelonitrile or similar naturally occurring hydroxynitriles.

  4. Susceptibility of Mycobacterium immunogenum and Pseudomonas fluorescens to formaldehyde and non-formaldehyde biocides in semi-synthetic metalworking fluids.

    Science.gov (United States)

    Selvaraju, Suresh B; Khan, Izhar U H; Yadav, Jagjit S

    2011-01-20

    Mycobacterium immunogenum, a newly identified member of the Mycobacterium chelonae_M. abscessus complex is considered a potential etiological agent for hypersensitivity pneumonitis (HP) in machine workers exposed to contaminated metalworking fluid (MWF). This study investigated the biocidal efficacy of the frequently applied commercial formaldehyde-releasing (HCHO) biocides Grotan and Bioban CS 1135 and non-HCHO type biocides Kathon 886 MW (isothiazolone) and Preventol CMK 40 (phenolic) toward this emerging mycobacterial species (M. immunogenum) in HP-linked MWFs, alone and in presence of a representative of the Gram-negative bacterial contaminants, Pseudomonas fluorescens, using two semi-synthetic MWF matrices (designated Fluid A and Fluid B). Relative biocide susceptibility analysis indicated M immunogenum to be comparatively more resistant (2-1600 fold) than P. fluorescens to the tested biocides under the varied test conditions. In terms of minimum inhibitory concentration, Kathon was the most effective biocide against M. immunogenum. Fluid factors had a major effect on the biocide susceptibility. Fluid A formulation provided greater protective advantage to the test organisms than Fluid B. Fluid dialysis (Fluid A) led to an increased biocidal efficacy of Grotan, Kathon and Preventol against M. immunogenum further implying the role of native fluid components. Used fluid matrix, in general, increased the resistance of the two test organisms against the biocides, with certain exceptions. M. immunogenum resistance increased in presence of the co-contaminant P. fluorescens. Collectively, the results show a multifactorial nature of the biocide susceptibility of MWF-colonizing mycobacteria and highlight the importance of more rigorous efficacy testing and validation of biocides prior to and during their application in metalworking fluid operations.

  5. Susceptibility of Mycobacterium immunogenum and Pseudomonas fluorescens to Formaldehyde and Non-Formaldehyde Biocides in Semi-Synthetic Metalworking Fluids

    Directory of Open Access Journals (Sweden)

    Suresh B. Selvaraju

    2011-01-01

    Full Text Available Mycobacterium immunogenum, a newly identified member of the Mycobacterium chelonae_M. abscessus complex is considered a potential etiological agent for hypersensitivity pneumonitis (HP in machine workers exposed to contaminated metalworking fluid (MWF. This study investigated the biocidal efficacy of the frequently applied commercial formaldehyde-releasing (HCHO biocides Grotan and Bioban CS 1135 and non-HCHO type biocides Kathon 886 MW (isothiazolone and Preventol CMK 40 (phenolic toward this emerging mycobacterial species (M. immunogenum in HP-linked MWFs, alone and in presence of a representative of the Gram-negative bacterial contaminants, Pseudomonas fluorescens, using two semi-synthetic MWF matrices (designated Fluid A and Fluid B. Relative biocide susceptibility analysis indicated M immunogenum to be comparatively more resistant (2–1600 fold than P. fluorescens to the tested biocides under the varied test conditions. In terms of minimum inhibitory concentration, Kathon was the most effective biocide against M. immunogenum. Fluid factors had a major effect on the biocide susceptibility. Fluid A formulation provided greater protective advantage to the test organisms than Fluid B. Fluid dialysis (Fluid A led to an increased biocidal efficacy of Grotan, Kathon and Preventol against M. immunogenum further implying the role of native fluid components. Used fluid matrix, in general, increased the resistance of the two test organisms against the biocides, with certain exceptions. M. immunogenum resistance increased in presence of the co-contaminant P. fluorescens. Collectively, the results show a multifactorial nature of the biocide susceptibility of MWF-colonizing mycobacteria and highlight the importance of more rigorous efficacy testing and validation of biocides prior to and during their application in metalworking fluid operations.

  6. 76 FR 52871 - Pseudomonas fluorescens Strain CL145A; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2011-08-24

    ... fluorescens-contaminated blood product was used for transfusions. In the past, EPA has registered several... and select ``Test Methods and Guidelines.'' C. How can I file an objection or hearing request? Under... following methods: Federal eRulemaking Portal: http://www.regulations.gov . Follow the on-line instructions...

  7. Photocatalytic disinfection of spoilage bacteria Pseudomonas fluorescens and Macrococcus caseolyticus by nano-TiO2

    Science.gov (United States)

    Photocatalytic disinfection of spoilage bacteria gram-negative (G-) P. fluorescens and gram-positive (G+) M. caseolyticus by nano-TiO2 under different experimental conditions and the disinfection mechanism were investigated. The experimental conditions included the initial bacterial populations, nan...

  8. Whole-genome sequence of Pseudomonas fluorescens EK007-RG4, a promising biocontrol agent against a broad range of bacteria, including the fire blight bacterium Erwinia amylovora

    DEFF Research Database (Denmark)

    Habibi, Roghayeh; Tarighi, Saeed; Behravan, Javad

    2017-01-01

    Here, we report the first draft whole-genome sequence of Pseudomonas fluorescens strain EK007-RG4, which was isolated from the phylloplane of a pear tree. P. fluorescens EK007-RG4 displays strong antagonism against Erwinia amylovora, the causal agent for fire blight disease, in addition to several...

  9. Mycorrhization between Cistus ladanifer L. and Boletus edulis Bull is enhanced by the mycorrhiza helper bacteria Pseudomonas fluorescens Migula.

    Science.gov (United States)

    Mediavilla, Olaya; Olaizola, Jaime; Santos-del-Blanco, Luis; Oria-de-Rueda, Juan Andrés; Martín-Pinto, Pablo

    2016-02-01

    Boletus edulis Bull. is one of the most economically and gastronomically valuable fungi worldwide. Sporocarp production normally occurs when symbiotically associated with a number of tree species in stands over 40 years old, but it has also been reported in 3-year-old Cistus ladanifer L. shrubs. Efforts toward the domestication of B. edulis have thus focused on successfully generating C. ladanifer seedlings associated with B. edulis under controlled conditions. Microorganisms have an important role mediating mycorrhizal symbiosis, such as some bacteria species which enhance mycorrhiza formation (mycorrhiza helper bacteria). Thus, in this study, we explored the effect that mycorrhiza helper bacteria have on the efficiency and intensity of the ectomycorrhizal symbiosis between C. ladanifer and B. edulis. The aim of this work was to optimize an in vitro protocol for the mycorrhizal synthesis of B. edulis with C. ladanifer by testing the effects of fungal culture time and coinoculation with the helper bacteria Pseudomonas fluorescens Migula. The results confirmed successful mycorrhizal synthesis between C. ladanifer and B. edulis. Coinoculation of B. edulis with P. fluorescens doubled within-plant mycorrhization levels although it did not result in an increased number of seedlings colonized with B. edulis mycorrhizae. B. edulis mycelium culture time also increased mycorrhization levels but not the presence of mycorrhizae. These findings bring us closer to controlled B. edulis sporocarp production in plantations.

  10. Biocontrol of avocado dematophora root rot by antagonistic Pseudomonas fluorescens PCL1606 correlates with the production of 2-hexyl 5-propyl resorcinol.

    Science.gov (United States)

    Cazorla, Francisco M; Duckett, Simon B; Bergström, Ed T; Noreen, Sadaf; Odijk, Roeland; Lugtenberg, Ben J J; Thomas-Oates, Jane E; Bloemberg, Guido V

    2006-04-01

    A collection of 905 bacterial isolates from the rhizospheres of healthy avocado trees was obtained and screened for antagonistic activity against Dematophora necatrix, the cause of avocado Dematophora root rot (also called white root rot). A set of eight strains was selected on the basis of growth inhibitory activity against D. necatrix and several other important soilborne phytopathogenic fungi. After typing of these strains, they were classified as belonging to Pseudomonas chlororaphis, Pseudomonas fluorescens, and Pseudomonas putida. The eight antagonistic Pseudomonas spp. were analyzed for their secretion of hydrogen cyanide, hydrolytic enzymes, and antifungal metabolites. P. chlororaphis strains produced the antibiotic phenazine-1-carboxylic acid and phenazine-1-carboxamide. Upon testing the biocontrol ability of these strains in a newly developed avocado-D. necatrix test system and in a tomato-F oxysporum test system, it became apparent that P. fluorescens PCL1606 exhibited the highest biocontrol ability. The major antifungal activity produced by strain P. fluorescens PCL1606 did not correspond to any of the major classes of antifungal antibiotics produced by Pseudomonas biocontrol strains. This compound was purified and subsequently identified as 2-hexyl 5-propyl resorcinol (HPR). To study the role of HPR in biocontrol activity, two Tn5 mutants of P. fluorescens PCL1606 impaired in antagonistic activity were selected. These mutants were shown to impair HRP production and showed a decrease in biocontrol activity. As far as we know, this is the first report of a Pseudomonas biocontrol strain that produces HPR in which the production of this compound correlates with its biocontrol activity.

  11. Exposure-related effects of Pseudomonas fluorescens, strain CL145A, on coldwater, coolwater, and warmwater fish

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Denise A. Mayer,

    2015-01-01

    The exposure-related effects of a commercially prepared spray-dried powder (SDP) formulation of Pseudomonas fluorescens, strain CL145A, were evaluated on coldwater, coolwater, and warmwater fish endemic to the Great Lakes and Upper Mississippi River Basins. Nine species of young-of-the-year fish were exposed to SDP for 24 hours by using continuous-flow, serial-dilution exposure systems at temperatures of 12 degrees Celsius (°C; 2 species; Oncorhynchus mykiss [rainbow trout] and Salvelinus fontinalis [brook trout]), 17 °C (3 species; Perca flavescens [yellow perch], Sander vitreus [walleye], and Acipenser fulvescens [lake sturgeon]), or 22 °C (4 species; Micropterus salmoides [largemouth bass], Micropterus dolomieu [smallmouth bass], Lepomis macrochirus [bluegill sunfish], and Ictalurus punctatus [channel catfish]).

  12. Adaptive Divergence in Experimental Populations of Pseudomonas fluorescens. V. Insight into the Niche Specialist Fuzzy Spreader Compels Revision of the Model Pseudomonas Radiation

    Science.gov (United States)

    Ferguson, Gayle C.; Bertels, Frederic; Rainey, Paul B.

    2013-01-01

    Pseudomonas fluorescens is a model for the study of adaptive radiation. When propagated in a spatially structured environment, the bacterium rapidly diversifies into a range of niche specialist genotypes. Here we present a genetic dissection and phenotypic characterization of the fuzzy spreader (FS) morphotype—a type that arises repeatedly during the course of the P. fluorescens radiation and appears to colonize the bottom of static broth microcosms. The causal mutation is located within gene fuzY (pflu0478)—the fourth gene of the five-gene fuzVWXYZ operon. fuzY encodes a β-glycosyltransferase that is predicted to modify lipopolysaccharide (LPS) O antigens. The effect of the mutation is to cause cell flocculation. Analysis of 92 independent FS genotypes showed each to have arisen as the result of a loss-of-function mutation in fuzY, although different mutations have subtly different phenotypic and fitness effects. Mutations within fuzY were previously shown to suppress the phenotype of mat-forming wrinkly spreader (WS) types. This prompted a reinvestigation of FS niche preference. Time-lapse photography showed that FS colonizes the meniscus of broth microcosms, forming cellular rafts that, being too flimsy to form a mat, collapse to the vial bottom and then repeatably reform only to collapse. This led to a reassessment of the ecology of the P. fluorescens radiation. Finally, we show that ecological interactions between the three dominant emergent types (smooth, WS, and FS), combined with the interdependence of FS and WS on fuzY, can, at least in part, underpin an evolutionary arms race with bacteriophage SBW25Φ2, to which mutation in fuzY confers resistance. PMID:24077305

  13. Influence of phenolic acids on indole acetic acid production and on the type III secretion system gene transcription in food-associated Pseudomonas fluorescens KM05.

    Science.gov (United States)

    Myszka, Kamila; Schmidt, Marcin T; Olejnik-Schmidt, Agnieszka K; Leja, Katarzyna; Czaczyk, Katarzyna

    2014-12-01

    The purpose of these investigations was to evaluate the reduction capability of phenolic acids (ferulic, chlorogenic, gallic, and p-coumaric acids) on indole acetic acid synthesis by food-associated Pseudomonas fluorescens KM05. Specific genetic primer for the type III secretion system (TTSS) in P. fluorescens KM05 was designed and the influence of phenolic acids on its expression was investigated. In the work the ferulic and chlorogenic acids at the concentration of 0.02 and 0.04 μg/ml affected on bacterial growth pattern and the signal molecules production. The phenolic acids, that were appreciable effective against P. fluorescens KM05 indole acetic acid production, significantly suppressed TTSS gene. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  14. Identification and characterization of the emhABC efflux system for polycyclic aromatic hydrocarbons in Pseudomonas fluorescens cLP6a.

    Science.gov (United States)

    Hearn, Elizabeth M; Dennis, Jonathan J; Gray, Murray R; Foght, Julia M

    2003-11-01

    The hydrocarbon-degrading environmental isolate Pseudomonas fluorescens LP6a possesses an active efflux mechanism for the polycyclic aromatic hydrocarbons phenanthrene, anthracene, and fluoranthene but not for naphthalene or toluene. PCR was used to detect efflux pump genes belonging to the resistance-nodulation-cell division (RND) superfamily in a plasmid-cured derivative, P. fluorescens cLP6a, which is unable to metabolize hydrocarbons. One RND pump, whose gene was identified in P. fluorescens cLP6a and was designated emhB, showed homology to the multidrug and solvent efflux pumps in Pseudomonas aeruginosa and Pseudomonas putida. The emhB gene is located in a gene cluster with the emhA and emhC genes, which encode the membrane fusion protein and outer membrane protein components of the efflux system, respectively. Disruption of emhB by insertion of an antibiotic resistance cassette demonstrated that the corresponding gene product was responsible for the efflux of polycyclic aromatic hydrocarbons. The emhB gene disruption did not affect the resistance of P. fluorescens cLP6a to tetracycline, erythromycin, trimethoprim, or streptomycin, but it did decrease resistance to chloramphenicol and nalidixic acid, indicating that the EmhABC system also functions in the efflux of these compounds and has an unusual selectivity. Phenanthrene efflux was observed in P. aeruginosa, P. putida, and Burkholderia cepacia but not in Azotobacter vinelandii. Polycyclic aromatic hydrocarbons represent a new class of nontoxic, highly hydrophobic compounds that are substrates of RND efflux systems, and the EmhABC system in P. fluorescens cLP6a has a narrow substrate range for these hydrocarbons and certain antibiotics.

  15. Impact of Medium on the Development and Physiology of Pseudomonas fluorescens Biofilms on Polyurethane Paint

    Science.gov (United States)

    2012-02-01

    1989). In Pseudomonas putida , increasing glucose and phosphate concentrations Approved for public release; distribution unlimited 14 increased...polyhydroxyalkanoates in Pseudomonas putida KT2442 and the fundamental role of PhaZ depolymerase for the metabolic balance. Environ Microbiol. 12(1...Effects of nutrients on biofilm formation and detachment of a Pseudomonas putida strain isolated from a paper machine. Water Res. 41(13):2885-2892

  16. Genome-wide analysis of bacterial determinants of plant growth promotion and induced systemic resistance by Pseudomonas fluorescens.

    Science.gov (United States)

    Cheng, Xu; Etalo, Desalegn W; van de Mortel, Judith E; Dekkers, Ester; Nguyen, Linh; Medema, Marnix H; Raaijmakers, Jos M

    2017-11-01

    Pseudomonas fluorescens strain SS101 (Pf.SS101) promotes growth of Arabidopsis thaliana, enhances greening and lateral root formation, and induces systemic resistance (ISR) against the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). Here, targeted and untargeted approaches were adopted to identify bacterial determinants and underlying mechanisms involved in plant growth promotion and ISR by Pf.SS101. Based on targeted analyses, no evidence was found for volatiles, lipopeptides and siderophores in plant growth promotion by Pf.SS101. Untargeted, genome-wide analyses of 7488 random transposon mutants of Pf.SS101 led to the identification of 21 mutants defective in both plant growth promotion and ISR. Many of these mutants, however, were auxotrophic and impaired in root colonization. Genetic analysis of three mutants followed by site-directed mutagenesis, genetic complementation and plant bioassays revealed the involvement of the phosphogluconate dehydratase gene edd, the response regulator gene colR and the adenylsulfate reductase gene cysH in both plant growth promotion and ISR. Subsequent comparative plant transcriptomics analyses strongly suggest that modulation of sulfur assimilation, auxin biosynthesis and transport, steroid biosynthesis and carbohydrate metabolism in Arabidopsis are key mechanisms linked to growth promotion and ISR by Pf.SS101. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  17. Proteomic detection of non-annotated protein-coding genes in Pseudomonas fluorescens Pf0-1.

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    Wook Kim

    Full Text Available Genome sequences are annotated by computational prediction of coding sequences, followed by similarity searches such as BLAST, which provide a layer of possible functional information. While the existence of processes such as alternative splicing complicates matters for eukaryote genomes, the view of bacterial genomes as a linear series of closely spaced genes leads to the assumption that computational annotations that predict such arrangements completely describe the coding capacity of bacterial genomes. We undertook a proteomic study to identify proteins expressed by Pseudomonas fluorescens Pf0-1 from genes that were not predicted during the genome annotation. Mapping peptides to the Pf0-1 genome sequence identified sixteen non-annotated protein-coding regions, of which nine were antisense to predicted genes, six were intergenic, and one read in the same direction as an annotated gene but in a different frame. The expression of all but one of the newly discovered genes was verified by RT-PCR. Few clues as to the function of the new genes were gleaned from informatic analyses, but potential orthologs in other Pseudomonas genomes were identified for eight of the new genes. The 16 newly identified genes improve the quality of the Pf0-1 genome annotation, and the detection of antisense protein-coding genes indicates the under-appreciated complexity of bacterial genome organization.

  18. Genetic responses induced in olive roots upon colonization by the biocontrol endophytic bacterium Pseudomonas fluorescens PICF7.

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    Elisabetta Schilirò

    Full Text Available Knowledge on the genetic basis underlying interactions between beneficial bacteria and woody plants is still very limited, and totally absent in the case of olive. We aimed to elucidate genetic responses taking place during the colonization of olive roots by the native endophyte Pseudomonas fluorescens PICF7, an effective biocontrol agent against Verticillium wilt of olive. Roots of olive plants grown under non-gnotobiotic conditions were collected at different time points after PICF7 inoculation. A Suppression Subtractive Hybridization cDNA library enriched in induced genes was generated. Quantitative real time PCR (qRT-PCR analysis validated the induction of selected olive genes. Computational analysis of 445 olive ESTs showed that plant defence and response to different stresses represented nearly 45% of genes induced in PICF7-colonized olive roots. Moreover, quantitative real-time PCR (qRT-PCR analysis confirmed induction of lipoxygenase, phenylpropanoid, terpenoids and plant hormones biosynthesis transcripts. Different classes of transcription factors (i.e., bHLH, WRKYs, GRAS1 were also induced. This work highlights for the first time the ability of an endophytic Pseudomonas spp. strain to mount a wide array of defence responses in an economically-relevant woody crop such as olive, helping to explain its biocontrol activity.

  19. Controlling instability in gacS-gacA regulatory genes during inoculant production of Pseudomonas fluorescens biocontrol strains.

    Science.gov (United States)

    Duffy, B K; Défago, G

    2000-08-01

    Secondary metabolism in fluorescent pseudomonads is globally regulated by gacS, which encodes a membrane-bound sensor kinase, and gacA, which encodes a transcriptional response regulator. Spontaneous mutation in either gene blocked biosynthesis of the antimicrobial compounds hydrogen cyanide, 2,4-diacetylphloroglucinol, pyoluteorin, and pyrrolnitrin by the model biocontrol strain Pseudomonas fluorescens CHA0. Spontaneous mutants also had altered abilities to utilize several carbon sources and to increase medium pH compared with the wild type, suggesting that gacS and gacA influence primary as well as secondary bacterial metabolism. Inoculant efficacy for biocontrol was significantly reduced by contamination with regulatory mutants which accumulated during inoculum production. Spontaneous mutants accumulated in all 192 separate liquid cultures examined, typically at a frequency of 1% or higher after 12 days. During scale-up in a simulated industrial fermentation process, mutants increased exponentially and accounted for 7, 23, and 61% of the total viable cells after transfer to 20-, 100-, and 500-ml preparations, respectively. GacS(-) and GacA(-) mutants had identical phenotypes and occurred at the same frequency, indicating that the selective pressures for the two mutants were similar. We developed a simple screening method for monitoring inoculant quality based on the distinctive appearance of mutant colonies (i.e., orange color, enlarged diameter, hyperfluorescence). Mutant competitiveness was favored in a nutrient-rich medium with a high electrolyte concentration (nutrient broth containing yeast extract). We were able to control mutant accumulation and to clean up contaminated cultures by using certain mineral amendments (i.e., zinc, copper, cobalt, manganese, and ammonium molybdate) or by diluting media 1/10. Spontaneous mutants and genetic constructs had the same response to culture conditions. Zinc and medium dilution were also effective for improving the

  20. Conversion of Sterically Demanding α,α-Disubstituted Phenylacetonitriles by the Arylacetonitrilase from Pseudomonas fluorescens EBC191

    Science.gov (United States)

    Baum, Stefanie; Williamson, Dael S.; Sewell, Trevor

    2012-01-01

    The nitrilase from Pseudomonas fluorescens EBC191 converted 2-methyl-2-phenylpropionitrile, which contains a quaternary carbon atom in the α-position toward the nitrile group, and also similar sterically demanding substrates, such as 2-hydroxy-2-phenylpropionitrile (acetophenone cyanohydrin) or 2-acetyloxy-2-methylphenylacetonitrile. 2-Methyl-2-phenylpropionitrile was hydrolyzed to almost stoichiometric amounts of the corresponding acid. Acetophenone cyanohydrin was transformed to the corresponding acid (atrolactate) and amide (atrolactamide) at a ratio of about 3.4:1. The (R)-acid and the (S)-amide were formed preferentially from acetophenone cyanohydrin. A homology model of the nitrilase suggested that steric hindrance with amino acid residue Tyr54 could impair the binding or conversion of sterically demanding substrates. Therefore, several enzyme variants that carried mutations in the respective residues were generated and subsequently analyzed for the substrate specificity and enantioselectivity of the reactions. Enzyme variants that demonstrated increased relative activities for the conversion of acetophenone cyanohydrin were identified. The chiral analysis of these reactions demonstrated peculiar reaction kinetics, which suggested that the enzyme variants converted the nonpreferred (S)-enantiomer of acetophenone cyanohydrin with a higher reaction rate than that of the (preferred) (R)-enantiomer. Recombinant whole-cell catalysts that simultaneously produced the nitrilase from P. fluorescens EBC191 and a plant-derived (S)-oxynitrilase from cassava (Manihot esculenta) converted acetophenone plus cyanide at pH 4.5 to (S)-atrolactate and (S)-atrolactamide. These recombinant cells are promising catalysts for the synthesis of stable chiral quaternary carbon centers from ketones. PMID:22020513

  1. Biological control of wheat root diseases by the CLP-producing strain Pseudomonas fluorescens HC1-07.

    Science.gov (United States)

    Yang, Ming-Ming; Wen, Shan-Shan; Mavrodi, Dmitri V; Mavrodi, Olga V; von Wettstein, Diter; Thomashow, Linda S; Guo, Jian-Hua; Weller, David M

    2014-03-01

    Pseudomonas fluorescens HC1-07, previously isolated from the phyllosphere of wheat grown in Hebei province, China, suppresses the soilborne disease of wheat take-all, caused by Gaeumannomyces graminis var. tritici. We report here that strain HC1-07 also suppresses Rhizoctonia root rot of wheat caused by Rhizoctonia solani AG-8. Strain HC1-07 produced a cyclic lipopeptide (CLP) with a molecular weight of 1,126.42 based on analysis by electrospray ionization mass spectrometry. Extracted CLP inhibited the growth of G. graminis var. tritici and R. solani in vitro. To determine the role of this CLP in biological control, plasposon mutagenesis was used to generate two nonproducing mutants, HC1-07viscB and HC1-07prtR2. Analysis of regions flanking plasposon insertions in HC1-07prtR2 and HC1-07viscB revealed that the inactivated genes were similar to prtR and viscB, respectively, of the well-described biocontrol strain P. fluorescens SBW25 that produces the CLP viscosin. Both genes in HC1-07 were required for the production of the viscosin-like CLP. The two mutants were less inhibitory to G. graminis var. tritici and R. solani in vitro and reduced in ability to suppress take-all. HC1-07viscB but not HC-07prtR2 was reduced in ability to suppress Rhizoctonia root rot. In addition to CLP production, prtR also played a role in protease production.

  2. An alternative physiological role for the EmhABC efflux pump in Pseudomonas fluorescens cLP6a

    Directory of Open Access Journals (Sweden)

    Adebusuyi Abigail A

    2011-11-01

    Full Text Available Abstract Background Efflux pumps belonging to the resistance-nodulation-division (RND superfamily in bacteria are involved in antibiotic resistance and solvent tolerance but have an unknown physiological role. EmhABC, a RND-type efflux pump in Pseudomonas fluorescens strain cLP6a, extrudes hydrophobic antibiotics, dyes and polycyclic aromatic hydrocarbons including phenanthrene. The effects of physico-chemical factors such as temperature or antibiotics on the activity and expression of EmhABC were determined in order to deduce its physiological role(s in strain cLP6a in comparison to the emhB disruptant strain, cLP6a-1. Results Efflux assays conducted with 14C-phenanthrene showed that EmhABC activity is affected by incubation temperature. Increased phenanthrene efflux was measured in cLP6a cells grown at 10°C and decreased efflux was observed at 35°C compared with cells grown at the optimum temperature of 28°C. Membrane fatty acids in cLP6a cells were substantially altered by changes in growth temperature and in the presence of tetracycline. Changed membrane fatty acids and increased membrane permeability were associated with ~30-fold increased expression of emhABC in cLP6a cells grown at 35°C, and with increased extracellular free fatty acids. Growth of P. fluorescens cLP6a at supra-optimal temperature was enhanced by the presence of EmhABC compared to strain cLP6a-1. Conclusions Combined, these observations suggest that the EmhABC efflux pump may be involved in the management of membrane stress effects such as those due to unfavourable incubation temperatures. Efflux of fatty acids replaced as a result of membrane damage or phospholipid turnover may be the primary physiological role of the EmhABC efflux pump in P. fluorescens cLP6a.

  3. The role of the antimicrobial compound 2,4-diacetylphloroglucinol in the impact of biocontrol Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators.

    Science.gov (United States)

    Couillerot, Olivier; Combes-Meynet, Emeline; Pothier, Joël F; Bellvert, Floriant; Challita, Elita; Poirier, Marie-Andrée; Rohr, René; Comte, Gilles; Moënne-Loccoz, Yvan; Prigent-Combaret, Claire

    2011-06-01

    Pseudomonads producing the antimicrobial metabolite 2,4-diacetylphloroglucinol (Phl) can control soil-borne phytopathogens, but their impact on other plant-beneficial bacteria remains poorly documented. Here, the effects of synthetic Phl and Phl(+) Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators were investigated. Most A. brasilense strains were moderately sensitive to Phl. In vitro, Phl induced accumulation of carotenoids and poly-β-hydroxybutyrate-like granules, cytoplasmic membrane damage and growth inhibition in A. brasilense Cd. Experiments with P. fluorescens F113 and a Phl(-) mutant indicated that Phl production ability contributed to in vitro growth inhibition of A. brasilense Cd and Sp245. Under gnotobiotic conditions, each of the three strains, P. fluorescens F113 and A. brasilense Cd and Sp245, stimulated wheat growth. Co-inoculation of A. brasilense Sp245 and Pseudomonas resulted in the same level of phytostimulation as in single inoculations, whereas it abolished phytostimulation when A. brasilense Cd was used. Pseudomonas Phl production ability resulted in lower Azospirillum cell numbers per root system (based on colony counts) and restricted microscale root colonization of neighbouring Azospirillum cells (based on confocal microscopy), regardless of the A. brasilense strain used. Therefore, this work establishes that Phl(+) pseudomonads have the potential to interfere with A. brasilense phytostimulators on roots and with their plant growth promotion capacity.

  4. PENGARUH APLIKASI PSEUDOMONAS FLUORESCENS P60 TERHADAP MUTU PATOLOGIS, MUTU FISIOLOGIS, DAN PERTUMBUHAN BIBIT PADI IR 64

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    Lisa Navitasari

    2014-08-01

    Full Text Available Effect of Pseudomonas fluorescens P60 on pathological and physiological quality and growth of rice IR 64  seedlings. The research objectives were (1 detection and identification of seed-borne pathogens of IR 64 rice, (2 testing Pseudomonas fluorescents P60 in inhibiting the in vitro growth of seed-borne pathogens colonies, (3 testing P. fluorescents P60 for pathological and physiological seed quality, and (4 testing P. fluorescents P60 on the growth of seedlings in the greenhouse. The results showed that some seed-borne pathogens can be found both on farmers’ IR 64 rice and factory’s; they were Aspergillus flavus, Alternaria padwickii, Pseudomonas glumae, and P. syringae. Application of P. flourescens P60 was able to inhibit the in vitrogrowth of colonies of all seed-borne pathogens, except P. syringae.  Related to pathological quality, the effect of P. flourescens P60 on percentage of seed-borne pathogens attack did not significantly different from that of benomil but smaller than distilled water. On the physiological quality of seeds, treatment of P. flourescens P60 has the same effect with benomil and distilled water, with  germination rate was more than 80%. In the greenhouse study,treatment of seed immersion time  in P. flourescens P60 suspension showed that the effect of immersion time as long as15 minutes and 25 minutes on  seedling height, root length, and seedling dry weightdid not significantly different. were. However, 25 minutes immersion time resulted in fresh seedling weight and root dry weight higher than that of 15 minutes immersion time.

  5. Structure Revision of N-Mercapto-4-formylcarbostyril Produced by Pseudomonas fluorescens G308 to 2-(2-Hydroxyphenyl)thiazole-4-carbaldehyde [aeruginaldehyde

    DEFF Research Database (Denmark)

    Guillemyn, Karel; Ballet, Steven; Ye, Lumeng

    2014-01-01

    An antibiotic substance isolated from Pseudomonas fluorescens strain G308 was earlier assigned the structure of N-mercapto-4-formylcarbostyril, but computational predictions of the H-1 and C-13 NMR magnetic shielding tensors show this structure to be incompatible with the published spectroscopic...... data. The same is true for six quinoline derivatives related to N-mercapto-4-formylcarbostyril by permutation of the O and S atoms. In contrast, 2-(2-hydroxyphenyl)thiazole-4-carbaldehyde [aeruginaldehyde], isolated from Pseudomonas protegens Pf-5, together with the reduced derivative aeruginol...

  6. Functional Characterization of the Mannitol Promoter of Pseudomonas fluorescens DSM 50106 and Its Application for a Mannitol-Inducible Expression System for Pseudomonas putida KT2440.

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    Jana Hoffmann

    Full Text Available A new pBBR1MCS-2-derived vector containing the Pseudomonas fluorescens DSM10506 mannitol promoter PmtlE and mtlR encoding its AraC/XylS type transcriptional activator was constructed and optimized for low basal expression. Mannitol, arabitol, and glucitol-inducible gene expression was demonstrated with Pseudomonas putida and eGFP as reporter gene. The new vector was applied for functional characterization of PmtlE. Identification of the DNA binding site of MtlR was achieved by in vivo eGFP measurement with PmtlE wild type and mutants thereof. Moreover, purified MtlR was applied for detailed in vitro investigations using electrophoretic mobility shift assays and DNaseI footprinting experiments. The obtained data suggest that MtlR binds to PmtlE as a dimer. The proposed DNA binding site of MtlR is AGTGC-N5-AGTAT-N7-AGTGC-N5-AGGAT. The transcription activation mechanism includes two binding sites with different binding affinities, a strong upstream binding site and a weaker downstream binding site. The presence of the weak downstream binding site was shown to be necessary to sustain mannitol-inducibility of PmtlE. Two possible functions of mannitol are discussed; the effector might stabilize binding of the second monomer to the downstream half site or promote transcription activation by inducing a conformational change of the regulator that influences the contact to the RNA polymerase.

  7. The Effect of Bacteria Colony Pseudomonas fluorescens (UB_Pf1 and Bacillus subtilis (UB_Bs1 on the Mortality of Pratylenchus coffeae (Tylenchida: Pratylenchidae

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    Presti Mardiyani Purwaningtyas

    2016-11-01

    Full Text Available Parasitic Root-Lession nematode of Pratylenchus coffeae can reduce the Indonesian coffee plants productivity. Several studies reported that Pseudomonas fluorescens and Bacillus subtilis endophytic bacteria were antagonistic bacteria to nematode. The objective of this research was to reveal the effectiveness of bacterial colonies density of P. fluorescens (UB_Pf1, B.subtilis (UB BS1, and a combination of both bacteria on nematode mortality using median lethal concentration (LC50 and median lethal time 50 (LT50. The densities of bacteria used in this study were 107, 109, 1011 and 1013 cfu/ml. 35 testing nematodes were used and the mortality was counted at 6, 12, 24, 36, and 48 hours after treatments. The results showed that LC50 values of P. fluorescens was (UB_Pf1 was 4,3x108 cfu/ml, LC50 B. subtilis (UB_Bs1 was 1,9x109cfu/ ml, and LC50 combination of both bacteria was, 8x107 cfu/ml. It implies that the application of the combination of both bacteria are more pathogenic than single bacterial treatment. The results also showed that the highest LT50 value was 13.21  hours combination of bacterial colonies with a density of 1013 cfu/ml and the lowest LT50 value was 52.00 hours on P. fluorescens (UB_Pf1 treatment with colonies density of 107 cfu/ml.How to CitePurwaningtyas, P. M., Rahardjo, B. T., & Tarno, H. (2016. The Effect of Bacteria Colony Pseudomonas fluorescens (UB_Pf1 and Bacillus subtilis (UB_Bs1 on the Mortality of Pratylenchus coffeae (Tylenchida: Pratylenchidae. Biosaintifika: Journal of Biology & Biology Education, 8(3, 286-293. 

  8. Effect of Two Biological Formulations Based on Bacillus subtilis and Pseudomonas fluorescens on Control of Didymella applanata, the Causal Agent of Red Raspberry Cane Spur Blight

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    Margarita Shternshis

    2016-01-01

    Full Text Available In vitro and in vivo studies were conducted to estimate the efficacy of the two microbial formulations based on Bacillus subtilis Cohn. and Pseudomonas fluorescens Mig. on the fungus Didymella applanata (Niessl. Sacc., the causal agent of red raspberry (Rubus idaeus L. spur blight. In vitro, both bacteria reduced the growth of D. applanata. In inoculation experiments with raspberry canes in two cultivars with different susceptibility to D. applanata, these antagonistic bacteria suppressed fungal development by reducing the lesions area and the number of D. applanata fruiting bodies. Field trials of two biological formulations under natural conditions showed a significant suppression of the disease. B. subtilis and P. fluorescens included in the formulations revealed antagonistic activity towards D. applanata that depended on the red raspberry cultivar and weather conditions. In all cases, B. subtilis showed better results than P. fluorescens in biocontrol of the raspberry spur blight. This study demonstrated for the first time the ability of the biocontrol agents B. subtilis and P. fluorescens to suppress red raspberry cane spur blight, a serious worldwide disease.

  9. Ability of the marine bacterium Pseudomonas fluorescens BA3SM1 to counteract the toxicity of CdSe nanoparticles.

    Science.gov (United States)

    Poirier, Isabelle; Kuhn, Lauriane; Demortière, Arnaud; Mirvaux, Boris; Hammann, Philippe; Chicher, Johana; Caplat, Christelle; Pallud, Marie; Bertrand, Martine

    2016-10-04

    In the marine environment, bacteria from estuarine and coastal sediments are among the first targets of nanoparticle pollution; it is therefore relevant to improve the knowledge of interactions between bacteria and nanoparticles. In this work, the response of the marine bacterium Pseudomonas fluorescens BA3SM1 to CdSe nanocrystals (CdSe NPs) of 3nm (NP3) and 8nm (NP8) in diameter was evaluated through microscopic, physiological, biochemical and proteomic approaches. Transmission electron microscopy images showed that NP3 were able to penetrate the bacteria, while NP8 were highly concentrated around the cells, embedded in large exopolysaccharides. In our experimental conditions, both CdSe NP sizes induced a decrease in respiration during the stationary growth phase, while only NP8 caused growth retardation and a decrease in pyoverdine production. Proteomic analyses highlighted that the strain responded to CdSe NP toxicity by inducing various defence mechanisms such as cell aggregation, extracellular CdSe NP sequestration, effective protection against oxidative stress, modifications of envelope organization and properties, and cadmium export. In addition, BA3SM1 presented a biosorption capacity of 1.6×10(16)NP3/g dry weight and 1.7×10(15)NP8/g dry weight. This strain therefore appears as a promising agent for NP bioremediation processes. Proteomic data are available via ProteomeXchange with identifier PXD004012. To the best of our knowledge, this is the first report focussing on the effects of CdSe colloidal nanocrystals (CdSe NPs) on a marine strain of Pseudomonas fluorescens. CdSe NPs are extensively used in the industry of renewable energies and it is regrettably expected that these pollutants will sometime soon appear in the marine environment through surface runoff, urban effluents and rivers. Bacteria living in estuarine and coastal sediments will be among the first targets of these new pollutants. The pseudomonads are frequently found in these ecosystems

  10. Metabolic and Transcriptomic Changes Induced in Arabidopsis by the Rhizobacterium Pseudomonas fluorescens SS1011[W][OA

    Science.gov (United States)

    van de Mortel, Judith E.; de Vos, Ric C.H.; Dekkers, Ester; Pineda, Ana; Guillod, Leandre; Bouwmeester, Klaas; van Loon, Joop J.A.; Dicke, Marcel; Raaijmakers, Jos M.

    2012-01-01

    Systemic resistance induced in plants by nonpathogenic rhizobacteria is typically effective against multiple pathogens. Here, we show that root-colonizing Pseudomonas fluorescens strain SS101 (Pf.SS101) enhanced resistance in Arabidopsis (Arabidopsis thaliana) against several bacterial pathogens, including Pseudomonas syringae pv tomato (Pst) and the insect pest Spodoptera exigua. Transcriptomic analysis and bioassays with specific Arabidopsis mutants revealed that, unlike many other rhizobacteria, the Pf.SS101-induced resistance response to Pst is dependent on salicylic acid signaling and not on jasmonic acid and ethylene signaling. Genome-wide transcriptomic and untargeted metabolomic analyses showed that in roots and leaves of Arabidopsis plants treated with Pf.SS101, approximately 1,910 genes and 50 metabolites were differentially regulated relative to untreated plants. Integration of both sets of “omics” data pointed to a prominent role of camalexin and glucosinolates in the Pf.SS101-induced resistance response. Subsequent bioassays with seven Arabidopsis mutants (myb51, cyp79B2cyp79B3, cyp81F2, pen2, cyp71A12, cyp71A13, and myb28myb29) disrupted in the biosynthesis pathways for these plant secondary metabolites showed that camalexin and glucosinolates are indeed required for the induction of Pst resistance by Pf.SS101. Also for the insect S. exigua, the indolic glucosinolates appeared to play a role in the Pf.SS101-induced resistance response. This study provides, to our knowledge for the first time, insight into the substantial biochemical and temporal transcriptional changes in Arabidopsis associated with the salicylic acid-dependent resistance response induced by specific rhizobacteria. PMID:23073694

  11. A novel psychrophilic lipase from Pseudomonas fluorescens with unique property in chiral resolution and biodiesel production via transesterification.

    Science.gov (United States)

    Luo, Yu; Zheng, Yitao; Jiang, Zhengbing; Ma, Yushu; Wei, Dongzhi

    2006-11-01

    A lipase-producing bacterium strain B68 screened from soil samples of China was identified as Pseudomonas fluorescens. With GenomeWalker, the open reading frame of lipase gene lipB68, encoding 476 amino acids, was cloned and expressed in Escherichia coli BL21 (DE3). By affinity chromatography, the recombinant LipB68 protein was purified to the purity of 95%. As a member of lipase subfamily I.3, LipB68 has a unique optimum temperature of 20 degrees C, which was the lowest in this subfamily. In chiral resolution, LipB68 effectively catalyzed the transesterification of both alpha-phenylethanol and alpha-phenylpropanol at 20 degrees C, achieving E values greater than 100 and 60 after 120 h, respectively. Among all the known catalysts in biodiesel production, LipB68 produced biodiesel with a yield of 92% after 12 h, at the lowest temperature of 20 degrees C, and is the first one of the I.3 lipase subfamily reported to be capable of catalyzing the transesterification reaction of biodiesel production. Since lipase-mediated biodiesel production is normally carried out at 35-50 degrees C, the availability of a highly active lipase with a low optimal temperature can provide substantial savings in energy consumption. Thus, this novel psychrophilic lipase (LipB68) may represent a highly competitive energy-saving biocatalyst.

  12. A novel psychrophilic lipase from Pseudomonas fluorescens with unique property in chiral resolution and biodiesel production via transesterification

    Energy Technology Data Exchange (ETDEWEB)

    Luo Yu; Zheng Yitao; Jiang Zhengbing; Ma Yushu; Wei Dongzhi [East China Univ. of Science and Tech., Shanghai (China). State Key Lab. of Bioreactor Engineering

    2006-11-15

    A lipase-producing bacterium strain B68 screened from soil samples of China was identified as Pseudomonas fluorescens. With GenomeWalker, the open reading frame of lipase gene lipB68, encoding 476 amino acids, was cloned and expressed in Escherichia coli BL21 (DE3). By affinity chromatography, the recombinant LipB68 protein was purified to the purity of 95%. As a member of lipase subfamily I.3, LipB68 has a unique optimum temperature of 20 C, which was the lowest in this subfamily. In chiral resolution, LipB68 effectively catalyzed the transesterification of both a-phenylethanol and a-phenylpropanol at 20 C, achieving E values greater than 100 and 60 after 120 h, respectively. Among all the known catalysts in biodiesel production, LipB68 produced biodiesel with a yield of 92% after 12 h, at the lowest temperature of 20 C, and is the first one of the I.3 lipase subfamily reported to be capable of catalyzing the transesterification reaction of biodiesel production. Since lipasemediated biodiesel production is normally carried out at 35-50 C, the availability of a highly active lipase with a low optimal temperature can provide substantial savings in energy consumption. Thus, this novel psychrophilic lipase (LipB68) may represent a highly competitive energy-saving biocatalyst. (orig.)

  13. Nematicidal protease genes screened from a soil metagenomic library to control Radopholus similis mediated by Pseudomonas fluorescens pf36.

    Science.gov (United States)

    Chen, Deqiang; Wang, Dongwei; Xu, Chunling; Chen, Chun; Li, Junyi; Wu, Wenjia; Huang, Xin; Xie, Hui

    2018-04-01

    Controlling Radopholus similis, an important phytopathogenic nematode, is a challenge worldwide. Herein, we constructed a metagenomic fosmid library from the rhizosphere soil of banana plants, and six clones with protease activity were obtained by functionally screening the library. Furthermore, subclones were constructed using the six clones, and three protease genes with nematicidal activity were identified: pase1, pase4, and pase6. The pase4 gene was successfully cloned and expressed, demonstrating that the protease PASE4 could effectively degrade R. similis tissues and result in nematode death. Additionally, we isolated a predominant R. similis-associated bacterium, Pseudomonas fluorescens (pf36), from 10 R. similis populations with different hosts. The pase4 gene was successfully introduced into the pf36 strain by vector transformation and conjugative transposition, and two genetically modified strains were obtained: p4MCS-pf36 and p4Tn5-pf36. p4MCS-pf36 had significantly higher protease expression and nematicidal activity (p < 0.05) than p4Tn5-pf36 in a microtiter plate assay, whereas p4Tn5-pf36 was superior to p4MCS-pf36 in terms of genetic stability and controlling R. similis in growth pot tests. This study confirmed that R. similis is inhibited by the associated bacterium pf36-mediated expression of nematicidal proteases. Herein, a novel approach is provided for the study and development of efficient, environmentally friendly, and sustainable biocontrol techniques against phytonematodes.

  14. The role of a P1-type ATPase from Pseudomonas fluorescens SBW25 in copper homeostasis and plant colonization.

    Science.gov (United States)

    Zhang, Xue-Xian; Rainey, Paul B

    2007-05-01

    The genome of the plant-colonizing bacterium Pseudomonas fluorescens SBW25 possesses a putative copper-transporting P1-type ATPase (CueA) that is induced on the plant surfaces. Using a chromosomally-integrated cueA-lacZ fusion, we show that transcription of cueA can be induced (in vitro) by ions of copper, silver, gold, and mercury. To investigate the biological significance of cueA, a nonpolar cueA deletion mutant (SBW25 delta cueA) was constructed. This mutant strain displayed a twofold reduction in its tolerance to copper compared with the wild-type strain; however, no change was observed in the sensitivity of the mutant strain to silver, gold, or mercury ions. To obtain insight into the ecological significance of cueA, the competitive ability of SBW25 delta cueA was determined relative to wild-type SBW25 in three environments (none contained added copper): minimal M9 medium, the root of sugar beet (Beta vulgaris), and the root of pea (Pisum sativum). Results showed that the fitness of SBW25 delta cueA was not different from the wild type in laboratory medium but was compromised in the two plant environments. Taken together, these data demonstrate a functional role for CueA in copper homeostasis and reveal an ecologically significant contribution to bacterial fitness in the plant rhizosphere. They also suggest that copper ions accumulate on plant surfaces.

  15. Measurement of glucose utilization by Pseudomonas fluorescens that are free-living and that are attached to surfaces

    International Nuclear Information System (INIS)

    Fletcher, M.

    1986-01-01

    The assimilation and respiration of glucose by attached and free-living Pseudomonas fluorescens were compared. The attachment surfaces were polyvinylidene fluoride, polyethylene, and glass. Specific uptake of [ 1 C]glucose was determined after bacterial biomass was measured by (1) microscopic counts or (2) prelabelling of cells by providing [ 3 H]leucine as substrate, followed by dual-labelling scintillation counting. The glucose concentration was 1.4, 3.5, 5.5, 7.6, or 9.7 μM. Glucose assimilation by cells which became detached from the surfaces during incubation with glucose was also measured after the detached cells were collected by filtration. The composition of the substratum had no effect on the amount of glucose assimilated by attached cells. Glucose assimilation by attached cells exceeded that by free-living cells by a factor of between 2 and 5 or more, and respiration of glucose by surface-associated cells was greater than that by free-living bacteria. Glucose assimilation by detached cells was greater than that by attached bacteria. Measurements of biomass by microscopic counts gave more consistent results than those obtained with dual-labelling, but in general, results obtained by both methods were corroborative

  16. Pseudomonas fluorescens C7R12 type III secretion system impacts mycorrhization of Medicago truncatula and associated microbial communities.

    Science.gov (United States)

    Viollet, Amandine; Pivato, Barbara; Mougel, Christophe; Cleyet-Marel, Jean-Claude; Gubry-Rangin, Cécile; Lemanceau, Philippe; Mazurier, Sylvie

    2017-01-01

    Type three secretion systems (T3SSs) mediate cell-to-cell interactions between Gram-negative bacteria and eukaryotes. We hypothesized that fluorescent pseudomonads harboring T3SS (T3SS+) would be beneficial to arbuscular mycorrhizal symbiosis because non-pathogenic fluorescent pseudomonads have been previously shown to be much more abundant in mycorrhizal than in non-mycorrhizal roots. We tested this hypothesis by comparing mycorrhization and the associated rhizosphere microbial communities of Medicago truncatula grown in a non-sterile soil inoculated with either the T3SS+ mycorrhiza helper bacterium Pseudomonas fluorescens (C7R12) or a T3SS- mutant of the strain. Results showed that the bacterial secretion system was responsible for the promotion of mycorrhization because root colonization by arbuscular mycorrhizal fungi was not promoted by the T3SS- mutant. The observed T3SS-mediated promotion of mycorrhization was associated with changes in the rhizosphere bacterial communities and the increased occurrence of Claroidoglomeraceae within the intraradical arbuscular mycorrhizal fungi. Furthermore, both pseudomonad strains promoted the host-free growth of a model arbuscular mycorrhizal fungus in vitro, suggesting that T3SS-mediated promotion of mycorrhization occurs during plant-fungal interactions rather than during the pre-symbiotic phase of fungal growth. Taken together, these data provide evidence for the involvement of T3SS in promoting arbuscular mycorrhization by a model fluorescent pseudomonad and suggest the implication of interactions between the bacterium and mycorrhizas.

  17. Identification of copper-induced genes in Pseudomonas fluorescens and use of a reporter strain to monitor bioavailable copper in soil

    DEFF Research Database (Denmark)

    Tom-Petersen, Andreas; Hosbond, Carsten; Nybroe, Ole

    2001-01-01

    negatively affect the bacterial soil community. In this study, our goal was to develop a specific and stable Cu reporter construction harboured by an indigenous soil bacterium to measure bioavailability of Cu in soil. Following mutagenesis of Pseudomonas fluorescens strain DF57 with a Tn5::luxAB promoter......-Cu15, the gene interrupted by the transposon encoded a protein carrying a Cu-binding domain but with low homology to known proteins. DF57-Cu15 was the most suitable Cu reporter due to its high specific response and tolerance to Cu in pure culture. DF57-Cu15 responded to soil solutions from Cu......-amended soil microcosms in a concentration-dependent manner. The chelator EDTA reduced the availability of Cu to P. fluorescens in soil. This showed that complex-bound Cu is not necessarily available to bacteria, We compared chemical analysis of soluble Cu and the reporter assay on soil solutions from Cu...

  18. Structure of a putative BenF-like porin from Pseudomonas fluorescens Pf-5 at 2.6 A resolution

    Energy Technology Data Exchange (ETDEWEB)

    Sampathkumar, P.; Swaminathan, S.; Lu, F.; Zhao, X.; Li, Z.; Gilmore, J.; Bain, K.; Rutter, M. E.; Gheyi, T.; Schwinn, D.; Bonanno, J. B.; Pieper, U.; Fajardo, J. E.; Fiser, A.; Almo, S. C.; Chance, M. R.; Baker, D.; Atwell, S.; Thompson, D. A.; Emtage, J. S.; Wasserman, S. R.; Sali, A.; Sauder, J. M.; Burley, S. K.

    2010-11-01

    Gram-negative bacteria typically overcome poor permeability of outer membranes through general porins like OmpF and OmpC, which form water-filled transmembrane pores permitting diffusion of hydrophilic molecules with no particular selectivity. Many bacteria lacking such general porins use substrate-specific porins to overcome growth-limiting conditions and facilitate selective transport of metabolites. Exclusive reliance on substrate-specific porins yields lower membrane permeability to small molecules (<600 Da) versus that seen for Escherichia coli. In Pseudomonads, transit of most small molecules across the cell membrane is thought to be mediated by substrate-specific channels of the OprD superfamily. This property explains, at least in part, the high incidence of Pseudomonas aeruginosa antibiotic resistance. High-throughput DNA sequencing of the P. aeruginosa chromosome revealed the presence of 19 genes encoding structurally related, substrate-specific porins (with 30-45% pairwise amino acid sequence identity) that mediate transmembrane passage of small, water-soluble compounds. The OprD superfamily encompasses the eponymous OprD subfamily, which includes 9 P. aeruginosa proteins that convey basic amino acids and carbapenem antibiotics, and the OpdK subfamily, which includes 11 P. aeruginosa proteins that convey aromatic acids and other small aromatic compounds. Genome sequencing of other gram-negative bacteria has revealed additional members of the OprD and OpdK subfamilies in various organisms, including other pseudomonads. Among the many bacteria in which OprD superfamily members have been identified are P. putida, P. fluorescens Pf-5, P. syringae, and Azotobacter vinelandii, all of which share closely related genes that encode the so-called BenF-like porins. In P. putida, benF is part of an operon involved in benzoate catabolism regulated by benR. Within this operon, benK, benE, and benF genes have been suggested to contribute toward either influx or efflux

  19. Reduction of Fusarium wilt in watermelon by Pseudomonas chlororaphis PCL1391 and P. fluorescens WCS365

    Directory of Open Access Journals (Sweden)

    G.T. Tziros

    2007-12-01

    Full Text Available Fusarium wilt of watermelon (Citrullus lanatus caused by Fusarium oxysporum f. sp. niveum is a devastatine soil-borne disease that causes extensive losses throughout the world. Two known Pseudomonas biocontrol strains were used separately and in combination to assess their antagonistic effectiveness against F. oxysporum f. sp. niveum in pot experiments. P. chlororaphis PCL1391 signifi cantly reduced disease severity. P. fl uorescens WCS365 was less effective in disease suppression, while a combination of the two bacteria had intermediate effects. The biological control of Fusarium wilt with P. chlororaphis offers a potentially useful tool in an integrated pest management program to control Fusarium wilt of watermelon.

  20. Supramolecular Structure and Functional Analysis of the Type III Secretion System in Pseudomonas fluorescens 2P24.

    Science.gov (United States)

    Liu, Ping; Zhang, Wei; Zhang, Li-Qun; Liu, Xingzhong; Wei, Hai-Lei

    2015-01-01

    The type III secretion system (T3SS) of plant and animal bacterial pathogens directs the secretion and injection of proteins into host cells. Some homologous genes of T3SS were found also in non-pathogenic bacteria, but the organization of its machinery and basic function are still unknown. In this study, we identified a T3SS gene cluster from the plant growth-promoting Pseudomonas fluorescens 2P24 and isolated the corresponding T3SS apparatus. The T3SS gene cluster of strain 2P24 is similar organizationally to that of pathogenic P. syringae, except that it lacks the regulator hrpR and the hrpK1 and hrpH genes, which are involved in translocation of proteins. Electron microscopy revealed that the T3SS supramolecular structure of strain 2P24 was comprised of two distinctive substructures: a long extracellular, filamentous pilus, and a membrane-embedded base. We show that strain 2P24 deploys a harpin homolog protein, RspZ1, to elicit a hypersensitive response when infiltrated into Nicotiana tabacum cv. xanthi leaves with protein that is partially purified, and by complementing the hrpZ1 mutation of pHIR11. The T3SS of strain 2P24 retained ability to secrete effectors, whereas its effector translocation activity appeared to be excessively lost. Mutation of the rscC gene from 2P24 T3SS abolished the secretion of effectors, but the general biocontrol properties were unaffected. Remarkably, strain 2P24 induced functional MAMP-triggered immunity that included a burst of reactive oxygen species, strong suppression of challenge cell death, and disease expansion, while it was not associated with the secretion functional T3SS.

  1. Supramolecular structure and functional analysis of the type III secretion system in Pseudomonas fluorescens 2P24

    Directory of Open Access Journals (Sweden)

    Ping eLiu

    2016-01-01

    Full Text Available The type III secretion system (T3SS of plant and animal bacterial pathogens directs the secretion and injection of proteins into host cells. Some homologous genes of T3SS were found also in nonpathogenic bacteria, but the organization of its machinery and basic function are still unknown. In this study, we identified a T3SS gene cluster from the plant growth-promoting Pseudomonas fluorescens 2P24 and isolated the corresponding T3SS apparatus. The T3SS gene cluster of strain 2P24 is similar organizationally to that of pathogenic P. syringae, except that it lacks the regulator hrpR and the hrpK1 and hrpH genes, which are involved in translocation of proteins. Electron microscopy revealed that the T3SS supramolecular structure of strain 2P24 was comprised of two distinctive substructures: a long extracellular, filamentous pilus and a membrane-embedded base. We show that strain 2P24 deploys a harpin homologue protein, RspZ1, to elicit a hypersensitive response when infiltrated into Nicotiana tabacum cv. xanthi leaves with protein that is partially purified, and by complementing the hrpZ1 mutation of pHIR11. The T3SS of strain 2P24 retained ability to secrete effectors, whereas its effector translocation activity appeared to be excessively lost. Mutation of the rscC gene from 2P24 T3SS abolished the secretion of effectors, but the general biocontrol properties were unaffected. Remarkably, strain 2P24 induced functional MAMP-triggered immunity that included a burst of reactive oxygen species, strong suppression of challenge cell death, and disease expansion, while it was not associated with the secretion functional T3SS.

  2. Pseudomonas fluorescens F113 Mutant with Enhanced Competitive Colonization Ability and Improved Biocontrol Activity against Fungal Root Pathogens ▿

    Science.gov (United States)

    Barahona, Emma; Navazo, Ana; Martínez-Granero, Francisco; Zea-Bonilla, Teresa; Pérez-Jiménez, Rosa María; Martín, Marta; Rivilla, Rafael

    2011-01-01

    Motility is one of the most important traits for efficient rhizosphere colonization by Pseudomonas fluorescens F113rif (F113). In this bacterium, motility is a polygenic trait that is repressed by at least three independent pathways, including the Gac posttranscriptional system, the Wsp chemotaxis-like pathway, and the SadB pathway. Here we show that the kinB gene, which encodes a signal transduction protein that together with AlgB has been implicated in alginate production, participates in swimming motility repression through the Gac pathway, acting downstream of the GacAS two-component system. Gac mutants are impaired in secondary metabolite production and are unsuitable as biocontrol agents. However, the kinB mutant and a triple mutant affected in kinB, sadB, and wspR (KSW) possess a wild-type phenotype for secondary metabolism. The KSW strain is hypermotile and more competitive for rhizosphere colonization than the wild-type strain. We have compared the biocontrol activity of KSW with those of the wild-type strain and a phenotypic variant (F113v35 [V35]) which is hypermotile and hypercompetitive but is affected in secondary metabolism since it harbors a gacS mutation. Biocontrol experiments in the Fusarium oxysporum f. sp. radicis-lycopersici/Lycopersicum esculentum (tomato) and Phytophthora cactorum/Fragaria vesca (strawberry) pathosystems have shown that the three strains possess biocontrol activity. Biocontrol activity was consistently lower for V35, indicating that the production of secondary metabolites was the most important trait for biocontrol. Strain KSW showed improved biocontrol compared with the wild-type strain, indicating that an increase in competitive colonization ability resulted in improved biocontrol and that the rational design of biocontrol agents by mutation is feasible. PMID:21685161

  3. Pseudomonas fluorescens F113 mutant with enhanced competitive colonization ability and improved biocontrol activity against fungal root pathogens.

    Science.gov (United States)

    Barahona, Emma; Navazo, Ana; Martínez-Granero, Francisco; Zea-Bonilla, Teresa; Pérez-Jiménez, Rosa María; Martín, Marta; Rivilla, Rafael

    2011-08-01

    Motility is one of the most important traits for efficient rhizosphere colonization by Pseudomonas fluorescens F113rif (F113). In this bacterium, motility is a polygenic trait that is repressed by at least three independent pathways, including the Gac posttranscriptional system, the Wsp chemotaxis-like pathway, and the SadB pathway. Here we show that the kinB gene, which encodes a signal transduction protein that together with AlgB has been implicated in alginate production, participates in swimming motility repression through the Gac pathway, acting downstream of the GacAS two-component system. Gac mutants are impaired in secondary metabolite production and are unsuitable as biocontrol agents. However, the kinB mutant and a triple mutant affected in kinB, sadB, and wspR (KSW) possess a wild-type phenotype for secondary metabolism. The KSW strain is hypermotile and more competitive for rhizosphere colonization than the wild-type strain. We have compared the biocontrol activity of KSW with those of the wild-type strain and a phenotypic variant (F113v35 [V35]) which is hypermotile and hypercompetitive but is affected in secondary metabolism since it harbors a gacS mutation. Biocontrol experiments in the Fusarium oxysporum f. sp. radicis-lycopersici/Lycopersicum esculentum (tomato) and Phytophthora cactorum/Fragaria vesca (strawberry) pathosystems have shown that the three strains possess biocontrol activity. Biocontrol activity was consistently lower for V35, indicating that the production of secondary metabolites was the most important trait for biocontrol. Strain KSW showed improved biocontrol compared with the wild-type strain, indicating that an increase in competitive colonization ability resulted in improved biocontrol and that the rational design of biocontrol agents by mutation is feasible.

  4. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots

    Directory of Open Access Journals (Sweden)

    Carmen eGómez-Lama Cabanás

    2014-09-01

    Full Text Available Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets, many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR experiments aiming to: (i validate the induction of these genes, and (ii shed light on their expression pattern along time (from 1 to 15 days. Induction of olive genes potentially coding for lypoxigenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e. jerf, bHLH, WRKYs, as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mount a wide array of systemic defense responses in distant tissues (stems, leaves. This sheds light on how olive plants respond to the ‘non-hostile’ colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.

  5. Accumulation of α-Keto Acids as Essential Components in Cyanide Assimilation by Pseudomonas fluorescens NCIMB 11764

    Science.gov (United States)

    Kunz, Daniel A.; Chen, Jui-Lin; Pan, Guangliang

    1998-01-01

    Pyruvate (Pyr) and α-ketoglutarate (αKg) accumulated when cells of Pseudomonas fluorescens NCIMB 11764 were cultivated on growth-limiting amounts of ammonia or cyanide and were shown to be responsible for the nonenzymatic removal of cyanide from culture fluids as previously reported (J.-L. Chen and D. A. Kunz, FEMS Microbiol. Lett. 156:61–67, 1997). The accumulation of keto acids in the medium paralleled the increase in cyanide-removing activity, with maximal activity (760 μmol of cyanide removed min−1 ml of culture fluid−1) being recovered after 72 h of cultivation, at which time the keto acid concentration was 23 mM. The reaction products that formed between the biologically formed keto acids and cyanide were unambiguously identified as the corresponding cyanohydrins by 13C nuclear magnetic resonance spectroscopy. Both the Pyr and α-Kg cyanohydrins were further metabolized by cell extracts and served also as nitrogenous growth substrates. Radiotracer experiments showed that CO2 (and NH3) were formed as enzymatic conversion products, with the keto acid being regenerated as a coproduct. Evidence that the enzyme responsible for cyanohydrin conversion is cyanide oxygenase, which was shown previously to be required for cyanide utilization, is based on results showing that (i) conversion occurred only when extracts were induced for the enzyme, (ii) conversion was oxygen and reduced-pyridine nucleotide dependent, and (iii) a mutant strain defective in the enzyme was unable to grow when it was provided with the cyanohydrins as a growth substrate. Pyr and αKg were further shown to protect cells from cyanide poisoning, and excretion of the two was directly linked to utilization of cyanide as a growth substrate. The results provide the basis for a new mechanism of cyanide detoxification and assimilation in which keto acids play an essential role. PMID:9797306

  6. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots.

    Science.gov (United States)

    Gómez-Lama Cabanás, Carmen; Schilirò, Elisabetta; Valverde-Corredor, Antonio; Mercado-Blanco, Jesús

    2014-01-01

    Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA) against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH) cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets), many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR) experiments aiming to: (1) validate the induction of these genes, and (2) shed light on their expression pattern along time (from 1 to 15 days). Induction of olive genes potentially coding for lipoxygenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase, and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e., JERF, bHLH, WRKY), as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mounts a wide array of systemic defense responses in distant tissues (stems, leaves). This sheds light on how olive plants respond to the "non-hostile" colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.

  7. Nunamycin and Nunapeptin: Two novel cyclic peptides are key components of the antimicrobial activity of the Greenlandic isolate Pseudomonas fluorescens In5

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian F.

    Pseudomonas spp. are a rich source of secondary metabolites including bioactive non-ribosomal peptides (NRPs) and polyketides. NRPs are synthesised in large assembly lines by multi-domain modular enzymes known as NRP-synthetases (NRPS). Nunamycin and nunapeptin are two cyclic NRPs synthesised...... by the Greenlandic isolate P. fluorescens In5. Nunamycin shows antifungal activity against the basidiomycete Rhizoctonia solani whereas the only partially structure elucidated nunapeptin appears most active against the ascomycete Fusarium graminearum and the oomycete Pythium aphanidermatum. Originally isolated from...

  8. Safety of spray-dried powder formulated Pseudomonas fluorescens strain CL145A exposure to subadult/adult unionid mussels during simulated open-water treatments

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Waller, Diane L.; Wise, Jeremy K.; Mayer, Denise A.; Aloisi, Douglas B.

    2015-01-01

    The exposure effects of a commercially prepared spray dried powder (SDP) formulation ofPseudomonas fluorescens (strain CL145A) on the survival of seven species of unionid mussels endemic to the Great Lakes and Mississippi River basins was evaluated in this study. The study exposures were completed within replicated 350-liter test tanks contained within a mobile bioassay laboratory sited on the shores of the Black River near La Crosse, Wisconsin. The test tanks were supplied with flowing, filtered river water which was interrupted during the exposure period.

  9. Efeito da pré-incubação de Pseudomonas fluorescens em diferentes temperaturas de refrigeração sobre a hidrólise de caseína no leite desnatado pasteurizado microfiltrado

    OpenAIRE

    Netto, Gabriel Gama

    2012-01-01

    Este trabalho objetivou avaliar o efeito da multiplicação de uma estirpe psicrotrófica proteolítica de Pseudomonas fluorescens nas temperaturas de 4, 7 e 10 ºC, estabelecidas pela Instrução Normativa 62 do MAPA, nas características físicoquímicas e proteólise do leite microfiltrado. Amostras leite desnatado e pasteurizado foram inoculadas com a cepa Pseudomonas fluorescens 041 em uma concentração de aproximadamente 104 UFC mL-1 e incubadas nas três temperaturas por 48 h e, posteriormente, ...

  10. Investigating the ability of Pseudomonas fluorescens UW4 to reduce cadmium stress in Lactuca sativa via an intervention in the ethylene biosynthetic pathway.

    Science.gov (United States)

    Albano, Lucas J; Macfie, Sheila M

    2016-12-01

    A typical plant response to any biotic or abiotic stress, including cadmium (Cd), involves increased ethylene synthesis, which causes senescence of the affected plant part. Stressed plants can experience reduced ethylene and improved growth if they are inoculated with bacteria that have the enzyme ACC deaminase, which metabolizes the ethylene precursor ACC (1-aminocyclopropane-1-carboxylate). We investigated whether one such bacterium, Pseudomonas fluorescens UW4, reduces the production of ethylene and improves the growth of lettuce (Lactuca sativa) sown in Cd-contaminated potting material (PRO-MIX® BX). Plants were inoculated with the wild-type P. fluorescens UW4 or a mutant strain that cannot produce ACC deaminase. Cadmium-treated plants contained up to 50 times more Cd than did control plants. In noninoculated plants, Cd induced a 5-fold increase in ethylene concentration. The wild-type bacterium prevented Cd-induced reductions in root biomass but there was no relationship between Cd treatment and ethylene production in inoculated plants. In contrast, when the concentration of ethylene was plotted against the extent of bacterial colonization of the roots, increased colonization with wild-type P. fluorescens UW4 was associated with 20% less ethylene production. Ours is the first study to show that the protective effect of this bacterium is proportional to the quantity of bacteria on the root surface.

  11. Quantitative and qualitative variability of the caseinolytic potential of different strains of Pseudomonas fluorescens: implications for the stability of casein micelles of UHT milks during their storage.

    Science.gov (United States)

    Baglinière, François; Tanguy, Gaëlle; Jardin, Julien; Matéos, Aurélie; Briard, Valérie; Rousseau, Florence; Robert, Benoît; Beaucher, Eric; Humbert, Gérard; Dary, Annie; Gaillard, Jean Luc; Amiel, Caroline; Gaucheron, Frédéric

    2012-12-15

    Pseudomonas fluorescens grows at low temperature and produces thermo-resistant protease(s) that can destabilize UHT (Ultra High Temperature) milk during its storage. The consequences of contamination of microfiltered milk with 9 strains of P. fluorescens on the stability of the corresponding UHT milk during storage had been investigated in this study. The strains were classified in two groups according to their ability to destabilize UHT milk. For the group of highly destabilizing strains, sedimentations of UHT milks, low values to phosphate test and the presence of aggregates were observed. Zeta potential and hydration of casein micelles decreased, whereas non casein nitrogen (NCN) and non protein nitrogen (NPN) contents increased. The analyses of NCN fraction by liquid chromatography coupled to mass spectrometry indicated that the different casein molecules were hydrolyzed in a similar way for the destabilizing strains suggesting that the same enzyme was implicated. For the group of slightly or not destabilizing strains no visual and biochemical alteration were found. This study showed that destabilization of UHT milk by P. fluorescens was highly variable and strain-dependent. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens is exempt from the...

  13. Identification of the minimal region in lipase ABC transporter recognition domain of Pseudomonas fluorescens for secretion and fluorescence of green fluorescent protein

    Directory of Open Access Journals (Sweden)

    Park Yeonwoo

    2012-05-01

    Full Text Available Abstract Background TliA is a thermostable lipase secreted by the type 1 secretion system (T1SS of Pseudomonas fluorescens. The secretion is promoted by its secretion/chaperone domain located near the C-terminus, which is composed mainly of four Repeat-in-Toxin (RTX repeats. In order to identify the minimal region of TliA responsible for its secretion, five different copies of the secretion/chaperone domain, each involving truncated N-terminal residues and a common C-terminus, were acquired and named as lipase ABC transporter recognition domains (LARDs. Each LARD was fused to epidermal growth factor (EGF or green fluorescent protein (GFP, and the secretion of EGF-LARD or GFP-LARD fusion proteins was assessed in Escherichia coli with ABC transporter. Results Among the fusion proteins, GFP or EGF with 105-residue LARD3 was most efficiently secreted. In addition, GFP-LARD3 emitted wild type GFP fluorescence. Structurally, LARD3 had the 4 RTX repeats exposed at the N-terminus, while other LARDs had additional residues prior to them or missed some of the RTX repeats. LARD3 was both necessary and sufficient for efficient secretion and maintenance of GFP fluorescence in E. coli, which was also confirmed in P. fluorescens and P. fluorescens ▵tliA, a knock-out mutant of tliA. Conclusion LARD3 was a potent secretion signal in T1SS for its fusion flanking RTX motif, which enhanced secretion and preserved the fluorescence of GFP. LARD3-mediated secretion in E. coli or P. fluorescens will enable the development of enhanced protein manufacturing factory and recombinant microbe secreting protein of interest in situ.

  14. PRODUCTION AND CHARACTERIZATION OF BIOSURFACTANT BY Pseudomonas fluorescens USING CASSAVA FLOUR WASTEWATER AS MEDIA

    Directory of Open Access Journals (Sweden)

    Venty Suryanti

    2013-12-01

    Full Text Available Biosurfactant with efficient emulsification properties could be produced by Pseudomonas flourescens using cassava flour wastewater (manipueira as media. The ability of P. flourescens to produce biosurfactant could suggest potential use in industrial and environmental applications. Media containing a mixture of natural manipueira and nutrient broth with 48 h fermentation was the optimum condition for the biosurfactant production. Based on UV-Vis and FT-IR spectra, the biosurfactant was indicated as rhamnolipids containing hydroxyl, ester, carboxylic and aliphatic carbon chain functional groups. Biosurfactant exhibited critical micelle concentration (CMC value of 715 mg/L and reduced the surface tension of the water from 80 mN/m to 59 mN/m. The biosurfactant was able to decrease the interfacial tension about 51-70% when benzyl chloride, palm oil and kerosene were used as water-immiscible compounds. The biosurfactant was able to form stable emulsion until 30 days when paraffin, soybean oil, lubricant oil and kerosene were used as water-immiscible compounds.

  15. Factors screening to statistical experimental design of racemic atenolol kinetic resolution via transesterification reaction in organic solvent using free Pseudomonas fluorescens lipase.

    Science.gov (United States)

    Agustian, Joni; Kamaruddin, Azlina Harun; Aboul-Enein, Hassan Y

    2017-07-01

    As the (R)-enantiomer of racemic atenolol has no β-blocking activity and no lack of side effects, switching from the racemate to the (S)-atenolol is more favorable. Transesterification of racemic atenolol using free enzymes investigated as a resource to resolve the racemate via this method is limited. Screenings of enzyme, medium, and acetyl donor were conducted first to give Pseudomonas fluorescens lipase, tetrahydrofuran, and vinyl acetate. A statistical design of the experiment was then developed using Central Composite Design on some operational factors, which resulted in the conversions of 11.70-61.91% and substrate enantiomeric excess (ee) of 7.31-100%. The quadratic models are acceptable with R 2 of 95.13% (conversion) and 89.63% (ee). The predicted values match the observed values reasonably well. Temperature, agitation speed, and substrate molar ratio factor have low effects on conversion and ee, but enzyme loading affects the responses highly. The interaction of temperature-agitation speed and temperature-substrate molar ratio show significant effects on conversion, while temperature-agitation speed, temperature-substrate molar ratio, and agitation speed-substrate molar ratio affect ee highly. Optimum conditions for the use of Pseudomonas fluorescens lipase, tetrahydrofuran, and vinyl acetate were found at 45°C, 175 rpm, 2000 U, and 1:3.6 substrate molar ratio. © 2017 Wiley Periodicals, Inc.

  16. Genomic analysis of the biocontrol strain Pseudomonas fluorescens Pf29Arp with evidence of T3SS and T6SS gene expression on plant roots.

    Science.gov (United States)

    Marchi, Muriel; Boutin, Morgane; Gazengel, Kévin; Rispe, Claude; Gauthier, Jean-Pierre; Guillerm-Erckelboudt, Anne-Yvonne; Lebreton, Lionel; Barret, Matthieu; Daval, Stéphanie; Sarniguet, Alain

    2013-06-01

    Several bacterial strains of the Pseudomonas genus provide plant growth stimulation, plant protection against pests or bioremediation. Among these bacteria, P. fluorescens Pf29Arp reduces the severity of take-all, a disease caused by the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) on wheat roots. In this study, we obtained a draft genome of Pf29Arp and subsequent comparative genomic analyses have revealed that this bacterial strain is closely related to strains of the 'P. brassicacearum-like' subgroup including P. brassicacearum ssp. brassicacearum NFM421 and P. fluorescens F113. Despite an overall chromosomal organization similar to these strains, a number of features including antibiotic synthesis gene clusters from secondary metabolism are not found in the Pf29Arp genome. But Pf29Arp possesses different protein secretion systems including type III (T3SS) and type VI (T6SS) secretion systems. Pf29Arp is the first Pseudomonas sp. strain described with four T6SS clusters (cluster I, II, III and IV). In addition, some protein-coding genes involved in the assembly of these secretion systems are basally expressed during Pf29Arp colonization of healthy wheat roots and display different expression patterns on necrotized roots caused by Ggt. These data suggest a role of T3SS and T6SS in the Pf29Arp adaptation to different root environments. © 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.

  17. 40 CFR 180.1108 - Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens is...

  18. Genome Analysis of Pseudomonas fluorescens PCL1751: A Rhizobacterium that Controls Root Diseases and Alleviates Salt Stress for Its Plant Host.

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    Shu-Ting Cho

    Full Text Available Pseudomonas fluorescens PCL1751 is a rod-shaped Gram-negative bacterium isolated from the rhizosphere of a greenhouse-grown tomato plant in Uzbekistan. It controls several plant root diseases caused by Fusarium fungi through the mechanism of competition for nutrients and niches (CNN. This mechanism does not rely on the production of antibiotics, so it avoids the concerns of resistance development and is environmentally safe. Additionally, this bacterium promotes plant growth by alleviating salt stress for its plant host. To investigate the genetic mechanisms that may explain these observations, we determined the complete genome sequence of this bacterium, examined its gene content, and performed comparative genomics analysis with other Pseudomonas strains. The genome of P. fluorescens PCL1751 consisted of one circular chromosome that is 6,143,950 base-pairs (bp in size; no plasmid was found. The annotation included 19 rRNA, 70 tRNA, and 5,534 protein-coding genes. The gene content analysis identified a large number of genes involved in chemotaxis and motility, colonization of the rhizosphere, siderophore biosynthesis, and osmoprotectant production. In contrast, the pathways involved in the biosynthesis of phytohormones or antibiotics were not found. Comparison with other Pseudomonas genomes revealed extensive variations in their genome size and gene content. The presence and absence of secretion system genes were highly variable. As expected, the synteny conservation among strains decreased as a function of phylogenetic divergence. The integration of prophages appeared to be an important driver for genome rearrangements. The whole-genome gene content analysis of this plant growth-promoting rhizobacterium (PGPR provided some genetic explanations to its phenotypic characteristics. The extensive and versatile substrate utilization pathways, together with the presence of many genes involved in competitive root colonization, provided further support

  19. Identification of Amino Acid Residues Responsible for the Enantioselectivity and Amide Formation Capacity of the Arylacetonitrilase from Pseudomonas fluorescens EBC191 ▿ †

    Science.gov (United States)

    Kiziak, Christoph; Stolz, Andreas

    2009-01-01

    The nitrilase from Pseudomonas fluorescens EBC191 converted (R,S)-mandelonitrile with a low enantioselectivity to (R)-mandelic acid and (S)-mandeloamide in a ratio of about 4:1. In contrast, the same substrate was hydrolyzed by the homologous nitrilase from Alcaligenes faecalis ATCC 8750 almost exclusively to (R)-mandelic acid. A chimeric enzyme between both nitrilases was constructed, which represented in total 16 amino acid exchanges in the central part of the nitrilase from P. fluorescens EBC191. The chimeric enzyme clearly resembled the nitrilase from A. faecalis ATCC 8750 in its turnover characteristics for (R,S)-mandelonitrile and (R,S)-2-phenylpropionitrile (2-PPN) and demonstrated an even higher enantioselectivity for the formation of (R)-mandelic acid than the nitrilase from A. faecalis. An alanine residue (Ala165) in direct proximity to the catalytically active cysteine residue was replaced in the nitrilase from P. fluorescens by a tryptophan residue (as found in the nitrilase from A. faecalis ATCC 8750 and most other bacterial nitrilases) and several other amino acid residues. Those enzyme variants that possessed a larger substituent in position 165 (tryptophan, phenylalanine, tyrosine, or histidine) converted racemic mandelonitrile and 2-PPN to increased amounts of the R enantiomers of the corresponding acids. The enzyme variant Ala165His showed a significantly increased relative activity for mandelonitrile (compared to 2-PPN), and the opposite was found for the enzyme variants carrying aromatic residues in the relevant position. The mutant forms carrying an aromatic substituent in position 165 generally formed significantly reduced amounts of mandeloamide from mandelonitrile. The important effect of the corresponding amino acid residue on the reaction specificity and enantiospecificity of arylacetonitrilases was confirmed by the construction of a Trp164Ala variant of the nitrilase from A. faecalis ATCC 8750. This point mutation converted the highly R

  20. The resistance-nodulation-division efflux pump EmhABC influences the production of 2,4-diacetylphloroglucinol in Pseudomonas fluorescens 2P24.

    Science.gov (United States)

    Tian, Tao; Wu, Xiao-Gang; Duan, Hui-Mei; Zhang, Li-Qun

    2010-01-01

    The polyketide metabolite 2,4-diacetylphloroglucinol (2,4-DAPG) plays a major role in the biological control of soil-borne plant diseases by Pseudomonas fluorescens 2P24. Two mutants (PM810 and PM820) with increased extracellular accumulation of 2,4-DAPG were isolated using transposon mutagenesis. The disrupted genes in these two mutants shared >80 % identity with the genes of the EmhR-EmhABC resistance-nodulation-division (RND) efflux system of P. fluorescens cLP6a. The deletion of emhA (PM802), emhB (PM803) or emhC (PM804) genes in strain 2P24 increased the extracellular accumulation of 2,4-DAPG, whereas the deletion of the emhR (PM801) gene decreased the biosynthesis of 2,4-DAPG. The promoter assay confirmed the elevated transcription of emhABC in the EmhR disrupted strain (PM801) and an indirect negative regulation of 2,4-DAPG biosynthetic locus transcription by the EmhABC efflux pump. Induction by exogenous 2,4-DAPG led to remarkable differences in transcription of chromosome-borne phlA : : lacZ fusion in PM901 and PM811 (emhA(-)) strains. Additionally, the EmhABC system in strain 2P24 was involved in the resistance to a group of toxic compounds, including ampicillin, chloramphenicol, tetracycline, ethidium bromide and crystal violet. In conclusion, our results suggest that the EmhABC system is an important element that influences the production of antibiotic 2,4-DAPG and enhances resistance to toxic compounds in P. fluorescens 2P24.

  1. Characterisation of the thermostable protease AprX in strains of Pseudomonas fluorescens and impact on the shelf-life of dairy products: preliminary results

    Directory of Open Access Journals (Sweden)

    Nadia Andrea Andreani

    2016-12-01

    Full Text Available Bacterial proteases are involved in food spoilage and shelf-life reduction. Among the bacterial proteases, a predominant role in spoilage of dairy products seems to be played by the thermostable metallo-protease AprX, which is produced by various strains of Pseudomonas fluorescens. Differences in AprX enzyme activity among different strains were highlighted, but the most proteolytic strains were not identified. In this study, the presence of the aprX gene was evaluated in 69 strains isolated from food matrices and 18 reference strains belonging to the P. fluorescens group, which had been previously typed by the multi locus sequence typing method. Subsequently, a subset of reference strains was inoculated in ultra-high temperature milk, and the expression of the aprX gene was evaluated at 22 and 6°C. On the same milk samples, the proteolytic activity was then evaluated through Azocasein and trinitrobenzenesulfonic acid solution assays. Finally, to assess the applicability of the former assay directly on dairy products the proteolityc activity was tested on industrial ricotta samples using the Azocasein assay. These results demonstrate the spread of aprX gene in most strains tested and the applicability of Azocasein assay to monitor the proteolytic activity in dairy products.

  2. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    Science.gov (United States)

    Maldonado-González, M. Mercedes; Bakker, Peter A. H. M.; Prieto, Pilar; Mercado-Blanco, Jesús

    2015-01-01

    The effective management of Verticillium wilts (VW), diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control VW of olive caused by the highly virulent, defoliating (D) pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V. dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i) olive D and non-defoliating V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii) strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii) strain PICF7 controls VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. A. thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7. PMID:25904904

  3. Non-target trials with Pseudomonas fluorescens strain CL145A, a lethal control agent of dreissenid mussels (Bivalvia: Dreissenidae

    Directory of Open Access Journals (Sweden)

    Daniel P. Molloy

    2013-01-01

    Full Text Available In an effort to develop an efficacious and environmentally safe method for managing zebra mussels (Dreissena polymorpha and quaggamussels (Dreissena rostriformis bugensis, we initiated a research project investigating the potential use of bacteria and their naturalmetabolic products as biocontrol agents. This project resulted in the discovery of an environmental isolate lethal to dreissenid mussels,Pseudomonas fluorescens strain CL145A (Pf-CL145A. In previous published reports we have demonstrated that: 1 Pf-CL145A’s mode ofaction is intoxication (not infection; 2 natural product within ingested bacterial cells lyse digestive tract epithelial cells leading to dreisseniddeath; and 3 high dreissenid kill rates (>90% are achievable following treatment with Pf-CL145A cells, irrespective of whether thebacterial cells are dead or alive. Investigating the environmental safety of Pf-CL145A was also a key element in our research efforts, andherein, we report the results of non-target trials demonstrating Pf-CL145A’s high specificity to dreissenids. These acute toxicity trials weretypically single-dose, short-term (24-72 h exposures to Pf-CL145A cells under aerated conditions at concentrations highly lethal todreissenids (100 or 200 mg/L. These trials produced no evidence of mortality among the ciliate Colpidium colpoda, the cladoceran Daphniamagna, three fish species (Pimephales promelas, Salmo trutta, and Lepomis macrochirus, and seven bivalve species (Mytilus edulis,Pyganodon grandis, Pyganodon cataracta, Lasmigona compressa, Strophitus undulatus, Lampsilis radiata, and Elliptio complanata. Lowmortality (3-27% was recorded in the amphipod Hyalella azteca, but additional trials suggested that most, if not all, of the mortality couldbe attributed to some other unidentified factor (e.g., possibly particle load or a water quality issue rather than Pf-CL145A’s dreissenidkillingnatural product. In terms of potential environmental safety, the results of

  4. Biodesel Production from Pseudomonas Fluorescens Lp1 Lipase Immobilized on Amino-silane Modified Super Paramagnetic Fe3O4 Nanoparticles

    Science.gov (United States)

    Kanimozhi, S.; Perinbam, K.

    2013-04-01

    An extracellular lipase from Pseudomonas fluorescens Lp1 isolated from oil contaminated soil was immobilized onto amino silane modified superparamagnetic Fe3O4 nanoparticles. The magnetic nanoparticles, magnetite was synthesized chemically by co-precipitation and characterized by Scanning Electron Microscopy (SEM), Fourier Transformed Infrared Spectroscopy (FT-IR) and Powder X-ray diffraction studies (XRD). The structure of the synthesized magnetic nanoparticles was uniform, spherical and the size was determined around 31 nm by powder XRD. The biodiesel production mixture was prepared by addition of waste cooking oil, lipase immobilized magnetite and methanol. The transesterified products were analyzed by Gas Liquid chromatography-Mass spectroscopy (GC-MS). The methyl esters such as Oxiraneundecanoic acid, 3-pentyl-methyl ester, Hexadecanoic acid, methyl ester and 10-Octadecenoic acid, methyl ester were obtained. The study experimentally proved the use of amino silane modified superparamagnetic Fe3O4 nanoparticles in biodiesel production from waste cooking oil.

  5. Biodesel Production from Pseudomonas Fluorescens Lp1 Lipase Immobilized on Amino-silane Modified Super Paramagnetic Fe3O4 Nanoparticles

    International Nuclear Information System (INIS)

    Kanimozhi, S; Perinbam, K

    2013-01-01

    An extracellular lipase from Pseudomonas fluorescens Lp1 isolated from oil contaminated soil was immobilized onto amino silane modified superparamagnetic Fe 3 O 4 nanoparticles. The magnetic nanoparticles, magnetite was synthesized chemically by co-precipitation and characterized by Scanning Electron Microscopy (SEM), Fourier Transformed Infrared Spectroscopy (FT-IR) and Powder X-ray diffraction studies (XRD). The structure of the synthesized magnetic nanoparticles was uniform, spherical and the size was determined around 31 nm by powder XRD. The biodiesel production mixture was prepared by addition of waste cooking oil, lipase immobilized magnetite and methanol. The transesterified products were analyzed by Gas Liquid chromatography-Mass spectroscopy (GC-MS). The methyl esters such as Oxiraneundecanoic acid, 3-pentyl-methyl ester, Hexadecanoic acid, methyl ester and 10-Octadecenoic acid, methyl ester were obtained. The study experimentally proved the use of amino silane modified superparamagnetic Fe 3 O 4 nanoparticles in biodiesel production from waste cooking oil.

  6. Proteomic analysis of elicitation of downy mildew disease resistance in pearl millet by seed priming with β-aminobutyric acid and Pseudomonas fluorescens.

    Science.gov (United States)

    Anup, Chandra Pal; Melvin, Prasad; Shilpa, Nataraj; Gandhi, Mayuri Nalin; Jadhav, Manali; Ali, Hussain; Kini, Kukkundoor Ramachandra

    2015-04-29

    Downy mildew is one of the severe diseases of pearl millet, globally affecting its commercial production. Priming of seeds of a susceptible cultivar of pearl millet with β-aminobutyric acid (BABA) and Pseudomonas fluorescens has reduced the downy mildew disease incidence level under field studies. In the current study, proteomic approach was used to elucidate the poorly studied resistance mechanism in these elicitor primed pearl millet seeds in response to Sclerospora graminicola infection. 2DE-MS/MS based proteomic approach revealed that majority of the 63 differentially accumulated (p≤0.05) proteins associated with energy and metabolism followed by stress and defense category. Multivariate statistics disclosed that infection caused by the pathogen rather than elicitor treatment had a major influence on the dynamics of protein abundance. Mechanism of priming mediated by BABA and P. fluorescens were different from each other as evident by the protein abundance profile of hierarchical clustering analysis. Over-representation of proteins pertaining to glucose metabolism suggests that seed priming ensures plant protection against disease without compromising its normal growth and development. In addition the study forms a basis for future investigation by functional analysis of these differentially accumulated proteins to further unravel the resistance mechanism of elicitor primed plant against the S. graminicola. The study is based on the comparative proteomic analysis between BABA and P. fluorescens mediated resistance in pearl millet, in response to downy mildew causing biotroph - S. graminicola. To our knowledge, this article is the first to report on seedling proteome of pearl millet whose genome is not yet sequenced. In addition, the study also provides clue for the plausible antagonistic cross-talk that might exist between jasmonic acid signaling and salicylic acid signaling in SAR and ISR mediated resistance by BABA and P. fluorescens against the downy

  7. Investigation of zoonotic disease pathogens (Aeromonas hydrophila, Pseudomonas fluorescens, Streptococcus iniae) seen in carp farms in the Northern Iraq-Erbil region by molecular methods

    Science.gov (United States)

    Ibraheem, Azad Saber; Önalan, Şükrü; Arabacı, Muhammed

    2017-04-01

    The aim of this study was to determine the zoonotic bacteria in carp farms in the Northern Iraq-Erbil region. Carp is the main fish species cultured in Erbil region. The most common zoonotic bacteria generally seen in carp farms are Aeromonas hydrophila, Pseudomonas fluorescens and Streptococcus iniae. Samples were collected from 25 carp farms in the Northern Iraq-Erbil region. Six carp samples were collected from each carp farm. Head kidney and intestine tissue samples were collected from each carp sample. Then head kidney and intestine tissue samples were pooled separately from each carp farm. Total bacterial DNA had been extracted from the 25 pooled head kidney and 25 intestinal tissue samples. The pathogen Primers were originally designed from 16S RNA gene region. Zoonotic bacteria were scanned in all tissue samples with absent/present analysis by RT-PCR. Furthermore, the capillary gel electrophoresis bands were used for confirmation of amplicon size which was planned during primer designing stage. As a result, thirteen carp farms were positive in the respect to Aeromonas hydrophila, eight carp farms were positive from head kidney and six carp farms were positive from the intestine, only one carp farm was positive from both head kidney and the intestine tissue samples. In the respect to Streptococcus iniae, four carp farms were positive from head kidney and two carp farms were positive from the intestine. Only one carp farm was positive in the respect to Pseudomonas fluorescens from the intestine. Totally, 9 of 25 carp farms were cleared (negative) the zoonotic bacteria. In conclusion, the zoonotic bacteria were high (64 %) in carp farms in the Northern Iraq-Erbil region.

  8. The periplasmic transaminase PtaA of Pseudomonas fluorescens converts the glutamic acid residue at the pyoverdine fluorophore to α-ketoglutaric acid.

    Science.gov (United States)

    Ringel, Michael T; Dräger, Gerald; Brüser, Thomas

    2017-11-10

    The periplasmic conversion of ferribactin to pyoverdine is essential for siderophore biogenesis in fluorescent pseudomonads, such as pathogenic Pseudomonas aeruginosa or plant growth-promoting Pseudomonas fluorescens The non-ribosomal peptide ferribactin undergoes cyclizations and oxidations that result in the fluorophore, and a strictly conserved fluorophore-bound glutamic acid residue is converted to a range of variants, including succinamide, succinic acid, and α-ketoglutaric acid residues. We recently discovered that the pyridoxal phosphate-containing enzyme PvdN is responsible for the generation of the succinamide, which can be hydrolyzed to succinic acid. Based on this, a distinct unknown enzyme was postulated to be responsible for the conversion of the glutamic acid to α-ketoglutaric acid. Here we report the identification and characterization of this enzyme in P. fluorescens strain A506. In silico analyses indicated a periplasmic transaminase in fluorescent pseudomonads and other proteobacteria that we termed PtaA for " p eriplasmic t ransaminase A " An in-frame-deleted ptaA mutant selectively lacked the α-ketoglutaric acid form of pyoverdine, and recombinant PtaA complemented this phenotype. The ptaA / pvdN double mutant produced exclusively the glutamic acid form of pyoverdine. PtaA is homodimeric and contains a pyridoxal phosphate cofactor. Mutation of the active-site lysine abolished PtaA activity and affected folding as well as Tat-dependent transport of the enzyme. In pseudomonads, the occurrence of ptaA correlates with the occurrence of α-ketoglutaric acid forms of pyoverdines. As this enzyme is not restricted to pyoverdine-producing bacteria, its catalysis of periplasmic transaminations is most likely a general tool for specific biosynthetic pathways. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. Influencia de la concentración salina y la temperatura en la composición de ácidos grasos de membrana de Pseudomonas fluorescens GNP-OHP-3 Influence of salinity and temperature on fatty acid composition of Pseudomonas fluorescens GNP-OHP-3 membrane

    Directory of Open Access Journals (Sweden)

    G. N. Pucci

    2004-03-01

    Full Text Available Las bacterias responden a los cambios ambientales modificando su composición, para evitar el daño que dichos cambios podrían ejercer. Una de las modificaciones más importantes es la variación de la composición de los ácidos grasos de las membranas celulares, que le permite mantener la homeoviscosidad ante situaciones de estrés. Trabajos previos han estudiado la acción de la temperatura, presión hidrostática y diferentes solventes sobre cepas de Pseudomonas putida. En este trabajo se estudió la acción conjunta de la temperatura y la salinidad sobre la composición de ácidos grasos de membranas celulares de Pseudomonas fluorescens GNP-OHP-3, una cepa bacteriana aislada de un hábitat contaminado con petróleo. Pseudomonas fluorescens GNP-OHP-3 respondió a las variaciones de temperatura modificando los ácidos grasos de sus membranas de manera similar a lo descripto en otros integrantes de su género: ante el aumento de temperatura se observó un incremento de ácidos grasos saturados y una disminución de los ácidos grasos insaturados. En el rango de concentraciones salinas ensayadas las variaciones de los ácidos grasos mayoritarios fueron en general erráticas. La respuesta de los ácidos grasos ciclo propano pudo expresarse con ecuaciones matemáticas que permitieron predecir el porcentaje de estos ácidos en relación a la concentración de cloruro de sodio.The bacteria respond to environmental changes modifying their composition. One of the most important modifications is the variation on fatty acid composition of cellular membranes to maintain the homeoviscosity. The action of temperature, hydrostatic pressure and solvents on Pseudomonas putida has been thoroughly studied. In this paper, the combined action of the temperature and salinity on fatty acid composition of cellular membranes of Pseudomonas fluorescens GNP-OHP-3, a bacterial strain isolated from a petroleum contaminated habitat, was studied. The modifications in the

  10. Development and validation of a real-time TaqMan assay for the detection and enumeration of Pseudomonas fluorescens ATCC 13525 used as a challenge organism in testing of food equipments.

    Science.gov (United States)

    Saha, Ratul; Bestervelt, Lorelle L; Donofrio, Robert S

    2012-02-01

    Pseudomonas fluorescens ATCC 13525 is used as the challenge organism to evaluate the efficacy of the clean-in-place (CIP) process of food equipment (automatic ice-maker) as per NSF/ANSI Standard 12. Traditional culturing methodology is presently used to determine the concentration of the challenge organism, which takes 48 h to confirm the cell density. Storage of the challenge preparation in the refrigerator might alter the cell density as P. fluorescens is capable of growing at 4 °C. Also, background organism can grow on the Pseudomonas F agar (PFA) used for the recovery of P. fluorescens thus affecting the results of the test. Real-time TaqMan assay targeting the cpn60 gene was developed for the enumeration and the identification of P. fluorescens because of its specificity, accuracy, and shorter turnaround time. The TaqMan primer-probe pair developed using the Allele ID® 7.0 probe design software was highly specific and sensitive for the target organism. The sensitivity of the assay was 10 colony forming units (CFU)/mL. The assay was also successful in determining the concentration of the challenge preparation within 2 h. Based on these observations, TaqMan assay targeting the cpn60 gene can be efficiently used for strain level identification and enumeration of bacteria. Pseudomonas fluorescens ATCC 13525 is used as a challenge organism in the efficacy testing of clean-in-place process of food equipments. Currently, culturing technique is used for its identification and estimation, which is not only time-consuming but also prone to error. Real-time TaqMan assay is more specific, sensitive, and accurate along with a shorter turnaround time compared to culturing techniques, thereby increasing the overall quality of the testing methodology to evaluate the clean-in-place process critical for the food industry to protect public health and safety. © 2012 Institute of Food Technologists®

  11. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

    Directory of Open Access Journals (Sweden)

    M. Mercedes eMaldonado-González

    2015-04-01

    Full Text Available The effective management of Verticillium wilts, diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control Verticillium wilt of olive caused by the highly-virulent, defoliating (D pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V.dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i olive D and non-defoliating (ND V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii strain PICF7 controls Verticillium wilt (VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. Arabidopsis thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7.

  12. Sequencing and Analysis of the Pseudomonas fluorescens GcM5-1A Genome: A Pathogen Living in the Surface Coat of Bursaphelenchus xylophilus.

    Directory of Open Access Journals (Sweden)

    Kai Feng

    Full Text Available It is known that several bacteria are adherent to the surface coat of pine wood nematode (Bursaphelenchus xylophilus, but their function and role in the pathogenesis of pine wilt disease remains debatable. The Pseudomonas fluorescens GcM5-1A is a bacterium isolated from the surface coat of pine wood nematodes. In previous studies, GcM5-1A was evident in connection with the pathogenicity of pine wilt disease. In this study, we report the de novo sequencing of the GcM5-1A genome. A 600-Mb collection of high-quality reads was obtained and assembled into sequence contigs spanning a 6.01-Mb length. Sequence annotation predicted 5,413 open reading frames, of which 2,988 were homologous to genes in the other four sequenced P. fluorescens isolates (SBW25, WH6, Pf0-1 and Pf-5 and 1,137 were unique to GcM5-1A. Phylogenetic studies and genome comparison revealed that GcM5-1A is more closely related to SBW25 and WH6 isolates than to Pf0-1 and Pf-5 isolates. Towards study of pathogenesis, we identified 79 candidate virulence factors in the genome of GcM5-1A, including the Alg, Fl, Waa gene families, and genes coding the major pathogenic protein fliC. In addition, genes for a complete T3SS system were identified in the genome of GcM5-1A. Such systems have proved to play a critical role in subverting and colonizing the host organisms of many gram-negative pathogenic bacteria. Although the functions of the candidate virulence factors need yet to be deciphered experimentally, the availability of this genome provides a basic platform to obtain informative clues to be addressed in future studies by the pine wilt disease research community.

  13. Application of Endophytic Pseudomonas fluorescens and a Bacterial Consortium to Brassica napus Can Increase Plant Height and Biomass under Greenhouse and Field Conditions

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    Richard D. Lally

    2017-12-01

    Full Text Available Plant associated bacteria with plant growth promotion (PGP properties have been proposed for use as environmentally friendly biofertilizers for sustainable agriculture; however, analysis of their efficacy in the field is often limited. In this study, greenhouse and field trials were carried out using individual endophytic Pseudomonas fluorescens strains, the well characterized rhizospheric P. fluorescens F113 and an endophytic microbial consortium of 10 different strains. These bacteria had been previously characterized with respect to their PGP properties in vitro and had been shown to harbor a range of traits associated with PGP including siderophore production, 1-aminocyclopropane-1-carboxylic acid (ACC deaminase activity, and inorganic phosphate solubilization. In greenhouse experiments individual strains tagged with gfp and Kmr were applied to Brassica napus as a seed coat and were shown to effectively colonize the rhizosphere and root of B. napus and in addition they demonstrated a significant increase in plant biomass compared with the non-inoculated control. In the field experiment, the bacteria (individual and consortium were spray inoculated to winter oilseed rape B. napus var. Compass which was grown under standard North Western European agronomic conditions. Analysis of the data provides evidence that the application of the live bacterial biofertilizers can enhance aspects of crop development in B. napus at field scale. The field data demonstrated statistically significant increases in crop height, stem/leaf, and pod biomass, particularly, in the case of the consortium inoculated treatment. However, although seed and oil yield were increased in the field in response to inoculation, these data were not statistically significant under the experimental conditions tested. Future field trials will investigate the effectiveness of the inoculants under different agronomic conditions.

  14. Endophytic Bacterium Pseudomonas fluorescens RG11 May Transform Tryptophan to Melatonin and Promote Endogenous Melatonin Levels in the Roots of Four Grape Cultivars.

    Science.gov (United States)

    Ma, Yaner; Jiao, Jian; Fan, Xiucai; Sun, Haisheng; Zhang, Ying; Jiang, Jianfu; Liu, Chonghuai

    2016-01-01

    Endophytes have been verified to synthesize melatonin in vitro and promote abiotic stress-induced production of endogenous melatonin in grape ( Vitis vinifera L.) roots. This study aimed to further characterize the biotransformation of tryptophan to melatonin in the endophytic bacterium Pseudomonas fluorescens RG11 and to investigate its capacity for enhancing endogenous melatonin levels in the roots of different grape cultivars. Using ultra performance liquid chromatography-tandem mass spectrometry combined with 15N double-labeled L -tryptophan as the precursor for melatonin, we detected isotope-labeled 5-hydroxytryptophan, serotonin, N -acetylserotonin, and melatonin, but tryptamine was not detected during the in vitro incubation of P. fluorescens RG11. Furthermore, the production capacity of these four compounds peaked during the exponential growth phase. RG11 colonization increased the endogenous levels of 5-hydroxytryptophan, N -acetylserotonin, and melatonin, but reduced those of tryptamine and serotonin, in the roots of the Red Globe grape cultivar under salt stress conditions. Quantitative real-time PCR revealed that RG11 reduced the transcription of grapevine tryptophan decarboxylase and serotonin N -acetyltransferase genes when compared to the un-inoculated control. These results correlated with decreased reactive oxygen species bursts and cell damage, which were alleviated by RG11 colonization under salt stress conditions. Additionally, RG11 promoted plant growth and enhanced the levels of endogenous melatonin in different grape cultivars. Intraspecific variation in the levels of melatonin precursors was found among four grape cultivars, and the associated root crude extracts appeared to significantly induce RG11 melatonin biosynthesis in vitro . Overall, this study provides useful information that enhances the existing knowledge of a potential melatonin synthesis pathway in rhizobacteria, and it reveals plant-rhizobacterium interactions that affect

  15. Exploiting genotypic diversity of 2,4-diacetylphloroglucinol-producing Pseudomonas spp.: characterization of superior root-colonizing P. fluorescens strain Q8r1-96.

    Science.gov (United States)

    Raaijmakers, J M; Weller, D M

    2001-06-01

    The genotypic diversity that occurs in natural populations of antagonistic microorganisms provides an enormous resource for improving biological control of plant diseases. In this study, we determined the diversity of indigenous 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas spp. occurring on roots of wheat grown in a soil naturally suppressive to take-all disease of wheat. Among 101 isolates, 16 different groups were identified by random amplified polymorphic DNA (RAPD) analysis. One RAPD group made up 50% of the total population of DAPG-producing Pseudomonas spp. Both short- and long-term studies indicated that this dominant genotype, exemplified by P. fluorescens Q8r1-96, is highly adapted to the wheat rhizosphere. Q8r1-96 requires a much lower dose (only 10 to 100 CFU seed(-1) or soil(-1)) to establish high rhizosphere population densities (10(7) CFU g of root(-1)) than Q2-87 and 1M1-96, two genotypically different, DAPG-producing P. fluorescens strains. Q8r1-96 maintained a rhizosphere population density of approximately 10(5) CFU g of root(-1) after eight successive growth cycles of wheat in three different, raw virgin soils, whereas populations of Q2-87 and 1M1-96 dropped relatively quickly after five cycles and were not detectable after seven cycles. In short-term studies, strains Q8r1-96, Q2-87, and 1M1-96 did not differ in their ability to suppress take-all. After eight successive growth cycles, however, Q8r1-96 still provided control of take-all to the same level as obtained in the take-all suppressive soil, whereas Q2-87 and 1M1-96 gave no control anymore. Biochemical analyses indicated that the superior rhizosphere competence of Q8r1-96 is not related to in situ DAPG production levels. We postulate that certain rhizobacterial genotypes have evolved a preference for colonization of specific crops. By exploiting diversity of antagonistic rhizobacteria that share a common trait, biological control can be improved significantly.

  16. Influence of Arbuscular Mycorrhizal Fungi and Pseudomonas fluorescens at Different Superphosphate Levels on Linseed (Linum usitatissimum L. Growth Response Influencia de Hongos Micorriza Arbusculares y Pseudomonas fluorescens con Diferentes Niveles de Superfosfato sobre la Respuesta al Crecimiento de Lino (Linum usitatissimum L.

    Directory of Open Access Journals (Sweden)

    Neetu Neetu

    2012-06-01

    Full Text Available The aim of this study was to investigate the influence of arbuscular mycorrhizal fungi (AMF Glomus mosseae (T.H. Nicolson & Gerd. Gerd. & Trappe and Acaulospora laevis (Gerd. & Trappe on linseed (Linum usitatissimum L. growth response with phosphate solubilizing bacteria Pseudomonas fluorescens; different doses of superphosphate were used: 20 kg ha4 (half recommended dose, 40 kg ha4 (recommended dose, and 80 kg ha4 (double recommended dose in earthen pots filled with sterilized soil under greenhouse conditions. Among all the growth parameters, the following were the highest in the G. mosseae + P. fluorescens combination at the medium concentration (recommended superphosphate dose: plant height (78.74 ± 1.8 cm, fresh shoot weight (3.45 ± 0.294 g, dry shoot weight (0.57 ± 0.007 g, fresh root weight (0.223 ± 0.023 g, dry root weight (0.036 ± 0.004 g, root length (17.67 ± 0.48 cm, AM spore number (94.4 ± 9.86, shoot (1.14 ± 0.115% and root (1.29 ± 0.110% P content, and acidic (0.447 ± 0.012 IU g-1 FW and alkaline phosphatase activity (0.119 ± 0.008 IU g-1 FW. The percentage mycorrhizal root colonization with the A. laevis + P. fluorescens (86.86 ± 2.17% combination and chlorophyll content with the G. mosseae + A. laevis + P. fluorescens (0.474 ± 0.009 mg g-1 FW combination recorded the highest values at the low concentration (half recommended superphosphate dose as compared with non-mycorrhizal plants (control. The high superphosphate dose clearly reduced or decreased all the growth parameters. Therefore, vigorous growth and maximum flax yield can be achieved by inoculating plants with AM fungi and P. fluorescens with the recommended dose or less than the recommended dose of superphosphate.La presente investigation tuvo como objetivo estudiar la influencia de hongos micorrícicos arbusculares (AMF, i.e., Glomus mosseae (T.H. Nicolson & Gerd. Gerd. & Trappey Acaulospora laevis (Gerd. & Trappe, en la respuesta de crecimiento de lino

  17. Pseudomonas fluorescens JH 70-4 promotes pb stabilization and early seedling growth of sudan grass in contaminated mining site soil.

    Science.gov (United States)

    Shim, Jaehong; Babu, A Giridhar; Velmurugan, Palanivel; Shea, Patrick J; Oh, Byung-Taek

    2014-01-01

    A bacterial strain (JH 70-4) exhibiting plant growth promoting characteristics (indoleacetic acid production and 1-aminocyclopropane-1-carboxylate deaminase activity), as well as heavy metal(loid) (HM) tolerance and Pb precipitation, was isolated from HM-contaminated soil at an abandoned mine site. The bacterium was identified as Pseudomonas fluorescens based on 16S rDNA sequencing. The JH 70-4 strain induced precipitation of Pb as PbS nanoparticles, confirmed by X-ray diffraction. Solution pH, incubation time, and Pb concentration influenced removal and PbS formation. Inoculating contaminated soil with JH 70-4 decreased Pb availability; exchangeable Pb decreased while organic- and sulphide-bound Pb increased. The toxicity characteristic leaching procedure showed a 65% decrease in Pb in leachate 60 d after inoculating soil with JH 70-4. Shoot and root lengths of Sudan grass grown in the inoculated soil were greater than in the uninoculated soil. Findings suggest that microbial Pb fixation is a viable strategy for remediating soil and promoting plant growth for phytostabilization of contaminated sites.

  18. Immobilization of Pseudomonas fluorescens lipase on hydrophobic supports and application in biodiesel synthesis by transesterification of vegetable oils in solvent-free systems.

    Science.gov (United States)

    Lima, Lionete N; Oliveira, Gladson C; Rojas, Mayerlenis J; Castro, Heizir F; Da Rós, Patrícia C M; Mendes, Adriano A; Giordano, Raquel L C; Tardioli, Paulo W

    2015-04-01

    This work describes the preparation of biocatalysts for ethanolysis of soybean and babassu oils in solvent-free systems. Polystyrene, Amberlite (XAD-7HP), and octyl-silica were tested as supports for the immobilization of Pseudomonas fluorescens lipase (PFL). The use of octyl-silica resulted in a biocatalyst with high values of hydrolytic activity (650.0 ± 15.5 IU/g), immobilization yield (91.3 ± 0.3 %), and recovered activity (82.1 ± 1.5 %). PFL immobilized on octyl-silica was around 12-fold more stable than soluble PFL, at 45 °C and pH 8.0, in the presence of ethanol at 36 % (v/v). The biocatalyst provided high vegetable oil transesterification yields of around 97.5 % after 24 h of reaction using babassu oil and around 80 % after 48 h of reaction using soybean oil. The PFL-octyl-silica biocatalyst retained around 90 % of its initial activity after five cycles of transesterification of soybean oil. Octyl-silica is a promising support that can be used to immobilize PFL for subsequent application in biodiesel synthesis.

  19. Multiple-level regulation of 2,4-diacetylphloroglucinol production by the sigma regulator PsrA in Pseudomonas fluorescens 2P24.

    Directory of Open Access Journals (Sweden)

    Xiaogang Wu

    Full Text Available BACKGROUND: Pseudomonas fluorescens 2P24 is a rhizospheric bacterium that aggressively colonizes the plant roots. It produces the antibiotic 2,4-diacetylphoroglucinol (2,4-DAPG, which contributes to the protection of various crop plants against soil borne diseases caused by bacterial and fungal pathogens. The biosynthesis of 2,4-DAPG is regulated at the transcriptional level in the expression of the phlACBD operon as well as at the posttranscriptional level by the Gac/Rsm signal transduction pathway. However, the detailed mechanism of such regulation is not clear. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we identified a binding site for the sigma regulator PsrA in the promoter region of the phlA gene. Electrophoretic mobility shift experiments revealed direct and specific binding of PsrA to the phlA promoter region. Consistent with the fact that its binding site locates within the promoter region of phlA, PsrA negatively regulates phlA expression, and its inactivation led to significant increase in 2,4-DAPG production. Interestingly, PsrA also activates the expression of the sigma factor RpoS, which negatively regulates 2,4-DAPG production by inducing the expression of the RNA-binding protein RsmA. CONCLUSIONS/SIGNIFICANCE: These results suggest that PsrA is an important regulator that modulates 2,4-DAPG biosynthesis at both transcriptional and posttranscriptional levels.

  20. Adaptive radiation of Pseudomonas fluorescens SBW25 in experimental microcosms provides an understanding of the evolutionary ecology and molecular biology of A-L interface biofilm formation.

    Science.gov (United States)

    Koza, Anna; Kusmierska, Anna; McLaughlin, Kimberley; Moshynets, Olena; Spiers, Andrew J

    2017-07-03

    Combined experimental evolutionary and molecular biology approaches have been used to investigate the adaptive radiation of Pseudomonas fluorescens SBW25 in static microcosms leading to the colonisation of the air-liquid interface by biofilm-forming mutants such as the Wrinkly Spreader (WS). In these microcosms, the ecosystem engineering of the early wild-type colonists establishes the niche space for subsequent WS evolution and colonisation. Random WS mutations occurring in the developing population that deregulate diguanylate cyclases and c-di-GMP homeostasis result in cellulose-based biofilms at the air-liquid interface. These structures allow Wrinkly Spreaders to intercept O2 diffusing into the liquid column and limit the growth of competitors lower down. As the biofilm matures, competition increasingly occurs between WS lineages, and niche divergence within the biofilm may support further diversification before system failure when the structure finally sinks. A combination of pleiotropic and epistasis effects, as well as secondary mutations, may explain variations in WS phenotype and fitness. Understanding how mutations subvert regulatory networks to express intrinsic genome potential and key innovations providing a selective advantage in novel environments is key to understanding the versatility of bacteria, and how selection and ecological opportunity can rapidly lead to substantive changes in phenotype and in community structure and function. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Polymorphism of a lipid extract from Pseudomonas fluorescens: Structure analysis of a hexagonal phase and of a novel cubic phase of extinction symbol Fd--

    International Nuclear Information System (INIS)

    Mariani, P.; Rivas, E.; Delacroix, H.; Luzzati, V.

    1990-01-01

    The phase diagram of the Pseudomonas fluorescens lipid extract is unusual, in the sense that it displays a cubic phase straddled by a hexagonal phase. The hexagonal phase was studied over an extended concentration range, and the reflections were phased on the assumption that the structure contains circular cylinders of known radius. The cubic phase, whose extinction symbol is Fd--, was analyzed by reference to space group No. 227 (Fd3m). The phases of the reflections were determined by using a novel pattern recognition approach, based upon the notion that the average fourth power of the electron density contrast 4 > is dependent on chemical composition but not on physical structure, provided that the function Δr(r) satisfies the constraints = 0 and 2 > = 1. The authors analyzed two cubic samples of different composition: for each of them they generated all the phase combinations compatible with the X-ray scattering data and they searched for those whose 4 > best agrees with the hexagonal phase. They concluded that the chemical composition of the phases being compared must be identical, that the X-ray scattering data should not be truncated artificially, and that the apodization must be mild so that the curvature takes a value intermediate between those corresponding to the raw data of the two phases. The structure may be visualized as a 3D generalization of the lipid monolayer. The structure, moreover, does not belong to the class of the infinite periodic surfaces without intersections

  2. Proteolytic activity of protease produced by Pseudomonas fluorescens IB 2312 in skimmed milk subject to the process of high pressure homogenization

    Directory of Open Access Journals (Sweden)

    Claudia Regina Gonçalves Pinho

    2014-08-01

    Full Text Available The presence of thermoresistant proteases produced by psychrotrophic microorganisms have been identified as a limiting factor of the UHT milk shelflife, causing undesirable changes in milk products. High pressure homogenization (HPH processing is a non-thermal method of food preservation, able to promotes the microbiological safety and inactivation of some enzymes. Thus, this work assessed the proteolytic activity of protease produced by Pseudomonas fluorescens in skim milk subjected to high pressure homogenization process. The milk samples were added by the protease enzymatic extract (10% v/v and subjected to pressures up to 300 MPa. The assays showed that pressures on the order of 300 MPa caused a 72.5% reduction in proteolytic activity. Therefore, the process at high pressures resulted in significant inactivation of this thermoresistent enzyme, which possibly favors the shelf-life extension of the UHT milk and also limits the yield and quality loss of cheeses due to undesirable sensory changes in flavor and texture caused by this enzyme.

  3. Efficacy of Pseudomonas fluorescens strain CL145A spray dried powder for controlling zebra mussels adhering to native unionid mussels within field enclosures

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Severson, Todd J.; Mayer, Denise A.

    2015-01-01

    The efficacy of a commercially prepared spray dried powder (SDP) formulation of Pseudomonas fluorescens (strain CL145A) was evaluated for removing zebra mussels (Dreissena polymorpha) adhering to a population of unionid mussels in Lake Darling (Alexandria, Minnesota). Two groups of unionid mussels were used in the study. Unionid mussels were collected near the test area, weighed, photographed, individually tagged, and randomly allocated to one of nine test enclosures in equal proportions and then divided into two groups. The first group of unionid mussels (Group 1, n = 5 per test enclosure) were indiscriminately selected from each test enclosure and used to estimate the number of zebra mussels adhering to unionid mussels prior to exposure. The second group of unionid mussels (Group 2, n = 22 per test enclosure) were used to evaluate the efficacy of SDP for removal of adhering zebra mussels. Both Group 1 and Group 2 mussels were used to evaluate the effects of SDP exposure on unionid mussel survival.

  4. Getting the ecology into the interactions between plants and the plant-growth promoting bacterium Pseudomonas fluorescens

    NARCIS (Netherlands)

    Hol, W.H.G.; Bezemer, T.M.; Biere, A.

    2013-01-01

    Plant growth-promoting rhizobacteria (PGPR) are increasingly appreciated for their contributions to primary productivity through promotion of growth and triggering of induced systemic resistance in plants. Here we focus on the beneficial effects of one particular species of PGPR (Pseudomonas

  5. Colonization of Arabidopsis roots by Pseudomonas fluorescens primes the plant to produce higher levels of ethylene upon pathogen infection

    NARCIS (Netherlands)

    Hase, S.; Pelt, J.A. van; Loon, L.C. van; Pieterse, C.M.J.

    2003-01-01

    Plants develop an enhanced defensive capacity against a broad spectrum of plant pathogens after colonization of the roots by selected strains of non-pathogenic, fluorescent Pseudomonas spp. In Arabidopsis thaliana, this rhizobacteria-induced systemic resistance (ISR) functions independently of

  6. Adaptive Divergence in Experimental Populations of Pseudomonas fluorescens. IV. Genetic Constraints Guide Evolutionary Trajectories in a Parallel Adaptive Radiation

    OpenAIRE

    McDonald, Michael J.; Gehrig, Stefanie M.; Meintjes, Peter L.; Zhang, Xue-Xian; Rainey, Paul B.

    2009-01-01

    The capacity for phenotypic evolution is dependent upon complex webs of functional interactions that connect genotype and phenotype. Wrinkly spreader (WS) genotypes arise repeatedly during the course of a model Pseudomonas adaptive radiation. Previous work showed that the evolution of WS variation was explained in part by spontaneous mutations in wspF, a component of the Wsp-signaling module, but also drew attention to the existence of unknown mutational causes. Here, we identify two new muta...

  7. Evidence for a role of biosurfactants produced by Pseudomonas fluorescens in the spoilage of fresh aerobically stored chicken meat.

    Science.gov (United States)

    Mellor, Glen E; Bentley, Jessica A; Dykes, Gary A

    2011-08-01

    Fresh chicken meat is a fat-rich environment and we therefore hypothesised that production of biosurfactants to increase bioavailability of fats may represent one way in which spoilage bacteria might enhance the availability of nutrients. Numbers of Pseudomonas were determined on a total of 20 fresh and 20 spoiled chicken thighs with skin. A total of 400 randomly isolated Pseudomonas colonies from fresh (200) and spoiled (200) chicken were screened for the presence of biosurfactant production. Biosurfactant producing strains represented 5% and 72% of the Pseudomonas spp. isolates from fresh (mean count 2.3 log(10) cfu g(-1)) and spoiled (mean count 7.4 log(10) cfu g(-1)) chicken skin, respectively. Partially-purified biosurfactants derived from a subgroup of four Pseudomonasfluorescens strains obtained through the screening process were subsequently used to investigate the role that the addition of these compounds plays in the spoilage of aerobically stored chicken. Emulsification potential of the four selected biosurfactants was measured against a range of hydrocarbons and oils. All four biosurfactants displayed a greater ability to emulsify rendered chicken fat than hydrocarbons (paraffin liquid, toluene and hexane) and oils (canola, olive, sunflower and vegetable). Storage trials (4 °C) of chicken meat treated with the four selected biosurfactants revealed a significantly greater (P biosurfactant treated samples, as compared to untreated samples on each day (0, 1, 2, 3) of storage. For biosurfactant treated samples the greatest increase in total aerobic count (1.3-1.7 log(10) cfu g(-1)) occurred following one day of incubation. These results indicate that biosurfactants produced by Pseudomonas spp. may play an important role in the spoilage of aerobically stored chicken meat by making nutrients more freely available and providing strains producing them with a competitive advantage. Copyright © 2011 Elsevier Ltd. All rights reserved.

  8. Enhanced biosorption of mercury(II) and cadmium(II) by cold-induced hydrophobic exobiopolymer secreted from the psychrotroph Pseudomonas fluorescens BM07

    Energy Technology Data Exchange (ETDEWEB)

    Zamil, Sheikh Shawkat; Choi, Mun Hwan; Song, Jung Hyun; Park, Hyunju; Xu, Ju; Yoon, Sung Chul [Gyeongsang National Univ., Jinju (Korea). Nano-Biomaterials Science Lab.; Chi, Ki-Whan [Ulsan Univ. (Korea). Dept. of Chemistry

    2008-09-15

    The cells of psychrotrophic Pseudomonas fluorescens BM07 were found to secrete large amounts of exobiopolymer (EBP) composed of mainly hydrophobic (water insoluble) polypeptide(s) (as contain {proportional_to}50 mol% hydrophobic amino acids, lacking cysteine residue) when grown on fructose containing limited M1 medium at the temperatures as low as 0-10 C but trace amount at high (30 C, optimum growth) temperature. Two types of nonliving BM07 cells (i.e., cells grown at 30 C and 10 C) as well as the freeze-dried EBP were compared for biosorption of mercury (Hg(II)) and cadmium (Cd(II)). The optimum adsorption pH was found 7 for Hg(II) but 6 for Cd(II), irrespective of the type of biomass. Equilibrium adsorption data well fitted the Langmuir adsorption model. The maximum adsorption (Q{sub max}) was 72.3, 97.4, and 286.2 mg Hg(II)/g dry biomass and 18.9, 27.0, and 61.5 mg Cd(II)/g dry biomass for cells grown at 30 C and 10 C and EBP, respectively, indicating major contribution of heavy metal adsorption by cold-induced EBP. Mercury(II) binding induced a significant shift of infrared (IR) amide I and II absorption of EBP whereas cadmium(II) binding showed only a very little shift. These IR shifts demonstrate that mercury(II) and cadmium(II) might have different binding sites in EBP, which was supported by X-ray diffraction and differential scanning calorimetric analysis and sorption results of chemically modified biomasses. This study implies that the psychrotrophs like BM07 strain may play an important role in the bioremediation of heavy metals in the temperate regions especially in the inactive cold season. (orig.)

  9. Role of the GacS sensor kinase in the regulation of volatile production by plant growth-promoting Pseudomonas fluorescens SBW25

    Directory of Open Access Journals (Sweden)

    Xu Cheng

    2016-11-01

    Full Text Available In plant-associated Pseudomonas species, the production of several secondary metabolites and exoenzymes is regulated by the GacS/GacA two-component regulatory system (the Gac-system. Here, we investigated if a mutation in the GacS sensor kinase affects the production of volatile organic compounds (VOCs in P. fluorescens SBW25 (Pf.SBW25 and how this impacts on VOCs-mediated growth promotion and induced systemic resistance of Arabidopsis and tobacco. A total of 205 VOCs were detected by Gas Chromatography Mass Spectrometry (GC-MS for Pf. SBW25 and the gacS-mutant grown on 2 different media for 3 and 6 days. Discriminant function analysis followed by hierarchical clustering revealed 24 VOCs that were significantly different in their abundance between Pf.SBW25 and the gacS-mutant, which included three acyclic alkenes (3-nonene, 4-undecyne, 1-undecene. These alkenes were significantly reduced by the gacS mutation independently of the growth media and of the incubation time. For Arabidopsis, both Pf.SBW25 and the gacS-mutant enhanced, via VOCs, root and shoot biomass, induced systemic resistance against leaf infections by P. syringae and rhizosphere acidification to the same extent. For tobacco, however, VOCs-mediated effects on shoot and root growth were significantly different between Pf.SBW25 and the gacS-mutant. While Pf.SBW25 inhibited tobacco root growth, the gacS-mutant enhanced root biomass and lateral root formation relative to the non-treated control plants. Collectively these results indicate that the sensor kinase GacS is involved in the regulation of VOCs production in Pf.SBW25, affecting plant growth in a plant species-dependent manner.

  10. Improvement of a dry formulation of Pseudomonas fluorescens EPS62e for fire blight disease biocontrol by combination of culture osmoadaptation with a freeze-drying lyoprotectant.

    Science.gov (United States)

    Cabrefiga, J; Francés, J; Montesinos, E; Bonaterra, A

    2014-10-01

    To study the effect of lyoprotectants and osmoadaptation on viability of Pseudomonas fluorescens EPS62e during freeze-drying and storage and to evaluate the formulation in terms of efficacy in biocontrol and fitness on pear flowers. A wettable powder formulation of a biocontrol agent of fire blight was optimized by means of lyoprotectants and culture osmoadaptation. Freeze-drying was used to obtain dehydrated cells, and the best viability (70% of survival) was obtained using lactose as lyoprotectant. Survival during lyophilization was additionally improved using physiological adaptation of cells during cultivation under salt-amended medium (osmoadaptation). The procedure increased the survival of cells after freeze-drying attaining viability values close to a 100% in the lactose-formulated product (3 × 10(11) CFU g(-1) ), and through the storage period of 1 year at 4°C. The dry formulation showed also an improved biocontrol efficacy and survival of EPS62e on pear flowers under low relative humidity conditions. Cell viability after freeze-drying was improved using lactose as lyoprotectant combined with a procedure of osmoadaptation during cultivation. The powder-formulated product remained active for 12 months and retained biocontrol levels similar to that of fresh cells. The formulation showed an improved survival of EPS62e on flowers and an increase of the efficacy of biocontrol of fire blight at low relative humidity. The results have a potential value for commercial application in biocontrol agents not only of fire blight but also of other plant diseases. © 2014 The Society for Applied Microbiology.

  11. Prokaryotic Homologs of the Eukaryotic 3-Hydroxyanthranilate 3,4-Dioxygenase and 2-Amino-3-Carboxymuconate-6-Semialdehyde Decarboxylase in the 2-Nitrobenzoate Degradation Pathway of Pseudomonas fluorescens Strain KU-7†

    OpenAIRE

    Muraki, Takamichi; Taki, Masami; Hasegawa, Yoshie; Iwaki, Hiroaki; Lau, Peter C. K.

    2003-01-01

    The 2-nitrobenzoic acid degradation pathway of Pseudomonas fluorescens strain KU-7 proceeds via a novel 3-hydroxyanthranilate intermediate. In this study, we cloned and sequenced a 19-kb DNA locus of strain KU-7 that encompasses the 3-hydroxyanthranilate meta-cleavage pathway genes. The gene cluster, designated nbaEXHJIGFCDR, is organized tightly and in the same direction. The nbaC and nbaD gene products were found to be novel homologs of the eukaryotic 3-hydroxyanthranilate 3,4-dioxygenase a...

  12. Salicylic acid degradation from aqueous solutions using Pseudomonas fluorescens HK44: parameters studies and application tools Degradação de ácido salicílico presente em soluções sintéticas utilizando Pseudomonas fluorescens HK44

    Directory of Open Access Journals (Sweden)

    Tatyane R. Silva

    2007-03-01

    Full Text Available The optimal conditions for salicylic acid biodegradation by Pseudomonas fluorescens HK44 were determined in this study with the intention to create a microbial sensor. Kinetic experiments permitted a definition of 60 and 30min the time needed to achieve the maximum degradation of salicylic acid presented in a medium with and without yeast extract, respectively. The degradation in medium without yeast extract and the quantification by spectrophotometry 230 nm were selected to be used in further tests. The use of preactivated cells or on the exponential growth phase showed better salicylic acid degradation percentages when compared to nonactivated cells or on the stationary growth state. Finally, the best cellular concentration used on the salicylic acid degradation was 0,1 g.L-1. Strain HK44 shows to be capable of degrade salicylic acid presented in simple aqueous systems, making this strain a promising tool for the application on a luminescent microbial sensor.Com a intenção de criar um sensor microbiano, as condições ótimas para a biodegradação de ácido salicílico por Pseudomonas fluorescens HK44 foram determinadas neste estudo. Os experimentos cinéticos permitiram a definição dos tempos de 60 e 30 minutos como necessários para atingir a máxima degradação de ácido salicílico presente em meio com ou sem extrato de lêvedo, respectivamente. A degradação no meio sem extrato de lêvedo e a quantificação através de espectrofotometria 230 nm foram selecionadas para serem utilizadas em testes posteriores. O uso de células pré-ativadas ou na fase exponencial de crescimento apresentou melhores porcentagens de degradação de ácido salicílico quando comparadas a células não-ativadas ou no estado estacionário de crescimento. Além disso, a melhor concentração celular utilizada nessa degradação foi 0,1 g.L¹. A cepa HK44 parece ser capaz de degradar o ácido salicílico presente em sistemas aquosos simples, tornando este

  13. From the root to the stem: interaction between the biocontrol root endophyte Pseudomonas fluorescens PICF7 and the pathogen Pseudomonas savastanoi NCPPB 3335 in olive knots

    Science.gov (United States)

    Maldonado-González, M Mercedes; Prieto, Pilar; Ramos, Cayo; Mercado-Blanco, Jesús

    2013-01-01

    Olive knot disease, caused by Pseudomonas savastanoi pv. savastanoi, is one of the most important biotic constraints for olive cultivation. Pseudomonas fluorescens PICF7, a natural colonizer of olive roots and effective biological control agent (BCA) against Verticillium wilt of olive, was examined as potential BCA against olive knot disease. Bioassays using in vitro-propagated olive plants were carried out to assess whether strain PICF7 controlled knot development either when co-inoculated with the pathogen in stems or when the BCA (in roots) and the pathogen (in stems) were spatially separated. Results showed that PICF7 was able to establish and persist in stem tissues upon artificial inoculation. While PICF7 was not able to suppress disease development, its presence transiently decreased pathogen population size, produced less necrotic tumours, and sharply altered the localization of the pathogen in the hyperplasic tissue, which may pose epidemiological consequences. Confocal laser scanning microscopy combined with fluorescent tagging of bacteria revealed that when PICF7 was absent the pathogen tended to be localized at the knot surface. However, presence of the BCA seemed to confine P. savastanoi at inner regions of the tumours. This approach has also enabled to prove that the pathogen can moved systemically beyond the hypertrophied tissue. PMID:23425069

  14. Annual changes in bioactive contents and production in field-grown blackberry after inoculation with Pseudomonas fluorescens.

    Science.gov (United States)

    Ramos-Solano, B; Garcia-Villaraco, A; Gutierrez-Mañero, F J; Lucas, J A; Bonilla, A; Garcia-Seco, D

    2014-01-01

    The aim of this study was two-fold: first, to characterize blackberry fruits from Rubus sp. var. Lochness along the year, and secondly, to evaluate the ability of a Pseudomonas strain (N21.4) to improve fruit yield and quality under field conditions in production greenhouses throughout the year. The strain was root or leaf inoculated to blackberry plants and fruits were harvested in each season. Nutritional parameters, antioxidant potential and bioactive contents were determined; total fruit yield was recorded. Blackberries grown under short day conditions (autumn and winter) showed significantly lower °Brix values than fruits grown under long day conditions. Interestingly, an increase in fruit °Brix, relevant for quality, was detected after bacterial challenge, together with significant and sustained increases in total phenolics and flavonoids. Improvements in inoculated fruits were more evident from October through early March, when environmental conditions are worse. In summary, N21.4 is an effective agent to increase fruit quality and production along the year in blackberry; this is an environmentally friendly approach to increase fruit quality. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  15. The interplay of StyR and IHF regulates substrate-dependent induction and carbon catabolite repression of styrene catabolism genes in Pseudomonas fluorescens ST

    Directory of Open Access Journals (Sweden)

    Leoni Livia

    2008-06-01

    Full Text Available Abstract Background In Pseudomonas fluorescens ST, the promoter of the styrene catabolic operon, PstyA, is induced by styrene and is subject to catabolite repression. PstyA regulation relies on the StyS/StyR two-component system and on the IHF global regulator. The phosphorylated response regulator StyR (StyR-P activates PstyA in inducing conditions when it binds to the high-affinity site STY2, located about -40 bp from the transcription start point. A cis-acting element upstream of STY2, named URE, contains a low-affinity StyR-P binding site (STY1, overlapping the IHF binding site. Deletion of the URE led to a decrease of promoter activity in inducing conditions and to a partial release of catabolite repression. This study was undertaken to assess the relative role played by IHF and StyR-P on the URE, and to clarify if PstyA catabolite repression could rely on the interplay of these regulators. Results StyR-P and IHF compete for binding to the URE region. PstyA full activity in inducing conditions is achieved when StyR-P and IHF bind to site STY2 and to the URE, respectively. Under catabolite repression conditions, StyR-P binds the STY1 site, replacing IHF at the URE region. StyR-P bound to both STY1 and STY2 sites oligomerizes, likely promoting the formation of a DNA loop that closes the promoter in a repressed conformation. We found that StyR and IHF protein levels did not change in catabolite repression conditions, implying that PstyA repression is achieved through an increase in the StyR-P/StyR ratio. Conclusion We propose a model according to which the activity of the PstyA promoter is determined by conformational changes. An open conformation is operative in inducing conditions when StyR-P is bound to STY2 site and IHF to the URE. Under catabolite repression conditions StyR-P cellular levels would increase, displacing IHF from the URE and closing the promoter in a repressed conformation. The balance between the open and the closed

  16. Efficacy of spray –Dried Pseudomonas fluorescens, strain CL145A (Zequanox®), for controlling Zebra Mussels (Dreissena polymorpha) within Lake Minnetonka, MN enclosures

    Science.gov (United States)

    Luoma, James A.; Severson, Todd J.

    2016-01-01

    The efficacy of whole water column and subsurface applications of the biopesticide Zequanox®, a commercially prepared spray-dried powder formulation of Pseudomonas fluorescens (strain CL145A), were evaluated for controlling zebra mussels (Dreissena polymorpha) within 27-m2 enclosures in Lake Minnetonka (Deephaven, Minnesota). Five treatments consisting of (1) two whole water column Zequanox applications, (2) two subsurface Zequanox applications, and (3) an untreated control were completed on each of three independent treatment days during September 2014. The two types of samplers used in the study were (1) type 1 samplers, which were custom built multi-plate samplers (wood, perforated aluminum, and tile substrates) that were placed into Robinson’s Bay in June of 2013 to allow for natural colonization by zebra mussels, and (2) type 2 samplers, which consisted of zebra mussels adhering to perforated aluminum trays that were placed into mesh containment bags. One day prior to treatment, three individual samplers of each type were distributed to test enclosures and exposed to a randomly assigned treatment. Sampling to determine the zebra mussel biomass adhering to type 1 samplers and the survival assessments for zebra mussels contained in type 2 samplers were completed ~40 days after exposure. The zebra mussel biomass adhering to type 1 samplers and the survival of zebra mussels contained in type 2 samplers were significantly less in groups treated with the highest Zequanox concentrations and in groups that received whole water column applications than comparable groups treated with lower Zequanox concentrations and subsurface applications. However, standardization of biomass and survival results to the amount of Zequanox applied showed that the lower concentrations and subsurface applications were more cost efficient, with respect to product used, at reducing zebra mussel biomass and for inducing zebra mussel mortality. Although the subsurface application methods

  17. Kinetics of biofilm formation and desiccation survival of Listeria monocytogenes in single and dual species biofilms with Pseudomonas fluorescens, Serratia proteamaculans or Shewanella baltica on food-grade stainless steel surfaces.

    Science.gov (United States)

    Daneshvar Alavi, Hessam Edin; Truelstrup Hansen, Lisbeth

    2013-01-01

    This study investigated the dynamics of static biofilm formation (100% RH, 15 °C, 48-72 h) and desiccation survival (43% RH, 15 °C, 21 days) of Listeria monocytogenes, in dual species biofilms with the common spoilage bacteria, Pseudomonas fluorescens, Serratia proteamaculans and Shewanella baltica, on the surface of food grade stainless steel. The Gram-negative bacteria reduced the maximum biofilm population of L. monocytogenes in dual species biofilms and increased its inactivation during desiccation. However, due to the higher desiccation resistance of Listeria relative to P. fluorescens and S. baltica, the pathogen survived in greater final numbers. In contrast, S. proteamaculans outcompeted the pathogen during the biofilm formation and exhibited similar desiccation survival, causing the N21 days of Serratia to be ca 3 Log10(CFU cm(-2)) greater than that of Listeria in the dual species biofilm. Microscopy revealed biofilm morphologies with variable amounts of exopolymeric substance and the presence of separate microcolonies. Under these simulated food plant conditions, the fate of L. monocytogenes during formation of mixed biofilms and desiccation depended on the implicit characteristics of the co-cultured bacterium.

  18. Characterization of the CrbS/R Two-Component System in Pseudomonas fluorescens Reveals a New Set of Genes under Its Control and a DNA Motif Required for CrbR-Mediated Transcriptional Activation

    Directory of Open Access Journals (Sweden)

    Edgardo Sepulveda

    2017-11-01

    Full Text Available The CrbS/R system is a two-component signal transduction system that regulates acetate utilization in Vibrio cholerae, P. aeruginosa, and P. entomophila. CrbS is a hybrid histidine kinase that belongs to a recently identified family, in which the signaling domain is fused to an SLC5 solute symporter domain through aSTAC domain. Upon activation by CrbS, CrbR activates transcription of the acs gene, which encodes an acetyl-CoA synthase (ACS, and the actP gene, which encodes an acetate/solute symporter. In this work, we characterized the CrbS/R system in Pseudomonas fluorescens SBW25. Through the quantitative proteome analysis of different mutants, we were able to identify a new set of genes under its control, which play an important role during growth on acetate. These results led us to the identification of a conserved DNA motif in the putative promoter region of acetate-utilization genes in the Gammaproteobacteria that is essential for the CrbR-mediated transcriptional activation of genes under acetate-utilizing conditions. Finally, we took advantage of the existence of a second SLC5-containing two-component signal transduction system in P. fluorescens, CbrA/B, to demonstrate that the activation of the response regulator by the histidine kinase is not dependent on substrate transport through the SLC5 domain.

  19. Quantification of Pseudomonas fluorescens strains F113, CHA0 and Pf153 in the rhizosphere of maize by strain-specific real-time PCR unaffected by the variability of DNA extraction efficiency.

    Science.gov (United States)

    Von Felten, Andreas; Défago, Geneviève; Maurhofer, Monika

    2010-05-01

    Pseudomonas fluorescens strains F113 and CHA0 are well-known plant growth-promoting rhizobacteria (PGPR) often used as model strains in biocontrol experiments. To monitor their persistence in large scale field experiments, culture-independent methods are needed. In this study, a strain-specific real-time PCR quantification tool was developed based on sequence-characterized amplified regions (SCAR) for P. fluorescens strains F113, CHA0 and Pf153. Differences in DNA extraction efficiencies from rhizosphere samples were circumvented using plasmid APA9 as internal standard to normalize C(T) values after real-time amplification. The detection limits of the real-time PCR assays for all three strains were approximately 10 cells for genomic DNA and 10(4)cells/g rhizosphere for maize samples grown in different natural soils. Population sizes of the three strains in the rhizosphere of maize measured by the new real-time PCR approaches were similar to those measured by most probable number (MPN)-PCR. A persistence study of the three strains indicated that the strains persisted differently over a period of 5weeks. In conclusion the newly developed real-time PCR approach is a fast and resource efficient method for monitoring individual biocontrol strains in natural soil, which makes it an apt quantification tool for future large-scale field experiments. Copyright 2010 Elsevier B.V. All rights reserved.

  20. Non-target effects of the microbial control agents Pseudomonas fluorescens DR54 and Clonostachys rosea IK726 in soils cropped with barley followed by sugar beet: a greenhouse assessment

    DEFF Research Database (Denmark)

    Johansen, A.; Knudsen, I. M. B.; Binnerup, S. J.

    2005-01-01

    on plant growth or soil microorganisms. Phospholipid fatty acid (PLFA) analysis detected the perturbations of the soil microbial community structure in the MCA treatments as well as the return to non- perturbed conditions reflecting the decline of inoculant populations. The PLFA technique appears......Non-target effects of a bacterial (Pseudomonas fluorescens DR54) and a fungal (Clonostachys rosea IK726) microbial control agent (MCA), on the indigenous microbiota in bulk soil and rhizosphere of barley, and subsequent a sugar beet crop, were studied in a greenhouse experiment. MCAs were...... introduced by seed and soil inoculation. Bulk and rhizosphere soils were sampled regularly during the growth of barley and sugar beet. The soils were assayed for the fate of MCAs and various features of the indigenous soil microbiota. At the end of the experiment (193 d), DR54 and IK726 had declined...

  1. INOCULACIÓN AL SUELO CON Pseudomonas fluorescens, Azospirillum oryzae, Bacillus subtilis Y MICROORGANISMOS DE MONTAÑA (MM Y SU EFECTO SOBRE UN SISTEMA DE ROTACIÓN SOYA-TOMATE BAJO CONDICIONES DE INVERNADERO

    Directory of Open Access Journals (Sweden)

    Leida Castro Barquero

    2015-01-01

    Full Text Available Se evaluó un sistema de rotación soya- tomate, con incorporación de biomasa verde y aplicación de inóculos microbianos individuales y en mezcla sobre el crecimiento de las plantas y propiedades edáficas; para ello se evaluaron en invernadero por 24 meses los siguientes 9 trata - mientos: solo tomate (T; rotación tomate-soya (TS; rotación tomate-soya con inoculaciones individuales de Azospirillum oryzae (A; de Pseudomonas fluorescens (P; de Bacillus sub - tilis (B; de microorganismos de montaña (MM; y las inoculaciones en mezcla de B. subtilis y P. fluorescens (BP; de B. subtilis, P. fluorescens y A. oryzae ( BPA; d e B. subtilis , P. fluorescens , Azospirillum sp. y MM (BPAMM. Se evalua - ron las variables físicas: densidad aparente y de partículas; conductividad hidráulica; poros totales; estabilidad de agregados; resistencia a la penetración (RP; las variables químicas: pH; conductividad eléctrica; contenido de N y C; relación C/N; contenido de nutrientes en suelos y foliares; las variables biológicas: respiración de suelos, unidades formadoras de colonias de hongos, bacterias y actinomicetos y el peso fres - co y seco foliar. Las variables físicas no fueron afectadas por los tratamientos, con excepción de RP, que fue mayor en el tratamiento T. Las varia - bles biológicas y químicas fueron sensibles a los tratamientos, con valores significativamente más altos en presencia de MM. Destaca el incremento del P en solución de suelo en tratamientos a los que se adicionó MM: pasó de 6 a 20 mg.l -1 ; esta condición se reflejó además en la cantidad de P en el tejido foliar al final del ensayo. Se determi - nó que el pH, CE y la respiración del suelo fueron afectados por la interacción entre los tratamientos aplicados y el tiempo transcurrido; los mayores valores se obtuvieron al final del ensayo y en los tratamientos con MM.

  2. Inoculación al suelo con Pseudomonas fluorescens, Azospirillum oryzae, Bacillus subtilis y microorganismos de montaña (mm y su efecto sobre un sistema de rotación soya-tomate bajo condiciones de invernadero

    Directory of Open Access Journals (Sweden)

    Leida Castro Barquero

    2015-11-01

    Full Text Available Se evaluó un sistema de rotación soyatomate, con incorporación de biomasa verde y aplicación de inóculos microbianos individuales y en mezcla sobre el crecimiento de las plantas y propiedades edáficas; para ello se evaluaron en invernadero por 24 meses los siguientes 9 tratamientos: solo tomate (T; rotación tomate-soya (TS; rotación tomate-soya con inoculaciones individuales de Azospirillum oryzae (A; de Pseudomonas fluorescens (P; de Bacillus subtilis (B; de microorganismos de montaña (MM; y las inoculaciones en mezcla de B. subtilis y P. fluorescens (BP; de B. subtilis, P. fluorescens y A. oryzae (BPA; de B. subtilis, P. fluorescens, Azospirillum sp. y MM (BPAMM. Se evaluaron las variables físicas: densidad aparente y de partículas; conductividad hidráulica; poros totales; estabilidad de agregados; resistencia a la penetración (RP; las variables químicas: pH; conductividad eléctrica; contenido de N y C; relación C/N; contenido de nutrientes en suelos y foliares; las variables biológicas: respiración de suelos, unidades formadoras de colonias de hongos, bacterias y actinomicetos y el peso fresco y seco foliar. Las variables físicas no fueron afectadas por los tratamientos, con excepción de RP, que fue mayor en el tratamiento T. Las variables biológicas y químicas fueron sensibles a los tratamientos, con valores significativamente más altos en presencia de MM. Destaca el incremento del P en solución de suelo en tratamientos a los que se adicionó MM: pasó de 6 a 20 mg.l-1; esta condición se reflejó además en la cantidad de P en el tejido foliar al final del ensayo. Se determinó que el pH, CE y la respiración del suelo fueron afectados por la interacción entre los tratamientos aplicados y el tiempo transcurrido; los mayores valores se obtuvieron al final del ensayo y en los tratamientos con MM.

  3. Binding of Pu(IV) to galacturonic acid and extracellular polymeric substances (EPS) from Shewanella putrefaciens, Clostridium sp. and Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Harper, R.M.; Kantar, C.; Honeyman, B.D.

    2008-01-01

    The conditional stability constants for trace-level concentrations of Pu(IV) complexing with galacturonic acid and EPS, isolated from axenic Clostridium sp., P. fluorescens and Shewanella putrefaciens CN32 cultures, were determined at pH 4 and an ionic strength of 0.1 M NaCl using an ion-exchange technique. The analysis of ion-exchange data with Schubert's technique indicates that the Pu binding by galacturonic acid and EPS from Clostridium sp. and S. putrefaciens can be described based on the formation of 1: 1 Pu(IV)-ligand complexes. However, the accurate description of Pu binding by EPS from P. fluorescens requires postulation of a mixture of 1: 1/1: 2 complexes between Pu(IV) and ligands under the experimental conditions studied. The results from the ion-exchange experiments were also modeled based on a non-electrostatic, discrete ligand approach in which bacterial EPS is conceptualized as being composed of a suite of monoprotic acids, HL 1 , of arbitrarily-assigned pK a (i) values (e.g., 4, 6 and 8). The examination of ion-exchange data in a chemical model suggested that only the pK a 4 (L 1 ) and 6 (L 2 ) ligands are sufficient to accurately simulate the Pu(IV)/EPS binding, implying that carboxylic groups in EPS are the primary binding sites for complexing with Pu(IV) under the experimental conditions examined. The affinity of EPS for complexing Pu(IV) decreases in the order of Clostridium sp. > S. putrefaciens > P. fluorescens although the concentrations of carboxylic groups in EPS decrease in the order of P. fluorescens > S. putrefaciens > Clostridium sp. This discrepancy may be due to differences in binding affinities between Na + ion in solution and EPS ligands. At I = 0.1 M, models demonstrated that the EPS from P. fluorescens exhibits a much stronger affinity for the Na + ion compared to ligands from other EPS; therefore, the deprotonated carboxylic sites of EPS from P. fluorescens are hypothesized to be mostly bound by Na + in solution. (orig.)

  4. Evaluation of gamma irradiation effect and Pseudomonas ...

    African Journals Online (AJOL)

    Antagonistic effect of Pseudomonas fluorescens and influence of gamma irradiation on the development of Penicillium expansum, the causal agent of postharvest disease on apple fruit was studied. P. fluorescens was originally isolated from rhizosphere of the apple trees. Suspension of P. fluorescens and P. expansum ...

  5. Efficiency of the formulated plant-growth promoting Pseudomonas fluorescens MC46 inoculant on triclocarban treatment in soil and its effect on Vigna radiata growth and soil enzyme activities.

    Science.gov (United States)

    Sipahutar, Merry Krisdawati; Piapukiew, Jittra; Vangnai, Alisa S

    2018-02-15

    For bioaugmentation-based treatment of triclocarban (TCC), an emerging soil pollutant that is recalcitrant to biodegradation and phytotransformation, efficient TCC-degrading bacteria with an effective soil-delivering means are required. This work developed the formulated bacterial inoculant, and successfully demonstrated its TCC removal and detoxification performance in pot soil experiment with Vigna radiata plants. The soil bacterium Pseudomonas fluorescens MC46 was isolated as TCC-degrading, plant-growth promoting bacterium. The characterizations were conducted in vitro revealing that it could utilize TCC as a sole carbon source, and at a wide and higher concentration range from 1.6-31.6mgkg -1 than those previously reported, while the detoxification was assessed by cytogenotoxicity and phytotoxicity tests. The developed sawdust-based inoculant formula combined with molasses (5% w/w), and either PEG or CMC-starch blend (1% w/w) could maintain a 20-week shelf-life inoculant stability in terms of cell viability, and TCC-degrading activity. Bioaugmentation of the formulated inoculants into TCC-contaminated soil efficiently removed TCC up to 74-76% of the initial concentration, mitigated toxicity, restored plant growth and health, and enhanced soil enzyme activities. This work is the first to demonstrate potential application of the formulated plant-growth promoting bacterial inoculant for the treatment and detoxification of a persistent TCC contaminated in soil. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Mutations in the central cavity and periplasmic domain affect efflux activity of the resistance-nodulation-division pump EmhB from Pseudomonas fluorescens cLP6a.

    Science.gov (United States)

    Hearn, Elizabeth M; Gray, Murray R; Foght, Julia M

    2006-01-01

    The EmhABC efflux system in Pseudomonas fluorescens cLP6a is homologous to the multidrug and solvent efflux systems belonging to the resistance-nodulation-division (RND) family and is responsible for polycyclic aromatic hydrocarbon transport, antibiotic resistance, and toluene efflux. To gain a better understanding of substrate transport in RND efflux pumps, the EmhB pump was subjected to mutational analysis. Mutagenesis of amino acids within the central cavity of the predicted three-dimensional structure of EmhB showed selective activity towards antibiotic substrates. An A384P/A385Y double mutant showed increased susceptibility toward rhodamine 6G compared to the wild type, and F386A and N99A single mutants showed increased susceptibility to dequalinium compared to the wild type. As well, the carboxylic acid side chain of D101, located in the central cavity region, was found to be essential for polycyclic aromatic hydrocarbon transport and resistance to all antibiotic substrates of EmhB. Phenylalanine residues located within the periplasmic pore domain were also targeted for mutagenesis, and the F325A and F281A mutations significantly impaired efflux activity for all EmhB substrates. One mutation (A206S) in the outer membrane protein docking domain increased antibiotic resistance and toluene tolerance, demonstrating the important role of this domain in transport activity. These data demonstrate the roles of the central cavity and periplasmic domains in the function of the RND efflux pump EmhB.

  7. Role of the GacS sensor kinase in the regulation of volatile production by plant growth-promoting Pseudomonas fluorescens SBW25

    NARCIS (Netherlands)

    Cheng, Xu; Cordovez, Viviane; Etalo, Desalegn W.; Voort, van der M.; Raaijmakers, Jos M.

    2016-01-01

    In plant-associated Pseudomonas species,the production of several secondary metabolites and exoenzymes is regulated by the GacS/GacA two-component regulatory system (the Gac-system). Here,we investigated if a mutation in the GacS sensor kinase affects the production of volatile organic compounds

  8. Role of the GacS Sensor Kinase in the Regulation of Volatile Production by Plant Growth-Promoting Pseudomonas fluorescens SBW25

    NARCIS (Netherlands)

    Cheng, Xu; Cordovez da Cunha, Viviane; Etalo, Desalegn W.; van der Voort, Menno; Raaijmakers, Jos M.

    2016-01-01

    In plant-associated Pseudomonas species, the production of several secondary metabolites and exoenzymes is regulated by the GacS/GacA two-component regulatory system (the Gac-system). Here, we investigated if a mutation in the GacS sensor kinase affects the production of volatile organic compounds

  9. Exploiting genotypic diversity of 2,4-Diacetylphloroglucinol-producing Pseudomonas spp.: characterization of superior root-colonizing P. fluorescens strain Q8r1-96

    NARCIS (Netherlands)

    Raaijmakers, J.M.; Weller, D.M.

    2001-01-01

    The genotypic diversity that occurs in natural populations of antagonistic microorganisms provides an enormous resource for improving biological control of plant diseases. In this study, we determined the diversity of indigenous 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas spp. occurring

  10. Effect of aluminium and copper on biofilm development of Pseudomonas pseudoalcaligenes KF707 and P. fluorescens as a function of different media compositions.

    Science.gov (United States)

    Booth, Sean C; George, Iain F S; Zannoni, Davide; Cappelletti, Martina; Duggan, Gavin E; Ceri, Howard; Turner, Raymond J

    2013-06-01

    Bioremediation efforts worldwide are faced with the problem of metals interfering with the degradation of organic pollutants. There has been little systematic investigation into how the important environmental factors of media composition, buffering agent, and carbon source affect the exertion of metal toxicity on bacteria. This study aimed to systematically separate and investigate the influence of these factors by examining planktonic and biofilm establishment and growth. Two Pseudomonads were chosen, the PCB degrader P. pseudoalcaligenes KF707 and P. fluorescens. The two strains were grown in the presence of Al(3+) and Cu(2+) under different media conditions of carbon source (Lysogeny broth, biphenyl, succinate, aspartic acid, butyric acid, oxaloacetic acid, putrescine and benzoic acid) and under different buffering conditions (high and low phosphate or MOPS). These experiments allowed for the elucidation of an effect of different metabolic conditions and metal speciation on planktonic bacteria growth and biofilm establishment and development under metal stress. Here we show that the nature of bacterial growth (planktonic and biofilm development) is dramatically affected by the interplay between toxic metals, carbon source and media composition. The capacity of a media to bind toxic metals as well as quality of carbon source greatly influences the amount of metal that bacteria can tolerate, depending on both the bacterium and metal. Future studies evaluating metal ion toxicity should consider these effects, as well as their interactions with specific environments into account in order to improve clean-up success.

  11. Evaluation of economically feasible, natural plant extract-based microbiological media for producing biomass of the dry rot biocontrol strain Pseudomonas fluorescens P22Y05 in liquid culture.

    Science.gov (United States)

    Khalil, Sadia; Ali, Tasneem Adam; Skory, Chris; Slininger, Patricia J; Schisler, David A

    2016-02-01

    The production of microbial biomass in liquid media often represents an indispensable step in the research and development of bacterial and fungal strains. Costs of commercially prepared nutrient media or purified media components, however, can represent a significant hurdle to conducting research in locations where obtaining these products is difficult. A less expensive option for providing components essential to microbial growth in liquid culture is the use of extracts of fresh or dried plant products obtained by using hot water extraction techniques. A total of 13 plant extract-based media were prepared from a variety of plant fruits, pods or seeds of plant species including Allium cepa (red onion bulb), Phaseolus vulgaris (green bean pods), and Lens culinaris (lentil seeds). In shake flask tests, cell production by potato dry rot antagonist Pseudomonas fluorescens P22Y05 in plant extract-based media was generally statistically indistinguishable from that in commercially produced tryptic soy broth and nutrient broth as measured by optical density and colony forming units/ml produced (P ≤ 0.05, Fisher's protected LSD). The efficacy of biomass produced in the best plant extract-based media or commercial media was equivalent in reducing Fusarium dry rot by 50-96% compared to controls. In studies using a high-throughput microbioreactor, logarithmic growth of P22Y05 in plant extract-based media initiated in 3-5 h in most cases but specific growth rate and the time of maximum OD varied as did the maximum pH obtained in media. Nutrient analysis of selected media before and after cell growth indicated that nitrogen in the form of NH4 accumulated in culture supernatants, possibly due to unbalanced growth conditions brought on by a scarcity of simple sugars in the media tested. The potential of plant extract-based media to economically produce biomass of microbes active in reducing plant disease is considerable and deserves further research.

  12. Complex regulation of AprA metalloprotease in Pseudomonas fluorescens M114: evidence for the involvement of iron, the ECF sigma factor, PbrA and pseudobactin M114 siderophore.

    Science.gov (United States)

    Maunsell, Bláithín; Adams, Claire; O'Gara, Fergal

    2006-01-01

    In the soil bacterium Pseudomonas fluorescens M114, extracellular proteolytic activity and fluorescent siderophore (pseudobactin M114) production were previously shown to be co-ordinately negatively regulated in response to environmental iron levels. An iron-starvation extracytoplasmic function sigma factor, PbrA, required for the transcription of siderophore biosynthetic genes, was also implicated in M114 protease regulation. The current study centred on the characterization and genetic regulation of the gene(s) responsible for protease production in M114. A serralysin-type metalloprotease gene, aprA, was identified and found to encode the major, if not only, extracellular protease produced by this strain. The expression of aprA and its protein product were found to be subject to complex regulation. Transcription analysis confirmed that PbrA was required for full aprA transcription under low iron conditions, while the ferric uptake regulator, Fur, was implicated in aprA repression under high iron conditions. Interestingly, the iron regulation of AprA was dependent on culture conditions, with PbrA-independent AprA-mediated proteolytic activity observed on skim milk agar supplemented with yeast extract, when supplied with iron or purified pseudobactin M114. These effects were not observed on skim milk agar without yeast extract. PbrA-independent aprA expression was also observed from a truncated transcriptional fusion when grown in sucrose asparagine tryptone broth supplied with iron or purified pseudobactin M114. Thus, experimental evidence suggested that iron mediated its effects via transcriptional activation by PbrA under low iron conditions, while an as-yet-unidentified sigma factor(s) may be required for the PbrA-independent aprA expression and AprA proteolytic activity induced by siderophore and iron.

  13. Explaining variation in nascent entrepreneurship

    NARCIS (Netherlands)

    A.J. van Stel (André); A.R.M. Wennekers (Sander); P. Reynolds (Paul); A.R. Thurik (Roy)

    2004-01-01

    textabstractThis paper aims at explaining cross-country variation in nascent entrepreneurship. Regression analysis is applied using various explanatory variables derived from three different approaches. We make use of the Global Entrepreneurship Monitor database, including nascent entrepreneurship

  14. Effectiveness of cleaning and sanitizing procedures in controlling the adherence of Pseudomonas fluorescens, Salmonella Enteritidis, and Staphylococcus aureus to domestic kitchen surfaces Eficiência dos procedimentos de limpeza e de sanitização no controle da adesão de Pseudomonas fluorescens, Salmonella Enteritidis e Staphylococcus aureus em superfícies usadas em cozinhas domésticas

    Directory of Open Access Journals (Sweden)

    Iara Dias Silva

    2010-03-01

    Full Text Available The effectiveness of cleaning and sanitizing procedures in controlling Staphylococcus aureus, Salmonella Enteritidis, and Pseudomonasfluorescens adhered to granite and stainless steel was evaluated. There was no significant difference (p > 0.05 in the adherence of pure cultures of these microorganisms to stainless steel. The numbers of P. fluorescens and S. Enteritidis adhered to granite were greater (p 0.05 between the surfaces. However, a significant difference was observed (p A eficiência dos procedimentos de limpeza e sanitização no controle de Staphylococcus aureus, Salmonella Enteritidis e Pseudomonasfluorescens aderidas em granito e aço inoxidável foi avaliada. Não houve diferença significativa (p > 0,05 na adesão destes microrganismos quando em cultura pura, em aço inoxidável. O número de células aderidas de P. fluorescens e S. Enteritidis foi maior (p 0,05 entre as superfícies. Entretanto, observou-se uma diferença (p < 0,05 entre as soluções sanitizantes utilizadas. Hipoclorito de sódio e ácido peracético apresentaram maior ação bactericida (p < 0,05 que o composto de amônia quaternária. Observou-se que S. aureus apresentou menor resistência à ação desses sanitizantes. Os resultados mostram a importância da adequada realização dos procedimentos de limpeza e sanitização para evitar a adesão bacteriana e formação de biofilme.

  15. Comparative evaluation of organic formulations of Pseudomonas ...

    African Journals Online (AJOL)

    Comparative evaluation of organic formulations of Pseudomonas fluorescens based biopesticides and their application in the management of bacterial wilt of brinjal ... Effective management of bacterial wilt of brinjal by P. fluorescens under local conditions signifies its potentiality and scope as a plant growth promoting ...

  16. Invasive behaviour and depolarization effect of Pseudomonas ...

    African Journals Online (AJOL)

    ... of a constitutive calcium dependent NOS and that of an inducible NOS activated by LPS. Ours results also suggest that P. fluorescens cytotoxicity and invasion are not mutually exclusive events. Key Words: Cytotoxicity, Lipopolysaccharide, Patch-clamp, Invasion, Pseudomonas fluorescens. Afr. J. Clin. Exper. Microbiol.

  17. Transesterification of Jatropha oil using immobilized Pseudomonas ...

    African Journals Online (AJOL)

    mild transesterification has become of much current interest for alternative fuel production. In the present study the ability of a commercial immobilized Pseudomonas fluorescens MTCC 103 to catalyze the transesterification of Jatropha oil and methanol was investigated. The cell of P. fluorescens was easily immobilized ...

  18. Carbazole-degradative IncP-7 plasmid pCAR1.2 is structurally unstable in Pseudomonas fluorescens Pf0-1, which accumulates catechol, the intermediate of the carbazole degradation pathway.

    Science.gov (United States)

    Takahashi, Yurika; Shintani, Masaki; Li, Li; Yamane, Hisakazu; Nojiri, Hideaki

    2009-06-01

    We determined the effect of the host on the function and structure of the nearly identical IncP-7 carbazole-degradative plasmids pCAR1.1 and pCAR1.2. We constructed Pseudomonas aeruginosa PAO1(pCAR1.2) and P. fluorescens Pf0-1Km(pCAR1.2) and compared their growth on carbazole- and succinate-containing media with that of P. putida KT2440(pCAR1.1). We also assessed the stability of the genetic structures of the plasmids in each of the three hosts. Pf0-1Km(pCAR1.2) showed dramatically delayed growth when carbazole was supplied as the sole carbon source, while the three strains grew at nearly the same rate on succinate. Among the carbazole-grown Pf0-1Km(pCAR1.2) cells, two types of deficient strains appeared and dominated the population; such dominance was not observed in the other two strains or for succinate-grown Pf0-1Km(pCAR1.2). Genetic analysis showed that the two deficient strains possessed pCAR1.2 derivatives in which the carbazole-degradative car operon was deleted or its regulatory gene, antR, was deleted by homologous recombination between insertion sequences. From genomic information and quantitative reverse transcription-PCR analyses of the genes involved in carbazole mineralization by Pf0-1Km(pCAR1.2), we found that the cat genes on the chromosome of Pf0-1Km, which are necessary for the degradation of catechol (a toxic intermediate in the carbazole catabolic pathway), were not induced in the presence of carbazole. The resulting accumulation of catechol may have enabled the strain that lost its carbazole-degrading ability to have overall higher fitness than the wild-type strain. These results suggest that the functions of the chromosomal genes contributed to the selection of plasmid derivatives with altered structures.

  19. Growth of Pseudomonas fluorescens on Cassava Starch ...

    African Journals Online (AJOL)

    Michael Horsfall

    al., 1998). The demand for the use of biopolymers involves their biodegradable and bio-compatible properties suitable for disposable products and their potential use as bio-materials for medical applications such as surgical sutures, long-term carriers for drugs, moulded plastics and films. (Holmes, 1985, Hahn et al., 1993).

  20. Characterization and identification of Pseudomonas fluorescens ...

    African Journals Online (AJOL)

    user

    2011-01-24

    Bangert et al., 1977; Bumpus and. Brock, 1988; Gregory, 1993). This dye has ... cetes and algae have been reported to degrade different pollutants (Bumpus and Brock, 1988; Leahy and Colwell,. 1990; Spain, 1995; Azmi et al., ...

  1. Interaction of bacteria-feeding soil flagellates and Pseudomonas spp

    DEFF Research Database (Denmark)

    Pedersen, Annette L; Ekelund, Flemming; Johansen, Anders

    2010-01-01

    Pseudomonas strains may be used as alternatives to fungicides as some of them produce secondary metabolites, which can inhibit growth of plant pathogenic fungi. Increased knowledge of non-target effects of the antagonistic bacteria on other soil organisms as well as of the survival and predation...... resistance of the antagonistic bacteria is necessary for risk assessment and increased performance of antagonistic bacteria as biological control agents. In the present study, we aimed to investigate the difference between Pseudomonas spp. with respect to their predation resistance to and effects...... on the three different and common soil flagellates Bodo caudatus, Cercomonas longicauda, and Neocercomonas jutlandica. Two antagonistic Pseudomonas: Pseudomonas fluorescens CHA0 and P. fluorescens DR54 and two positive control strains: P. fluorescens DSM 50090T and Pseudomonas chlororaphis ATCC 43928 were...

  2. Control biológico del marchitamiento vascular causado por Fusarium oxysporum f. sp. phaseoli en fríjol Phaseolus vulgaris L., mediante la acción combinada de Entrophospora colombiana, Trichoderma sp. Y Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Avendaño Camila

    2006-06-01

    Hidden="false" UnhideWhenUsed="false" QFormat="true" Name="Intense Emphasis" /> Entrophospora colombiana, Trichoderma sp., Pseudomonas fluorescens y una combinación de estos antagonistas fueron evaluados como biocontroladores de Fusarium oxysporumf. sp. Phaseoli en plantas de fríjol de la variedad ‘ICA Tundama’. El ensayo se estableció en una casa de malla del Programa nacional de manejo integrado de plagas (MIP de la Corporación Colombiana de Investigación Agropecuaria (Corpoica, en el Centro de Investigación Tibaitat

  3. Optimization of alkaline protease production from Pseudomonas ...

    African Journals Online (AJOL)

    A protease producing bacteria was isolated from meat waste contaminated soil and identified as Pseudomonas fluorescens. Optimization of the fermentation medium for maximum protease production was carried out. The culture conditions like inoculum concentration, incubation time, pH, temperature, carbon sources, ...

  4. Optimization of alkaline protease production from Pseudomonas ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-12-15

    Dec 15, 2009 ... the metal ions tested. Key words: Alkaline protease, casein agar, meat waste contaminated soil, Pseudomonas fluorescens. INTRODUCTION. Proteases are the most important industrial enzymes that execute a wide variety of functions and have various important biotechnological applications (Mohen et al.,.

  5. The vectorial release of nascent immunoglobulin peptides.

    Science.gov (United States)

    Bevan, M J

    1971-03-01

    A microsomal preparation from a mouse plasmacytoma, MOPC 47A, that secretes immunoglobulin A was used to study the release of nascent immunoglobulin peptides in vitro. Nascent chains were released with puromycin and characterized with specific antiserum against the immunoglobulin product of the tumour. When the tissue had been prelabelled with [(3)H]leucine the experiments were complicated by the large background of completed radioactive polypeptides in the microsomal preparation. Up to one-third of the released radioactivity in the microsomal preparation could be recognized as immunoglobulin. With [(3)H]-puromycin as the radioactive label, however, the results are much easier to interpret, although the proportion of released radioactivity that can be identified as immunoglobulin is lower (up to one-tenth). Both types of experiment demonstrate that all of the recognizable nascent immunoglobulin chains remain in association with the microsomal vesicles after release from the ribosomes.

  6. Bioreporter Pseudomonas fluorescens HK44 Immobilized in a Silica Matrix

    Czech Academy of Sciences Publication Activity Database

    Trögl, Josef; Ripp, S.; Kuncová, Gabriela; Sayler, G. S.; Demnerová, K.

    2003-01-01

    Roč. 57, č. 12 (2003), s. 596-599 ISSN 0354-7531 R&D Projects: GA ČR GA104/01/0461 Institutional research plan: CEZ:AV0Z4072921 Keywords : bioluminescence bioreporter * optical sensor * naphthalene detection Subject RIV: CE - Biochemistry

  7. Novel Pseudomonas fluorescens Septic Sacroiliitis in a Healthy Soldier

    Science.gov (United States)

    2013-08-01

    laparotomy in 39% of patients in 1 series.18 As in our case, physical findings consistent with SI joint involvement include maximal tenderness over... surgery may be indicated for abscess, osteo- myelitis, a sequestrum of necrotic bone, or failure to show clinical improvement with intravenous...18. Cohn SM, Schoetz DJ Jr: Pyogenic sacroiliitis: another imitator of the acute abdomen. Surgery 1986; 100(1): 95–8. 19. Doita M, Yoshiya S

  8. Pseudomonas fluorescens strains selectively suppress annual bluegrass (Poa annua L.)

    Science.gov (United States)

    Annual bluegrass (Poa annua L.) is a cool-season annual grass that is a major weed species in turf, turfgrass-seed production, sod production, and golf courses of the western United States. There are few selective herbicides available for the management of annual bluegrass. While the life cycles o...

  9. Interactions between biosurfactant-producing Pseudomonas and Phytophthora species

    OpenAIRE

    Tran, H.

    2007-01-01

    Fluorescent Pseudomonas bacteria produce a wide variety of antimicrobial metabolites, including soap-like compounds referred to as biosurfactants. The results of this thesis showed that biosurfactant-producing Pseudomonas bacteria are effective in controlling Phytophthora foot rot disease of black pepper in Vietnam and promote root and shoot development of the ‘King of Spices’. Biosurfactant-producing P. fluorescens strain SS101 was also effective in controlling tomato late blight caused by P...

  10. The Biogenesis of Nascent Circular RNAs

    Directory of Open Access Journals (Sweden)

    Yang Zhang

    2016-04-01

    Full Text Available Steady-state circular RNAs (circRNAs have been mapped to thousands of genomic loci in mammals. We studied circRNA processing using metabolic tagging of nascent RNAs with 4-thiouridine (4sU. Strikingly, the efficiency of circRNA processing from pre-mRNA is extremely low endogenously. Additional studies revealed that back-splicing outcomes correlate with fast RNA Polymerase II elongation rate and are tightly controlled by cis-elements in vivo. Additionally, prolonged 4sU labeling in cells shows that circRNAs are largely processed post-transcriptionally and that circRNAs are stable. Circular RNAs that are abundant at a steady-state level tend to accumulate. This is particularly true in cells, such as neurons, that have slow division rates. This study uncovers features of circRNA biogenesis by investigating the link between nascent circRNA processing and transcription.

  11. Adipose Tissue Dysfunction in Nascent Metabolic Syndrome

    Directory of Open Access Journals (Sweden)

    Andrew A. Bremer

    2013-01-01

    Full Text Available The metabolic syndrome (MetS confers an increased risk for both type 2 diabetes mellitus (T2DM and cardiovascular disease (CVD. Moreover, studies on adipose tissue biology in nascent MetS uncomplicated by T2DM and/or CVD are scanty. Recently, we demonstrated that adipose tissue dysregulation and aberrant adipokine secretion contribute towards the syndrome’s low-grade chronic proinflammatory state and insulin resistance. Specifically, we have made the novel observation that subcutaneous adipose tissue (SAT in subjects with nascent MetS has increased macrophage recruitment with cardinal crown-like structures. We have also shown that subjects with nascent MetS have increased the levels of SAT-secreted adipokines (IL-1, IL-6, IL-8, leptin, RBP-4, CRP, SAA, PAI-1, MCP-1, and chemerin and plasma adipokines (IL-1, IL-6, leptin, RBP-4, CRP, SAA, and chemerin, as well as decreased levels of plasma adiponectin and both plasma and SAT omentin-1. The majority of these abnormalities persisted following correction for increased adiposity. Our data, as well as data from other investigators, thus, highlight the importance of subcutaneous adipose tissue dysfunction in subjects with MetS and its contribution to the proinflammatory state and insulin resistance. This adipokine profile may contribute to increased insulin resistance and low-grade inflammation, promoting the increased risk of T2DM and CVD.

  12. Disruption of transporters affiliated with enantio-pyochelin biosynthesis gene cluster of Pseudomonas protegens Pf-5 has pleiotropic effects

    Science.gov (United States)

    Pseudomonas protegens Pf-5 (formerly Pseudomonas fluorescens) is a biocontrol bacterium that produces the siderophore enantio-pyochelin under conditions of iron starvation in a process that is often accompanied by the secretion of its biosynthesis intermediates, salicylic acid and dihydroaeruginoic ...

  13. Study on the spoilage potential ofPseudomonas fluorescenson salmon stored at different temperatures.

    Science.gov (United States)

    Xie, Jing; Zhang, Zhen; Yang, Sheng-Ping; Cheng, Ying; Qian, Yun-Fang

    2018-01-01

    The bacterial kinetics and quality indexes [sensory quality, total volatile basic nitrogen (TVB-N), thiobarbituric acid value, biogenic amine, and amino acids] were analyzed on salmon inoculated with Pseudomonas fluorescens during storage under different temperatures (30, 10, and 4 °C). The bacterial kinetics revealed that P. fluorescens showed a steady growth at low temperatures (10 and 4 °C). The TVB-N yield factors of the sample stored at 4 °C indicated that each bacterial cell of P. fluorescens displayed greater spoilage activity at low temperatures. A remarkable correlation was found between the production of biogenic amines and bacterial counts. The results also highlighted that P. fluorescens cultured at 4 °C had higher demand for amino acids.

  14. Antibiotic and biosurfactant properties of cyclic lipopeptides produced by fluorescent Pseudomonas spp. from the sugar beet rhizosphere

    DEFF Research Database (Denmark)

    Nielsen, T H; Sørensen, D; Tobiasen, C

    2002-01-01

    Cyclic lipopeptides (CLPs) with antibiotic and biosurfactant properties are produced by a number of soil bacteria, including fluorescent Pseudomonas spp. To provide new and efficient strains for the biological control of root-pathogenic fungi in agricultural crops, we isolated approximately 600...... in the peptide moiety. Production of specific CLPs could be affiliated with Pseudomonas fluorescens strain groups belonging to biotype I, V, or VI. In vitro analysis using both purified CLPs and whole-cell P. fluorescens preparations demonstrated that all CLPs exhibited strong biosurfactant properties...

  15. Chemical sanitizers to control biofilms formed by two Pseudomonas species on stainless steel surface

    Directory of Open Access Journals (Sweden)

    Danila Soares Caixeta

    2012-03-01

    Full Text Available The biofilm formation of Pseudomonas aeruginosa and Pseudomonas fluorescens on AISI 304 stainless steel in the presence of reconstituted skim milk under different temperatures was conducted, and the potential of three chemical sanitizers in removing the mono-species biofilms formed was compared. Pseudomonas aeruginosa cultivated in skim milk at 28 °C presented better growth rate (10.4 log CFU.mL-1 when compared with 3.7 and 4.2 log CFU.mL-1 for P. aeruginosa and P. fluorescens cultivated at 7 °C, respectively. Pseudomonas aeruginosa formed biofilm when cultivated at 28 °C. However, only the adhesion of P. aeruginosa and P. fluorescens was observed when incubated at 7 °C. The sodium dichloroisocyanurate was the most efficient sanitizer in the reduction of the adhered P. aeruginosa cells at 7 and 28 °C and those on the biofilm, respectively. The hydrogen peroxide was more effective in the reduction of adhered cells of P. fluorescens at 7 °C.

  16. KAJIAN PEMANFAATAN LIMBAH ORGANIK CAIR UNTUK PEMBIAKAN MASAL AGENS ANTAGONIS PSEUDOMONAS FLOURESCENS SERTA UJI POTENSINYA SEBAGAI BIO-PESTISIDA

    Directory of Open Access Journals (Sweden)

    Giyanto

    2009-08-01

    Full Text Available Pseudomonas fluorescens has been well known as biological control agent for plant diseases control. The problem to apply the agents widely in the field or in the level of farmer is limited technology of mass production with low cost, due to the simple technology of propagation has not been yet available. The objective of this research is to study the potency of liquid organic wastes as media for mass production of P. fluorescens and to formulate them as bio-pesticide. The results showed that modification of coconut water to pH of 7.0 could be used as media for growing P. flourescens. The P. fluorescens also could grow well in livestock liquid waste by adding 10% meat extract. On the other hand, the liquid tofu waste and liquid compost waste became good media for growing of P. fluorescens by addition of 10% meat extract and 1.25% sugar. Tetes tebu will be very good media for P. fluorescens at 5% final concentration and by adding of 10% meat extract and 2.5% of sugar. The P. fluorescens showed high antagonistic effect to Ralstonia solanacearum and Sclerotium rolfsii in all of modified liquid organic wastes media. Survival and antagonisctic activity of P. fluorescens in modified organic liquid wastes stored at 5oC or room temperature were 12 weeks. In vivo antagonistic and plant growth promoting activity showed that P. fluorescens grown in liquid organic waste suppressed the incidence of stem rot diseases caused by Sclerotium rolfsii and increased the vigor of plant growth on watermelon. Formulation of the P. fluorescens grown in modified coconut water gave the best performance of P. fluorescens in supppressing of plant diseases and inducing plant growth. The product of BeMOR(e (beneficial microorganism from the result of this research will be proposed to be patented

  17. The Fluid Dynamics of Nascent Biofilms

    Science.gov (United States)

    Farthing, Nicola; Snow, Ben; Wilson, Laurence; Bees, Martin

    2017-11-01

    Many anti-biofilm approaches target mature biofilms with biochemical or physio-chemical interventions. We investigate the mechanics of interventions at an early stage that aim to inhibit biofilm maturation, focusing on hydrodynamics as cells transition from planktonic to surface-attached. Surface-attached cells generate flow fields that are relatively long-range compared with cells that are freely-swimming. We look at the effect of these flows on the biofilm formation. In particular, we use digital inline holographic microscopy to determine the three-dimensional flow due to a surface-attached cell and the effect this flow has on both tracers and other cells in the fluid. We compare experimental data with two models of cells on boundaries. The first approach utilizes slender body theory and captures many of the features of the experimental field. The second model develops a simple description in terms of singularity solutions of Stokes' flow, which produces qualitatively similar dynamics to both the experiments and more complex model but with significant computational savings. The range of validity of multiple cell arrangements is investigated. These two descriptions can be used to investigate the efficacy of actives developed by Unilever on nascent biofilms.

  18. Differentiation of isolated native of desulphurizators Pseudomonas by means of the study of the profile of fatty acids

    International Nuclear Information System (INIS)

    Silva Gomez, Edelberto; Morales P, Alicia Lucia; Callejas Castro Solange; Florez Sandoval Maria Cecilia; Rodriguez Wilson

    2000-01-01

    The content of cellular fatty acids was determined by HRGC of twelve Colombian isolated Pseudomonas aeruginosa 17,18, 19, 20, 21, 22 and 103 pseudomonas sp 23,24,25,26 and 27 with desulphurization capacity, pseudomonas aeruginosa ATCC 9027 and 10145, pseudomonas sp ATCC 39327 and pseudomonas fluorescens. Fifty-three different types of fatty acids were found, among saturated and unsaturated of lineal chain, and mainly hydroxy acids and ramified. Of these, 17 have not been described in the literature for this genus. A group of 6 acids was presented with more frequency (15:0; 16:0; 16:1; 17:1; 3-OH 16:0 and 2-OH 15:0) in more than 75% of the pseudomonas studied. The studied microorganisms are related because they share the presence of some characteristic fatty acids, that which allows assuring that they belong to same taxonomic unit. The analysis cluster developed by means of plotting in dendrograms of the qualitative and quantitative contents of the acids fatty totals showed the formation of two attaches, the I conformed by ATCC 39327 and the isolated 17 and 25, and the II by Ps. Fluorescens and the isolated one 27. The dendrogram of the hydroxy acids shows the formation of four attaches, the I attache, (isolated 17 and 20), the II A (isolated 22 and 103), the II B (isolated 27 and ps. fluorescens) and the attache III (isolated 18 and 19). In that of the ramified fatty acids the formation of a main attache is observed conformed by four sub-groups, the IA (isolated 17), the I B (isolated 18 and 24), the I C (isolated 19 and 25 and Ps. fluorescens) and I D (isolated 27). These results show that the isolated 27,25,24, 19, 18 and 17, in their order, have narrow relationship to Ps. fluorescens

  19. Trigger Factor-Induced Nascent Chain Dynamics Changes Suggest Two Different Chaperone-Nascent Chain Interactions during Translation.

    Science.gov (United States)

    Koubek, Jiří; Chang, Yi-Che; Yang, Sunny Yao-Chen; Huang, Joseph Jen-Tse

    2017-06-02

    Protein biogenesis is poorly understood due to the ribosome that perturbs measurement attempted on the ribosome-bound nascent chain (RNC). Investigating nascent chain dynamics may provide invaluable insight into the co-translational processes such as structure formation or interaction with a chaperone [e.g., the bacterial trigger factor (TF)]. In this study, we aim to establish a platform for studying nascent chain dynamics by exploring the local environment near the fluorescent dye on site-specifically labeled RNCs with time-resolved fluorescence anisotropy. To prepare a quantitative model of fluorescence depolarization, we utilized intrinsically disordered protein bound to ribosome, which helped us couple the sub-nanosecond depolarization with the motion of the nascent chain backbone. This was consistent with zinc-finger-domain-containing RNCs, where the extent of sub-nanosecond motion decreased upon the addition of zinc when the fluorophore was in close proximity of the domain. After the characterization of disordered nascent chain dynamics, we investigated the synthesis of a model cytosolic protein, Entner-Doudoroff aldolase, labeled at different sites during various stages of translation. Depending on the stage of translation, the addition of the TF to the nascent chain led to two different responses in the nascent chain dynamics serendipitously, suggesting steric hindrance between the nascent chain and the chaperone as a mechanism for TF dissociation from the ribosome during translation. Overall, our study demonstrates the possible use of site-specific labeling and time-resolved anisotropy to gain insight on chaperone binding event at various stages of translation and hints on TF co-translational mechanism. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Diversity of Pseudomonas Genomes, Including Populus-Associated Isolates, as Revealed by Comparative Genome Analysis.

    Science.gov (United States)

    Jun, Se-Ran; Wassenaar, Trudy M; Nookaew, Intawat; Hauser, Loren; Wanchai, Visanu; Land, Miriam; Timm, Collin M; Lu, Tse-Yuan S; Schadt, Christopher W; Doktycz, Mitchel J; Pelletier, Dale A; Ussery, David W

    2016-01-01

    The Pseudomonas genus contains a metabolically versatile group of organisms that are known to occupy numerous ecological niches, including the rhizosphere and endosphere of many plants. Their diversity influences the phylogenetic diversity and heterogeneity of these communities. On the basis of average amino acid identity, comparative genome analysis of >1,000 Pseudomonas genomes, including 21 Pseudomonas strains isolated from the roots of native Populus deltoides (eastern cottonwood) trees resulted in consistent and robust genomic clusters with phylogenetic homogeneity. All Pseudomonas aeruginosa genomes clustered together, and these were clearly distinct from other Pseudomonas species groups on the basis of pangenome and core genome analyses. In contrast, the genomes of Pseudomonas fluorescens were organized into 20 distinct genomic clusters, representing enormous diversity and heterogeneity. Most of our 21 Populus-associated isolates formed three distinct subgroups within the major P. fluorescens group, supported by pathway profile analysis, while two isolates were more closely related to Pseudomonas chlororaphis and Pseudomonas putida. Genes specific to Populus-associated subgroups were identified. Genes specific to subgroup 1 include several sensory systems that act in two-component signal transduction, a TonB-dependent receptor, and a phosphorelay sensor. Genes specific to subgroup 2 contain hypothetical genes, and genes specific to subgroup 3 were annotated with hydrolase activity. This study justifies the need to sequence multiple isolates, especially from P. fluorescens, which displays the most genetic variation, in order to study functional capabilities from a pangenomic perspective. This information will prove useful when choosing Pseudomonas strains for use to promote growth and increase disease resistance in plants. Copyright © 2015 Jun et al.

  1. The characterization and metabolism of rat hepatic nascent HLD subfractions

    International Nuclear Information System (INIS)

    Winkler, K.E.

    1988-01-01

    Nascent HDL was isolated from recirculating rat liver perfusates and separated by heparin-sepharose chromatography into a non-retained fraction (A) and a fraction (B) that eluted with 0.5 M NaCl. Fractions A and B contained 70% and 30% of the nascent HDL protein, respectively. Livers perfused by the single-pass technique produced fractions A and B in the same ratio as livers perfused by recirculation. The apolipoprotein compositions were similar to those in the recirculating perfusion; however, both fractions A and B had more triglyceride (greater than 50% of total lipid). In a preliminary study designed to investigate whether nascent HDL-apo E was secreted by Kupffer cells or hepatocytes, label was targeted to Kupffer cells by perfusing rat livers with 3 H-acetylated LDL or 3 H-amino acids incorporated into large multilamellar vesicles. For metabolic studies, nascent HDL and nascent VLDL were isolated from rat livers that had been perfused with 3 H-glycerol to label the triglyceride

  2. Chemical sanitizers to control biofilms formed by two Pseudomonas species on stainless steel surface Sanificantes químicos no controle de biofilmes formados por duas espécies de Pseudomonas em superfície de aço inoxidável

    Directory of Open Access Journals (Sweden)

    Danila Soares Caixeta

    2012-03-01

    Full Text Available The biofilm formation of Pseudomonas aeruginosa and Pseudomonas fluorescens on AISI 304 stainless steel in the presence of reconstituted skim milk under different temperatures was conducted, and the potential of three chemical sanitizers in removing the mono-species biofilms formed was compared. Pseudomonas aeruginosa cultivated in skim milk at 28 °C presented better growth rate (10.4 log CFU.mL-1 when compared with 3.7 and 4.2 log CFU.mL-1 for P. aeruginosa and P. fluorescens cultivated at 7 °C, respectively. Pseudomonas aeruginosa formed biofilm when cultivated at 28 °C. However, only the adhesion of P. aeruginosa and P. fluorescens was observed when incubated at 7 °C. The sodium dichloroisocyanurate was the most efficient sanitizer in the reduction of the adhered P. aeruginosa cells at 7 and 28 °C and those on the biofilm, respectively. The hydrogen peroxide was more effective in the reduction of adhered cells of P. fluorescens at 7 °C.A capacidade de adesão e formação de biofilme por Pseudomonas aeruginosa e Pseudomonas fluorescens em aço inoxidável AISI 304, na presença de leite desnatado resconstituído sobre diferentes temperaturas foi conduzido e o potencial de três sanificantes químicos na remoção de biofilmes monoespécies foi comparado. Pseudomonas aeruginosa cultivada em leite desnatado a 28 °C apresentou melhor crescimento (10,4 log UFC.mL-1 quando comparado com 3,7 and 4,2 log UFC.mL-1 para P. aeruginosa e P. fluorescens cultivadas a 7 °C, respectivamente. Pseudomonas aeruginosa formou biofilme quando cultivada a 28 °C. Contudo foi observado somente adesão de P. aeruginosa e P. fluorescens quando incubada a 7 °C. O dicloroisocianurato de sódio foi o sanificante mais eficiente na redução de células aderidas e em biofilme de P. aeruginosa a 7 e 28 °C, respectivamente. O peróxido de hidrogênio foi o mais eficiente na redução de células aderidas de P. fluorescens a 7 °C.

  3. Entry and Exit Dynamics of Nascent Business Owners

    DEFF Research Database (Denmark)

    Rocha, Vera; Carneiro, Anabela; Varum, Celeste

    2015-01-01

    This paper reports a comprehensive study on the dynamics of nascent business owners using a unique longitudinal matched employer–employee dataset. We follow over 157,000 individuals who leave paid employment and become business owners during the period 1992–2007. The contributions of this paper...... are twofold. First, we analyze both entry and exit, identifying and characterizing different profiles of individuals leaving paid employment to become business owners, and distinguishing exits by dissolution from exits by ownership transfer. Second, we provide new evidence on how particular experiences...... in the labor market and entry modes shape the post-entry dynamics of nascent business owners. By differentiating between different entry and exit routes, this paper provides new evidence on different human capital patterns among nascent business owners and on key determinants of entrepreneurial survival. Our...

  4. Pseudomonas - Fact Sheet

    OpenAIRE

    Public Health Agency

    2012-01-01

    Fact sheet on Pseudomonas, including:What is Pseudomonas?What infections does it cause?Who is susceptible to pseudomonas infection?How will I know if I have pseudomonas infection?How can Pseudomonas be prevented from spreading?How can I protect myself from Pseudomonas?How is Pseudomonas infection treated?

  5. Nigeria's Nascent Democracy and 'WAR' Against Corruption: A Rear ...

    African Journals Online (AJOL)

    One of the problems facing the nascent democracy in Nigeria which is more pressing than economic development is the high rate of brazen corruption in virtually all facets of the polity's national life. Thus, the thrust of this paper is a review of the recent 'WAR' against corruption in Nigeria. The paper surveys a number of ...

  6. From nascent to actual entrepreneurship: the effect of entry barriers

    NARCIS (Netherlands)

    A.J. van Stel (André); D. Storey (David); A.R.M. Wennekers (Sander); A.R. Thurik (Roy)

    2005-01-01

    textabstractThis exploratory study focuses on the conversion from nascent to actual entrepreneurship and the role of entry barriers in this process. Using data for a sample of countries participating in the Global Entrepreneurship Monitor between 2002 and 2004, we estimate a twoequation model

  7. Accelerators as Authentic Training Experiences for Nascent Entrepreneurs

    Science.gov (United States)

    Miles, Morgan P.; de Vries, Huibert; Harrison, Geoff; Bliemel, Martin; de Klerk, Saskia; Kasouf, Chick J.

    2017-01-01

    Purpose: The purpose of this paper is to address the role of accelerators as authentic learning-based entrepreneurial training programs. Accelerators facilitate the development and assessment of entrepreneurial competencies in nascent entrepreneurs through the process of creating a start-up venture. Design/methodology/approach: Survey data from…

  8. Chromosomal insertion of the entire Escherichia coli lactose operon, into two strains of Pseudomonas, using a modified mini-Tn5 delivery system

    DEFF Research Database (Denmark)

    Hansen, L. H.; Sørensen, S. J.; Jensen, Lars Bogø

    1997-01-01

    A 12-kb PstI fragment including the entire E. coli lactose operon (lacIPOZYA) was inserted in one copy into the chromosome of Pseudomonas putida, Pseudomonas fluorescens and an E. coli strain with lac(-) phenotype. This was made possible by improvements of an already existing mini-Tn5 transposon...... flanked by NotI sites needed in the mini-Tn5 delivery system; (b) the generation of E. coli nonlysogenic strains expressing the pi protein thus being capable of maintaining and delivering R6K-based mini-Tn5 vectors to other E. coli strains; (c) the successful insertion of the E. coli lactose operon...... into the P. fluorescens chromosome giving P. fluorescens the ability to grow on lactose; (d) evidence from Southern blotting that contradicts the assumption that the mini-Tn5 delivery system always creates one-copy inserts. These improvements allow insertion of large DNA fragments encoding highly expressed...

  9. Variation of the Pseudomonas community structure on oak leaf lettuce during storage detected by culture-dependent and -independent methods.

    Science.gov (United States)

    Nübling, Simone; Schmidt, Herbert; Weiss, Agnes

    2016-01-04

    The genus Pseudomonas plays an important role in the lettuce leaf microbiota and certain species can induce spoilage. The aim of this study was to investigate the occurrence and diversity of Pseudomonas spp. on oak leaf lettuce and to follow their community shift during a six day cold storage with culture-dependent and culture-independent methods. In total, 21 analysed partial Pseudomonas 16S rRNA gene sequences matched closely (> 98.3%) to the different reference strain sequences, which were distributed among 13 different phylogenetic groups or subgroups within the genus Pseudomonas. It could be shown that all detected Pseudomonas species belonged to the P. fluorescens lineage. In the culture-dependent analysis, 73% of the isolates at day 0 and 79% of the isolates at day 6 belonged to the P. fluorescens subgroup. The second most frequent group, with 12% of the isolates, was the P. koreensis subgroup. This subgroup was only detected at day 0. In the culture-independent analysis the P. fluorescens subgroup and P. extremaustralis could not be differentiated by RFLP. Both groups were most abundant and amounted to approximately 46% at day 0 and 79% at day 6. The phytopathogenic species P. salmonii, P. viridiflava and P. marginalis increased during storage. Both approaches identified the P. fluorescens group as the main phylogenetic group. The results of the present study suggest that pseudomonads found by plating methods indeed represent the most abundant part of the Pseudomonas community on oak leaf lettuce. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Characterisation of the nascent polypeptide-associated complex in fission yeast

    DEFF Research Database (Denmark)

    Andersen, Katrine M; Semple, Colin A; Hartmann-Petersen, Rasmus

    2007-01-01

    The nascent polypeptide-associated complex (NAC) is an abundant and phylogenetically conserved protein complex. It is composed of two subunits and interacts with nascent polypeptide chains emerging from the ribosome. It has been proposed to protect the nascent chains from premature interaction...

  11. Transcription arrest caused by long nascent RNA chains

    DEFF Research Database (Denmark)

    Bentin, Thomas; Cherny, Dmitry; Larsen, H Jakob

    2004-01-01

    The transcription process is highly processive. However, specific sequence elements encoded in the nascent RNA may signal transcription pausing and/or termination. We find that under certain conditions nascent RNA chains can have a strong and apparently sequence-independent inhibitory effect...... on transcription. Using phage T3 RNA polymerase (T3 RNAP) and covalently closed circular (cccDNA) DNA templates that did not contain any strong termination signal, transcription was severely inhibited after a short period of time. Less than approximately 10% residual transcriptional activity remained after 10 min...... of incubation. The addition of RNase A almost fully restored transcription in a dose dependent manner. Throughout RNase A rescue, an elongation rate of approximately 170 nt/s was maintained and this velocity was independent of RNA transcript length, at least up to 6 kb. Instead, RNase A rescue increased...

  12. Identifying entrepreneurial opportunities by nascent entrepreneurs in Sfax Region

    OpenAIRE

    Borchani, Mariem; Aloulou, Wassim; Beujelbene, Younès; Liñán, Francisco (Coordinador); Guzmán Cuevas, Joaquín J. (Coordinador)

    2011-01-01

    The purpose of this paper is to identify variables influencing the identification of entrepreneurial opportunity by nascent entrepreneurs in Sfax region. These variables included the personality of entrepreneur, social network and prior knowledge. Theoretically, we have a conceptual framework privileged in recent entrepreneurship research (Shane & Venkataraman, 2000). Empirically, our research is based on an exploratory study, while adopting a deductive approach type. We used the questionnair...

  13. Nascent RNA sequencing reveals distinct features in plant transcription

    OpenAIRE

    Hetzel, Jonathan; Duttke, Sascha H.; Benner, Christopher; Chory, Joanne

    2016-01-01

    Transcription is a fundamental and dynamic step in the regulation of gene expression, but the characteristics of plant transcription are poorly understood. We adapted the global nuclear run-on sequencing (GRO-seq) and 5′GRO-seq methods for plants and provide a plant version of the next-generation sequencing software HOMER (homer.ucsd.edu/homer/plants) to facilitate data analysis. Mapping nascent transcripts in Arabidopsis thaliana seedlings enabled identification of known and novel transcript...

  14. Kinetics of Enantiomerically Enriched Synthesis of Solketal Esters Using Native and SBA-15 supported P. Fluorescens Lipase

    Directory of Open Access Journals (Sweden)

    Zniszczoł Aurelia

    2017-06-01

    Full Text Available The studies showed that alkaline lipase from Pseudomonas fluorescens enables an irreversible transesterification of vinyl esters to give enantiomeric excess (eeR of about 80% using vinyl butyrate as acyl donor and diisopropyl ether as a solvent, at partially optimized conditions. For the native lipase the process was adequately described by a five-parameter Ping-Pong Bi Bi model for both enantiomers plus expression accounting for the formation of enzyme-acyl donor complex, but for the same lipase supported on mesoporous materials of SBA-15-Oc type, R-product inhibition also had to be taken into account. The use of hydrophobic support increased by more than two-fold the rate of the S-solketal conversion but even more that of R-solketal. Thus the immobilization of lipase had very positive effect on the process kinetics but decreased its enantioselectivity.

  15. Nascent RNA sequencing reveals distinct features in plant transcription.

    Science.gov (United States)

    Hetzel, Jonathan; Duttke, Sascha H; Benner, Christopher; Chory, Joanne

    2016-10-25

    Transcriptional regulation of gene expression is a major mechanism used by plants to confer phenotypic plasticity, and yet compared with other eukaryotes or bacteria, little is known about the design principles. We generated an extensive catalog of nascent and steady-state transcripts in Arabidopsis thaliana seedlings using global nuclear run-on sequencing (GRO-seq), 5'GRO-seq, and RNA-seq and reanalyzed published maize data to capture characteristics of plant transcription. De novo annotation of nascent transcripts accurately mapped start sites and unstable transcripts. Examining the promoters of coding and noncoding transcripts identified comparable chromatin signatures, a conserved "TGT" core promoter motif and unreported transcription factor-binding sites. Mapping of engaged RNA polymerases showed a lack of enhancer RNAs, promoter-proximal pausing, and divergent transcription in Arabidopsis seedlings and maize, which are commonly present in yeast and humans. In contrast, Arabidopsis and maize genes accumulate RNA polymerases in proximity of the polyadenylation site, a trend that coincided with longer genes and CpG hypomethylation. Lack of promoter-proximal pausing and a higher correlation of nascent and steady-state transcripts indicate Arabidopsis may regulate transcription predominantly at the level of initiation. Our findings provide insight into plant transcription and eukaryotic gene expression as a whole.

  16. Screening for Pseudomonas and Bacillus antagonistic rhizobacteria strains for the biocontrol of Fusarium wilt of chickpea

    Directory of Open Access Journals (Sweden)

    Hannane Abed

    2016-07-01

    Full Text Available The aim of this work is to study the ability of several isolates belonging to Rhizobacteria (Pseudomonas and Bacillus collected from several chickpea growing areas in Algeria, to control the mycelium growth of Fusarium oxysporum f. sp. ciceris. Interesting isolates were characterized for their morphological characteristics, physiological and biochemical activities as potential bio-control agent. Fungal inhibition tests were performed using plate assay and each isolate were tested for the production of protease, cyanide hydrogen, indole acetic acid, antifungal volatile and extracellular compound. According to API 50 CH, we are able to identify six Bacillus species (B. subtilis, B. circulans, B. lentus, B. aneurinilyticus, B. firmus, B. licheniformis; and with API 20NE test we have identified three Pseudomonas species (P. aeruginosa, P. luteola, P. fluorescens. The ability of bacterial isolates was varied in production of Protease, Gelatinase, Amylase, Cellulase, Acid Indole acetic, Lipase, Catalase and Cyanid Hydrogen. This is traduced in different rate of inhibition growth due to various extracellular compounds, where B61 (Bacillus aneurinilyticus and P39 (Pseudomonas luteola and P70 (Pseudomonas fluorescens were the most efficient with 77 and 55.5% respectively, while B39 (Bacillus firmus and P41 (Pseudomonas luteola were the most efficient by volatile compounds with 70.5 and 77.5% respectively. Our results indicate that these bacteria isolates can be used in the biocontrol of Fusarium oxysporum f. sp. ciceris.

  17. Efficacy of lactoferricin B in controlling ready-to-eat vegetable spoilage caused by Pseudomonas spp.

    Science.gov (United States)

    Federico, Baruzzi; Pinto, Loris; Quintieri, Laura; Carito, Antonia; Calabrese, Nicola; Caputo, Leonardo

    2015-12-23

    The microbial content of plant tissues has been reported to cause the spoilage of ca. 30% of chlorine-disinfected fresh vegetables during cold storage. The aim of this work was to evaluate the efficacy of antimicrobial peptides in controlling microbial vegetable spoilage under cold storage conditions. A total of 48 bacterial isolates were collected from ready-to-eat (RTE) vegetables and identified as belonging to Acinetobacter calcoaceticus, Aeromonas media, Pseudomonas cichorii, Pseudomonas fluorescens, Pseudomonas jessenii, Pseudomonas koreensis, Pseudomonas putida, Pseudomonas simiae and Pseudomonas viridiflava species. Reddish or brownish pigmentation was found when Pseudomonas strains were inoculated in wounds on leaves of Iceberg and Trocadero lettuce and escarole chicory throughout cold storage. Bovine lactoferrin (BLF) and its hydrolysates (LFHs) produced by pepsin, papain and rennin, were assayed in vitro against four Pseudomonas spp. strains selected for their heavy spoiling ability. As the pepsin-LFH showed the strongest antimicrobial effect, subsequent experiments were carried out using the peptide lactoferricin B (LfcinB), well known to be responsible for its antimicrobial activity. LfcinB significantly reduced (P ≤ 0.05) spoilage by a mean of 36% caused by three out of four inoculated spoiler pseudomonads on RTE lettuce leaves after six days of cold storage. The reduction in the extent of spoilage was unrelated to viable cell density in the inoculated wounds. This is the first paper providing direct evidence regarding the application of an antimicrobial peptide to control microbial spoilage affecting RTE leafy vegetables during cold storage.

  18. Iron-regulated salicylate synthesis by Pseudomonas spp.

    Science.gov (United States)

    Visca, P; Ciervo, A; Sanfilippo, V; Orsi, N

    1993-09-01

    Two iron-regulated compounds have been found in acidified ethyl acetate extracts from culture supernatants of Pseudomonas aeruginosa and Pseudomonas cepacia type-strains. Synthesis of both compounds paralleled iron-deficient growth, and was repressed in the presence of 100 microM-FeCl3. Yields of these substances varied among different strains and attained maximum levels during stationary phase. Thin layer chromatographic analysis in five different solvent systems revealed that the slower-moving compound chromatographed as two distinct bands, and showed RF values and spectral properties similar to pyochelin. The faster-moving compound co-migrated as a single band with a standard of commercial salicylic acid in each of the chromatographic systems tested. Moreover, a molecule with an identical RF was also produced by Pseudomonas fluorescens CHA401, which is known to synthesize salicylic acid as the only siderophore during iron-limited growth. Spectrophotometric and spectrofluorometric titrations led to the identification of this iron-regulated compound as salicylic acid, in agreement with the structure deduced from 1H-NMR and mass spectroscopy. The identity of the P. cepacia siderophore azurechelin as salicylic acid was also conclusively demonstrated. Salicylic acid, like pyochelin and pyoverdin, promoted P. aeruginosa growth in an iron-depleted medium. These results are consistent with a putative siderophore activity for salicylic acid, i.e. azurechelin, as has been demonstrated for P. aeruginosa, P. fluorescens and P. cepacia. Thus, salicylic acid is likely to act as a siderophore in more than one species belonging to the genus Pseudomonas.

  19. The Pseudomonas community in metal-contaminated sediments as revealed by quantitative PCR: a link with metal bioavailability.

    Science.gov (United States)

    Roosa, Stéphanie; Wauven, Corinne Vander; Billon, Gabriel; Matthijs, Sandra; Wattiez, Ruddy; Gillan, David C

    2014-10-01

    Pseudomonas bacteria are ubiquitous Gram-negative and aerobic microorganisms that are known to harbor metal resistance mechanisms such as efflux pumps and intracellular redox enzymes. Specific Pseudomonas bacteria have been quantified in some metal-contaminated environments, but the entire Pseudomonas population has been poorly investigated under these conditions, and the link with metal bioavailability was not previously examined. In the present study, quantitative PCR and cell cultivation were used to monitor and characterize the Pseudomonas population at 4 different sediment sites contaminated with various levels of metals. At the same time, total metals and metal bioavailability (as estimated using an HCl 1 m extraction) were measured. It was found that the total level of Pseudomonas, as determined by qPCR using two different genes (oprI and the 16S rRNA gene), was positively and significantly correlated with total and HCl-extractable Cu, Co, Ni, Pb and Zn, with high correlation coefficients (>0.8). Metal-contaminated sediments featured isolates of the Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas lutea and Pseudomonas aeruginosa groups, with other bacterial genera such as Mycobacterium, Klebsiella and Methylobacterium. It is concluded that Pseudomonas bacteria do proliferate in metal-contaminated sediments, but are still part of a complex community. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  20. Antimicrobial resistance of Pseudomonas spp. isolated from wastewater and wastewater-impacted marine coastal zone.

    Science.gov (United States)

    Luczkiewicz, Aneta; Kotlarska, Ewa; Artichowicz, Wojciech; Tarasewicz, Katarzyna; Fudala-Ksiazek, Sylwia

    2015-12-01

    In this study, species distribution and antimicrobial susceptibility of cultivated Pseudomonas spp. were studied in influent (INF), effluent (EFF), and marine outfall (MOut) of wastewater treatment plant (WWTP). The susceptibility was tested against 8 antimicrobial classes, active against Pseudomonas spp.: aminoglycosides, carbapenems, broad-spectrum cephalosporins from the 3rd and 4th generation, extended-spectrum penicillins, as well as their combination with the β-lactamase inhibitors, monobactams, fluoroquinolones, and polymyxins. Among identified species, resistance to all antimicrobials but colistin was shown by Pseudomonas putida, the predominant species in all sampling points. In other species, resistance was observed mainly against ceftazidime, ticarcillin, ticarcillin-clavulanate, and aztreonam, although some isolates of Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas pseudoalcaligenes, and Pseudomonas protegens showed multidrug-resistance (MDR) phenotype. Among P. putida, resistance to β-lactams and to fluoroquinolones as well as multidrug resistance become more prevalent after wastewater treatment, but the resistance rate decreased in marine water samples. Obtained data, however, suggests that Pseudomonas spp. are equipped or are able to acquire a wide range of antibiotic resistance mechanisms, and thus should be monitored as possible source of resistance genes.

  1. Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis

    Czech Academy of Sciences Publication Activity Database

    Grosskinsky, D. K.; Tafner, R.; Moreno, M. V.; Stenglein, S. A.; Garcia de Salamone, I. E.; Nelson, L. M.; Novák, Ondřej; Strnad, Miroslav; van der Graaff, E.; Roitsch, Thomas

    2016-01-01

    Roč. 6, MAR 17 (2016), s. 23310 ISSN 2045-2322 R&D Projects: GA MŠk(CZ) LO1204; GA ČR GA15-22322S; GA MŠk(CZ) LO1415 Institutional support: RVO:61389030 ; RVO:67179843 Keywords : GROWTH-PROMOTING RHIZOBACTERIA * PLANT-GROWTH * SALICYLIC-ACID Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.259, year: 2016

  2. Mechanisms of nascent fiber formation during avian skeletal muscle hypertrophy

    Science.gov (United States)

    McCormick, K. M.; Schultz, E.

    1992-01-01

    This study examined two putative mechanisms of new fiber formation in postnatal skeletal muscle, namely longitudinal fragmentation of existing fibers and de novo formation. The relative contributions of these two mechanisms to fiber formation in hypertrophying anterior latissimus dorsi (ALD) muscle were assessed by quantitative analysis of their nuclear populations. Muscle hypertrophy was induced by wing-weighting for 1 week. All nuclei formed during the weighting period were labeled by continuous infusion of 5-bromo-2'-deoxyuridine (BrdU), a thymidine analog, and embryonic-like fibers were identified using an antibody to ventricular-like embryonic (V-EMB) myosin. The number of BrdU-labeled and unlabeled nuclei in V-EMB-positive fibers were counted. Wing-weighting resulted in significant muscle enlargement and the appearance of many V-EMB+ fibers. The majority of V-EMB+ fibers were completely independent of mature fibers and had a nuclear density characteristics of developing fibers. Furthermore, nearly 100% of the nuclei in independent V-EMB+ fibers were labeled. These findings strongly suggest that most V-EMB+ fibers were nascent fibers formed de novo during the weighting period by satellite cell activation and fusion. Nascent fibers were found primarily in the space between fascicles where they formed a complex anastomosing network of fibers running at angles to one another. Although wing-weighting induced an increase in the number of branched fibers, there was no evidence that V-EMB+ fibers were formed by longitudinal fragmentation. The location of newly formed fibers in wing-weighted and regenerating ALD muscle was compared to determine whether satellite cells in the ALD muscle were unusual in that, if stimulated to divide, they would form fibers in the inter- and intrafascicular space. In contrast to wing-weighted muscle, nascent fibers were always found closely associated with necrotic fibers. These results suggest that wing-weighting is not simply another

  3. Anti-pseudomona and Anti-bacilli Activity of Some Medicinal Plants of Iran

    Directory of Open Access Journals (Sweden)

    Gholam Hosein Shahidi Bonjar

    2003-10-01

    Full Text Available The use of plants in treatment of burns, dermatophytes, and infectious diseases is common in traditional medicine of Iran. Based on ethno pharmacological and taxonomic information, antibacterial activities of methanol extracts of some medicinal plants of Iran were determined by In Vitro bioassays using agar diffusion-method against standard strains of Pseudomonas aeruginosa, P. fluorescens, Bacillus subtilis, B. cereus and B. pumilis at 20 mg/ml. From 180 plant species of 72 families, 78 species (43.3% in 42 families (58.3% showed antibacterial activities against B. cereus (88.4%, B. subtilis (39.7%, B. pumilis (37.1%, P. fluorescens (37.1% and P. aeruginos (10.2%. The most active plant families were Apiaceae, Compositae and Labiatae with 9, 8 and 7 active plant species respectively. Minimum inhibitory concentrations (MIC of the active plants were determined using two fold serial dilutions. Most active plant against Bacilli was Myrtus communis L. with MIC of 1.87 mg/ml. For Pseudomonas species, Dianthus caryophyllus L. and Terminalia chebula (Gaertner Retz. were more active with the MIC of 0.46 mg/ml for P. fluorescens and of 1.87 mg/ml for P. aeruginosa respectively.

  4. The results of a nascent language emancipation in France

    DEFF Research Database (Denmark)

    Nolan, John Shaun

    2013-01-01

    This paper sheds light on the attitudes that eastern or Upper Breton school pupils have toward their heritage language, Gallo, in the context of recent favourable changes to its socio-political status. Gallo is Brittany's Romance language and, more precisely, its Oïl language variety. Situated...... in the application of language emancipation and in considerations regarding attitudes and ideology and their importance to language policy, this is an analysis which provides an example of how the Gallo sociolinguistic situation is a case of 'nascent' language emancipation. This is done through a study of how...... the status and the future of Gallo are perceived in the context of its inclusion in Brittany's language education policy. This comparative attitudinal study suggests that although there continues to be negative language attitudes towards Gallo, there are clear indications of a favourable generational...

  5. Nascent Connections: R-Loops and Chromatin Patterning.

    Science.gov (United States)

    Chédin, Frédéric

    2016-12-01

    RNA molecules, such as long noncoding RNAs (lncRNAs), have critical roles in regulating gene expression, chromosome architecture, and the modification states of chromatin. Recent developments suggest that RNA also influences gene expression and chromatin patterns through the interaction of nascent transcripts with their DNA template via the formation of co-transcriptional R-loop structures. R-loop formation over specific, conserved, hotspots occurs at thousands of genes in mammalian genomes and represents an important and dynamic feature of mammalian chromatin. Here, focusing primarily on mammalian systems, I describe the accumulating connections and possible mechanisms linking R-loop formation and chromatin patterning. The possible contribution of aberrant R-loops to pathological conditions is also discussed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Organizational Knowledge Communication – a Nascent 3rd Order Disciplinarity

    DEFF Research Database (Denmark)

    Kastberg, Peter

    2014-01-01

    disciplinarity. Whereas each discipline is a strand in its own right in the helix, these strands, nevertheless, also allow for disciplinary integration, albeit punctually and dynamically. And it is exactly in such trilateral punctual and dynamic integrations that Organizational Knowledge Communication becomes...... visible, becomes a disciplinarity. I theoretically present an example of such a punctual integration and point to some of the immediate research promises that it holds. This theoretical account ends by describing Organizational Knowledge Communication as a nascent 3rd order disciplinarity.......There is an emerging tendency that the organizational communication functions of larger companies enter into a symbiotic relationship with the companies’ Knowledge Management function. A tendency this journal has labelled Organizational Knowledge Communication. This should come as no surprise...

  7. Bistability in a Metabolic Network Underpins the De Novo Evolution of Colony Switching in Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Gallie, Jenna; Libby, Eric; Bertels, Frederic

    2015-01-01

    Phenotype switching is commonly observed in nature. This prevalence has allowed the elucidation of a number of underlying molecular mechanisms. However, little is known about how phenotypic switches arise and function in their early evolutionary stages. The first opportunity to provide empirical ...

  8. Structural and Mechanistic Insights into the Pseudomonas fluorescens 2-Nitrobenzoate 2-Nitroreductase NbaA.

    Science.gov (United States)

    Kim, Yong-Hak; Song, Wooseok; Kim, Jin-Sik; Jiao, Li; Lee, Kangseok; Ha, Nam-Chul

    2015-08-01

    The bacterial 2-nitroreductase NbaA is the primary enzyme initiating the degradation of 2-nitrobenzoate (2-NBA), and its activity is controlled by posttranslational modifications. To date, the structure of NbaA remains to be elucidated. In this study, the crystal structure of a Cys194Ala NbaA mutant was determined to a 1.7-Å resolution. The substrate analog 2-NBA methyl ester was used to decipher the substrate binding site by inhibition of the wild-type NbaA protein. Tandem mass spectrometry showed that 2-NBA methyl ester produced a 2-NBA ester bond at the Tyr193 residue in the wild-type NbaA but not residues in the Tyr193Phe mutant. Moreover, covalent binding of the 2-NBA methyl ester to Tyr193 reduced the reactivity of the Cys194 residue on the peptide link. The Tyr193 hydroxyl group was shown to be essential for enzyme catalysis, as a Tyr193Phe mutant resulted in fast dissociation of flavin mononucleotide (FMN) from the protein with the reduced reactivity of Cys194. FMN binding to NbaA varied with solution NaCl concentration, which was related to the catalytic activity but not to cysteine reactivity. These observations suggest that the Cys194 reactivity is negatively affected by a posttranslational modification of the adjacent Tyr193 residue, which interacts with FMN and the substrate in the NbaA catalytic site. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms

    DEFF Research Database (Denmark)

    Bonnichsen, Lise; Svenningsen, Nanna Bygvraa; Rybtke, Morten Levin

    2015-01-01

    Pseudomonads produce several lipopeptide biosurfactants that have antimicrobial properties but that also facilitate surface motility and influence biofilm formation. Detailed studies addressing the significance of lipopeptides for biofilm formation and architecture are rare. Hence, the present...

  10. Evolution of fitness trade-offs in locally adapted populations of Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Schick, Alana; Bailey, Susan; Kassen, Rees

    2015-01-01

    , consistent with a multioptima version of Fisher’s geometric model of adaptation. We also find that fitness of pairs of beneficial mutations are consistently less than additive in selection environments, producing a pattern of diminishing returns, but are more variable in alternative environments, being...

  11. Specific and nonspecific immunostimulation study of Euphorbia hirta on Pseudomonas fluorescens-infected Cyprinus carpio.

    Science.gov (United States)

    Pratheepa, V; Sukumaran, N

    2011-05-01

    Infectious diseases are one of the major factors affecting the production of fish worldwide. The pathogens (especially bacteria) affect the immune system of fish and the administration of immunostimulants can increase resistance to infectious diseases by enhancing both specific and nonspecific defense mechanisms. In the present study, we have conducted an experiment on the pathogen-infected Cyprinus carpio Linn. (Cyprinidae), using Euphorbia hirta Linn. (Euphorbiaceae) plant leaves as immunostimulants. The aqueous extract of the leaves was prepared and the immunostimulant action was recorded by giving different concentrations of plant extract supplemented diet. The results obtained from the studies show that the higher concentration of the extract (50 g/kg diet) provided significant immune response (specific and nonspecific) on the fish. The 50 g/kg leaf extract of E. hirta enhanced the phagocytic ratio on 10th and 15th day after the infection. The results of the specific and nonspecific immunostimulation studies are statistically significant. This work will guide the researchers for the discovery of significant aquaculture nutrients to improve the immunostimulant action on fish.

  12. You Cannot Live of Love Alone – The Interrelation of Legitimacy and Effectuation in Nascent Markets

    DEFF Research Database (Denmark)

    Günzel-Jensen, Franziska; Rask, Morten

    2015-01-01

    This paper explores how success in legitimacy building can create restrictions and problems for new venture’s development in highly volatile settings. Through a longitudinal single in-depth case study in the nascent e-mobility market, we uncover unwanted effects of this process. In a nascent market...

  13. Behavioral attributes : A comparison between nascent entrepreneurs, established entrepreneurs and managers

    NARCIS (Netherlands)

    Nandram, S.S.; Born, M.Ph.

    2009-01-01

    This paper describes entrepreneurs, managers and nascent entrepreneurs and their entrepreneurial attributes in the context of their life stages in a sample of 275 entrepreneurs, 111 managers and 76 nascent entrepreneurs. Findings are much more in favor of the homogeneity perspective derived from the

  14. Antimicrobial properties of Pseudomonas strains producing the antibiotic mupirocin.

    Science.gov (United States)

    Matthijs, Sandra; Vander Wauven, Corinne; Cornu, Bertrand; Ye, Lumeng; Cornelis, Pierre; Thomas, Christopher M; Ongena, Marc

    2014-10-01

    Mupirocin is a polyketide antibiotic with broad antibacterial activity. It was isolated and characterized about 40 years ago from Pseudomonas fluorescens NCIMB 10586. To study the phylogenetic distribution of mupirocin producing strains in the genus Pseudomonas a large collection of Pseudomonas strains of worldwide origin, consisting of 117 Pseudomonas type strains and 461 strains isolated from different biological origins, was screened by PCR for the mmpD gene of the mupirocin gene cluster. Five mmpD(+) strains from different geographic and biological origin were identified. They all produced mupirocin and were strongly antagonistic against Staphylococcus aureus. Phylogenetic analysis showed that mupirocin production is limited to a single species. Inactivation of mupirocin production leads to complete loss of in vitro antagonism against S. aureus, except on certain iron-reduced media where the siderophore pyoverdine is responsible for the in vitro antagonism of a mupirocin-negative mutant. In addition to mupirocin some of the strains produced lipopeptides of the massetolide group. These lipopeptides do not play a role in the observed in vitro antagonism of the mupirocin producing strains against S. aureus. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  15. Start-Up Funding Intentions Among Nascent Nonprofit Entrepreneurs: An Exploratory Investigation

    Directory of Open Access Journals (Sweden)

    Fredrik O. Andersson

    2018-04-01

    Full Text Available This paper explores the start-up funding intentions of nascent nonprofit entrepreneurs, i.e., individuals in the process of creating a new formal nonprofit organization. The main questions being examined are from which sources nascent nonprofit entrepreneurs anticipate to obtain start-up funding from, how much start-up funding nascent nonprofit entrepreneurs anticipate they will need to formally launch their new nonprofit, and if there are any differences in funding intentions among nascent nonprofit entrepreneurs with and without previous start-up experience. The results from a survey of 103 nascent nonprofit entrepreneurs in Kansas City are presented and contrasted with existing research on funding of new nonprofit organizations. The results show an apparent preference for start-up funding from philanthropic grants and private donations, along with personal contributions of the founder(s.

  16. Metabolism of triglyceride-rich nascent rat hepatic high density lipoproteins

    International Nuclear Information System (INIS)

    Winkler, K.E.; Marsh, J.B.

    1989-01-01

    Nascent high density lipoprotein (HDL) and nascent very low density lipoprotein (VLDL) were isolated from rat livers that had been perfused with [3H]glycerol to label the triglyceride. When injected into intact rats, the labeled HDL-triglyceride disappeared as rapidly as the VLDL-triglyceride, with only 10% of the injected label remaining in the plasma after 30 min. The protein moiety of nascent HDL was labeled with [35S]methionine in a similar fashion and the labeled nascent HDL was separated into nonretained (NR) and retained (R) fractions by heparin-Sepharose affinity chromatography. When injected into rats, 55% of the injected label in nascent fraction NR and 72% of that in nascent fraction R was recovered from plasma at 30 min, compared to only 10% of the triglyceride label from unfractionated nascent HDL, indicating dissociation of triglyceride and apolipoprotein clearance. The plasma decay curves for both triglyceride and protein were biexponential. By 5 min, 15% of the 35S label remaining in plasma represented apoE and apoC that had been transferred from nascent HDL fractions NR and R to the d less than 1.063 g/ml fraction of plasma. Plasma HDL was labeled in vivo with [35S]methionine, separated into fractions NR and R, and the clearance of the two plasma HDL fractions was compared with that of the corresponding nascent HDL fractions. Except for a faster rate of removal of the nascent HDL fractions during the first 5 min, the serum decay curves were very similar

  17. Changes in the lung microbiome following lung transplantation include the emergence of two distinct Pseudomonas species with distinct clinical associations.

    Directory of Open Access Journals (Sweden)

    Robert P Dickson

    Full Text Available Multiple independent culture-based studies have identified the presence of Pseudomonas aeruginosa in respiratory samples as a positive risk factor for bronchiolitis obliterans syndrome (BOS. Yet, culture-independent microbiological techniques have identified a negative association between Pseudomonas species and BOS. Our objective was to investigate whether there may be a unifying explanation for these apparently dichotomous results.We performed bronchoscopies with bronchoalveolar lavage (BAL on lung transplant recipients (46 procedures in 33 patients and 26 non-transplant control subjects. We analyzed bacterial communities in the BAL fluid using qPCR and pyrosequencing of 16S rRNA gene amplicons and compared the culture-independent data with the clinical metadata and culture results from these subjects.Route of bronchoscopy (via nose or via mouth was not associated with changes in BAL microbiota (p = 0.90. Among the subjects with positive Pseudomonas bacterial culture, P. aeruginosa was also identified by culture-independent methods. In contrast, a distinct Pseudomonas species, P. fluorescens, was often identified in asymptomatic transplant subjects by pyrosequencing but not detected via standard bacterial culture. The subject populations harboring these two distinct pseudomonads differed significantly with respect to associated symptoms, BAL neutrophilia, bacterial DNA burden and microbial diversity. Despite notable differences in culturability, a global database search of UM Hospital Clinical Microbiology Laboratory records indicated that P. fluorescens is commonly isolated from respiratory specimens.We have reported for the first time that two prominent and distinct Pseudomonas species (P. fluorescens and P. aeruginosa exist within the post-transplant lung microbiome, each with unique genomic and microbiologic features and widely divergent clinical associations, including presence during acute infection.

  18. Characterization of the ability to form biofilms by plant-associated Pseudomonas species.

    Science.gov (United States)

    Ueda, Akihiro; Saneoka, Hirofumi

    2015-04-01

    Successful colonization is the initial step for plant-bacteria interactions; therefore, the development of strategies to improve adherence to plant surfaces is critically important for environmental bacteria. Biofilm formation is thought to be one such strategy for bacteria to establish stable colonization on inert and living surfaces. Although biofilms play potential roles in enabling persistent bacterial colonization, little attention has been paid to biofilms formed by plant-associated bacteria. In this study, we characterized the biofilm-forming ability of 6 species of bacteria from the family Pseudomonadaceae: Pseudomonas protegens, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas mendocina, and Pseudomonas syringae. These strains exhibit different degrees of biofilm formation depending on incubation time and nutrient availability. Distinct preferences for growth media were observed, as biofilms were formed by P. protegens with rich nutrients and by P. fluorescens and P. putida with poor nutrients. Likewise, P. stutzeri did not form biofilms with rich nutrients but did form biofilms under nutrient-poor conditions. These observations indicate that particular components in media may influence biofilm formation. P. putida, one of the strains with high biofilm-forming ability, showed the highest ability for initial attachment, which may be mediated by the hydrophobicity of its cell surface. P. mendocina also has high ability for initial attachment, and this strain produces cell surface-attached extracellular polysaccharides that promote cell aggregation. Thus, each strain possesses different properties that facilitate biofilm formation. Shedding light on bacterial strategies for colonization via biofilm formation would enable a better understanding of plant-bacteria interactions.

  19. Spoilage potential of Pseudomonas species isolated from goat milk.

    Science.gov (United States)

    Scatamburlo, T M; Yamazi, A K; Cavicchioli, V Q; Pieri, F A; Nero, L A

    2015-02-01

    Pseudomonas spp. are usually associated with spoilage microflora of dairy products due to their proteolytic potential. This is of particular concern for protein-based products, such as goat milk cheeses and fermented milks. Therefore, the goal of the present study was to characterize the proteolytic activity of Pseudomonas spp. isolated from goat milk. Goat milk samples (n=61) were obtained directly from bulk tanks on dairy goat farms (n=12), and subjected to a modified International Organization for Standardization (ISO) protocol to determine the number and proteolytic activity of Pseudomonas spp. Isolates (n=82) were obtained, identified by PCR, and subjected to pulsed-field gel electrophoresis with XbaI macro-restriction. Then, the isolates were subjected to PCR to detect the alkaline protease gene (apr), and phenotypic tests were performed to check proteolytic activity at 7°C, 25°C, and 35°C. Mean Pseudomonas spp. counts ranged from 2.9 to 4.8 log cfu/mL, and proteolytic Pseudomonas spp. counts ranged from 1.9 to 4.6 log cfu/mL. All isolates were confirmed to be Pseudomonas spp., and 41 were identified as Pseudomonas fluorescens, which clustered into 5 groups sharing approximately 82% similarity. Thirty-six isolates (46.9%) were positive for the apr gene; and 57 (69.5%) isolates presented proteolytic activity at 7°C, 82 (100%) at 25°C, and 64 (78%) at 35°C. The isolates were distributed ubiquitously in the goat farms, and no relationship among isolates was observed when the goat farms, presence of apr, pulsotypes, and proteolytic activity were taken into account. We demonstrated proteolytic activity of Pseudomonas spp. present in goat milk by phenotypic and genotypic tests and indicated their spoilage potential at distinct temperatures. Based on these findings and the ubiquity of Pseudomonas spp. in goat farm environments, proper monitoring and control of Pseudomonas spp. during production are critical. Copyright © 2015 American Dairy Science Association

  20. Nascent transcription affected by RNA polymerase IV in Zea mays.

    Science.gov (United States)

    Erhard, Karl F; Talbot, Joy-El R B; Deans, Natalie C; McClish, Allison E; Hollick, Jay B

    2015-04-01

    All eukaryotes use three DNA-dependent RNA polymerases (RNAPs) to create cellular RNAs from DNA templates. Plants have additional RNAPs related to Pol II, but their evolutionary role(s) remain largely unknown. Zea mays (maize) RNA polymerase D1 (RPD1), the largest subunit of RNA polymerase IV (Pol IV), is required for normal plant development, paramutation, transcriptional repression of certain transposable elements (TEs), and transcriptional regulation of specific alleles. Here, we define the nascent transcriptomes of rpd1 mutant and wild-type (WT) seedlings using global run-on sequencing (GRO-seq) to identify the broader targets of RPD1-based regulation. Comparisons of WT and rpd1 mutant GRO-seq profiles indicate that Pol IV globally affects transcription at both transcriptional start sites and immediately downstream of polyadenylation addition sites. We found no evidence of divergent transcription from gene promoters as seen in mammalian GRO-seq profiles. Statistical comparisons identify genes and TEs whose transcription is affected by RPD1. Most examples of significant increases in genic antisense transcription appear to be initiated by 3'-proximal long terminal repeat retrotransposons. These results indicate that maize Pol IV specifies Pol II-based transcriptional regulation for specific regions of the maize genome including genes having developmental significance. Copyright © 2015 by the Genetics Society of America.

  1. [Non-nascent hydrogen mechanism of plumbane generation].

    Science.gov (United States)

    Zou, Yan; Jin, Fu-xia; Chen, Zhi-jiang; Qiu, De-ren; Yang, Peng-yuan

    2005-10-01

    The mechanism of plumbane generation in dichromate system was studied via investigation of the relationship between the plumbane yield and the molar number of the reactants. A flow injection hydride generator was used in the study. Reactant moler number was calculated by the injected volume and the reactant concentration, and the plumbane yield was measured via an AAS spectrometer equipped with an electrothermal quartz tube atomizer. Experimental results show that the acid was first used for the neutralization of NaOH and successively participated in the redox reaction of borohydride with dichromate with a constant molar ratio of 9.95 +/- 0.42 (expressed in terms of mean +/- standard deviation). At the same time, plumbane generation was displayed as synchronously taking place with the redox reaction, and the yield increased with the increase of acid. The mechanism of plumbane generation was thus deduced as an induced reaction or a catalytic reaction by the redox reaction. Up to this end, the non-nascent hydrogen mechanism of hydride generation has been verified for all the IVA elements.

  2. NASCENTES ANTROPOGÊNICAS: PROCESSOS TECNOGÊNICOS E HIDROGEOMORFOLÓGICOS

    Directory of Open Access Journals (Sweden)

    Miguel Fernandes Felippe

    2014-04-01

    Full Text Available Nascentes de cursos d’água são sistemas ambientais em que a água emerge naturalmente do meio subterrâneo integrando a rede de drenagem superficial. Porém, no Tecnógeno, as intervenções humanas podem criar condições necessárias para a exfiltração da água onde originalmente isso não ocorreria. Se essa nova zona de descarga se conecta com a rede de drenagem via fluxos superficiais, ela pode ser considerada uma nascente de acordo com os conceitos acadêmicos mais aceitos. Entretanto, por sua origem ter sido induzida pelos seres humanos, esta deve ser considerada uma nascente antropogênica. Esse trabalho aborda conceitualmente as nascentes antropogênicas baseado em trabalhos de campo realizados no espaço metropolitano de Belo Horizonte-MG, explorando suas características principais. Além disso, são evidenciadas as diferenças entre a influência humana sobre as nascentes e a antropogenia desses sistemas, identificando os processos antrópicos que podem originar nascentes e discutindo as possíveis conseqüências dessas intervenções. Por fim, seis casos ilustrativos são apresentados, demonstrando como as nascentes antropogênicas podem ser formadas e suas conseqüências para a dinâmica hidrogeomorfológica.

  3. Experimental Investigation of Nascent Soot Physical Properties and The Influence on Particle Morphology and Growth

    Science.gov (United States)

    Lieb, Sydnie Marie

    Soot released to the atmosphere is a dangerous pollutant for human health and the environment. Understanding the physical properties and surface properties of these particles is important to properly explaining the growth of soot particles in flames as well as their interactions with other particles and gases in the environment. Particles below 15 nm in diameter, nascent soot particles, dominate the early growth stages of soot formation; previously these particles were characterized as hard graphitic spheres. New evidence derived from the current dissertation work, to a large extent, challenges this prior characterization. This dissertation study begins by revisiting the use of atomic force microscope (AFM) as a tool to investigate the structural properties of nascent soot. The impact of tip artifacts, which are known to complicate measurements of features below 10 nm in diameter, are carefully considered so as to provide a concise interpretation of the morphology of nascent soot as seen by AFM. The results of the AFM morphology collaborate with earlier photo- and thermal-fragmentation particle mass spectrometry and Fourier transform infrared spectroscopy that nascent soot is not a graphitized carbon material and that they are not spherical. Furthermore, phase mode imaging is introduced as a method to investigate the physical properties of nascent soot particles in a greater detail and finer resolution. The helium ion microscope (HIM) has been identified as a useful technique for the imaging of nascent soot. Using this imaging method nascent soot particles were imaged with a high resolution that had not been obtained by prior techniques. The increased contrast provides a closer look at the nascent soot particles and further suggested that these particles are not as structurally homogeneous as previously thought. Geometric shape analysis was performed to characterize the particles in terms of sphericity, circularity, and fractal dimension. The geometric analysis

  4. An Institutional Perspective on Business Planning Activities for Nascent Entrepreneurs in Sweden and the US

    Directory of Open Access Journals (Sweden)

    Benson Honig

    2013-11-01

    Full Text Available We compared nascent characteristics and behaviors leading to business planning activities in Sweden with the United States (US, examining the effects of institutional pressures exerted in the different countries. We analyzed institutional factors leading 362 Swedish and 347 US entrepreneurs to write plans during a two-year period. We show that national cultures moderate how institutional pressures influence nascent behaviors, questioning generic applications of institutional theory. We found business planning behaviors moderated by nationality, showing significant and negative effects for business classes in the US. Implications are drawn for institutional theory and the study of nascent businesses, as well as for normative business planning literature and practice of nascent businesses.

  5. Signal Recognition Particle-ribosome Binding Is Sensitive to Nascent Chain Length*

    Science.gov (United States)

    Noriega, Thomas R.; Tsai, Albert; Elvekrog, Margaret M.; Petrov, Alexey; Neher, Saskia B.; Chen, Jin; Bradshaw, Niels; Puglisi, Joseph D.; Walter, Peter

    2014-01-01

    The signal recognition particle (SRP) directs ribosome-nascent chain complexes (RNCs) displaying signal sequences to protein translocation channels in the plasma membrane of prokaryotes and endoplasmic reticulum of eukaryotes. It was initially proposed that SRP binds the signal sequence when it emerges from an RNC and that successful binding becomes impaired as translation extends the nascent chain, moving the signal sequence away from SRP on the ribosomal surface. Later studies drew this simple model into question, proposing that SRP binding is unaffected by nascent chain length. Here, we reinvestigate this issue using two novel and independent fluorescence resonance energy transfer assays. We show that the arrival and dissociation rates of SRP binding to RNCs vary according to nascent chain length, resulting in the highest affinity shortly after a functional signal sequence emerges from the ribosome. Moreover, we show that SRP binds RNCs in multiple and interconverting conformations, and that conversely, RNCs exist in two conformations distinguished by SRP interaction kinetics. PMID:24808175

  6. The ribosomal protein uL22 modulates the shape of the nascent protein exit tunnel

    DEFF Research Database (Denmark)

    Wekselman, I.; Zimmerman, E.; Davidovich, C.

    2017-01-01

    Nascent proteins progress through an elongated tunnel until theyexit from the ribosome. Biochemical, genetic and structural stud-ies have shown that the tunnel is not just a passive path, but alsohas regulatory properties. Erythromycin is a clinically usefulantibiotic that binds to an rRNA pocket...... and specific sequences within nascent chains trigger confor-mational rearrangements in the exit tunnel that are essential forthe translation of specific genes...

  7. Study of the biological impact of Pseudomonas spp.fluoresents on hemolyphatic metabolites and histology of the digestive tract of larvae 15 migratory locust Locusta migratoria

    International Nuclear Information System (INIS)

    Oulebsir- Mohan, H.; Doumandji-Mitiche, B.

    2012-01-01

    This study allows to test the effect of entomopathogenic bacteria of Pseudomonas fluorescens bv III and Pseudomonas fluorescence bv V on the haemolymph of Locusta migratoria metabolites, namely proteins and carbohydrates as well as on the histology of the digestive system of fifth stage larvae of migratory locust Locusta migratoria. The results show an important decrease of haemolymph protein concentration compared to controls with an increase in carbohydrate concentration. Examination of histological sections of various parts of the digestive tract showed some changes in treated. (author)

  8. Metallo-β-Lactamases in Clinical Pseudomonas Isolates in Taiwan and Identification of VIM-3, a Novel Variant of the VIM-2 Enzyme

    OpenAIRE

    Yan, Jing-Jou; Hsueh, Po-Ren; Ko, Wen-Chien; Luh, Kwen-Tay; Tsai, Shu-Huei; Wu, Hsiu-Mei; Wu, Jiunn-Jong

    2001-01-01

    A total of 209 clinical isolates of Pseudomonas (193 Pseudomonas aeruginosa, 10 P. putida, 4 P. stutzeri, and 2 P. fluorescens isolates) with reduced susceptibilities to imipenem and/or ceftazidime were subjected to PCR assays with primers specific for blaIMP-1, blaIMP-2, blaVIM-1, and blaVIM-2 and sequence analysis to identify the metallo-β-lactamases (MBLs) prevalent among these organisms in Taiwan; and 21 isolates gave positive results. Five isolates including two P. putida and three P. st...

  9. Different Ancestries of R Tailocins in Rhizospheric Pseudomonas Isolates

    Science.gov (United States)

    Ghequire, Maarten G.K.; Dillen, Yörg; Lambrichts, Ivo; Proost, Paul; Wattiez, Ruddy; De Mot, René

    2015-01-01

    Bacterial genomes accommodate a variety of mobile genetic elements, including bacteriophage-related clusters that encode phage tail-like protein complexes playing a role in interactions with eukaryotic or prokaryotic cells. Such tailocins are unable to replicate inside target cells due to the lack of a phage head with associated DNA. A subset of tailocins mediate antagonistic activities with bacteriocin-like specificity. Functional characterization of bactericidal tailocins of two Pseudomonas putida rhizosphere isolates revealed not only extensive similarity with the tail assembly module of the Pseudomonas aeruginosa R-type pyocins but also differences in genomic integration site, regulatory genes, and lytic release modules. Conversely, these three features are quite similar between strains of the P. putida and Pseudomonas fluorescens clades, although phylogenetic analysis of tail genes suggests them to have evolved separately. Unlike P. aeruginosa R pyocin elements, the tailocin gene clusters of other pseudomonads frequently carry cargo genes, including bacteriocins. Compared with P. aeruginosa, the tailocin tail fiber sequences that act as specificity determinants have diverged much more extensively among the other pseudomonad species, mostly isolates from soil and plant environments. Activity of the P. putida antibacterial particles requires a functional lipopolysaccharide layer on target cells, but contrary to R pyocins from P. aeruginosa, strain susceptibilities surpass species boundaries. PMID:26412856

  10. DESENVOLVIMENTO DE UMA TIPOLOGIA HIDROGEOMORFOLÓGICA DE NASCENTES BASEADA EM ESTATÍSTICA NEBULOSA MULTIVARIADA

    Directory of Open Access Journals (Sweden)

    Miguel Fernandes Felippe

    2014-09-01

    Full Text Available As nascentes de cursos d’água são sistemas hidrogeomorfológicos complexos, caracterizados por uma grande variabilidade de seus elementos estruturantes. Esta heterogeneidade ainda pouco conhecida cria dificuldades para os processos de gestão ambiental, pois, em teoria, cada nascente responde a perturbações de maneira diferente. Nesse sentido, este trabalho tem como objetivo desenvolver uma tipologia de nascentes com base em suas características hidrogeomorfológicas que sistematize sua heterogeneidade, sem perder seus aspectos individuais. Para atingir esta meta um método de máxima verossimilhança baseado na teoria dos conjuntos nebulosos foi usado para criar perfis multivariados para 79 nascentes catalogadas em três parques urbanos de Belo Horizonte - MG. Após uma série de testes estatísticos, propuseram-se seis tipos básicos no qual as nascentes podem ser enquadradas. No entanto, o principal avanço desta técnica é permitir a comparação entre os diferentes graus de pertencimento de cada nascente em cada tipo criado, auxiliando na compreensão da sua diversidade, sem esconder a sua heterogeneidade.

  11. Biosurfactant production by Pseudomonas strains isolated from floral nectar.

    Science.gov (United States)

    Ben Belgacem, Z; Bijttebier, S; Verreth, C; Voorspoels, S; Van de Voorde, I; Aerts, G; Willems, K A; Jacquemyn, H; Ruyters, S; Lievens, B

    2015-06-01

    To screen and identify biosurfactant-producing Pseudomonas strains isolated from floral nectar; to characterize the produced biosurfactants; and to investigate the effect of different carbon sources on biosurfactant production. Four of eight nectar Pseudomonas isolates were found to produce biosurfactants. Phylogenetic analysis based on three housekeeping genes (16S rRNA gene, rpoB and gyrB) classified the isolates into two groups, including one group closely related to Pseudomonas fluorescens and another group closely related to Pseudomonas fragi and Pseudomonas jessenii. Although our nectar pseudomonads were able to grow on a variety of water-soluble and water-immiscible carbon sources, surface active agents were only produced when using vegetable oil as sole carbon source, including olive oil, sunflower oil or waste frying sunflower oil. Structural characterization based on thin layer chromatography (TLC) and ultra high performance liquid chromatography-accurate mass mass spectrometry (UHPLC-amMS) revealed that biosurfactant activity was most probably due to the production of fatty acids (C16:0; C18:0; C18:1 and C18:2), and mono- and diglycerides thereof. Four biosurfactant-producing nectar pseudomonads were identified. The active compounds were identified as fatty acids (C16:0; C18:0; C18:1 and C18:2), and mono- and diglycerides thereof, produced by hydrolysis of triglycerides of the feedstock. Studies on biosurfactant-producing micro-organisms have mainly focused on microbes isolated from soils and aquatic environments. Here, for the first time, nectar environments were screened as a novel source for biosurfactant producers. As nectars represent harsh environments with high osmotic pressure and varying pH levels, further screening of nectar habitats for biosurfactant-producing microbes may lead to the discovery of novel biosurfactants with broad tolerance towards different environmental conditions. © 2015 The Society for Applied Microbiology.

  12. A comparative analysis of metal transportomes from metabolically versatile Pseudomonas

    Directory of Open Access Journals (Sweden)

    Rodrigue Agnes

    2008-09-01

    Full Text Available Abstract Background The availability of complete genome sequences of versatile Pseudomonas occupying remarkably diverse ecological niches enabled to gain insights into their adaptative assets. The objective of this study was to analyze the complete genetic repertoires of metal transporters (metal transportomes from four representative Pseudomonas species and to identify metal transporters with "Genomic Island" associated features. Methods A comparative metal transporter inventory was built for the following four Pseudomonas species: P.putida (Ppu KT2440, P.aeruginosa (Pae PA01, P.fluorescens (Pfl Pf-5 and P.syringae (Psypv.tomato DC3000 using TIGR-CMR and Transport DB. Genomic analysis of essential and toxic metal ion transporters was accomplished from the above inventory. Metal transporters with "Genomic Island" associated features were identified using Islandpath analysis. Results Dataset cataloguing has been executed for 262 metal transporters from the four spp. Additional metal ion transporters belonging to NiCoT, Ca P-type ATPase, Cu P-type ATPases, ZIP and MgtC families were identified. In Psy DC3000, 48% of metal transporters showed strong GI features while it was 45% in Ppu KT2440. In Pfl Pf-5 and Pae PA01 only 26% of their metal transporters exhibited GI features. Conclusion Our comparative inventory of 262 metal transporters from four versatile Pseudomonas spp is the complete suite of metal transportomes analysed till date in a prokaryotic genus. This study identified differences in the basic composition of metal transportomes from Pseudomonas occupying diverse ecological niches and also elucidated their novel features. Based on this inventory we analysed the role of horizontal gene transfer in expansion and variability of metal transporter families.

  13. Pseudomonas aeruginosa in Healthcare Settings

    Science.gov (United States)

    ... Sepsis Sharps Safety - CDC Transplant Safety Vaccine Safety Pseudomonas aeruginosa in Healthcare Settings Recommend on Facebook Tweet ... and/or help treat infections? What is a Pseudomonas infection? Pseudomonas infection is caused by strains of ...

  14. Antibiotic resistance patterns of Pseudomonas spp. isolated from the River Danube

    Directory of Open Access Journals (Sweden)

    Clemens eKittinger

    2016-05-01

    Full Text Available Spread and persistence of antibiotic resistance pose a severe threat to human health, yet there is still lack of knowledge about reservoirs of antibiotic resistant bacteria in the environment. We took the opportunity of the Joint Danube Survey 3 (JDS3, the world's biggest river research expedition of its kind in 2013, to analyse samples originating from different sampling points along the whole length of the river. Due to its high clinical relevance, we concentrated on the characterization of Pseudomonas spp. and evaluated the resistance profiles of Pseudomonas spp. which were isolated from eight sampling points. In total, 520 Pseudomonas isolates were found, 344 (66.0% isolates were identified as Pseudomonas putida, and 141 (27.1% as Pseudomonas fluorescens, all other Pseudomonas species were represented by less than five isolates, among those two P. aeruginosa isolates. Thirty seven percent (37% of all isolated Pseudomonas species showed resistance to at least one out of eleven tested antibiotics. The most common resistance was against meropenem (30.4% / 158 isolates piperacillin/tazobactam (10.6% / 55 isolates and ceftazidime (4.2% / 22 isolates. 16 isolates (3.1% / 16 isolates were multi-resistant. For each tested antibiotic at least one resistant isolate could be detected. Sampling points from the upper stretch of the River Danube showed more resistant isolates than downriver. Our results suggest that antibiotic resistance can be acquired by and persists even in Pseudomonas species that are normally not in direct contact with humans. A possible scenario is that these bacteria provide a reservoir of antibiotic resistance genes that can spread to related human pathogens by horizontal gene transfer.

  15. Antifungal Activity of Selected Indigenous Pseudomonas and Bacillus from the Soybean Rhizosphere

    Directory of Open Access Journals (Sweden)

    A. F. García

    2009-01-01

    Full Text Available The purpose of this study was to isolate and select indigenous soil Pseudomonas and Bacillus bacteria capable of developing multiple mechanisms of action related to the biocontrol of phytopathogenic fungi affecting soybean crops. The screening procedure consisted of antagonism tests against a panel of phytopathogenic fungi, taxonomic identification, detection by PCR of several genes related to antifungal activity, in vitro detection of the antifungal products, and root colonization assays. Two isolates, identified and designated as Pseudomonas fluorescens BNM296 and Bacillus amyloliquefaciens BNM340, were selected for further studies. These isolates protected plants against the damping-off caused by Pythium ultimum and were able to increase the seedling emergence rate after inoculation of soybean seeds with each bacterium. Also, the shoot nitrogen content was higher in plants when seeds were inoculated with BNM296. The polyphasic approach of this work allowed us to select two indigenous bacterial strains that promoted the early development of soybean plants.

  16. Nascent RNA kinetics: Transient and steady state behavior of models of transcription

    Science.gov (United States)

    Choubey, Sandeep

    2018-02-01

    Regulation of transcription is a vital process in cells, but mechanistic details of this regulation still remain elusive. The dominant approach to unravel the dynamics of transcriptional regulation is to first develop mathematical models of transcription and then experimentally test the predictions these models make for the distribution of mRNA and protein molecules at the individual cell level. However, these measurements are affected by a multitude of downstream processes which make it difficult to interpret the measurements. Recent experimental advancements allow for counting the nascent mRNA number of a gene as a function of time at the single-inglr cell level. These measurements closely reflect the dynamics of transcription. In this paper, we consider a general mechanism of transcription with stochastic initiation and deterministic elongation and probe its impact on the temporal behavior of nascent RNA levels. Using techniques from queueing theory, we derive exact analytical expressions for the mean and variance of the nascent RNA distribution as functions of time. We apply these analytical results to obtain the mean and variance of nascent RNA distribution for specific models of transcription. These models of initiation exhibit qualitatively distinct transient behaviors for both the mean and variance which further allows us to discriminate between them. Stochastic simulations confirm these results. Overall the analytical results presented here provide the necessary tools to connect mechanisms of transcription initiation to single-cell measurements of nascent RNA.

  17. Do Business Demographics Matter to Nascent Venture Performance? A Longitudinal Evidence from the Kauffman Firm Survey.

    Directory of Open Access Journals (Sweden)

    Augustine Yao Dzathor

    2013-08-01

    Full Text Available Scant literature exists on impact of organizational demographics on nascent venture performance. Business demographics were looked at through the lenses of organizational traits, resource-based view and population ecology theories. The combined as well as the  individual longitudinal impact of five business demographics (nature of product, primary location of business, number of active owner-operators, business legal status and technological orientation on nascent venture performance was examined. 754 out of the 863 businesses that survived cleaning of the first four years of the Kauffman Firm Survey (KFS dataset were retained in the analysis sample to maintain the original sample stratification of the data set. Results of multiple regression revealed that business demographics globally had significant effect on nascent venture performance in year-2 and year-4 of first four years of existence of the cohort of businesses studied. Three of the individual business demographics indicators, namely: nature of product, business legal status and technological orientation also had significant but inconsistent effects on nascent venture performance across the four years under study. The study threw light on the longitudinal effects of business demographics on nascent venture performance and drew attention to the fact that business demographics can theoretically be envisioned through the lenses of trait theory, resource based view and population ecology theory.

  18. Pseudomonas screening assay

    Science.gov (United States)

    Margalit, Ruth (Inventor)

    1993-01-01

    A method for the detection of Pseudomonas bacteria is described where an Azurin-specific antibody is employed for detecting the presence of Azurin in a test sample. The detection of the presence of Azurin in the sample is a conclusive indicator of the presence of the Pseudomonas bacteria since the Azurin protein is a specific marker for this bacterial strain.

  19. Comparative genomic and functional analyses: unearthing the diversity and specificity of nematicidal factors in Pseudomonas putida strain 1A00316

    Science.gov (United States)

    Guo, Jing; Jing, Xueping; Peng, Wen-Lei; Nie, Qiyu; Zhai, Yile; Shao, Zongze; Zheng, Longyu; Cai, Minmin; Li, Guangyu; Zuo, Huaiyu; Zhang, Zhitao; Wang, Rui-Ru; Huang, Dian; Cheng, Wanli; Yu, Ziniu; Chen, Ling-Ling; Zhang, Jibin

    2016-01-01

    We isolated Pseudomonas putida (P. putida) strain 1A00316 from Antarctica. This bacterium has a high efficiency against Meloidogyne incognita (M. incognita) in vitro and under greenhouse conditions. The complete genome of P. putida 1A00316 was sequenced using PacBio single molecule real-time (SMRT) technology. A comparative genomic analysis of 16 Pseudomonas strains revealed that although P. putida 1A00316 belonged to P. putida, it was phenotypically more similar to nematicidal Pseudomonas fluorescens (P. fluorescens) strains. We characterized the diversity and specificity of nematicidal factors in P. putida 1A00316 with comparative genomics and functional analysis, and found that P. putida 1A00316 has diverse nematicidal factors including protein alkaline metalloproteinase AprA and two secondary metabolites, hydrogen cyanide and cyclo-(l-isoleucyl-l-proline). We show for the first time that cyclo-(l-isoleucyl-l-proline) exhibit nematicidal activity in P. putida. Interestingly, our study had not detected common nematicidal factors such as 2,4-diacetylphloroglucinol (2,4-DAPG) and pyrrolnitrin in P. putida 1A00316. The results of the present study reveal the diversity and specificity of nematicidal factors in P. putida strain 1A00316. PMID:27384076

  20. Potencial da nascente do Rio Vieira e entorno: uma interpretação da geodiversidade

    OpenAIRE

    Rachel Inêz Castro de Oliveira

    2017-01-01

    Este artigo contempla o aproveitamento dos elementos geológicos, geomorfológicos, espeleológicos, pedológicos e hidrográficos que compõem a geodiversidade como recurso educativo, turístico e interpretativo, em uma das principais nascentes do Rio Vieira e entorno, no município de Montes Claros, Norte de Minas. Considerando a especificidade desse campo da geodiversidade, o objetivo foi investigar as potencialidades da nascente do Rio Vieira e entorno, por meio do inventário dos elementos repres...

  1. Learning Entrepreneurial Leadership among Nascent Food Entrepreneurs in Denmark and New Zealand

    DEFF Research Database (Denmark)

    Ramsgaard, Michael Breum; Warren, Lorraine

    This paper examines the process of learning entrepreneurial leadership in small food businesses in the early stages. It draws on the growing body of research around entrepreneurial leadership that deals with leadership in ventures but overlooks the importance of entrepreneurial leadership...... in the nascent phases. We analyse the factors constituting the processes of learning entrepreneurial leadership among nascent food entrepreneurs. The primary source of data derives from four in-depth interviews in each country with founders of SME food businesses with no more than 5 years of company history...

  2. Pseudomonas granadensis sp. nov., a new bacterial species isolated from the Tejeda, Almijara and Alhama Natural Park, Granada, Spain.

    Science.gov (United States)

    Pascual, Javier; García-López, Marina; Bills, Gerald F; Genilloud, Olga

    2015-02-01

    During the course of screening bacterial isolates as sources of as-yet unknown bioactive compounds with pharmaceutical applications, a chemo-organotrophic, Gram-negative bacterium was isolated from a soil sample taken from the Tejeda, Almijara and Alhama Natural Park, Granada, Spain. Strain F-278,770(T) was oxidase- and catalase-positive, aerobic, with a respiratory type of metabolism with oxygen as the terminal electron acceptor, non-spore-forming and motile by one polar flagellum, although some cells had two polar flagella. Phylogenetic analysis of the 16S rRNA, gyrB, rpoB and rpoD genes revealed that strain F-278,770(T) belongs to the Pseudomonas koreensis subgroup (Pseudomonas fluorescens lineage), with Pseudomonas moraviensis, P. koreensis, P. baetica and P. helmanticensis as its closest relatives. Chemotaxonomic traits such as polar lipid and fatty acid compositions and G+C content of genomic DNA corroborated the placement of strain F-278,770(T) in the genus Pseudomonas. DNA-DNA hybridization assays and phenotypic traits confirmed that this strain represents a novel species of the genus Pseudomonas, for which the name Pseudomonas granadensis sp. nov. is proposed. The type strain is F-278,770(T) ( = DSM 28040(T) = LMG 27940(T)). © 2015 Fundacion MEDINA, Centro de Excelencia en Investigacion de Medicamentos Innovadores en Andalucia.

  3. Production of the Antibiotic Phenazine-1-Carboxylic Acid by Fluorescent Pseudomonas Species in the Rhizosphere of Wheat

    OpenAIRE

    Thomashow, Linda S.; Weller, David M.; Bonsall, Robert F.; Pierson, Leland S.

    1990-01-01

    Pseudomonas fluorescens 2-79 and P. aureofaciens 30-84 produce the antibiotic phenazine-1-carboxylic acid and suppress take-all, an important root disease of wheat caused by Gaeumannomyces graminis var. tritici. To determine whether the antibiotic is produced in situ, wheat seeds were treated with strain 2-79 or 30-84 or with phenazine-nonproducing mutants or were left untreated and then were sown in natural or steamed soil in the field or growth chamber. The antibiotic was isolated only from...

  4. Nascent ventures competing for start-up capital: matching reputations and investors

    NARCIS (Netherlands)

    Ebbers, J.J.; Wijnberg, N.M.

    2012-01-01

    Although nascent ventures have not yet developed a performance-based reputation, the individual reputations of their founders, based on the performance of their earlier ventures, can function as important signals to investors. Selection system theory distinguishes between different types of

  5. The effect of human capital, social capital, and perceptual values on nascent entrepreneurs' export intentions

    DEFF Research Database (Denmark)

    Christensen, Poul Rind; Evald, Majbritt Rostgaard; Klyver, Kim

    2011-01-01

    This study investigates the influence of human capital, social capital, and cognition on nascent entrepreneurs' export intentions. The results indicate that while human capital and social capital influence the level of intended export, cognitive characteristics, such as self-efficacy and risk...... aversion, do not seem to influence entrepreneurs' intended level of export...

  6. Nascent chromatin capture proteomics determines chromatin dynamics during DNA replication and identifies unknown fork components

    DEFF Research Database (Denmark)

    Alabert, Constance; Bukowski-Wills, Jimi-Carlo; Lee, Sung-Po

    2014-01-01

    replication in human cells. NCC relies on biotin-dUTP labelling of replicating DNA, affinity purification and quantitative proteomics. Comparing nascent chromatin with mature post-replicative chromatin, we provide association dynamics for 3,995 proteins. The replication machinery and 485 chromatin factors...

  7. The effect of human capital, social capital, and perceptual values on nascent entrepreneurs' export intentions

    DEFF Research Database (Denmark)

    Christensen, Poul Rind; Evald, Majbritt Rostgaard; Klyver, Kim

    2011-01-01

    This study investigates the influence of human capital, social capital, and cognition on nascent entrepreneurs' export intentions. The results indicate that while human capital and social capital influence the level of intended export, cognitive characteristics, such as self-efficacy and risk...

  8. Initial characterization of the nascent polypeptide-associated complex in yeast.

    Science.gov (United States)

    Reimann, B; Bradsher, J; Franke, J; Hartmann, E; Wiedmann, M; Prehn, S; Wiedmann, B

    1999-03-30

    The three subunits of the nascent polypeptide-associated complex (alpha, beta1, beta3) in Saccharomyces cerevisiae are encoded by three genes (EGD2, EGD1, BTT1). We found the complex bound to ribosomes via the beta-subunits in a salt-sensitive manner, in close proximity to nascent polypeptides. Estimation of the molecular weight of the complex of wild-type cells and cells lacking one or two subunits revealed that the composition of the complex is variable and that as yet unknown proteins might be included. Regardless of the variability, a certain balance of the subunits has to be maintained: the deletion of one subunit causes downregulation of the remaining subunits at physiological growth temperature. Cells lacking both beta-subunits are unable to grow at 37 degrees C, most likely due to a toxic effect of the alpha-subunit. Based on in vitro experiments, it has been proposed that the function of mammalian nascent-polypeptide associated complexes (NAC) is to prevent inappropriate targeting of non-secretory nascent polypeptides. In vivo, however, the lack of NAC does not cause secretion of signal-less invertase in yeast. This result and the lack of a drastic phenotype of cells missing one, two or three subunits at optimal conditions (28 degrees C, YPD-medium) suggest either the existence of a substitute for NAC or that cells tolerate or 'repair' the damage caused by the absence of NAC.

  9. Identification of a protein linked to nascent poliovirus RNA and to the polyuridylic acid of negative-strand RNA.

    Science.gov (United States)

    Pettersson, R F; Ambros, V; Baltimore, D

    1978-08-01

    A protein similar to that previously demonstrated on poliovirus RNA and replicative intermediate RNA (VPg) was found on all sizes of nascent viral RNA molecules and on the polyuridylic acid isolated from negative-strand RNA. 32P-labeled nascent chains were released from their template RNA and fractionated by exclusion chromatography on agarose. Fingerprint analysis using two-dimensional polyacrylamide gels of RNase T1 oligonucleotides derived from nascent chains of different lengths showed that a size fractionation of nascent chains was achieved. VPg was recovered from nascent chains varying in length from 7,500 nucleotides (full-sized RNA) to about 500 nucleotides. No other type of 5' terminus could be demonstrated on nascent RNA, and the yield of VPg was consistent with one molecule of the protein on each nascent chain. These results are consistent with the concept that the protein is added to the 5' end of the growing RNA chains at a very early stage, possibly as a primer of RNA synthesis. Analysis of the polyuridylic acid tract isolated from the replicative intermediate and double-stranded RNAs indicated that a protein of the same size as that found on the nascent chains and virion RNA is also linked to the negative-strand RNAs. It is likely that a similar mechanism is responsible for initiation of synthesis of both plus- and minus-strand RNAs.

  10. Protracted and asynchronous accumulation of PSD95-family MAGUKs during maturation of nascent dendritic spines.

    Science.gov (United States)

    Lambert, Jason T; Hill, Travis C; Park, Deborah K; Culp, Julie H; Zito, Karen

    2017-10-01

    The formation and stabilization of new dendritic spines is a key component of the experience-dependent neural circuit plasticity that supports learning, but the molecular maturation of nascent spines remains largely unexplored. The PSD95-family of membrane-associated guanylate kinases (PSD-MAGUKs), most notably PSD95, has a demonstrated role in promoting spine stability. However, nascent spines contain low levels of PSD95, suggesting that other members of the PSD-MAGUK family might act to stabilize nascent spines in the early stages of spiny synapse formation. Here, we used GFP-fusion constructs to quantitatively define the molecular composition of new spines, focusing on the PSD-MAGUK family. We found that PSD95 levels in new spines were as low as those previously associated with rapid subsequent spine elimination, and new spines did not achieve mature levels of PSD95 until between 12 and 20 h following new spine identification. Surprisingly, we found that the PSD-MAGUKs PSD93, SAP97, and SAP102 were also substantially less enriched in new spines. However, they accumulated in new spines more quickly than PSD95: SAP102 enriched to mature levels within 3 h, SAP97 and PSD93 enriched gradually over the course of 6 h. Intriguingly, when we restricted our analysis to only those new spines that persisted, SAP97 was the only PSD-MAGUK already present at mature levels in persistent new spines when first identified. Our findings uncover a key structural difference between nascent and mature spines, and suggest a mechanism for the stabilization of nascent spines through the sequential arrival of PSD-MAGUKs. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1161-1174, 2017. © 2017 Wiley Periodicals, Inc.

  11. The ribosome quality control pathway can access nascent polypeptides stalled at the Sec61 translocon.

    Science.gov (United States)

    von der Malsburg, Karina; Shao, Sichen; Hegde, Ramanujan S

    2015-06-15

    Cytosolic ribosomes that stall during translation are split into subunits, and nascent polypeptides trapped in the 60S subunit are ubiquitinated by the ribosome quality control (RQC) pathway. Whether the RQC pathway can also target stalls during cotranslational translocation into the ER is not known. Here we report that listerin and NEMF, core RQC components, are bound to translocon-engaged 60S subunits on native ER membranes. RQC recruitment to the ER in cultured cells is stimulated by translation stalling. Biochemical analyses demonstrated that translocon-targeted nascent polypeptides that subsequently stall are polyubiquitinated in 60S complexes. Ubiquitination at the translocon requires cytosolic exposure of the polypeptide at the ribosome-Sec61 junction. This exposure can result from either failed insertion into the Sec61 channel or partial backsliding of translocating nascent chains. Only Sec61-engaged nascent chains early in their biogenesis were relatively refractory to ubiquitination. Modeling based on recent 60S-RQC and 80S-Sec61 structures suggests that the E3 ligase listerin accesses nascent polypeptides via a gap in the ribosome-translocon junction near the Sec61 lateral gate. Thus the RQC pathway can target stalled translocation intermediates for degradation from the Sec61 channel. © 2015 von der Malsburg et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  12. Pseudomonas Lipopeptide Biosurfactants

    DEFF Research Database (Denmark)

    Bonnichsen, Lise

    Pseudomonas lipopetide biosurfactants are amphiphilic molecules with a broad range of natural functions. Due to their surface active properties, it has been suggested that Pseudomonas lipopetides potentially play a role in biodegradation of hydrophobic compounds and have essential functions...... lipopetide biosurfactants in pollutant biodegradation and natural roles in biofilm formation. The work presented is a combination of environmental microbiology and exploiting genetic manipulation of pure cultures to achieve insightinto the effects and mechanisms of lipopeptides on microbial processes...

  13. Influence of volatile organic compounds emitted by Pseudomonas and Serratia strains on Agrobacterium tumefaciens biofilms.

    Science.gov (United States)

    Plyuta, Vladimir; Lipasova, Valentina; Popova, Alexandra; Koksharova, Olga; Kuznetsov, Alexander; Szegedi, Erno; Chernin, Leonid; Khmel, Inessa

    2016-07-01

    The ability to form biofilms plays an important role in bacteria-host interactions, including plant pathogenicity. In this work, we investigated the action of volatile organic compounds (VOCs) produced by rhizospheric strains of Pseudomonas chlororaphis 449, Pseudomonas fluorescens B-4117, Serratia plymuthica IC1270, as well as Serratia proteamaculans strain 94, isolated from spoiled meat, on biofilms formation by three strains of Agrobacterium tumefaciens which are causative agents of crown-gall disease in a wide range of plants. In dual culture assays, the pool of volatiles emitted by the tested Pseudomonas and Serratia strains suppressed the formation of biofilms of A. tumefaciens strains grown on polycarbonate membrane filters and killed Agrobacterium cells in mature biofilms. The individual VOCs produced by the tested Pseudomonas strains, that is, ketones (2-nonanone, 2-heptanone, 2-undecanone), and dimethyl disulfide (DMDS) produced by Serratia strains, were shown to kill A. tumefaciens cells in mature biofilms and suppress their formation. The data obtained in this study suggest an additional potential of some ketones and DMDS as protectors of plants against A. tumefaciens strains, whose virulence is associated with the formation of biofilms on the infected plants. © 2016 APMIS. Published by John Wiley & Sons Ltd.

  14. Antimicrobial Activity of Various Plant Extracts on Pseudomonas Species Associated with Spoilage of Chilled Fish

    Directory of Open Access Journals (Sweden)

    Osan Bahurmiz

    2016-11-01

    Full Text Available The antimicrobial activity of various plant extracts on Pseudomonas bacteria isolated from spoiled chilled tilapia (Oreochromis sp. was evaluated in this study. In the first stage of this study, red tilapia was subjected to chilled storage (4°C for 3 weeks, and spoilage bacteria were isolated and identified from the spoiled fish. Pseudomonas was the dominant bacteria isolated from the spoiled fish and further identification revealed that P. putida, P. fluorescens and Pseudomonas spp. were the main species of this group. In the second stage, methanolic extracts of 15 selected plant species were screened for their antimicrobial activity, by agar disc diffusion method, against the Pseudomonas isolates. Results indicated that most of the extracts had different degrees of activity against the bacterial isolates. The strongest activity was exhibited by bottlebrush flower (Callistemon viminalis extract. This was followed by extracts from guava bark (Psidium guajava and henna leaf (Lawsonia inermis. Moderate antimicrobial activities were observed in extracts of clove (Syzygium aromaticum, leaf and peel of tamarind (Tamarindus indica, cinnamon bark (Cinnamomum zeylanicum, wild betel leaf (Piper sarmentosum and fresh thyme (Thymus spp.. Weak or no antimicrobial activity was observed from the remaining extracts. The potential antimicrobial activity shown by some plant extracts in this study could significantly contribute to the fish preservation.

  15. Ferric-Pyoverdine Recognition by Fpv Outer Membrane Proteins of Pseudomonas protegens Pf-5

    Science.gov (United States)

    Hartney, Sierra L.; Mazurier, Sylvie; Girard, Maëva K.; Mehnaz, Samina; Davis, Edward W.; Gross, Harald; Lemanceau, Philippe

    2013-01-01

    The soil bacterium Pseudomonas protegens Pf-5 (previously called P. fluorescens Pf-5) produces two siderophores, enantio-pyochelin and a compound in the large and diverse pyoverdine family. Using high-resolution mass spectroscopy, we determined the structure of the pyoverdine produced by Pf-5. In addition to producing its own siderophores, Pf-5 also utilizes ferric complexes of some pyoverdines produced by other strains of Pseudomonas spp. as sources of iron. Previously, phylogenetic analysis of the 45 TonB-dependent outer membrane proteins in Pf-5 indicated that six are in a well-supported clade with ferric-pyoverdine receptors (Fpvs) from other Pseudomonas spp. We used a combination of phylogenetics, bioinformatics, mutagenesis, pyoverdine structural determinations, and cross-feeding bioassays to assign specific ferric-pyoverdine substrates to each of the six Fpvs of Pf-5. We identified at least one ferric-pyoverdine that was taken up by each of the six Fpvs of Pf-5. Functional redundancy of the Pf-5 Fpvs was also apparent, with some ferric-pyoverdines taken up by all mutants with a single Fpv deletion but not by a mutant having deletions in two of the Fpv-encoding genes. Finally, we demonstrated that phylogenetically related Fpvs take up ferric complexes of structurally related pyoverdines, thereby establishing structure-function relationships that can be employed in the future to predict the pyoverdine substrates of Fpvs in other Pseudomonas spp. PMID:23222724

  16. Dissemination and persistence of Pseudomonas spp. in small-scale dairy farms

    Directory of Open Access Journals (Sweden)

    Daniele Michele Nucera

    2016-05-01

    Full Text Available This study was aimed at collecting data on presence, dissemination and persistence of Pseudomonas in small-scale dairy farms. Six farms (located in Piedmont were visited three times over 2014: 116 waters (wells and different faucets/pipes and 117 environmental samples (milking equipments and drains were collected. Enumeration of Pseudomonadaceae was performed, 3-5 colonies/samples were selected for identification via 16SrDNA/oprI polymerase chain reaction (PCR, and typed by enterobacterial-repetitive- intergenic-consensus (ERIC-PCR. Pseudomonadaceae were detected in 77% of samples. No statistical differences were found among proportions of positives across farms, sample typologies and seasons. Most isolates were Pseudomonas fluorescens (45%, and ERIC-PCR showed 32 persistent types diffused across farms. All in all, Pseudomonas spp. represents a challenge, considering its presence over time in water as well as in teat cups, indicating a continuous source of contamination. Moreover, persistency of strains may indicate biofilm-formation and/or sanitisers resistance, therefore emphasising the role of primary production for preventing milk contamination by Pseudomonas spp.

  17. Comparative genomic analysis of four representative plant growth-promoting rhizobacteria in Pseudomonas

    Science.gov (United States)

    2013-01-01

    Background Some Pseudomonas strains function as predominant plant growth-promoting rhizobacteria (PGPR). Within this group, Pseudomonas chlororaphis and Pseudomonas fluorescens are non-pathogenic biocontrol agents, and some Pseudomonas aeruginosa and Pseudomonas stutzeri strains are PGPR. P. chlororaphis GP72 is a plant growth-promoting rhizobacterium with a fully sequenced genome. We conducted a genomic analysis comparing GP72 with three other pseudomonad PGPR: P. fluorescens Pf-5, P. aeruginosa M18, and the nitrogen-fixing strain P. stutzeri A1501. Our aim was to identify the similarities and differences among these strains using a comparative genomic approach to clarify the mechanisms of plant growth-promoting activity. Results The genome sizes of GP72, Pf-5, M18, and A1501 ranged from 4.6 to 7.1 M, and the number of protein-coding genes varied among the four species. Clusters of Orthologous Groups (COGs) analysis assigned functions to predicted proteins. The COGs distributions were similar among the four species. However, the percentage of genes encoding transposases and their inactivated derivatives (COG L) was 1.33% of the total genes with COGs classifications in A1501, 0.21% in GP72, 0.02% in Pf-5, and 0.11% in M18. A phylogenetic analysis indicated that GP72 and Pf-5 were the most closely related strains, consistent with the genome alignment results. Comparisons of predicted coding sequences (CDSs) between GP72 and Pf-5 revealed 3544 conserved genes. There were fewer conserved genes when GP72 CDSs were compared with those of A1501 and M18. Comparisons among the four Pseudomonas species revealed 603 conserved genes in GP72, illustrating common plant growth-promoting traits shared among these PGPR. Conserved genes were related to catabolism, transport of plant-derived compounds, stress resistance, and rhizosphere colonization. Some strain-specific CDSs were related to different kinds of biocontrol activities or plant growth promotion. The GP72 genome

  18. Use of Antimicrobial Food Additives as Potential Dipping Solutions to Control Pseudomonas spp. Contamination in the Frankfurters and Ham.

    Science.gov (United States)

    Oh, Mi-Hwa; Park, Beom-Young; Jo, Hyunji; Lee, Soomin; Lee, Heeyoung; Choi, Kyoung-Hee; Yoon, Yohan

    2014-01-01

    This study evaluated the effect of sodium diacetate and sodium lactate solutions for reducing the cell count of Pseudomonas spp. in frankfurters and hams. A mixture of Pseudomonas aeruginosa (NCCP10338, NCCP10250, and NCCP11229), and Pseudomonas fluorescens (KACC10323 and KACC10326) was inoculated on cooked frankfurters and ham. The inoculated samples were immersed into control (sterile distilled water), sodium diacetate (5 and 10%), sodium lactate (5 and 10%), 5% sodium diacetate + 5% sodium lactate, and 10% sodium diacetate + 10% sodium lactate for 0-10 min. Inoculated frankfurters and ham were also immersed into acidified (pH 3.0) solutions such as acidified sodium diacetate (5 and 10%), and acidified sodium lactate (5 and 10%) in addition to control (acidified distilled water) for 0-10 min. Total aerobic plate counts for Pseudomonas spp. were enumerated on Cetrimide agar. Significant reductions (ca. 2 Log CFU/g) in Pseudomonas spp. cells on frankfurters and ham were observed only for a combination treatment of 10% sodium lactate + 10% sodium diacetate. When the solutions were acidified to pH 3.0, the total reductions of Pseudomonas spp. were 1.5-4.0 Log CFU/g. The order of reduction amounts of Pseudomonas spp. cell counts was 10% sodium lactate > 5% sodium lactate ≥ 10% sodium diacetate > 5% sodium diacetate > control for frankfurters, and 10% sodium lactate > 5% sodium lactate > 10% sodium diacetate > 5% sodium diacetate > control for ham. The results suggest that using acidified food additive antimicrobials, as dipping solutions, should be useful in reducing Pseudomonas spp. on frankfurters and ham.

  19. Presence of rhlAB, rhlR and rhlR genes in Pseudomonas aeruginosa natives overproductors of ramnolipids

    Directory of Open Access Journals (Sweden)

    Roger A. Palomino

    2017-10-01

    Full Text Available Genes associated to rhamnolipids production were molecularly characterized in 61 bacterial strains from LAMYBIM bacterial collection (Laboratorio de Microbiología y Biotecnología Microbiana, Universidad Nacional Mayor de San Marcos, Perú. Strains were isolated from peruvian environments hydrocarbons polluted and were classified as RL overproducers (n= 21, RL producers (n = 20 and non-producers (n = 20 producers. Molecular identification using the 16S rRNA gene was preceded by the biochemical identification of 61 strains selected with the API 20 NE system. Pseudomonas aeruginosa was the most prevalent strain of the RL overproducers and RL producers. Species such as Burkholderia cepacea, Pseudomonas fluorescens, Aeromonas hydrophila and Chryseobacterium indologenes, were found too. In the same way, non-producers microorganisms were also characterized. The PCR amplification and agarose gel electrophoresis techniques, standarized by the UNAM laboratory, showed that the selected strains had the genes: rhlA, rhlB, rhlR and rhlC. For the sequencing of the rhLABR gene region, four strains were selected: Pseudomonas aeruginosa T2K2, Pseudomonas aeruginosa III T1P2, Pseudomonas aeruginosa 6K-11 and Pseudomonas aeruginosa ATCC 9027, applying the methodology standardized by the UNAM and were compared with Pseudomonas aeruginosa PAO1. Our results show that the genes studied in the selected strains are synonymous with their homologues in Pseudomonas aeruginosa PAO1 standard strain. Therefore, genotypical differences that explain the overproduction of rhamnolipid might be found in other molecular markers not covered in this study.

  20. In-silico Taxonomic Classification of 373 Genomes Reveals Species Misidentification and New Genospecies within the Genus Pseudomonas

    Directory of Open Access Journals (Sweden)

    Phuong N. Tran

    2017-07-01

    Full Text Available The genus Pseudomonas has one of the largest diversity of species within the Bacteria kingdom. To date, its taxonomy is still being revised and updated. Due to the non-standardized procedure and ambiguous thresholds at species level, largely based on 16S rRNA gene or conventional biochemical assay, species identification of publicly available Pseudomonas genomes remains questionable. In this study, we performed a large-scale analysis of all Pseudomonas genomes with species designation (excluding the well-defined P. aeruginosa and re-evaluated their taxonomic assignment via in silico genome-genome hybridization and/or genetic comparison with valid type species. Three-hundred and seventy-three pseudomonad genomes were analyzed and subsequently clustered into 145 distinct genospecies. We detected 207 erroneous labels and corrected 43 to the proper species based on Average Nucleotide Identity Multilocus Sequence Typing (MLST sequence similarity to the type strain. Surprisingly, more than half of the genomes initially designated as Pseudomonas syringae and Pseudomonas fluorescens should be classified either to a previously described species or to a new genospecies. Notably, high pairwise average nucleotide identity (>95% indicating species-level similarity was observed between P. synxantha-P. libanensis, P. psychrotolerans–P. oryzihabitans, and P. kilonensis- P. brassicacearum, that were previously differentiated based on conventional biochemical tests and/or genome-genome hybridization techniques.

  1. Antibiotic and biosurfactant properties of cyclic lipopeptides produced by fluorescent Pseudomonas spp. from the sugar beet rhizosphere

    DEFF Research Database (Denmark)

    Nielsen, T H; Sørensen, D; Tobiasen, C

    2002-01-01

    Cyclic lipopeptides (CLPs) with antibiotic and biosurfactant properties are produced by a number of soil bacteria, including fluorescent Pseudomonas spp. To provide new and efficient strains for the biological control of root-pathogenic fungi in agricultural crops, we isolated approximately 600...... fluorescent Pseudomonas spp. from two different agricultural soils by using three different growth media. CLP production was observed in a large proportion of the strains (approximately 60%) inhabiting the sandy soil, compared to a low proportion (approximately 6%) in the loamy soil. Chemical structure...... and that some also had antibiotic properties towards root-pathogenic microfungi. The CLP-producing P. fluorescens strains provide a useful resource for selection of biological control agents, whether a single strain or a consortium of strains was used to maximize the synergistic effect of multiple antagonistic...

  2. Biological control of Sclerotinia sclerotiorum (Lib.) de Bary, the causal agent of white mold, by Pseudomonas species on canola petals.

    Science.gov (United States)

    Behnam, S; Ahmadzadeh, M; Sharifi Tehrani, A; Hedjaroude, Gh A; Farzaneh, M

    2007-01-01

    Sclerotinia sclerotiorum is an important pathogen on canola. Due to the public concern over pesticide use, alternative methods of disease control, such as biological control, should be considered. Several bacterial strains were isolated from canola and soja plants. Inhibition of S. sclerotiorum by bacterial strains in vitro was assayed on PDA medium in dual culture test. Eight Pseudomonas sp. strains (PB-3, PB-4, PB-5, PB-6, PB-7, PB-8, PB-10 and PB-11) caused inhibition zone against 5. sclerotiorum hyphal growth. The biocontrol potential of the bacteria was tested in a plant assay. Disease suppression was investigated using a petal inoculation technique. Canola petals were pretreated with bacteria, and then inoculated with 5. sclerotiorum ascospores 24 h later. Greenhouse experiment showed that application of Pseudomonas sp. strains (1 x 10(8) cfu ml(-1)) effectively suppressed S. sclerotiorum (1 x 10(5) ascospores ml(-1)) on petals and all of them achieved significant (P<0.01) disease suppression. Fourteen days after inoculation, strain PB-3 had 88/7% disease control and strain PB-4 had 69/9% disease control. Result from all studies indicates PB-3 to be effective biocontrol against S. sclerotiorum of canola. PB-3, PB-4, PB-7, PB-8, PB-10 and PB-11 were identified as Pseudomonas fluorescens biovar III. PB-5 and PB-6 was identified as Pseudomonas fluorescens biovar II. Strains PB-3, PB-4, PB-6, PB-10 and PB-11 produced protease and HCN. Strain PB-5 produce protease; no HCN.

  3. Generating social capital in globalised networks for growth and development in nascent entrepreneurial ventures

    OpenAIRE

    Simba, A

    2016-01-01

    Global markets are no longer dominated by multinational enterprises (MNEs) alone, international new ventures (INVs) or born globals are increasingly becoming serious contenders in terms of employment & wealth creation as well as revenue generation. In seeking to penetrate global markets they often rely on their entrepreneurial behaviours. Specifically, these nascent but entrepreneurial firms often take advantage of existing and newly developed networks, both domestically and internationally, ...

  4. Glyphosate-Induced Specific and Widespread Perturbations in the Metabolome of Soil Pseudomonas Species

    Directory of Open Access Journals (Sweden)

    Ludmilla Aristilde

    2017-06-01

    Full Text Available Previous studies have reported adverse effects of glyphosate on crop-beneficial soil bacterial species, including several soil Pseudomonas species. Of particular interest is the elucidation of the metabolic consequences of glyphosate toxicity in these species. Here we investigated the growth and metabolic responses of soil Pseudomonas species grown on succinate, a common root exudate, and glyphosate at different concentrations. We conducted our experiments with one agricultural soil isolate, P. fluorescens RA12, and three model species, P. putida KT2440, P. putida S12, and P. protegens Pf-5. Our results demonstrated both species- and strain-dependent growth responses to glyphosate. Following exposure to a range of glyphosate concentrations (up to 5 mM, the growth rate of both P. protegens Pf-5 and P. fluorescens RA12 remained unchanged whereas the two P. putida strains exhibited from 0 to 100% growth inhibition. We employed a 13C-assisted metabolomics approach using liquid chromatography-mass spectrometry to monitor disruptions in metabolic homeostasis and fluxes. Profiling of the whole-cell metabolome captured deviations in metabolite levels involved in the tricarboxylic acid cycle, ribonucleotide biosynthesis, and protein biosynthesis. Altered metabolite levels specifically in the biosynthetic pathway of aromatic amino acids (AAs, the target of toxicity for glyphosate in plants, implied the same toxicity target in the soil bacterium. Kinetic flux experiments with 13C-labeled succinate revealed that biosynthetic fluxes of the aromatic AAs were not inhibited in P. fluorescens Pf-5 in the presence of low and high glyphosate doses but these fluxes were inhibited by up to 60% in P. putida KT2440, even at sub-lethal glyphosate exposure. Notably, the greatest inhibition was found for the aromatic AA tryptophan, an important precursor to secondary metabolites. When the growth medium was supplemented with aromatic AAs, P. putida S12 exposed to a lethal

  5. Antagonism between two root-associated beneficial Pseudomonas strains does not affect plant growth promotion and induced resistance against a leaf-chewing herbivore.

    Science.gov (United States)

    Pangesti, Nurmi; Vandenbrande, Simon; Pineda, Ana; Dicke, Marcel; Raaijmakers, Jos M; Van Loon, Joop J A

    2017-04-01

    Plant growth-promoting microbes residing on the roots may cooperate or compete, thereby affecting their collective benefit to the host plant. Pseudomonas simiae WCS417r (formerly known as P. fluorescens WCS417r) and Pseudomonas fluorescens SS101 are well known for their ability to induce systemic resistance in Arabidopsis. Here, we evaluate how these species interact on the roots of Arabidopsis thaliana Col-0 and how their co-inoculation affects plant defense to the leaf-chewing herbivore Mamestra brassicae and plant growth promotion. WCS417r and SS101, applied individually to root tips or at two different positions along the roots, established similar population densities on Arabidopsis roots. When co-inoculated at the same position on the roots, however, WCS417r established significantly higher population densities than SS101. Both upon single inoculation and co-inoculation, the two pseudomonads induced the same level of induced systemic resistance against the caterpillar M. brassicae and the same increase in plant biomass. These results suggest that combined inoculation of both Pseudomonas strains does not significantly modify the plant's defensive capacity compared to individual inoculation, resulting in a similar effect on performance of the generalist herbivore M. brassicae. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Elastic coupling of nascent apCAM adhesions to flowing actin networks.

    Science.gov (United States)

    Mejean, Cecile O; Schaefer, Andrew W; Buck, Kenneth B; Kress, Holger; Shundrovsky, Alla; Merrill, Jason W; Dufresne, Eric R; Forscher, Paul

    2013-01-01

    Adhesions are multi-molecular complexes that transmit forces generated by a cell's acto-myosin networks to external substrates. While the physical properties of some of the individual components of adhesions have been carefully characterized, the mechanics of the coupling between the cytoskeleton and the adhesion site as a whole are just beginning to be revealed. We characterized the mechanics of nascent adhesions mediated by the immunoglobulin-family cell adhesion molecule apCAM, which is known to interact with actin filaments. Using simultaneous visualization of actin flow and quantification of forces transmitted to apCAM-coated beads restrained with an optical trap, we found that adhesions are dynamic structures capable of transmitting a wide range of forces. For forces in the picoNewton scale, the nascent adhesions' mechanical properties are dominated by an elastic structure which can be reversibly deformed by up to 1 µm. Large reversible deformations rule out an interface between substrate and cytoskeleton that is dominated by a number of stiff molecular springs in parallel, and favor a compliant cross-linked network. Such a compliant structure may increase the lifetime of a nascent adhesion, facilitating signaling and reinforcement.

  7. Elastic coupling of nascent apCAM adhesions to flowing actin networks.

    Directory of Open Access Journals (Sweden)

    Cecile O Mejean

    Full Text Available Adhesions are multi-molecular complexes that transmit forces generated by a cell's acto-myosin networks to external substrates. While the physical properties of some of the individual components of adhesions have been carefully characterized, the mechanics of the coupling between the cytoskeleton and the adhesion site as a whole are just beginning to be revealed. We characterized the mechanics of nascent adhesions mediated by the immunoglobulin-family cell adhesion molecule apCAM, which is known to interact with actin filaments. Using simultaneous visualization of actin flow and quantification of forces transmitted to apCAM-coated beads restrained with an optical trap, we found that adhesions are dynamic structures capable of transmitting a wide range of forces. For forces in the picoNewton scale, the nascent adhesions' mechanical properties are dominated by an elastic structure which can be reversibly deformed by up to 1 µm. Large reversible deformations rule out an interface between substrate and cytoskeleton that is dominated by a number of stiff molecular springs in parallel, and favor a compliant cross-linked network. Such a compliant structure may increase the lifetime of a nascent adhesion, facilitating signaling and reinforcement.

  8. Potencial da nascente do Rio Vieira e entorno: uma interpretação da geodiversidade

    Directory of Open Access Journals (Sweden)

    Rachel Inêz Castro de Oliveira

    2017-12-01

    Full Text Available Este artigo contempla o aproveitamento dos elementos geológicos, geomorfológicos, espeleológicos, pedológicos e hidrográficos que compõem a geodiversidade como recurso educativo, turístico e interpretativo, em uma das principais nascentes do Rio Vieira e entorno, no município de Montes Claros, Norte de Minas. Considerando a especificidade desse campo da geodiversidade, o objetivo foi investigar as potencialidades da nascente do Rio Vieira e entorno, por meio do inventário dos elementos representativos da geodiversidade. A abordagem metodológica consistiu em fases concomitantes: levantamento bibliográfico, trabalhos de campo e elaboração de mapas. A área de estudo foi inventariada, caracterizando a geodiversidade local. As inferências e constatações produzidas possibilitaram compreender que existe potencial da nascente do Rio Vieira e entorno para a interpretação da geodiversidade. Espera-se que este estudo contribua fornecendo subsídios para que mais pesquisas sejam realizadas na área.

  9. Bioluminescence of Pseudomonas Fluorescens HK44 in the Course of Encapsulation into Silica Gel. Effect of Methanol

    Czech Academy of Sciences Publication Activity Database

    Trögl, J.; Kuncová, Gabriela; Kuráň, P.

    2010-01-01

    Roč. 55, č. 6 (2010), s. 569-575 ISSN 0015-5632 R&D Projects: GA MŠk ME 893 Institutional research plan: CEZ:AV0Z40720504 Keywords : catabolic reporter bacterium * recombinant escheria-coli * optical biosensor Subject RIV: CE - Biochemistry Impact factor: 0.977, year: 2010

  10. Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

    NARCIS (Netherlands)

    Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

    1998-01-01

    The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative. denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing bacteria,

  11. Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

    NARCIS (Netherlands)

    Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

    1998-01-01

    The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative, denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing

  12. Structure-function relationship of flavoproteins : with special reference to p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens

    NARCIS (Netherlands)

    Berkel, van W.J.H.

    1989-01-01

    In this thesis different studies probing the structurefunction relationship of some flavoproteins are dealt with. The attention has been focused on two central themes:
    The first part of the thesis deals with studies concerning the application of affinity

  13. Effect of organic management of soils on suppressiveness to Gaeumannomyces graminis var. tritici and its antagonist, Pseudomonas fluorescens

    NARCIS (Netherlands)

    Hiddink, G.A.; Bruggen, van A.H.C.; Termorshuizen, A.J.; Raaijmakers, J.M.; Semenov, A.V.

    2005-01-01

    Organic management of soils is generally considered to reduce the incidence and severity of plant diseases caused by soil-borne pathogens. In this study, take-all severity on roots of barley and wheat, caused by Gaeumannomyces graminis var. tritici, was significantly lower in organically-managed

  14. No apparent costs for facultative antibiotic production by the soil bacterium Pseudomonas fluorescens Pf0-1

    NARCIS (Netherlands)

    Garbeva, P.V.; Tyc, O.; Remus-Emsermann, M.N.P.; Van der Wal, A.; Vos, M.; Silby, M.W.; De Boer, W.

    2011-01-01

    Background: Many soil-inhabiting bacteria are known to produce secondary metabolites that can suppress microorganisms competing for the same resources. The production of antimicrobial compounds is expected to incur fitness costs for the producing bacteria. Such costs form the basis for models on the

  15. Pseudomonas fluorescens HK44: Lessons Lerned from a Model Whole-Cell Bioreporter with a Broad Application History

    Czech Academy of Sciences Publication Activity Database

    Trögl, J.; Chauhan, A.; Ripp, S.; Layton, A.C.; Kuncová, Gabriela; Sayler, G.S.

    2012-01-01

    Roč. 12, č. 2 (2012), s. 1544-1571 ISSN 1424-8220 R&D Projects: GA MŠk ME 892; GA MŠk ME 893 Grant - others:USDA(US) 2009-39210-20230 Institutional research plan: CEZ:AV0Z40720504 Keywords : bioluminiscence * bioreporter * biosensors Subject RIV: EI - Biotechnology ; Bionics Impact factor: 1.953, year: 2012

  16. No Apparent Costs for Facultative Antibiotic Production by the Soil Bacterium Pseudomonas fluorescens Pf0-1

    OpenAIRE

    Garbeva, Paolina; Tyc, Olaf; Remus-Emsermann, Mitja N. P.; van der Wal, Annemieke; Vos, Michiel; Silby, Mark; de Boer, Wietse

    2011-01-01

    BACKGROUND: Many soil-inhabiting bacteria are known to produce secondary metabolites that can suppress microorganisms competing for the same resources. The production of antimicrobial compounds is expected to incur fitness costs for the producing bacteria. Such costs form the basis for models on the co-existence of antibiotic-producing and non-antibiotic producing strains. However, so far studies quantifying the costs of antibiotic production by bacteria are scarce. The current study reports ...

  17. Polychlorinated biphenyl rhizoremediation by Pseudomonas fluorescens F113 derivatives, using a Sinorhizobium meliloti nod system to drive bph gene expression

    Czech Academy of Sciences Publication Activity Database

    Villacieros, M.; Whelan, C.; Macková, Martina; Molgaard, J.; Sánchez-Contreras, M.; Lloret, J.; Cárcer de, D.; Oruezábal, R. I.; Bolanos, L.; Macek, Tomáš; Karlson, U.; Dowling, D. N.; Martín, M.; Rivilla, R.

    2005-01-01

    Roč. 71, č. 5 (2005), 2687-2694 ISSN 0099-2240 Grant - others:5thFW(XE) QLK3-CT-2001-00101; European Commission(XE) BIO4-CT97-2227 Institutional research plan: CEZ:AV0Z4055905 Keywords : polychlorinated biphenyls * rhizoremediation Subject RIV: CC - Organic Chemistry Impact factor: 3.818, year: 2005

  18. Protozoan-induced regulation of cyclic lipopeptide biosynthesis Is an effective predation defense mechanism for Pseudomonas fluorescens

    NARCIS (Netherlands)

    Mazzola, M.; Bruijn, de I.; Cohen, M.F.; Raaijmakers, J.M.

    2009-01-01

    Environmental bacteria are exposed to a myriad of biotic interactions that influence their function and survival. The grazing activity of protozoan predators significantly impacts the dynamics, diversification, and evolution of bacterial communities in soil ecosystems. To evade protozoan predation,

  19. Lógica Fuzzy e processo analítico hierárquico – AHP na avaliação da qualidade ambiental de nascentes

    OpenAIRE

    Silva Junior, Celso Henrique Leite; Freire, Ana Talita Galvão; Mendes, Jonas Jansen

    2015-01-01

    O presente trabalho objetivou a avaliação da qualidade ambiental das nascentes do rio Bacanga utilizando uma metodologia pouco empregada nesses tipos de estudos, a Teoria Fuzzy. No cálculo do Índice de Qualidade Ambiental das Nascentes, para a ponderação dos parâmetros foi utilizado o AHP (Analytic Hierarchy Process - Processo Analítico Hierárquico). Como resultado, das 11 nascentes analisadas 5 nascentes foram enquadradas na classe A com Qualidade Ambiental Ótima, 3 nascentes foram enquadrad...

  20. Bacterial Feeders, the Nematode Caenorhabditis elegans and the Flagellate Cercomonas longicauda, have different Effects on Outcome of Competition among the Pseudomonas Biocontrol Strains CHA0 and DSS73

    DEFF Research Database (Denmark)

    Pedersen, Annette; Nybroe, Ole; Winding, Anne

    2009-01-01

    50090 or one of two biocontrol strains P. fluorescens CHA0 or Pseudomonas sp. DSS73) or combinations of two bacterial strains. DSM50090 is a suitable food bacterium, DSS73 is of intermediate food quality, and CHA0 is inedible to the bacterial feeders. Bacterial and protozoan cell numbers were measured......How bacterial feeding fauna affects colonization and survival of bacteria in soil is not well understood, which constrains the applicability of bacterial inoculants in agriculture. This study aimed to unravel how food quality of bacteria and bacterial feeders with different feeding habits (the...

  1. Gentamicin in Pseudomonas aeruginosa

    African Journals Online (AJOL)

    infections by Ps. aeruginosa is contra-indicated. In our study only 2,3 % of the Ps. aeruginosa strains were resistant to gentamicin (MIC 25 Ilg/ml). In view of the synergy reported for combined gentamicin and carbeni- cillin therapy," a combination of these two drugs may be recommended in the treatment of all Pseudomonas.

  2. Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Alhede, Maria; Bjarnsholt, Thomas; Givskov, Michael

    2014-01-01

    The opportunistic gram-negative bacterium Pseudomonas aeruginosa is implicated in many chronic infections and is readily isolated from chronic wounds, medical devices, and the lungs of cystic fibrosis patients. P. aeruginosa is believed to persist in the host organism due to its capacity to form...

  3. Pseudomonas aeruginosa biofilm infections

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2014-01-01

    the use of conventional antimicrobial compounds in many cases cannot eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections. The present review is focussed on the important opportunistic pathogen and biofilm model organism Pseudomonas aeruginosa. Initially...

  4. Uptake of Hydrocarbon by Pseudomonas fluorescens (P1) and Pseudomonas putida (K1) Strains in the Presence of Surfactants: A Cell Surface Modification

    OpenAIRE

    Kaczorek, Ewa; Olszanowski, Andrzej

    2010-01-01

    The objective of this research was the evaluation of the effects of exogenous added surfactants on hydrocarbon biodegradation and on cell surface properties. Crude oil hydrocarbons are often difficult to remove from the environment because of their insolubility in water. The addition of surfactants enhances the removal of hydrocarbons by raising the solubility of these compounds. These surfactants cause them to become more vulnerable to degradation, thereby facilitating transportation across ...

  5. The complete genome sequence of the plant growth-promoting bacterium Pseudomonas sp. UW4.

    Directory of Open Access Journals (Sweden)

    Jin Duan

    Full Text Available The plant growth-promoting bacterium (PGPB Pseudomonas sp. UW4, previously isolated from the rhizosphere of common reeds growing on the campus of the University of Waterloo, promotes plant growth in the presence of different environmental stresses, such as flooding, high concentrations of salt, cold, heavy metals, drought and phytopathogens. In this work, the genome sequence of UW4 was obtained by pyrosequencing and the gaps between the contigs were closed by directed PCR. The P. sp. UW4 genome contains a single circular chromosome that is 6,183,388 bp with a 60.05% G+C content. The bacterial genome contains 5,423 predicted protein-coding sequences that occupy 87.2% of the genome. Nineteen genomic islands (GIs were predicted and thirty one complete putative insertion sequences were identified. Genes potentially involved in plant growth promotion such as indole-3-acetic acid (IAA biosynthesis, trehalose production, siderophore production, acetoin synthesis, and phosphate solubilization were determined. Moreover, genes that contribute to the environmental fitness of UW4 were also observed including genes responsible for heavy metal resistance such as nickel, copper, cadmium, zinc, molybdate, cobalt, arsenate, and chromate. Whole-genome comparison with other completely sequenced Pseudomonas strains and phylogeny of four concatenated "housekeeping" genes (16S rRNA, gyrB, rpoB and rpoD of 128 Pseudomonas strains revealed that UW4 belongs to the fluorescens group, jessenii subgroup.

  6. Comparison of prominent Azospirillum strains in Azospirillum-Pseudomonas-Glomus consortia for promotion of maize growth.

    Science.gov (United States)

    Couillerot, Olivier; Ramírez-Trujillo, Augusto; Walker, Vincent; von Felten, Andreas; Jansa, Jan; Maurhofer, Monika; Défago, Geneviève; Prigent-Combaret, Claire; Comte, Gilles; Caballero-Mellado, Jesus; Moënne-Loccoz, Yvan

    2013-05-01

    Azospirillum are prominent plant growth-promoting rhizobacteria (PGPR) extensively used as phytostimulatory crop inoculants, but only few studies are dealing with Azospirillum-containing mixed inocula involving more than two microorganisms. We compared here three prominent Azospirillum strains as part of three-component consortia including also the PGPR Pseudomonas fluorescens F113 and a mycorrhizal inoculant mix composed of three Glomus strains. Inoculant colonization of maize was assessed by quantitative PCR, transcription of auxin synthesis gene ipdC (involved in phytostimulation) in Azospirillum by RT-PCR, and effects on maize by secondary metabolic profiling and shoot biomass measurements. Results showed that phytostimulation by all the three-component consortia was comparable, despite contrasted survival of the Azospirillum strains and different secondary metabolic responses of maize to inoculation. Unexpectedly, the presence of Azospirillum in the inoculum resulted in lower phytostimulation in comparison with the Pseudomonas-Glomus two-component consortium, but this effect was transient. Azospirillum's ipdC gene was transcribed in all treatments, especially with three-component consortia, but not with all plants and samplings. Inoculation had no negative impact on the prevalence of mycorrhizal taxa in roots. In conclusion, this study brought new insights in the functioning of microbial consortia and showed that Azospirillum-Pseudomonas-Glomus three-component inoculants may be useful in environmental biotechnology for maize growth promotion.

  7. Virulence Attributes and Host Response Assays for Determining Pathogenic Potential of Pseudomonas Strains Used in Biotechnology.

    Directory of Open Access Journals (Sweden)

    Azam F Tayabali

    Full Text Available Pseudomonas species are opportunistically pathogenic to humans, yet closely related species are used in biotechnology applications. In order to screen for the pathogenic potential of strains considered for biotechnology applications, several Pseudomonas strains (P.aeruginosa (Pa, P.fluorescens (Pf, P.putida (Pp, P.stutzeri (Ps were compared using functional virulence and toxicity assays. Most Pa strains and Ps grew at temperatures between 28°C and 42°C. However, Pf and Pp strains were the most antibiotic resistant, with ciprofloxacin and colistin being the most effective of those tested. No strain was haemolytic on sheep blood agar. Almost all Pa, but not other test strains, produced a pyocyanin-like chromophore, and caused cytotoxicity towards cultured human HT29 cells. Murine endotracheal exposures indicated that the laboratory reference strain, PAO1, was most persistent in the lungs. Only Pa strains induced pro-inflammatory and inflammatory responses, as measured by elevated cytokines and pulmonary Gr-1 -positive cells. Serum amyloid A was elevated at ≥ 48 h post-exposure by only some Pa strains. No relationship was observed between strains and levels of peripheral leukocytes. The species designation or isolation source may not accurately reflect pathogenic potential, since the clinical strain Pa10752 was relatively nonvirulent, but the industrial strain Pa31480 showed comparable virulence to PAO1. Functional assays involving microbial growth, cytotoxicity and murine immunological responses may be most useful for identifying problematic Pseudomonas strains being considered for biotechnology applications.

  8. Virulence Attributes and Host Response Assays for Determining Pathogenic Potential of Pseudomonas Strains Used in Biotechnology

    Science.gov (United States)

    Tayabali, Azam F.; Coleman, Gordon; Nguyen, Kathy C.

    2015-01-01

    Pseudomonas species are opportunistically pathogenic to humans, yet closely related species are used in biotechnology applications. In order to screen for the pathogenic potential of strains considered for biotechnology applications, several Pseudomonas strains (P.aeruginosa (Pa), P.fluorescens (Pf), P.putida (Pp), P.stutzeri (Ps)) were compared using functional virulence and toxicity assays. Most Pa strains and Ps grew at temperatures between 28°C and 42°C. However, Pf and Pp strains were the most antibiotic resistant, with ciprofloxacin and colistin being the most effective of those tested. No strain was haemolytic on sheep blood agar. Almost all Pa, but not other test strains, produced a pyocyanin-like chromophore, and caused cytotoxicity towards cultured human HT29 cells. Murine endotracheal exposures indicated that the laboratory reference strain, PAO1, was most persistent in the lungs. Only Pa strains induced pro-inflammatory and inflammatory responses, as measured by elevated cytokines and pulmonary Gr-1 -positive cells. Serum amyloid A was elevated at ≥ 48 h post-exposure by only some Pa strains. No relationship was observed between strains and levels of peripheral leukocytes. The species designation or isolation source may not accurately reflect pathogenic potential, since the clinical strain Pa10752 was relatively nonvirulent, but the industrial strain Pa31480 showed comparable virulence to PAO1. Functional assays involving microbial growth, cytotoxicity and murine immunological responses may be most useful for identifying problematic Pseudomonas strains being considered for biotechnology applications. PMID:26619347

  9. The Complete Genome Sequence of the Plant Growth-Promoting Bacterium Pseudomonas sp. UW4

    Science.gov (United States)

    Duan, Jin; Jiang, Wei; Cheng, Zhenyu; Heikkila, John J.; Glick, Bernard R.

    2013-01-01

    The plant growth-promoting bacterium (PGPB) Pseudomonas sp. UW4, previously isolated from the rhizosphere of common reeds growing on the campus of the University of Waterloo, promotes plant growth in the presence of different environmental stresses, such as flooding, high concentrations of salt, cold, heavy metals, drought and phytopathogens. In this work, the genome sequence of UW4 was obtained by pyrosequencing and the gaps between the contigs were closed by directed PCR. The P. sp. UW4 genome contains a single circular chromosome that is 6,183,388 bp with a 60.05% G+C content. The bacterial genome contains 5,423 predicted protein-coding sequences that occupy 87.2% of the genome. Nineteen genomic islands (GIs) were predicted and thirty one complete putative insertion sequences were identified. Genes potentially involved in plant growth promotion such as indole-3-acetic acid (IAA) biosynthesis, trehalose production, siderophore production, acetoin synthesis, and phosphate solubilization were determined. Moreover, genes that contribute to the environmental fitness of UW4 were also observed including genes responsible for heavy metal resistance such as nickel, copper, cadmium, zinc, molybdate, cobalt, arsenate, and chromate. Whole-genome comparison with other completely sequenced Pseudomonas strains and phylogeny of four concatenated “housekeeping” genes (16S rRNA, gyrB, rpoB and rpoD) of 128 Pseudomonas strains revealed that UW4 belongs to the fluorescens group, jessenii subgroup. PMID:23516524

  10. 40 CFR 180.1154 - CryIA(c) and CryIC derived delta-endotoxins of Bacillus thuringiensis var. kurstaki encapsulated...

    Science.gov (United States)

    2010-07-01

    ...-endotoxins of Bacillus thuringiensis var. kurstaki encapsulated in killed Pseudomonas fluorescens, and the... Bacillus thuringiensis var. kurstaki encapsulated in killed Pseudomonas fluorescens, and the expression... thuringiensis var. kurstaki encapsulated in killed Pseudomonas fluorescens and the expression plasmid and...

  11. Probing the role of nascent helicity in p27 function as a cell cycle regulator.

    Directory of Open Access Journals (Sweden)

    Steve Otieno

    Full Text Available p27 regulates the activity of Cdk complexes which are the principal governors of phase transitions during cell division. Members of the p27 family of proteins, which also includes p21 and p57, are called the Cip/Kip cyclin-dependent kinase regulators (CKRs. Interestingly, the Cip/Kip CKRs play critical roles in cell cycle regulation by being intrinsically unstructured, a characteristic contrary to the classical structure-function paradigm. They exhibit nascent helicity which has been localized to a segment referred to as sub-domain LH. The nascent helicity of this sub-domain is conserved and we hypothesize that it is an important determinant of their functional properties. To test this hypothesis, we successfully designed and prepared p27 variants in which domain LH was either more or less helical with respect to the wild-type protein. Thermal denaturation experiments showed that the ternary complexes of the p27 variants bound to Cdk2/Cyclin A were less stable compared to the wild-type complex. Isothermal titration calorimetry experiments showed a decrease in the enthalpy of binding for all the mutants with respect to p27. The free energies of binding varied within a much narrower range. In vitro Cdk2 inhibition assays showed that the p27 variants exhibited disparate inhibitory potencies. Furthermore, when over-expressed in NIH 3T3 mouse fibroblast cells, the less helical p27 variants were less effective in causing cell cycle arrest relative to the wild-type p27. Our results indicate that the nascent helicity of sub-domain LH plays a key role mediating the biological function of p27.

  12. Nascent histamine induces α-synuclein and caspase-3 on human cells

    Energy Technology Data Exchange (ETDEWEB)

    Caro-Astorga, Joaquín; Fajardo, Ignacio; Ruiz-Pérez, María Victoria; Sánchez-Jiménez, Francisca; Urdiales, José Luis, E-mail: jlurdial@uma.es

    2014-09-05

    Highlights: • Nascent histamine alters cyclin expression pattern. • Nascent histamine increases expression of α-synuclein. • Nascent histamine activates caspase-3. - Abstract: Histamine (Hia) is the most multifunctional biogenic amine. It is synthetized by histidine decarboxylase (HDC) in a reduced set of mammalian cell types. Mast cells and histaminergic neurons store Hia in specialized organelles until the amine is extruded by exocytosis; however, other immune and cancer cells are able to produce but not store Hia. The intracellular effects of Hia are still not well characterized, in spite of its physiopathological relevance. Multiple functional relationships exist among Hia metabolism/signaling elements and those of other biogenic amines, including growth-related polyamines. Previously, we obtained the first insights for an inhibitory effect of newly synthetized Hia on both growth-related polyamine biosynthesis and cell cycle progression of non-fully differentiated mammalian cells. In this work, we describe progress in this line. HEK293 cells were transfected to express active and inactive versions of GFP-human HDC fusion proteins and, after cell sorting by flow cytometry, the relative expression of a large number of proteins associated with cell signaling were measured using an antibody microarray. Experimental results were analyzed in terms of protein–protein and functional interaction networks. Expression of active HDC induced a cell cycle arrest through the alteration of the levels of several proteins such as cyclin D1, cdk6, cdk7 and cyclin A. Regulation of α-synuclein and caspase-3 was also observed. The analyses provide new clues on the molecular mechanisms underlying the regulatory effects of intracellular newly synthetized Hia on cell proliferation/survival, cell trafficking and protein turnover. This information is especially interesting for emergent and orphan immune and neuroinflammatory diseases.

  13. On the Response of Nascent Soot Nanostructure and Oxidative Reactivity to Photoflash Exposure

    Directory of Open Access Journals (Sweden)

    Wei Wang

    2017-07-01

    Full Text Available Soot particles are a kind of major pollutant from fuel combustion. To enrich the understanding of soot, this work focuses on investigating detailed influences of instantaneous external irradiation (conventional photoflash exposure on nanostructure as well as oxidation reactivity of nascent soot particles. By detailed soot characterizations flash can reduce the mass of soot and soot nanostructure can be reconstructed substantially without burning. After flash, the degree of soot crystallization increases while the soot reactive rate decreases and the activation energy increases. In addition, nanostructure and oxidative reactivity of soot in air and Ar after flash are different due to their different thermal conductivities.

  14. Radial glial neural progenitors regulate nascent brain vascular network stabilization via inhibition of Wnt signaling.

    Directory of Open Access Journals (Sweden)

    Shang Ma

    Full Text Available The cerebral cortex performs complex cognitive functions at the expense of tremendous energy consumption. Blood vessels in the brain are known to form stereotypic patterns that facilitate efficient oxygen and nutrient delivery. Yet little is known about how vessel development in the brain is normally regulated. Radial glial neural progenitors are well known for their central role in orchestrating brain neurogenesis. Here we show that, in the late embryonic cortex, radial glial neural progenitors also play a key role in brain angiogenesis, by interacting with nascent blood vessels and regulating vessel stabilization via modulation of canonical Wnt signaling. We find that ablation of radial glia results in vessel regression, concomitant with ectopic activation of Wnt signaling in endothelial cells. Direct activation of Wnt signaling also results in similar vessel regression, while attenuation of Wnt signaling substantially suppresses regression. Radial glial ablation and ectopic Wnt pathway activation leads to elevated endothelial expression of matrix metalloproteinases, while inhibition of metalloproteinase activity significantly suppresses vessel regression. These results thus reveal a previously unrecognized role of radial glial progenitors in stabilizing nascent brain vascular network and provide novel insights into the molecular cascades through which target neural tissues regulate vessel stabilization and patterning during development and throughout life.

  15. High Efficient Reduction of Graphene Oxide via Nascent Hydrogen at Room Temperature

    Directory of Open Access Journals (Sweden)

    Qiqi Zhuo

    2018-02-01

    Full Text Available To develop a green and efficient method to synthesize graphene in relative milder conditions is prerequisite for graphene applications. A chemical reducing method has been developed to high efficiently reduce graphene oxide (GO using Fe2O3 and NH3BH3 as catalyst and reductants, respectively. During the process, environmental and strong reductive nascent hydrogen were generated surrounding the surface of GO sheets by catalyst hydrolysis reaction of NH3BH3 and were used for reduction of GO. The reduction process was studied by ultraviolet absorption spectroscopy, Raman spectroscopy, and Fourier transform infrared spectrum. The structure and morphology of the reduced GO were characterized with scanning electron microscopy and transmission electron microscopy. Compared to metal (Mg/Fe/Zn/Al particles and acid system which also use nascent hydrogen to reduce GO, this method exhibited higher reduction efficiency (43.6%. Also the reduction was carried out at room temperature condition, which is environmentally friendly. As a supercapacitor electrode, the reversible capacity of reduced graphene oxide was 113.8 F g−1 at 1 A g−1 and the capacitance retention still remained at 90% after 200 cycles. This approach provides a new method to reduce GO with high reduction efficiency by green reductant.

  16. Nanostructure and Oxidation Reactivity of Nascent Soot Particles in Ethylene/Pentanol Flames

    Directory of Open Access Journals (Sweden)

    Yaoyao Ying

    2017-01-01

    Full Text Available As byproducts of the combustion process of hydrocarbon fuels, soot particles are difficult to remove, and they can greatly harm human health and pollute the environment. Therefore, the formation and growth processes of the soot particles has become a study focus of researchers. In this paper, the nanostructure and oxidation reactivity of carbonaceous particles collected from ethylene inverse diffusion flames with or without the additions of three pentanol isomers (1-pentanol, 3-methyl-1-butanol, and 2-methyl-1-butanol were investigated in detail. The nanostructure and oxidation characteristics of nascent soot particles were characterized using high resolution transmission electron microscopy (HRTEM, X-ray diffractometry (XRD and thermogravimetric analysis (TGA. It was found that the nascent soot cluster of pure ethylene flame had a loose structure, while the additions of pentanol isomers made the soot agglomerates more compact and delayed the growth of graphitic structures. The pentanol isomer additions also contributed to a higher disorder of the crystallite arrangement in the soot nanostructure. According to the TGA experiments, the results showed that the addition of pentanol isomers enhanced the oxidation reactivity of soot particles, which could help to reduce soot particle emissions.

  17. An Annotation Agnostic Algorithm for Detecting Nascent RNA Transcripts in GRO-Seq.

    Science.gov (United States)

    Azofeifa, Joseph G; Allen, Mary A; Lladser, Manuel E; Dowell, Robin D

    2017-01-01

    We present a fast and simple algorithm to detect nascent RNA transcription in global nuclear run-on sequencing (GRO-seq). GRO-seq is a relatively new protocol that captures nascent transcripts from actively engaged polymerase, providing a direct read-out on bona fide transcription. Most traditional assays, such as RNA-seq, measure steady state RNA levels which are affected by transcription, post-transcriptional processing, and RNA stability. GRO-seq data, however, presents unique analysis challenges that are only beginning to be addressed. Here, we describe a new algorithm, Fast Read Stitcher (FStitch), that takes advantage of two popular machine-learning techniques, hidden Markov models and logistic regression, to classify which regions of the genome are transcribed. Given a small user-defined training set, our algorithm is accurate, robust to varying read depth, annotation agnostic, and fast. Analysis of GRO-seq data without a priori need for annotation uncovers surprising new insights into several aspects of the transcription process.

  18. Role of the visual experience-dependent nascent proteome in neuronal plasticity.

    Science.gov (United States)

    Liu, Han-Hsuan; McClatchy, Daniel B; Schiapparelli, Lucio; Shen, Wanhua; Yates, John R; Cline, Hollis T

    2018-02-07

    Experience-dependent synaptic plasticity refines brain circuits during development. To identify novel protein synthesis-dependent mechanisms contributing to experience-dependent plasticity, we conducted a quantitative proteomic screen of the nascent proteome in response to visual experience in Xenopus optic tectum using bio-orthogonal metabolic labeling (BONCAT). We identified 83 differentially synthesized candidate plasticity proteins (CPPs). The CPPs form strongly interconnected networks and are annotated to a variety of biological functions, including RNA splicing, protein translation, and chromatin remodeling. Functional analysis of select CPPs revealed the requirement for eukaryotic initiation factor three subunit A (eIF3A), fused in sarcoma (FUS), and ribosomal protein s17 (RPS17) in experience-dependent structural plasticity in tectal neurons and behavioral plasticity in tadpoles. These results demonstrate that the nascent proteome is dynamic in response to visual experience and that de novo synthesis of machinery that regulates RNA splicing and protein translation is required for experience-dependent plasticity. © 2018, Liu et al.

  19. Antimicrobial activity of gallic acid against food-related Pseudomonas strains and its use as biocontrol tool to improve the shelf life of fresh black truffles.

    Science.gov (United States)

    Sorrentino, Elena; Succi, Mariantonietta; Tipaldi, Luca; Pannella, Gianfranco; Maiuro, Lucia; Sturchio, Marina; Coppola, Raffaele; Tremonte, Patrizio

    2018-02-02

    Refrigeration alone or in combination with other technologies represents the main tool used in the last decades to preserve the freshness of black truffles. This is principally due to the delicateness and vulnerability of this edible hypogeous fungus, so that other invasive preservation practices cannot be adopted. However, the proliferation of some microbial species during the cold storage still represents an unsolved problem. Pseudomonads are among the main spoiler bacteria responsible for the deterioration of refrigerated black truffles. Their growth ability at low temperatures requires the use of additional hurdles to prolong the shelf-life of truffles without altering their major features. The use of natural compounds may represent an alternative system for the biocontrol of this kind of product. Specifically, gallic acid (GA) is a phenolic acid naturally present in different foods, whose effectiveness was in vitro demonstrated against Pseudomonas spp. In our study, we reported the antimicrobial activity expressed by GA not only in vitro, using as target bacteria Pseudomonas putida DSMZ 291 T , P. fluorescens DSMZ 50090 T , P. fragi DSMZ 3456 T and Pseudomonas spp. P30-4, previously isolated from black truffles, but also in situ on fresh black truffles stored at 4°C for 28days. Our results showed Minimum Inhibitory Concentrations (MIC) of 2.5mg/mL GA for all tested strains, except for P. fluorescens DSMZ 50090 T , having a MIC corresponding to 5mg/mL GA. The Minimum Bactericidal Concentration (MBC) was 10mg/mL for all strains. The analysis of kinetic parameters showed that the survival declined passing from 2.5 to 10mg/mL GA concentrations, with P. fluorescens confirmed to be the most resistant strain. Moreover, images obtained from Scanning Electron Microscopy revealed that Pseudomonas cells were strongly injured by the treatment with GA at 2.5mg/mL concentration, displaying visible pores on the cellular surfaces, absence of flagella and lysis with loss of

  20. Modeling carbon dioxide effect in a controlled atmosphere and its interactions with temperature and pH on the growth of L. monocytogenes and P. fluorescens.

    Science.gov (United States)

    Couvert, Olivier; Guégan, Stéphanie; Hézard, Bernard; Huchet, Véronique; Lintz, Adrienne; Thuault, Dominique; Stahl, Valérie

    2017-12-01

    The effect of carbon dioxide, temperature, and pH on growth of Listeria monocytogenes and Pseudomonas fluorescens was studied, following a protocol to monitor microbial growth under a constant gas composition. In this way, the CO 2 dissolution didn't modify the partial pressures in the gas phase. Growth curves were acquired at different temperatures (8, 12, 22 and 37 °C), pH (5.5 and 7) and CO 2 concentration in the gas phase (0, 20, 40, 60, 80, 100% of the atmospheric pressure, and over 1 bar). These three factors greatly influenced the growth rate of L. monocytogenes and P. fluorescens, and significant interactions have been observed between the carbon dioxide and the temperature effects. Results showed no significant effect of the CO 2 concentration at 37 °C, which may be attributed to low CO2 solubility at high temperature. An inhibitory effect of CO 2 appeared at lower temperatures (8 and 12 °C). Regardless of the temperature, the gaseous CO 2 is sparingly soluble at acid pH. However, the CO 2 inhibition was not significantly different between pH 5.5 and pH 7. Considering the pKa of the carbonic acid, these results showed the dissolved carbon under HCO 3 - form didn't affect the bacterial inhibition. Finally, a global model was proposed to estimate the growth rate vs. CO 2 concentration in the aqueous phase. This dissolved concentration is calculated according to the physical equations related to the CO 2 equilibriums, involving temperature and pH interactions. This developed model is a new tool available to manage the food safety of MAP. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. The Pseudomonas secondary metabolite 2,4-diacetylphloroglucinol is a signal inducing rhizoplane expression of Azospirillum genes involved in plant-growth promotion.

    Science.gov (United States)

    Combes-Meynet, Emeline; Pothier, Joël F; Moënne-Loccoz, Yvan; Prigent-Combaret, Claire

    2011-02-01

    During evolution, plants have become associated with guilds of plant-growth-promoting rhizobacteria (PGPR), which raises the possibility that individual PGPR populations may have developed mechanisms to cointeract with one another on plant roots. We hypothesize that this has resulted in signaling phenomena between different types of PGPR colonizing the same roots. Here, the objective was to determine whether the Pseudomonas secondary metabolite 2,4-diacetylphloroglucinol (DAPG) can act as a signal on Azospirillum PGPR and enhance the phytostimulation effects of the latter. On roots, the DAPG-producing Pseudomonas fluorescens F113 strain but not its phl-negative mutant enhanced the phytostimulatory effect of Azospirillum brasilense Sp245-Rif on wheat. Accordingly, DAPG enhanced Sp245-Rif traits involved in root colonization (cell motility, biofilm formation, and poly-β-hydroxybutyrate production) and phytostimulation (auxin production). A differential fluorescence induction promoter-trapping approach based on flow cytometry was then used to identify Sp245-Rif genes upregulated by DAPG. DAPG enhanced expression of a wide range of Sp245-Rif genes, including genes involved in phytostimulation. Four of them (i.e., ppdC, flgE, nirK, and nifX-nifB) tended to be upregulated on roots in the presence of P. fluorescens F113 compared with its phl-negative mutant. Our results indicate that DAPG can act as a signal by which some beneficial pseudomonads may stimulate plant-beneficial activities of Azospirillum PGPR.

  2. ANTAGONISTIC POTENTIAL OF FLUORESCENT Pseudomonas ...

    African Journals Online (AJOL)

    Prof. Adipala Ekwamu

    This study focused on the antagonistic potential of fluorescent Pseudomonas in vitro, and its inoculation effect on growth performance of Lycopersicon esculentum in Fusarium oxysporum and Rhizoctonia solani infested soil. Biochemical characteristics of fluorescent Pseudomonas showed that all ten isolates were positive ...

  3. Comparative genomics of plant-associated Pseudomonas spp.: insights into diversity and inheritance of traits involved in multitrophic interactions.

    Directory of Open Access Journals (Sweden)

    Joyce E Loper

    2012-07-01

    Full Text Available We provide here a comparative genome analysis of ten strains within the Pseudomonas fluorescens group including seven new genomic sequences. These strains exhibit a diverse spectrum of traits involved in biological control and other multitrophic interactions with plants, microbes, and insects. Multilocus sequence analysis placed the strains in three sub-clades, which was reinforced by high levels of synteny, size of core genomes, and relatedness of orthologous genes between strains within a sub-clade. The heterogeneity of the P. fluorescens group was reflected in the large size of its pan-genome, which makes up approximately 54% of the pan-genome of the genus as a whole, and a core genome representing only 45-52% of the genome of any individual strain. We discovered genes for traits that were not known previously in the strains, including genes for the biosynthesis of the siderophores achromobactin and pseudomonine and the antibiotic 2-hexyl-5-propyl-alkylresorcinol; novel bacteriocins; type II, III, and VI secretion systems; and insect toxins. Certain gene clusters, such as those for two type III secretion systems, are present only in specific sub-clades, suggesting vertical inheritance. Almost all of the genes associated with multitrophic interactions map to genomic regions present in only a subset of the strains or unique to a specific strain. To explore the evolutionary origin of these genes, we mapped their distributions relative to the locations of mobile genetic elements and repetitive extragenic palindromic (REP elements in each genome. The mobile genetic elements and many strain-specific genes fall into regions devoid of REP elements (i.e., REP deserts and regions displaying atypical tri-nucleotide composition, possibly indicating relatively recent acquisition of these loci. Collectively, the results of this study highlight the enormous heterogeneity of the P. fluorescens group and the importance of the variable genome in tailoring

  4. Comparative Genomics of Plant-Associated Pseudomonas spp.: Insights into Diversity and Inheritance of Traits Involved in Multitrophic Interactions

    Science.gov (United States)

    Loper, Joyce E.; Hassan, Karl A.; Mavrodi, Dmitri V.; Davis, Edward W.; Lim, Chee Kent; Shaffer, Brenda T.; Elbourne, Liam D. H.; Stockwell, Virginia O.; Hartney, Sierra L.; Breakwell, Katy; Henkels, Marcella D.; Tetu, Sasha G.; Rangel, Lorena I.; Kidarsa, Teresa A.; Wilson, Neil L.; van de Mortel, Judith E.; Song, Chunxu; Blumhagen, Rachel; Radune, Diana; Hostetler, Jessica B.; Brinkac, Lauren M.; Durkin, A. Scott; Kluepfel, Daniel A.; Wechter, W. Patrick; Anderson, Anne J.; Kim, Young Cheol; Pierson, Leland S.; Pierson, Elizabeth A.; Lindow, Steven E.; Kobayashi, Donald Y.; Raaijmakers, Jos M.; Weller, David M.; Thomashow, Linda S.; Allen, Andrew E.; Paulsen, Ian T.

    2012-01-01

    We provide here a comparative genome analysis of ten strains within the Pseudomonas fluorescens group including seven new genomic sequences. These strains exhibit a diverse spectrum of traits involved in biological control and other multitrophic interactions with plants, microbes, and insects. Multilocus sequence analysis placed the strains in three sub-clades, which was reinforced by high levels of synteny, size of core genomes, and relatedness of orthologous genes between strains within a sub-clade. The heterogeneity of the P. fluorescens group was reflected in the large size of its pan-genome, which makes up approximately 54% of the pan-genome of the genus as a whole, and a core genome representing only 45–52% of the genome of any individual strain. We discovered genes for traits that were not known previously in the strains, including genes for the biosynthesis of the siderophores achromobactin and pseudomonine and the antibiotic 2-hexyl-5-propyl-alkylresorcinol; novel bacteriocins; type II, III, and VI secretion systems; and insect toxins. Certain gene clusters, such as those for two type III secretion systems, are present only in specific sub-clades, suggesting vertical inheritance. Almost all of the genes associated with multitrophic interactions map to genomic regions present in only a subset of the strains or unique to a specific strain. To explore the evolutionary origin of these genes, we mapped their distributions relative to the locations of mobile genetic elements and repetitive extragenic palindromic (REP) elements in each genome. The mobile genetic elements and many strain-specific genes fall into regions devoid of REP elements (i.e., REP deserts) and regions displaying atypical tri-nucleotide composition, possibly indicating relatively recent acquisition of these loci. Collectively, the results of this study highlight the enormous heterogeneity of the P. fluorescens group and the importance of the variable genome in tailoring individual strains

  5. Avaliação do Potencial de Conservação de Nascentes na Zona da Mata Pernambucana

    Directory of Open Access Journals (Sweden)

    Lívia Câmara Machado

    2017-12-01

    Full Text Available As nascentes são importantes fontes de água para a realização das atividades e sobrevivência de agricultores familiares nos espaços rurais. Devido a isso, estas fontes possuem relevância para o desenvolvimento e dinâmica de áreas de produção agrícola e para o consumo humano. Em decorrência dos problemas vivenciados pelos usos e apropriações inadequadas de áreas de nascentes, impactos negativos são visíveis e dificultam a conservação destas fontes que são importantes formas de acesso à água no meio rural. A realidade aponta alguns conflitos para a conservação de nascentes, que vai desde a forma de uso e ocupação do território até as práticas realizadas pelos usuários. O estudo propõe indicar o potencial de conservação de nascentes utilizadas para o abastecimento humano, a partir da ferramenta metodológica interdisciplinar para avaliação do potencial de conservação de nascentes no Assentamento Caricé, em Vitória de Santo Antão, Pernambuco. As técnicas adotadas no estudo foram de documentação indireta, (pesquisa documental e bibliográfica; e de documentação direta (pesquisa de campo, pesquisa de laboratório, observação assistemática, entrevistas estruturadas e roteiro de campo. Os parâmetros de análise das nascentes foram agrupados em três eixos distintos, o ecológico, o socioterritorial e o político, conforme prevê a metodologia interdisciplinar para avaliação do potencial de conservação das nascentes. De acordo com a ferramenta metodológica interdisciplinar, a maioria das nascentes do Assentamento Caricé, localizado no município de Vitória de Santo Antão em Pernambuco se encontram com o potencial de conservação no nível intermediário. As análises realizadas para o eixo político neste trabalho confirmam a realidade no meio rural, que não dispõe de assistência técnica, apoio e iniciativas para conservar os recursos naturais.

  6. Abalou Bangu! a Fábrica Bangu e a Indústria Nascente (1889-1914)

    OpenAIRE

    FREITAS, A. O.

    2005-01-01

    Esta pesquisa analisa o papel da Companhia da Indústria do Brasil, popularmente conhecida como fábrica Bangu no processo de industrialização e de construção do espaço urbano no Brasil, mais especificamente no Distrito Federal, no período entre 1884 e 1914. Tomamos como ponto de referência a proposta de urbanização e industrialização, a idéia de progresso e modernidade, bem como da mudança do conceito de trabalho promovidos pela República nascente. Percebemos, então, que a Fábrica Bangu reflet...

  7. X chromosome reactivation initiates in nascent primordial germ cells in mice.

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    Michihiko Sugimoto

    2007-07-01

    Full Text Available During primordial germ cell (PGC development, epigenetic reprogramming events represented by X chromosome reactivation and erasure of genomic imprinting are known to occur. Although precise timing is not given, X reactivation is thought to take place over a short period of time just before initiation of meiosis. Here, we show that the cessation of Xist expression commences in nascent PGCs, and re-expression of some X-linked genes begins in newly formed PGCs. The X reactivation process was not complete in E14.5 PGCs, indicating that X reactivation in developing PGCs occurs over a prolonged period. These results set the reactivation timing much earlier than previously thought and suggest that X reactivation may involve slow passive steps.

  8. A Two-Way Street: Regulatory Interplay between RNA Polymerase and Nascent RNA Structure.

    Science.gov (United States)

    Zhang, Jinwei; Landick, Robert

    2016-04-01

    The vectorial (5'-to-3' at varying velocity) synthesis of RNA by cellular RNA polymerases (RNAPs) creates a rugged kinetic landscape, demarcated by frequent, sometimes long-lived, pauses. In addition to myriad gene-regulatory roles, these pauses temporally and spatially program the co-transcriptional, hierarchical folding of biologically active RNAs. Conversely, these RNA structures, which form inside or near the RNA exit channel, interact with the polymerase and adjacent protein factors to influence RNA synthesis by modulating pausing, termination, antitermination, and slippage. Here, we review the evolutionary origin, mechanistic underpinnings, and regulatory consequences of this interplay between RNAP and nascent RNA structure. We categorize and rationalize the extensive linkage between the transcriptional machinery and its product, and provide a framework for future studies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Efecto de la aplicación de pseudomonas fluorescentes en las propiedades físico-químicas y biológicas de suelo : balance nutricional y estimulación de hormonas

    OpenAIRE

    Martínez Moreno, Joaquín Ignacio

    2015-01-01

    El objetivo de esta propuesta es el estudio del efecto de la aplicación de un fitofortificante (bacterias, Pseudomonas fluorescens) en la evolución tanto de las propiedades físico-químicas y microbiológicas/bioquímicas del suelo como en la estimulación de hormonas del crecimiento vegetal, así como estudiar su efecto en el balance nutricional, la calidad y la producción de los cultivos de maíz y melón. Escuela Técnica superior de Ingeniería Agronómica Universidad Politécnica ...

  10. Selective inhibition of Erwinia amylovora by the herbicidally active germination-arrest factor (GAF) produced by Pseudomonas bacteria.

    Science.gov (United States)

    Halgren, A; Azevedo, M; Mills, D; Armstrong, D; Thimmaiah, M; McPhail, K; Banowetz, G

    2011-10-01

     The germination-arrest factor (GAF) produced by Pseudomonas fluorescens WH6, and identified as 4-formylaminooxyvinylglycine, specifically inhibits the germination of a wide range of grassy weeds. This study was undertaken to determine whether GAF has antimicrobial activity in addition to its inhibitory effects on grass seed germination. Culture filtrate from Ps. fluorescens WH6 had little or no effect on 17 species of bacteria grown in Petri dish lawns, but the in vitro growth of Erwinia amylovora, the causal agent of the disease of orchard crops known as fire blight, was strongly inhibited by the filtrate. The anti-Erwinia activity of WH6 culture filtrate was shown to be due to its GAF content, and a commercially available oxyvinylglycine, 4-aminoethoxyvinylglycine (AVG), exhibited anti-Erwinia activity similar to that of GAF. The effects of GAF on Erwinia were reversed by particular amino acids. The biological properties of GAF include a rather specific antimicrobial activity against Erw. amylovora. This may be a general property of oxyvinylglycines as AVG exhibited similar activity. The ability of particular amino acids to reverse GAF inhibition is consistent with a potential effect of this compound on the activity of aminotransferases. The results presented here demonstrate a novel antimicrobial activity of oxyvinylglycines and suggest that GAF and/or GAF-producing bacteria may have potential for the control of fire blight. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology. No claim to US Government works.

  11. Acyl-Homoserine Lactone Production Is More Common among Plant-Associated Pseudomonas spp. than among Soilborne Pseudomonas spp.†

    Science.gov (United States)

    Elasri, Miena; Delorme, Sandrine; Lemanceau, Philippe; Stewart, Gordon; Laue, Bridget; Glickmann, Eric; Oger, Phil M.; Dessaux, Yves

    2001-01-01

    A total of 137 soilborne and plant-associated bacterial strains belonging to different Pseudomonas species were tested for their ability to synthesize N-acyl-homoserine lactones (NAHL). Fifty-four strains synthesized NAHL. Interestingly, NAHL production appears to be more common among plant-associated than among soilborne Pseudomonas spp. Indeed, 40% of the analyzed Pseudomonas syringae strains produced NAHL which were identified most often as the short-chain NAHL, N-hexanoyl-l-homoserine lactone, N-(3-oxo-hexanoyl)-homoserine lactone, and N-(3-oxo-octanoyl)-l-homoserine lactone (no absolute correlation between genomospecies of P. syringae and their ability to produce NAHL could be found). Six strains of fluorescent pseudomonads, belonging to the species P. chlororaphis, P. fluorescens, and P. putida, isolated from the plant rhizosphere produced different types of NAHL. In contrast, none of the strains isolated from soil samples were shown to produce NAHL. The gene encoding the NAHL synthase in P. syringae pv. maculicola was isolated by complementation of an NAHL-deficient Chromobacterium mutant. Sequence analysis revealed the existence of a luxI homologue that we named psmI. This gene is sufficient to confer NAHL synthesis upon its bacterial host and has strong homology to psyI and ahlI, two genes involved in NAHL production in P. syringae pv. tabaci and P. syringae pv. syringae, respectively. We identified another open reading frame that we termed psmR, transcribed convergently in relation to psmI and partly overlapping psmI; this gene encodes a putative LuxR regulatory protein. This gene organization, with luxI and luxR homologues facing each other and overlapping, has been found so far only in the enteric bacteria Erwinia and Pantoea and in the related species P. syringae pv. tabaci. PMID:11229911

  12. The clinical characteristics of adult bacterial meningitis caused by non-Pseudomonas (Ps.) aeruginosa Pseudomonas species: A clinical comparison with Ps. aeruginosa meningitis.

    Science.gov (United States)

    Huang, Chi-Ren; Lien, Chia-Yi; Tsai, Wan-Chen; Lai, Wei-An; Hsu, Che-Wei; Tsai, Nai-Wen; Chang, Chiung-Chih; Lu, Cheng-Hsien; Chien, Chun-Chih; Chang, Wen-Neng

    2018-01-01

    Adult bacterial meningitis (ABM) caused by non-Pseudomonas (Ps.) aeruginosa Pseudomonas (NPAP) species infection has rarely been reported. The clinical characteristics of 52 cases of Pseudomonas ABM (11 NPAP- and 41 Ps. aeruginosa-related meningitis) collected during a 30-year study period (1986-2015) were included. Eleven cases of NPAP ABM were identified in the literature, and their clinical data were also collected. Therefore, a total of 22 NPAP ABM cases were enrolled. The clinical characteristics of the NPAP ABM and Ps. aeruginosa ABM groups were compared. Of the implicated NPAP strains, Ps. putida and Ps. stutzeri were the most common (7 cases each), followed by Ps. mendocina in 4, Ps. fluorescens in 1, Ps. fulva in 1, Ps. alcaligenes in 1, and Ps. mosselii in 1. Of the 22 cases, 50% (11/22) had an underlying postneurosurgical state. Fever (77.3%, 17/22) and altered consciousness (45.5%, 10/22) were the most common clinical presentations. Antibiotic non-susceptibility was found in 3 strains of Ps. putida and 1 Ps. mosselii strain. Compared to the patients with Ps. aeruginosa ABM, those with NPAP ABM had a higher incidence of spontaneous infections and a better survival rate. In conclusion, although Ps. putida, Ps. stutzeri and Ps. mendocina were the major implicated strains of NPAP ABM, the clinical characteristics of this specific group of ABM demonstrated marked heterogeneity. Even though the cases with NPAP ABM had better therapeutic results than those with Ps. aeruginosa ABM, further large-scale studies are needed to better delineate this specific group of ABM. Copyright © 2017. Published by Elsevier Taiwan.

  13. The clinical characteristics of adult bacterial meningitis caused by non-Pseudomonas (Ps. aeruginosa Pseudomonas species: A clinical comparison with Ps. aeruginosa meningitis

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    Chi-Ren Huang

    2018-01-01

    Full Text Available Adult bacterial meningitis (ABM caused by non-Pseudomonas (Ps. aeruginosa Pseudomonas (NPAP species infection has rarely been reported. The clinical characteristics of 52 cases of Pseudomonas ABM (11 NPAP- and 41 Ps. aeruginosa-related meningitis collected during a 30-year study period (1986–2015 were included. Eleven cases of NPAP ABM were identified in the literature, and their clinical data were also collected. Therefore, a total of 22 NPAP ABM cases were enrolled. The clinical characteristics of the NPAP ABM and Ps. aeruginosa ABM groups were compared. Of the implicated NPAP strains, Ps. putida and Ps. stutzeri were the most common (7 cases each, followed by Ps. mendocina in 4, Ps. fluorescens in 1, Ps. fulva in 1, Ps. alcaligenes in 1, and Ps. mosselii in 1. Of the 22 cases, 50% (11/22 had an underlying postneurosurgical state. Fever (77.3%, 17/22 and altered consciousness (45.5%, 10/22 were the most common clinical presentations. Antibiotic non-susceptibility was found in 3 strains of Ps. putida and 1 Ps. mosselii strain. Compared to the patients with Ps. aeruginosa ABM, those with NPAP ABM had a higher incidence of spontaneous infections and a better survival rate. In conclusion, although Ps. putida, Ps. stutzeri and Ps. mendocina were the major implicated strains of NPAP ABM, the clinical characteristics of this specific group of ABM demonstrated marked heterogeneity. Even though the cases with NPAP ABM had better therapeutic results than those with Ps. aeruginosa ABM, further large-scale studies are needed to better delineate this specific group of ABM.

  14. Accelerated storage testing of freeze-dried Pseudomonas ...

    African Journals Online (AJOL)

    The viability of freeze-dried P. fluorescens strains (BTP1, PI9 and BB2) was evaluated by using the Arrhenius model. This model was described by measuring the reaction rate constants (D or k) and temperature sensitivity of rate constant (z or Ea). The freeze-dried P. fluorescens strains were stored in glass tubes at 60, ...

  15. Escherichia coli BdcA controls biofilm dispersal in Pseudomonas aeruginosa and Rhizobium meliloti

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    Wood Thomas K

    2011-10-01

    Full Text Available Abstract Background Previously we showed that BdcA controls Escherichia coli biofilm dispersal by binding the ubiquitous bacterial signal cyclic diguanylate (c-di-GMP; upon reducing the concentration of c-di-GMP, the cell shifts to the planktonic state by increasing motility, decreasing aggregation, and decreasing production of biofilm adhesins. Findings Here we report that BdcA also increases biofilm dispersal in other Gram-negative bacteria including Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhizobium meliloti. BdcA binds c-di-GMP in these strains and thereby reduces the effective c-di-GMP concentrations as demonstrated by increases in swimming motility and swarming motility as well as by a reduction in extracellular polysaccharide production. We also develop a method to displace existing biofilms by adding BdcA via conjugation from E. coli in mixed-species biofilms. Conclusion Since BdcA shows the ability to control biofilm dispersal in diverse bacteria, BdcA has the potential to be used as a tool to disperse biofilms for engineering and medical applications.

  16. PSEUDOMONAS FLUORESCENTES ASSOCIADAS À CULTURA DE COUVE: INFLUÊNCIA DA ADUBAÇÃO

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    VALÉRIA CRISTINA PALMEIRA ZAGO

    2011-01-01

    Full Text Available The microbiota soil has an important role as an indicator of the sustainability of agroecosystems, reflecting the environmental changes, particularly the antrophic actions. To evaluate the influence of different fertilizers in populations of Pseudomonas spp, in the common kale was conducted a field experiment with kale at the Agrobiology Embrapa National Center, in Seropédica, RJ, on a Argisol. The experimental design was random blocks in factorial 3 x 4, with treatments (home biosolid, cattle manure and urea fertilization, four dose levels (0, 100, 200 and 400 kg de N.ha-1 and four replicates. The amount of fertilizer applied was given according to the dosage of nitrogen.ha-1 desired. From the rhizosphere, at 15 and 30 days after transplanting the seedlings to the field, we selected strains showing fluorescence under UV light with a wavelength of 366 nm. For grouping the isolates were considered the main morphological characteristics. The majority being identified as Pseudomonas putida (54% and P. fluorescens (14%, by API 20NE System (bioMérieux, Analytab Products. The results obtained from the reactions of the API 20NE test kit showed a wide variation in the utilization of carbon compounds and enzymatic inter-and intraspecific. Some groups of isolates colonized preferentially the cabbage rhizosphere of plants fertilized with biosolids and different doses of fertilizers used. There were a smaller number of groups present in treatments with urea.

  17. Characterization of Extracellular Polymeric Substances Produced by Pseudomonas fragi Under Air and Modified Atmosphere Packaging.

    Science.gov (United States)

    Wang, Guang-Yu; Ma, Fang; Wang, Hu-Hu; Xu, Xing-Lian; Zhou, Guang-Hong

    2017-09-01

    Extracellular polymeric substances (EPS) play an important role in bacterial biochemical properties. The characteristics of EPS from 2 strains of Pseudomonas fragi cultured in meat aerobically (control) and in modified atmosphere packaging (MAP) were studied. The amount and components of EPS, the surface properties, and the effect on biofilm formation of several spoilage organisms were evaluated. The results showed that MAP inhibited the growth of the P. fragi strains. Compared with the control, more loose and less bound EPS (containing protein and carbohydrate) were produced by P. fragi in MAP samples. MAP also caused increased cell autoaggregation and surface hydrophobicity. After the removal of the EPS, the surface property changes were strain-dependent, suggesting that membrane compositions were also changed. In addition, the EPS displayed significant antibiofilm activity on Pseudomonas fluorescens and Serratia liquefaciens. In conclusion, P. fragi strains not only modified the amount, components, and surface properties of EPS but also changed the cell membrane compositions to adapt to MAP stress. Moreover, EPS may play an important role in microbial community competitions. © 2017 Institute of Food Technologists®.

  18. Binding of Signal Recognition Particle Gives Ribosome/Nascent Chain Complexes a Competitive Advantage in Endoplasmic Reticulum Membrane Interaction

    Science.gov (United States)

    Neuhof, Andrea; Rolls, Melissa M.; Jungnickel, Berit; Kalies, Kai-Uwe; Rapoport, Tom A.

    1998-01-01

    Most secretory and membrane proteins are sorted by signal sequences to the endoplasmic reticulum (ER) membrane early during their synthesis. Targeting of the ribosome-nascent chain complex (RNC) involves the binding of the signal sequence to the signal recognition particle (SRP), followed by an interaction of ribosome-bound SRP with the SRP receptor. However, ribosomes can also independently bind to the ER translocation channel formed by the Sec61p complex. To explain the specificity of membrane targeting, it has therefore been proposed that nascent polypeptide-associated complex functions as a cytosolic inhibitor of signal sequence- and SRP-independent ribosome binding to the ER membrane. We report here that SRP-independent binding of RNCs to the ER membrane can occur in the presence of all cytosolic factors, including nascent polypeptide-associated complex. Nontranslating ribosomes competitively inhibit SRP-independent membrane binding of RNCs but have no effect when SRP is bound to the RNCs. The protective effect of SRP against ribosome competition depends on a functional signal sequence in the nascent chain and is also observed with reconstituted proteoliposomes containing only the Sec61p complex and the SRP receptor. We conclude that cytosolic factors do not prevent the membrane binding of ribosomes. Instead, specific ribosome targeting to the Sec61p complex is provided by the binding of SRP to RNCs, followed by an interaction with the SRP receptor, which gives RNC–SRP complexes a selective advantage in membrane targeting over nontranslating ribosomes. PMID:9436994

  19. Does Ethicality Wane with Adulthood? A Study of the Ethical Values of Entrepreneurship Students and Nascent Entrepreneurs

    Science.gov (United States)

    Lourenço, Fernando; Sappleton, Natalie; Cheng, Ranis

    2015-01-01

    The authors examined the following questions: Does gender influence the ethicality of enterprise students to a greater extent than it does nascent entrepreneurs? If this is the case, then is it due to factors associated with adulthood such as age, work experience, marital status, and parental status? Sex-role socialization theory and moral…

  20. Tomato seed and root exudate sugars: composition, utilization by Pseudomonas biocontrol strains and role in rhizosphere colonization.

    Science.gov (United States)

    Lugtenberg, B J; Kravchenko, L V; Simons, M

    1999-10-01

    The role of tomato seed and root exudate sugars as nutrients for Pseudomonas biocontrol bacteria was studied. To this end, the major exudate sugars of tomato seeds, seedlings and roots were identified and quantified using high-performance liquid chromatographic (HPLC) analysis. Glucose, fructose and maltose were present in all studied growth stages of the plant, but the ratios of these sugars were strongly dependent on the developmental stage. In order to study the putative role of exudate sugar utilization in rhizosphere colonization, two approaches were adopted. First, after co-inoculation on germinated tomato seeds, the root-colonizing ability of the efficient root-colonizing P. fluorescens strain WCS365 in a gnotobiotic quartz sand-plant nutrient solution system was compared with that of other Pseudomonas biocontrol strains. No correlation was observed between the colonizing ability of a strain and its ability to use the major exudate sugars as the only carbon and energy source. Secondly, a Tn5lacZ mutant of P. fluorescens strain WCS365, strain PCL1083, was isolated, which is impaired in its ability to grow on simple sugars, including those found in exudate. The mutation appeared to reside in zwf, which encodes glucose-6-phosphate dehydrogenase. The mutant grows as well as the parental strain on other media, including tomato root exudate. After inoculation of germinated sterile tomato seeds, the mutant cells reached the same population levels at the root tip as the wild-type strain, both alone and in competition, indicating that the ability to use exudate sugars does not play a major role in tomato root colonization, despite the fact that sugars have often been reported to represent the major exudate carbon source. This conclusion is supported by the observation that the growth of mutant PCL1083 in vitro is inhibited by glucose, a major exudate sugar, at a concentration of 0.001%, which indicates that the glucose concentration in the tomato rhizosphere is very

  1. A study on a nascent entomopathogenic association between caenorhabditis briggsae and serratia sp.SCBI

    Science.gov (United States)

    Abebe-Akele, Feseha

    Life is inconceivable in the absence of interactions which could be cooperative, antagonistic or neutral. Interactions are in constant flux because on one hand it is often difficult to demarcate where one form of interaction ends and the other begins on the other hand what is cooperative at one point in time could evolve into antagonistic or neutral or vice versa. Thus, organisms, as a consequence of mutation, adaptation and natural selection would inevitably enter into natural associations from which they emerge as mutual partners, inveterate enemies or passive cohabitants. Entomopathogenic nematode (EPN) partnerships are tripartite interactions where a nematode-bacteria symbiont duo attacks a third organism -an insect or insect larva-for the mutual benefit of the attacking partners and the detriment of the insect they invade. All three participants in the interaction---the nematode worms with their symbiont bacteria and the target insect host-are among the most ancient, diverse and abundant species on earth, however, these EPN partnerships are not as common as circumstances would suggest. EPN associations, which are arguably at the peak of evolutionary co adaptations, where two primitive forms of life cooperate to take advantage of a larger species are not only fascinating but immensely important for humans. The biological and molecular mechanisms underlying entomopathogenesis have been studied in great detail for decades for their potential as biological control agents against invasive insects. In spite of intense research in The EPN field, the evolutionary history of EPN associations are largely unknown because there are no known intermediate forms. In this thesis, a nascent EPN partnership is described between Caenorhabditid nematodes and Serratia sp. SCBI. Comparative analysis of this association with other EPNs suggests that crucial aspect of EPN associations may be the ability of partners to co-exist without killing each other and that the end results of

  2. Chronic Pseudomonas aeruginosa cervical osteomyelitis

    Directory of Open Access Journals (Sweden)

    Sujeet Kumar Meher

    2016-01-01

    Full Text Available Pseudomonas aeruginosa is a rare cause of osteomyelitis of the cervical spine and is usually seen in the background of intravenous drug use and immunocompromised state. Very few cases of osteomyelitis of the cervical spine caused by pseudomonas aeruginosa have been reported in otherwise healthy patients. This is a case presentation of a young female, who in the absence of known risk factors for cervical osteomyelitis presented with progressively worsening neurological signs and symptoms.

  3. Bacteriocinogenicity and production of pyocins from Pseudomonas ...

    African Journals Online (AJOL)

    ), P. fluorescens (13) and P. stutzeri (3) were isolated from Nigerian patients with burns, wound and skin infections, UTI, diarrhoea and eye infections. 81.5, 55.6, 76.9 and 33.3% of these species produced pyocins at a range of 410 – 670 g/ml.

  4. Phosphorylcholine Phosphatase: A Peculiar Enzyme of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Domenech

    2011-01-01

    Full Text Available Pseudomonas aeruginosa synthesizes phosphorylcholine phosphatase (PchP when grown on choline, betaine, dimethylglycine or carnitine. In the presence of Mg2+ or Zn2+, PchP catalyzes the hydrolysis of p-nitrophenylphosphate (p-NPP or phosphorylcholine (Pcho. The regulation of pchP gene expression is under the control of GbdR and NtrC; dimethylglycine is likely the metabolite directly involved in the induction of PchP. Therefore, the regulation of choline metabolism and consequently PchP synthesis may reflect an adaptive response of P. aeruginosa to environmental conditions. Bioinformatic and biochemistry studies shown that PchP contains two sites for alkylammonium compounds (AACs: one in the catalytic site near the metal ion-phosphoester pocket, and another in an inhibitory site responsible for the binding of the alkylammonium moiety. Both sites could be close to each other and interact through the residues 42E, 43E and 82YYY84. Zn2+ is better activator than Mg2+ at pH 5.0 and it is more effective at alleviating the inhibition produced by the entry of Pcho or different AACs in the inhibitory site. We postulate that Zn2+ induces at pH 5.0 a conformational change in the active center that is communicated to the inhibitory site, producing a compact or closed structure. However, at pH 7.4, this effect is not observed because to the hydrolysis of the [Zn2+L2−1L20(H2O2] complex, which causes a change from octahedral to tetrahedral in the metal coordination geometry. This enzyme is also present in P. fluorescens, P. putida, P. syringae, and other organisms. We have recently crystallized PchP and solved its structure.

  5. The ribosome destabilizes native and non-native structures in a nascent multidomain protein.

    Science.gov (United States)

    Liu, Kaixian; Rehfus, Joseph E; Mattson, Elliot; Kaiser, Christian M

    2017-07-01

    Correct folding is a prerequisite for the biological activity of most proteins. Folding has largely been studied using in vitro refolding assays with isolated small, robustly folding proteins. A substantial fraction of all cellular proteomes is composed of multidomain proteins that are often not amenable to this approach, and their folding remains poorly understood. These large proteins likely begin to fold during their synthesis by the ribosome, a large molecular machine that translates the genetic code. The ribosome affects how folding proceeds, but the underlying mechanisms remain largely obscure. We have utilized optical tweezers to study the folding of elongation factor G, a multidomain protein composed of five domains. We find that interactions among unfolded domains interfere with productive folding in the full-length protein. The N-terminal G-domain constitutes an independently folding unit that, upon in vitro refolding, adopts two similar states that correspond to the natively folded and a non-native, possibly misfolded structure. The ribosome destabilizes both of these states, suggesting a mechanism by which terminal misfolding into highly stable, non-native structures is avoided. The ribosome may thus directly contribute to efficient folding by modulating the folding of nascent multidomain proteins. © 2017 The Protein Society.

  6. Introduction of subsidisation in nascent climate-friendly learning technologies and evaluation of its effectiveness

    International Nuclear Information System (INIS)

    Rout, Ullash K.; Akimoto, Keigo; Sano, Fuminori; Tomoda, Toshimasa

    2010-01-01

    Given its importance as a practical phenomenon underlying the progress of learning technologies, attention should be paid to the role of subsidisation in learning theory, particularly in the case of nascent climate-related sociable learning technologies, in order to examine its benefits. Thus, this study focuses on subsidy procurement of energy technologies in several economies in the context of the component learning track in endogenous global clusters in order to suggest improvements to the adoption mechanism and examine the climate stabilization constraint. At the same time, the study attempts to determine the global progress ratio of the lithium-ion battery in order to analyse various endogenous learning scenarios for hybrid technologies. An integrated energy system model with highly disaggregated global regions (DNE21+) is used to execute this research in a medium time frame. Subsidisation of the learning track of battery technology encourages greater development of plug-in hybrid vehicles, promotes early diffusion of hybrid technologies, and relieves heavy dependency on crude oil and biofuels. The subsidies in the common learning domains in few economies benefit the nearby economies because of the technology spillover that occurs through numerous cross-feedback learning mechanisms. Endogenous learning with subsidies augments diffusion potentials, abates emissions, and shifts sectoral emissions.

  7. Entrepreneurial Judgment and Value Capture, the Case of the Nascent Offshore Renewable Industry

    Directory of Open Access Journals (Sweden)

    Truls Erikson

    2015-11-01

    Full Text Available Entrepreneurship may be regarded as the mechanism of change towards sustainability. Any entrepreneur that seeks to start a new venture in an emerging industry will face resource and time constraints. The question we raise here is how the entrepreneur should prioritize use of time and resources to increase likeliness of success. To address this question we depart from a theoretical perspective of entrepreneurship seen as judgment, and bridges it over to entrepreneurship seen as co-creation. In other words, we combine the subjective with the intersubjective, and explore the effects of the actions successful green technology entrepreneurs in the emerging offshore renewable energy industry make in building their new ventures in nascent markets. Inspired by earlier studies on market entry, combined with new ways to understand new venture emergence, we find that independent entrepreneurs benefit from leapfrogging typical stages in the technology development process and rather devote time and efforts on resource acquisition. We also find that the most important value-capturing, decision-making heuristics are those related to “hybrid governance”. We discuss implications for theory, practice, and policy.

  8. Initial characterization of a bolA homologue from Pseudomonas fluorescens indicates different roles for BolA-like proteins in P. fluorescens and Escherichia coli

    DEFF Research Database (Denmark)

    Koch, Birgit; Nybroe, Ole

    2006-01-01

    A expression. The mutant grew slower than the wild-type strain in minimal medium with L-serine as the sole nitrogen source, while growth rates were similar on a mixture of L-serine and L-cysteine. Reverse transcriptase polymerase chain reaction analysis indicated that the bolA homologue is the second gene...

  9. Hot Tub Rash (Pseudomonas Dermatitis/Folliculitis)

    Science.gov (United States)

    ... page] Español [PDF – 1 page] “Hot Tub Rash” ( Pseudomonas Dermatitis / Folliculitis) If contaminated water comes in contact ... is often caused by infection with the germ Pseudomonas aeruginosa . This germ is common in the environment ( ...

  10. Pseudomonas yamanorum sp. nov., a psychrotolerant bacterium isolated from a subantarctic environment.

    Science.gov (United States)

    Arnau, Víctor Gonzalo; Sánchez, Leandro Arturo; Delgado, Osvaldo Daniel

    2015-02-01

    A psychrotolerant strain, 8H1(T), was isolated from soil samples collected in Isla de los Estados, Ushuaia, Argentina. Cells were Gram-negative, aerobic, straight rods, occurring singly or in pairs, non-spore-forming and motile by means of two polar flagella. The isolate was able to grow in the range 4-35 °C, with optimum growth at 28 °C. The predominant cellular fatty acids were summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), C16 : 0 and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The polar lipid pattern of strain 8H1(T) comprised phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unknown phospholipid. Ubiquinone 9 (Q-9) was the predominant lipoquinone. The DNA G+C content was 59.8 mol%. 16S rRNA gene sequence-based phylogeny suggested the affiliation of strain 8H1(T) to the 'Pseudomonas fluorescens group', displaying ≥98.5 % sequence similarity to 29 type strains. A multilocus sequence analysis (MLSA) study performed by concatenating 16S rRNA, gyrB, rpoD and rpoB gene sequences showed that isolate 8H1(T) could be discriminated from closely related species of the genus Pseudomonas and placed in the 'Pseudomonas gessardii subgroup', including the species with the highest MLSA sequence similarities: Pseudomonas brenneri (96.2 %), P. gessardii (96.1 %), P. proteolytica (96.0 %), P. meridiana (96.0 %) and P. mucidolens (95.4 %). DNA-DNA hybridization analysis between 8H1(T) and the type strains of these closely related species revealed relatedness values of 27.0, 8.8, 41.2, 39.7 and 46.1 %, respectively. These results, together with differences in several phenotypic features, support the classification of a novel species, for which the name Pseudomonas yamanorum sp. nov. is proposed. The type strain is 8H1(T) ( = DSM 26522(T) = CCUG 63249(T) = LMG 27247(T)). © 2015 IUMS.

  11. Genome-Wide Dynamics of Nascent Noncoding RNA Transcription in Porcine Heart After Myocardial Infarction.

    Science.gov (United States)

    Kaikkonen, Minna U; Halonen, Paavo; Liu, Oscar Hsin-Fu; Turunen, Tiia A; Pajula, Juho; Moreau, Pierre; Selvarajan, Ilakya; Tuomainen, Tomi; Aavik, Einari; Tavi, Pasi; Ylä-Herttuala, Seppo

    2017-06-01

    Microarrays and RNA sequencing are widely used to profile transcriptome remodeling during myocardial ischemia. However, the steady-state RNA analysis lacks in sensitivity to detect all noncoding RNA species and does not provide separation between transcriptional and post-transcriptional regulations. Here, we provide the first comprehensive analysis of nascent RNA profiles of mRNAs, primary micro-RNAs, long noncoding RNAs, and enhancer RNAs in a large animal model of acute infarction. Acute infarction was induced by cardiac catheterization of domestic swine. Nuclei isolated from healthy, border zone, and ischemic regions of the affected heart were subjected to global run-on sequencing. Global run-on sequencing analysis indicated that half of affected genes are regulated at the level of transcriptional pausing. A gradient of induction of inflammatory mediators and repression of peroxisome proliferator-activated receptor signaling and oxidative phosphorylation was detected when moving from healthy toward infarcted area. In addition, we interrogated the transcriptional regulation of primary micro-RNAs and provide evidence that several arrhythmia-related target genes exhibit repression at post-transcriptional level. We identified 450 long noncoding RNAs differently regulated by ischemia, including novel conserved long noncoding RNAs expressed in antisense orientation to myocardial transcription factors GATA-binding protein 4, GATA-binding protein 6, and Krüppel-like factor 6. Finally, characterization of enhancers exhibiting differential expression of enhancer RNAs pointed a central role for Krüppel-like factor, MEF2C, ETS, NFY, ATF, E2F2, and NRF1 transcription factors in determining transcriptional responses to ischemia. Global run-on sequencing allowed us to follow the gradient of gene expression occurring in the ischemic heart and identify novel noncoding RNAs regulated by oxygen deprivation. These findings highlight potential new targets for diagnosis and

  12. Mechanical modulation of nascent stem cell lineage commitment in tissue engineering scaffolds.

    Science.gov (United States)

    Song, Min Jae; Dean, David; Knothe Tate, Melissa L

    2013-07-01

    Taking inspiration from tissue morphogenesis in utero, this study tests the concept of using tissue engineering scaffolds as delivery devices to modulate emergent structure-function relationships at early stages of tissue genesis. We report on the use of a combined computational fluid dynamics (CFD) modeling, advanced manufacturing methods, and experimental fluid mechanics (micro-piv and strain mapping) for the prospective design of tissue engineering scaffold geometries that deliver spatially resolved mechanical cues to stem cells seeded within. When subjected to a constant magnitude global flow regime, the local scaffold geometry dictates the magnitudes of mechanical stresses and strains experienced by a given cell, and in a spatially resolved fashion, similar to patterning during morphogenesis. In addition, early markers of mesenchymal stem cell lineage commitment relate significantly to the local mechanical environment of the cell. Finally, by plotting the range of stress-strain states for all data corresponding to nascent cell lineage commitment (95% CI), we begin to "map the mechanome", defining stress-strain states most conducive to targeted cell fates. In sum, we provide a library of reference mechanical cues that can be delivered to cells seeded on tissue engineering scaffolds to guide target tissue phenotypes in a temporally and spatially resolved manner. Knowledge of these effects allows for prospective scaffold design optimization using virtual models prior to prototyping and clinical implementation. Finally, this approach enables the development of next generation scaffolds cum delivery devices for genesis of complex tissues with heterogenous properties, e.g., organs, joints or interface tissues such as growth plates. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. "Hot Tub Rash" and "Swimmer's Ear" (Pseudomonas)

    Science.gov (United States)

    Facts About “Hot Tub Rash” and “Swimmer’s Ear” (Pseudomonas) What is Pseudomonas and how can it affect me? Pseudomonas (sue-doh- ... a major cause of infections commonly known as “hot tub rash” and “swimmer’s ear.” This germ is ...

  14. The Pseudomonas Quinolone Signal (PQS)

    DEFF Research Database (Denmark)

    Sams, Thomas; Baker, Ysobel; Hodgkinson, James

    2015-01-01

    Pseudomonas aeruginosa is an opportunistichuman pathogen that routinely appears near the top ofpublic health threat lists worldwide. P. aeruginosa causes in-fections by secreting a wealth of exceptionally active exo-products, leading to tissue damage. The synthesis of manyof these virulence factors...... is now known to be under the con-trol of the quorum sensing (QS) system. Over the last15 years, the Pseudomonas quinolone signal (PQS) has beenfound to play a crucial role in QS by linking the two seg-ments (las and rhl) of the P. aeruginosa N-acylhomoserinelactone-dependent QS signaling pathways. Herein...

  15. A Longitudinal Examination of The Impact of Founding Owner Operator Characteristics on Nascent Venture Performance: Evidence from the Kauffman Firm Survey

    Directory of Open Access Journals (Sweden)

    Augustine Y. Dzathor

    2013-07-01

    Full Text Available This study attempts to revive and clarify the debate on ‘the entrepreneurial man’. We longitudinally examined the effects of seven founding-owner-operator characteristics (prior industry experience, level of formal education, age, gender, ethnicity and time committed to business operations on nascent venture performance. Our results indicate that owner work experience, level of education and hours worked in the business have significant effect on nascent venture performance, while inadequate owner reputation and luck of ethnic social capital may negatively affect nascent venture performance. Our findings also suggest that characteristic of the ‘entrepreneurial man’ are dynamic and leans towards a temporal contingency model. Different entrepreneur characteristics seem to assume prominence in firm performance at different times in a nascent venture’s life trajectory.

  16. Defining the specificity of cotranslationally acting chaperones by systematic analysis of mRNAs associated with ribosome-nascent chain complexes.

    Directory of Open Access Journals (Sweden)

    Marta del Alamo

    2011-07-01

    Full Text Available Polypeptides exiting the ribosome must fold and assemble in the crowded environment of the cell. Chaperones and other protein homeostasis factors interact with newly translated polypeptides to facilitate their folding and correct localization. Despite the extensive efforts, little is known about the specificity of the chaperones and other factors that bind nascent polypeptides. To address this question we present an approach that systematically identifies cotranslational chaperone substrates through the mRNAs associated with ribosome-nascent chain-chaperone complexes. We here focused on two Saccharomyces cerevisiae chaperones: the Signal Recognition Particle (SRP, which acts cotranslationally to target proteins to the ER, and the Nascent chain Associated Complex (NAC, whose function has been elusive. Our results provide new insights into SRP selectivity and reveal that NAC is a general cotranslational chaperone. We found surprising differential substrate specificity for the three subunits of NAC, which appear to recognize distinct features within nascent chains. Our results also revealed a partial overlap between the sets of nascent polypeptides that interact with NAC and SRP, respectively, and showed that NAC modulates SRP specificity and fidelity in vivo. These findings give us new insight into the dynamic interplay of chaperones acting on nascent chains. The strategy we used should be generally applicable to mapping the specificity, interplay, and dynamics of the cotranslational protein homeostasis network.

  17. Disruption of Transporters Affiliated with Enantio-Pyochelin Biosynthesis Gene Cluster of Pseudomonas protegens Pf-5 Has Pleiotropic Effects.

    Science.gov (United States)

    Lim, Chee Kent; Penesyan, Anahit; Hassan, Karl A; Loper, Joyce E; Paulsen, Ian T

    2016-01-01

    Pseudomonas protegens Pf-5 (formerly Pseudomonas fluorescens) is a biocontrol bacterium that produces the siderophore enantio-pyochelin under conditions of iron starvation in a process that is often accompanied by the secretion of its biosynthesis intermediates, salicylic acid and dihydroaeruginoic acid. In this study, we investigated whether several transporters that are encoded by genes within or adjacent to the enantio-pyochelin biosynthetic cluster, serve as efflux systems for enantio-pyochelin and/or its intermediates. In addition, we determined whether these transporters have broad substrates range specificity using a Phenotype Microarray system. Intriguingly, knockouts of the pchH and fetF transporter genes resulted in mutant strains that secrete higher levels of enantio-pyochelin as well as its intermediates salicylic acid and dihydroaeruginoic acid. Analyses of these mutants did not indicate significant change in transcription of biosynthetic genes involved in enantio-pyochelin production. In contrast, the deletion mutant of PFL_3504 resulted in reduced transcription of the biosynthetic genes as well as decreased dihydroaeruginoic acid concentrations in the culture supernatant, which could either point to regulation of gene expression by the transporter or its role in dihydroaeruginoic acid transport. Disruption of each of the transporters resulted in altered stress and/or chemical resistance profile of Pf-5, which may reflect that these transporters could have specificity for rather a broad range of substrates.

  18. Disruption of Transporters Affiliated with Enantio-Pyochelin Biosynthesis Gene Cluster of Pseudomonas protegens Pf-5 Has Pleiotropic Effects.

    Directory of Open Access Journals (Sweden)

    Chee Kent Lim

    Full Text Available Pseudomonas protegens Pf-5 (formerly Pseudomonas fluorescens is a biocontrol bacterium that produces the siderophore enantio-pyochelin under conditions of iron starvation in a process that is often accompanied by the secretion of its biosynthesis intermediates, salicylic acid and dihydroaeruginoic acid. In this study, we investigated whether several transporters that are encoded by genes within or adjacent to the enantio-pyochelin biosynthetic cluster, serve as efflux systems for enantio-pyochelin and/or its intermediates. In addition, we determined whether these transporters have broad substrates range specificity using a Phenotype Microarray system. Intriguingly, knockouts of the pchH and fetF transporter genes resulted in mutant strains that secrete higher levels of enantio-pyochelin as well as its intermediates salicylic acid and dihydroaeruginoic acid. Analyses of these mutants did not indicate significant change in transcription of biosynthetic genes involved in enantio-pyochelin production. In contrast, the deletion mutant of PFL_3504 resulted in reduced transcription of the biosynthetic genes as well as decreased dihydroaeruginoic acid concentrations in the culture supernatant, which could either point to regulation of gene expression by the transporter or its role in dihydroaeruginoic acid transport. Disruption of each of the transporters resulted in altered stress and/or chemical resistance profile of Pf-5, which may reflect that these transporters could have specificity for rather a broad range of substrates.

  19. Expression of the diguanylate cyclase GcbA is regulated by FleQ in response to cyclic di-GMP in Pseudomonas putida KT2440.

    Science.gov (United States)

    Xiao, Yujie; Nie, Hailing; Liu, Huizhong; Chen, Wenli; Huang, Qiaoyun

    2016-12-01

    Cyclic di-GMP (c-di-GMP), a ubiquitous bacterial second messenger that regulates diverse cellular processes, is synthesized by diguanylate cyclase (DGC) and degraded by phosphodiesterase (PDE). GcbA is a well conserved DGC among Pseudomonas species, and has been reported to influence biofilm formation and flagellar motility in Pseudomonas fluorescens and Pseudomonas aeruginosa. Here we confirm the function of GcbA in Pseudomonas putida and reveal that expression of GcbA is regulated by FleQ in response to c-di-GMP. GcbA deletion impaired initial biofilm formation and enhanced swimming motility, but showed no influence on biofilm maturation in Pseudomonas putida. Deletion of the c-di-GMP effector FleQ led to a significant decrease in transcription of gcbA. Moreover, reducing c-di-GMP levels promoted gcbA transcription in a FleQ dependent way, while enhancing c-di-GMP levels abolished the promotion. In in vitro experiments we found that FleQ bound to gcbA promoter DNA and the binding was inhibited by c-di-GMP. Besides, FleN, an anti-activator of FleQ, and the sigma factor RpoN also participated in transcription of gcbA. Our finding expands the complexity of FleQ-dependent regulation and reveals a self-regulation function of c-di-GMP by regulating GcbA expression via FleQ. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  20. SWI/SNF associates with nascent pre-mRNPs and regulates alternative pre-mRNA processing.

    Directory of Open Access Journals (Sweden)

    Anu Tyagi

    2009-05-01

    Full Text Available The SWI/SNF chromatin remodeling complexes regulate the transcription of many genes by remodeling nucleosomes at promoter regions. In Drosophila, SWI/SNF plays an important role in ecdysone-dependent transcription regulation. Studies in human cells suggest that Brahma (Brm, the ATPase subunit of SWI/SNF, regulates alternative pre-mRNA splicing by modulating transcription elongation rates. We describe, here, experiments that study the association of Brm with transcribed genes in Chironomus tentans and Drosophila melanogaster, the purpose of which was to further elucidate the mechanisms by which Brm regulates pre-mRNA processing. We show that Brm becomes incorporated into nascent Balbiani ring pre-mRNPs co-transcriptionally and that the human Brm and Brg1 proteins are associated with RNPs. We have analyzed the expression profiles of D. melanogaster S2 cells in which the levels of individual SWI/SNF subunits have been reduced by RNA interference, and we show that depletion of SWI/SNF core subunits changes the relative abundance of alternative transcripts from a subset of genes. This observation, and the fact that a fraction of Brm is not associated with chromatin but with nascent pre-mRNPs, suggest that SWI/SNF affects pre-mRNA processing by acting at the RNA level. Ontology enrichment tests indicate that the genes that are regulated post-transcriptionally by SWI/SNF are mostly enzymes and transcription factors that regulate postembryonic developmental processes. In summary, the data suggest that SWI/SNF becomes incorporated into nascent pre-mRNPs and acts post-transcriptionally to regulate not only the amount of mRNA synthesized from a given promoter but also the type of alternative transcript produced.

  1. Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique.

    Science.gov (United States)

    Bhaskara, Srividya

    2016-01-14

    Histone deacetylases 1 and 2 (HDAC1,2) localize to the sites of DNA replication. In the previous study, using a selective inhibitor and a genetic knockdown system, we showed novel functions for HDAC1,2 in replication fork progression and nascent chromatin maintenance in mammalian cells. Additionally, we used a BrdU-ChIP-Slot-Western technique that combines chromatin immunoprecipitation (ChIP) of bromo-deoxyuridine (BrdU)-labeled DNA with slot blot and Western analyses to quantitatively measure proteins or histone modification associated with nascent DNA. Actively dividing cells were treated with HDAC1,2 selective inhibitor or transfected with siRNAs against Hdac1 and Hdac2 and then newly synthesized DNA was labeled with the thymidine analog bromodeoxyuridine (BrdU). The BrdU labeling was done at a time point when there was no significant cell cycle arrest or apoptosis due to the loss of HDAC1,2 functions. Following labeling of cells with BrdU, chromatin immunoprecipitation (ChIP) of histone acetylation marks or the chromatin-remodeler was performed with specific antibodies. BrdU-labeled input DNA and the immunoprecipitated (or ChIPed) DNA was then spotted onto a membrane using the slot blot technique and immobilized using UV. The amount of nascent DNA in each slot was then quantitatively assessed using Western analysis with an anti-BrdU antibody. The effect of loss of HDAC1,2 functions on the levels of newly synthesized DNA-associated histone acetylation marks and chromatin remodeler was then determined by normalizing the BrdU-ChIP signal obtained from the treated samples to the control samples.

  2. Variable Copy Number, Intra-Genomic Heterogeneities and Lateral Transfers of the 16S rRNA Gene in Pseudomonas

    Science.gov (United States)

    Bodilis, Josselin; Nsigue-Meilo, Sandrine; Besaury, Ludovic; Quillet, Laurent

    2012-01-01

    Even though the 16S rRNA gene is the most commonly used taxonomic marker in microbial ecology, its poor resolution is still not fully understood at the intra-genus level. In this work, the number of rRNA gene operons, intra-genomic heterogeneities and lateral transfers were investigated at a fine-scale resolution, throughout the Pseudomonas genus. In addition to nineteen sequenced Pseudomonas strains, we determined the 16S rRNA copy number in four other Pseudomonas strains by Southern hybridization and Pulsed-Field Gel Electrophoresis, and studied the intra-genomic heterogeneities by Denaturing Gradient Gel Electrophoresis and sequencing. Although the variable copy number (from four to seven) seems to be correlated with the evolutionary distance, some close strains in the P. fluorescens lineage showed a different number of 16S rRNA genes, whereas all the strains in the P. aeruginosa lineage displayed the same number of genes (four copies). Further study of the intra-genomic heterogeneities revealed that most of the Pseudomonas strains (15 out of 19 strains) had at least two different 16S rRNA alleles. A great difference (5 or 19 nucleotides, essentially grouped near the V1 hypervariable region) was observed only in two sequenced strains. In one of our strains studied (MFY30 strain), we found a difference of 12 nucleotides (grouped in the V3 hypervariable region) between copies of the 16S rRNA gene. Finally, occurrence of partial lateral transfers of the 16S rRNA gene was further investigated in 1803 full-length sequences of Pseudomonas available in the databases. Remarkably, we found that the two most variable regions (the V1 and V3 hypervariable regions) had probably been laterally transferred from another evolutionary distant Pseudomonas strain for at least 48.3 and 41.6% of the 16S rRNA sequences, respectively. In conclusion, we strongly recommend removing these regions of the 16S rRNA gene during the intra-genus diversity studies. PMID:22545126

  3. Apolipoprotein B-containing lipoprotein particle assembly: Lipid capacity of the nascent lipoprotein particle

    Energy Technology Data Exchange (ETDEWEB)

    Manchekar, Medha; Forte, Trudy M.; Datta, Geeta; Richardson, Paul E.; Segrest, Jere P.; Dashti, Nassrin

    2003-12-01

    '' without a structural requirement for MTP; (2) amino acids between 931 to 1000 of apoB-100 are critical for the formation of a nascent lipoprotein particle, and (3) the ''lipid pocket'' created by the first 1000 amino acid residues of apoB-100 is PL-rich, suggesting a small bilayer type organization and has a maximum capacity on the order of 70 molecules of lipid. This model is supported by the allatom molecular model of the {beta}{alpha}{sub 1} lipid pocket presented in the accompanying paper.

  4. Pathogenic variation in isolates of Pseudomonas causing the brown blotch of cultivated mushroom, Agaricus bisporus

    Directory of Open Access Journals (Sweden)

    Mohamed A. Abou-Zeid

    2012-09-01

    Full Text Available Twenty seven bacterial isolates were isolated from superficial brown discolorations on the caps of cultivated Agaricus bisporus. After White Line Assay (WLA and the assist of Biolog computer-identification system, isolates were divided into groups: (I comprised ninteen bacterial isolates that positively responded to a Pseudomonas "reactans" reference strain (NCPPB1311 in WLA and were identified as Pseudomonas tolaasii, (II comprised two isolates which were WLA+ towards the reference strain (JCM21583 of P. tolaasii and were proposed to be P. "reactans". The third group comprised six isolates, two of which weakly responded to the strain of P. tolaasii and were identified as P. gingeri whereas the other four were WLA- and identified as P. fluorescens (three isolates and P. marginalis (one isolate. Isolates of P. tolaasii showed high aggressiveness compared with those of P. "reactans" in pathogenicity tests. Cubes of 1 cm³ of A. bisporus turned brown and decreased in size when were inoculated with 10 µl of P. tolaasii suspension containing 10(8 CFU ml-1, whereas a similar concentration of P. "reactans" caused only light browning. Fifty µl of the same concentration of P. tolaasii isolates gave typical brown blotch symptoms on fresh mushroom sporophores whereas the two P. "reactans" isolates caused superficial light discoloration only after inoculation with 100 µl of the same concentration. Mixture from both bacterial suspensions increased the brown areas formed on the pileus. This is the first pathogenicity report of P. tolasii and P. "reactans" isolated from cultivated A. bisporus in Egypt.

  5. Hydrocarbonoclastic bacteria of the genus Pseudomonas in Samanea saman (Jacq. Merr. rhizosphere

    Directory of Open Access Journals (Sweden)

    Juliana Coromoto Mayz

    2017-01-01

    Full Text Available The research goal includes isolation, characterization and identification of Pseudomonas species existing in the rhizosphere of a legume present (colonizing or survivor in a savanna soil polluted by an oil spill, in order to explain the support of the growth of this leguminous plant through the reduction of the toxicity of spilled oil (hydrocarbonoclastic effects. The site is located at Amana del Tamarindo village entrance, Monagas State, Venezuela (9 ° 38' 52 "N, 63 ° 7' 20" E, 46 masl. An area of 50 m2 was sampled.  In concordance to the descriptions, keys, and comparison with the UOJ Herbarium exsiccatae, the legume collected was identified as Samanea saman (Jacq. Merr., which belongs to the Fabaceae family. The results of the biochemical characterization and the production of pyocyanine and fluorescein pigments allowed identifying 10 isolates as P. fluorescens, 5 as P. putida and 5 as P. aeruginosa. Samanea saman is recommended for re-vegetation of the contaminated area.

  6. Loss of Apc heterozygosity and abnormal tissue building in nascent intestinal polyps in mice carrying a truncated Apc gene.

    Science.gov (United States)

    Oshima, M; Oshima, H; Kitagawa, K; Kobayashi, M; Itakura, C; Taketo, M

    1995-05-09

    Mutations in the APC (adenomatous polyposis coli) gene appear to be responsible for not only familial adenomatous polyposis but also many sporadic cases of gastrointestinal cancers. Using homologous recombination in mouse embryonic stem cells, we constructed mice that contained a mutant gene encoding a product truncated at a 716 (Apc delta 716). Mendelian transmission of the gene caused most homozygous mice to die in utero before day 8 of gestation. The heterozygotes developed multiple polyps throughout the intestinal tract, mostly in the small intestine. The earliest polyps arose multifocally during the third week after birth, and new polyps continued to appear thereafter. Surprisingly, every nascent polyp consisted of a microadenoma covered with a layer of the normal villous epithelium. These microadenomas originated from single crypts by forming abnormal outpockets into the inner (lacteal) side of the neighboring villi. We carefully dissected such microadenomas from nascent polyps by peeling off the normal epithelium and determined their genotype by PCR: all microadenomas had already lost the wild-type Apc allele, whereas the mutant allele remained unchanged. These results indicate that loss of heterozygosity followed by formation of intravillous microadenomas is responsible for polyposis in Apc delta 716 intestinal mucosa. It is therefore unlikely that the truncated product interacts directly with the wild-type protein and causes the microadenomas by a dominant negative mechanism.

  7. How Business Idea Fit Affects Sustainability and Creates Opportunities for Value Co-Creation in Nascent Firms

    Directory of Open Access Journals (Sweden)

    Gian Luca Casali

    2018-01-01

    Full Text Available A well-defined business idea is essential for nascent business sustainability in the future. The business idea must fit firm knowledge and resources to a profitable business opportunity. This work adopts the framework of value co-creation, strongly related to the service-dominant logic paradigm. We ask how does business idea fit affect new venture sustainability and create opportunities for value co-creation. We propose that a business idea that lacks fit is less sustainable, but it could create opportunities for value co-creation. This study develops and validates an empirically grounded taxonomy of business idea fit based on 729 Australian nascent firms using quantitative data generated from the results of a large study called CAUSEE (Comprehensive Australian Study of Entrepreneurial Emergence. A cluster analysis is used to identify distinct patterns of business idea fit. The empirical taxonomy developed in this study found four distinct clusters of firms, which were distinguished by the fit of their new business idea to knowledge, resources and market profitability: very good fit, low knowledge fit, low profit fit and low fit. Results show how these different patterns of fit create opportunities for value co-creation to create business future sustainability.

  8. Dual binding mode of the nascent polypeptide-associated complex reveals a novel universal adapter site on the ribosome.

    Science.gov (United States)

    Pech, Markus; Spreter, Thomas; Beckmann, Roland; Beatrix, Birgitta

    2010-06-18

    Nascent polypeptide-associated complex (NAC) was identified in eukaryotes as the first cytosolic factor that contacts the nascent polypeptide chain emerging from the ribosome. NAC is present as a homodimer in archaea and as a highly conserved heterodimer in eukaryotes. Mutations in NAC cause severe embryonically lethal phenotypes in mice, Drosophila melanogaster, and Caenorhabditis elegans. In the yeast Saccharomyces cerevisiae NAC is quantitatively associated with ribosomes. Here we show that NAC contacts several ribosomal proteins. The N terminus of betaNAC, however, specifically contacts near the tunnel exit ribosomal protein Rpl31, which is unique to eukaryotes and archaea. Moreover, the first 23 amino acids of betaNAC are sufficient to direct an otherwise non-associated protein to the ribosome. In contrast, alphaNAC (Egd2p) contacts Rpl17, the direct neighbor of Rpl31 at the ribosomal tunnel exit site. Rpl31 was also recently identified as a contact site for the SRP receptor and the ribosome-associated complex. Furthermore, in Escherichia coli peptide deformylase (PDF) interacts with the corresponding surface area on the eubacterial ribosome. In addition to the previously identified universal adapter site represented by Rpl25/Rpl35, we therefore refer to Rpl31/Rpl17 as a novel universal docking site for ribosome-associated factors on the eukaryotic ribosome.

  9. Exploiting translational stalling peptides in an effort to extend azithromycin interaction within the prokaryotic ribosome nascent peptide exit tunnel.

    Science.gov (United States)

    Washington, Arren Z; Tapadar, Subhasish; George, Alex; Oyelere, Adegboyega K

    2015-08-15

    The ribosome is the primary protein synthesis machine in the cell and is a target for treatment of a variety of diseases including bacterial infection and cancer. The ribosomal peptide exit tunnel, the route of egress for the nascent peptide, is an inviting site for drug design. Toward a rational engagement of the nascent peptide components for the design of small molecule inhibitors of ribosome function, we designed and disclosed herein a set of N-10 indole functionalized azithromycin analogs. The indole moiety of these compounds is designed to mimic the translation stalling interaction of SecM W155 side-chain with the prokaryotic (Escherichia coli) ribosome A751 residue. Many of these N-10 functionalized compounds have enhanced translation inhibition activities against E. coli ribosome relative to azithromycin while a subset inhibited the growth of representative susceptible bacteria strains to about the same extent as azithromycin. Moreover, the inclusion of bovine serum in the bacterial growth media enhanced the anti-bacterial potency of the N-10 functionalized azithromycin analogs by as high as 10-fold. Published by Elsevier Ltd.

  10. Metabolic and biofungicidal properties of maize rhizobacteria for ...

    African Journals Online (AJOL)

    Pseudomonas putida, Pseudomonas fluorescens and Azospirillum lipoferum have highly produced many of the investigated metabolites. Streptomyces hygroscopicus, Streptomyces fasciculatus, Pseudomonas aeruginosa, P. putida, P. fluorescens and A. lipoferum inhibited mycelial growth of F. verticillioides and ...

  11. Elucidation of the Vibrio anguillarum genetic response to the potential fish probiont Pseudomonas fluorescens AH2, using RNA-arbitrarily primed PCR

    DEFF Research Database (Denmark)

    Holmstrøm, Kim; Gram, Lone

    2003-01-01

    with the iron chelator 2,2-dipyridyl. A chromosomal transcript homologous to vibE that participates in vibriobactin synthesis in Vibrio cholerae was also upregulated during AH2 exposure. This transcript could represent a functionally active gene in V. anguillarum involved in biosynthesis of anguibactin...

  12. A combined microcosm and mesocosm approach to examine factors affecting survival and mortality of Pseudomonas fluorescens Ag1 in seawater

    DEFF Research Database (Denmark)

    Ahl, Thomas; Christoffersen, K.; Riemann, B.

    1995-01-01

    .5‰). In microcosms with 0.2 µm-filtered seawater, addition of a carbon + nitrogen + phosphorus nutrient mixture was needed to induce proliferation of Ag1. In nutrient-amended microcosms the Ag1 population maintained viability, as determined by the direct viable counts method, at a level close to 100%. In natural...

  13. Sulfur-34S Stable Isotope Labeling of Amino Acids for Quantification (SULAQ34) of Proteomic Changes in Pseudomonas fluorescens during Naphthalene Degradation

    OpenAIRE

    Herbst, Florian-Alexander; Taubert, Martin; Jehmlich, Nico; Behr, Tobias; Schmidt, Frank; von Bergen, Martin; Seifert, Jana

    2013-01-01

    The relative quantification of proteins is one of the major techniques used to elucidate physiological reactions. Because it allows one to avoid artifacts due to chemical labeling, the metabolic introduction of heavy isotopes into proteins and peptides is the preferred method for relative quantification. For eukaryotic cells, stable isotope labeling by amino acids in cell culture (SILAC) has become the gold standard and can be readily applied in a vast number of scenarios. In the microbial re...

  14. Automatic Formation of Hypotheses on the Relationships between Structure of Naphthalene Analogs and Bioluminescence Response of Bioreporter Pseudomonas fluorescens HK44

    Czech Academy of Sciences Publication Activity Database

    Trögl, J.; Hálová, J.; Kuncová, Gabriela; Pařík, P.

    2010-01-01

    Roč. 55, č. 5 (2010), s. 411-417 ISSN 0015-5632 R&D Projects: GA MŠk OC 121 Institutional research plan: CEZ:AV0Z40720504 Keywords : substituted naphthalenes * bioconversion * optical biosensor Subject RIV: CE - Biochemistry Impact factor: 0.977, year: 2010

  15. Effects of spray-dried Pseudomonas fluorescens, strain CL145A (Zequanox®) on reproduction and early development of the fathead minnow (Pimephales promelas).

    Science.gov (United States)

    Waller, Diane L.; Luoma, James A.

    2016-01-01

    The biopesticide, Zequanox®, is registered for dreissenid mussel control in open water systems. Previous toxicity trials with nontarget organisms, including young-of-the year of several fish species and invertebrates, demonstrated selectivity of Zequanox for dreissenids. However, data are lacking on its safety to reproductive and early life stages of fish. The present study evaluated the effects of Zequanox on spawning and early life stages of the fathead minnow, Pimephales promelas, at the maximum approved concentration (100 mg Zequanox active ingredient /L) and exposure duration (8 h) for open water application. The results showed no significant effect of Zequanox on survival, condition, or cumulative egg deposition (21 d) in adult fathead minnow. Eggs (<24-h old) exposed to Zequanox developed to the eyed-stage at a similar rate to that of unexposed eggs. Additionally, Zequanox did not have a significant effect on survival and growth (90 d) of newly hatched fry (<24-h old). The results indicate that Zequanox treatment will not affect survival, spawning, and early life development of fathead minnows when applied at the recommended treatment regime.

  16. Identification of copper-induced genes in Pseudomonas fluorescens and use of a reporter strain to monitor bioavailable copper in soil

    DEFF Research Database (Denmark)

    Tom-Petersen, Andreas; Hosbond, Carsten; Nybroe, Ole

    2001-01-01

    Copper (Cu) is an essential trace metal required by living organisms but it is potentially toxic at higher concentrations. Cu salts are used as bactericides in agriculture and as growth promoters in pig production. They are consequently introduced to the environment, e.g. through manure, and might......-containing soil microcosms supplemented with either manure or straw. Organic matter increased the amount of soluble Cu but not the amount of bioavailable Cu. Probably, Cu binds with high affinity to organic constituents in pig manure or barley straw. Hence, determination of soluble Cu by chemical analysis cannot...

  17. A New Twist to the Kirby-Bauer Antibiotic Susceptibility Test Activity?Increasing Antibiotic Sensitivity of Pseudomonas fluorescens through Thermal Stress

    OpenAIRE

    Gerbig, Donald G.; Engohang-Ndong, Jean; Aubihl, Heather

    2013-01-01

    Antibiotic sensitivity and the effect of temperature on microbial growth are two standard laboratory activities found in most microbial laboratory manuals. We have found a novel way to combine the two activities to demonstrate how temperature can influence antibiotic sensitivity using a standard incubator in instructional laboratory settings. This activity reinforces the important concepts of microbial growth and temperature along with Kirby-Bauer antibiotic susceptibility testing. We found t...

  18. Direct imaging of RAB27B-enriched secretory vesicle biogenesis in lacrimal acinar cells reveals origins on a nascent vesicle budding site.

    Directory of Open Access Journals (Sweden)

    Lilian Chiang

    Full Text Available This study uses YFP-tagged Rab27b expression in rabbit lacrimal gland acinar cells, which are polarized secretory epithelial cells, to characterize early stages of secretory vesicle trafficking. Here we demonstrate the utility of YFP-Rab27b to delineate new perspectives on the mechanisms of early vesicle biogenesis in lacrimal gland acinar cells, where information is significantly limited. Protocols were developed to deplete the mature YFP-Rab27b-enriched secretory vesicle pool in the subapical region of the cell, and confocal fluorescence microscopy was used to track vesicle replenishment. This analysis revealed a basally-localized organelle, which we termed the "nascent vesicle site," from which nascent vesicles appeared to emerge. Subapical vesicular YFP-Rab27b was co-localized with p150(Glued, a component of the dynactin cofactor of cytoplasmic dynein. Treatment with the microtubule-targeted agent, nocodazole, did not affect release of mature secretory vesicles, although during vesicle repletion it significantly altered nascent YFP-Rab27b-enriched secretory vesicle localization. Instead of moving to the subapical region, these vesicles were trapped at the nascent vesicle site which was adjacent to, if not a sub-compartment of, the trans-Golgi network. Finally, YFP-Rab27b-enriched secretory vesicles which reached the subapical cytoplasm appeared to acquire the actin-based motor protein, Myosin 5C. Our findings show that Rab27b enrichment occurs early in secretory vesicle formation, that secretory vesicles bud from a visually discernable nascent vesicle site, and that transport from the nascent vesicle site to the subapical region requires intact microtubules.

  19. Pyoverdine synthesis by the Mn(II-oxidizing bacterium Pseudomonas putida GB-1

    Directory of Open Access Journals (Sweden)

    Dorothy Lundquist Parker

    2014-05-01

    Full Text Available When iron-starved, the Mn(II-oxidizing bacteria Pseudomonas putida strains GB-1 and MnB1 produce pyoverdines (PVDGB-1 and PVDMnB1, siderophores that both influence iron uptake and inhibit manganese(II oxidation by these strains. To explore the properties and genetics of a PVD that can affect manganese oxidation, LC-MS/MS and various siderotyping techniques were used to identify the peptides of PVDGB-1 and PVDMnB1 as being (for both PVDs: chromophore-Asp-Lys-OHAsp-Ser-Gly-aThr-Lys-cOHOrn, resembling a structure previously reported for P. putida CFML 90-51, which does not oxidize Mn. All three strains also produced an azotobactin and a sulfonated PVD, each with the peptide sequence above, but with unknown regulatory or metabolic effects. Bioinformatic analysis of the sequenced genome of P. putida GB-1 suggested that a particular non-ribosomal peptide synthetase, coded by the operon PputGB1_4083-4086, could produce the peptide backbone of PVDGB-1. To verify this prediction, plasmid integration disruption of PputGB1_4083 was performed and the resulting mutant failed to produce detectable PVD. In silico analysis of the modules in PputGB1_4083-4086 predicted a peptide sequence of Asp-Lys-Asp-Ser-Ala-Thr-Lsy-Orn, which closely matches the peptide determined by MS/MS. To extend these studies to other organisms, various Mn(II-oxidizing and non-oxidizing isolates of P. putida, P. fluorescens, P. marincola, P. fluorescens-syringae group, P. mendocina-resinovorans group and P. stutzerii group were screened for PVD synthesis. The PVD producers (12 out of 16 tested strains were siderotyped and placed into four sets of differing PVD structures, some corresponding to previously characterized PVDs and some to novel PVDs. These results combined with previous studies suggested that the presence of OHAsp or the flexibility of the pyoverdine polypeptide may enable efficient binding of Mn(III.

  20. Pyoverdine synthesis by the Mn(II)-oxidizing bacterium Pseudomonas putida GB-1

    Science.gov (United States)

    Parker, Dorothy L.; Lee, Sung-Woo; Geszvain, Kati; Davis, Richard E.; Gruffaz, Christelle; Meyer, Jean-Marie; Torpey, Justin W.; Tebo, Bradley M.

    2014-01-01

    When iron-starved, the Mn(II)-oxidizing bacteria Pseudomonas putida strains GB-1 and MnB1 produce pyoverdines (PVDGB-1 and PVDMnB1), siderophores that both influence iron uptake and inhibit manganese(II) oxidation by these strains. To explore the properties and genetics of a PVD that can affect manganese oxidation, LC-MS/MS, and various siderotyping techniques were used to identify the peptides of PVDGB-1 and PVDMnB1 as being (for both PVDs): chromophore-Asp-Lys-OHAsp-Ser-Gly-aThr-Lys-cOHOrn, resembling a structure previously reported for P. putida CFML 90-51, which does not oxidize Mn. All three strains also produced an azotobactin and a sulfonated PVD, each with the peptide sequence above, but with unknown regulatory or metabolic effects. Bioinformatic analysis of the sequenced genome of P. putida GB-1 suggested that a particular non-ribosomal peptide synthetase (NRPS), coded by the operon PputGB1_4083-4086, could produce the peptide backbone of PVDGB-1. To verify this prediction, plasmid integration disruption of PputGB1_4083 was performed and the resulting mutant failed to produce detectable PVD. In silico analysis of the modules in PputGB1_4083-4086 predicted a peptide sequence of Asp-Lys-Asp-Ser-Ala-Thr-Lsy-Orn, which closely matches the peptide determined by MS/MS. To extend these studies to other organisms, various Mn(II)-oxidizing and non-oxidizing isolates of P. putida, P. fluorescens, P. marincola, P. fluorescens-syringae group, P. mendocina-resinovorans group, and P. stutzerii group were screened for PVD synthesis. The PVD producers (12 out of 16 tested strains) were siderotyped and placed into four sets of differing PVD structures, some corresponding to previously characterized PVDs and some to novel PVDs. These results combined with previous studies suggested that the presence of OHAsp or the flexibility of the pyoverdine polypeptide may enable efficient binding of Mn(III). PMID:24847318

  1. Pseudomonas-follikulitis efter badning i spabad

    DEFF Research Database (Denmark)

    Uldall Pallesen, Kristine Appel; Andersen, Klaus Ejner; Mørtz, Charlotte Gotthard

    2012-01-01

    . We describe a 23-year-old healthy woman who developed a pustular rash and general malaise after using a spa bath contaminated with Pseudomonas aeruginosa. Bacterial culture from a pustule confirmed Pseudomonas folliculitis and the patient was treated with ciprofloxacin with rapid good effect....

  2. Petroleum-hydrocarbons biodegradation by Pseudomonas strains ...

    African Journals Online (AJOL)

    Many indigenous microorganisms in water and soil are capable of degrading hydrocarbon contaminants. In this study, two bacterial strains were isolated from a contaminated soil of a refinery of Arzew (Oran). The isolated strains were identified as Pseudomonas aeruginosa (P3) and Pseudomonas fluoresens (P4).

  3. Pseudomonas Septic Arthritis | Thanni | Nigerian Journal of ...

    African Journals Online (AJOL)

    BACKGROUND: Septic arthritis due to pseudomonas species is unusual and when it occurs, there is often an underlying cause like immune depression, intravenous drug abuse or a penetrating injury. PATIENT AND METHOD: We report a case of pseudomonas septic arthritis complicating cannulation of a leg vein following ...

  4. Combined effects of NaCl, NaOH, and biocides (monolaurin or lauric acid) on inactivation of Listeria monocytogenes and Pseudomonas spp.

    Science.gov (United States)

    Vasseur, C; Rigaud, N; Hébraud, M; Labadie, J

    2001-09-01

    This study highlighted combinations of chemical stresses that could decrease or eliminate Listeria monocytogenes and Pseudomonas spp. surviving in food processing plants. Strains of L. monocytogenes, Pseudomonas fragi, and Pseudomonas fluorescens isolated from processing environments (meat and milk) were grown at 20 degrees C up to the early stationary phase. The strains were then subjected to 30 min of physicochemical treatments. These treatments included individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (fatty acids) challenges. Survival of the strains was studied after individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (monolaurin, lauric acid) challenges. Individual pH shocks had lower efficiencies than those used in combinations with other parameters. The treatment pH 5.4 followed by pH 10.5 had a low efficiency against L. monocytogenes. The opposite combination, pH 10.5 followed by pH 5.4, led to a 3-log reduction of the L. monocytogenes population. Pseudomonas spp. strains were much more sensitive than L. monocytogenes, and population reductions of 5 and 8 log (total destruction), respectively, were observed after the same treatments. As for L. monocytogenes, the combination pH 10.5 followed by pH 5.4 is more deleterious than the opposite. Whatever the bacterial species, the most efficient treatments were combinations of alkaline, osmotic, and biocide shocks. For instance, the combination pH 10.5 and 10% NaCl plus biocides showed reductions of 5 to 8 log for both bacteria. The origins of the observed lethal effects are discussed.

  5. Multi-imaging analysis of nascent surface structures generated during femtosecond laser irradiation of silicon in high vacuum

    Science.gov (United States)

    Gesuele, F.; JJ Nivas, J.; Fittipaldi, R.; Altucci, C.; Bruzzese, R.; Maddalena, P.; Amoruso, S.

    2018-02-01

    We report a correlative imaging analysis of a crystalline silicon target after irradiation with a low number of 1055 nm, 850 fs laser pulses with several microscopy techniques (e.g., scanning electron microscopy, atomic force microscopy, Raman micro-imaging and confocal optical microscopy). The analysis is carried out on samples irradiated both in high vacuum and at atmospheric pressure conditions, evidencing interesting differences induced by the ambient environment. In high-vacuum conditions, the results evidence the formation of a halo, which is constituted by alternate stripes of amorphous and crystalline silicon, around the nascent ablation crater. In air, such an effect is drastically reduced, due to the significant back-deposition of nanoparticulate material induced by the larger ambient pressure.

  6. Buildup of aerosol precursor gases and sulfur-induced activation of soot in nascent jet aircraft exhaust plumes

    Energy Technology Data Exchange (ETDEWEB)

    Kaercher, B.; Hirschberg, M.M.; Fabian, P. [Muenchen Univ. (Germany). Lehrstuhl fuer Bioklimatologie und Immissionsforschung; Gerz, T. [Deutsche Forschungsanstalt fuer Luft- und Raumfahrt e.V. (DLR), Oberpfaffenhofen (Germany). Inst. fuer Physik der Atmosphaere

    1997-12-31

    Research issues concerning the chemical transformation of exhaust trace gases are summarized. The photochemical evolution of NO{sub x} early in the plume is strongly coupled to plume mixing. Substantial amounts of HNO{sub 3} are generated in nascent plumes even if no NO{sub 2} is emitted. The production of H{sub 2}SO{sub 4} becomes very efficient if part of the fuel sulfur is emitted as SO{sub 3}. Each emitted soot particle can acquire 1-10% by mass fully oxidized sulfur molecules prior to binary homogeneous nucleation, if a few percent of the exhaust SO{sub x} are emitted as SO{sub 3}, indicating an important activation pathway for soot, and leading to a marked enhancement of new aerosol formation and growth rates. (author) 11 refs.

  7. Effects of carcinogen treatment on rat liver DNA synthesis in vivo and on nascent DNA synthesis and elongation in cultured hepatocytes

    International Nuclear Information System (INIS)

    Zurlo, J.; Mignano, J.E.; Eustice, D.C.; Poirier, M.C.; Yager, J.D.

    1986-01-01

    One objective of this study was to determine the effects of N-hydroxy-2-acetylaminofluorene (N-OH-AAF) treatment on DNA synthesis in regenerating rat liver. It was found that N-OH-AAF caused a dose-dependent inhibition of [ 3 H]thymidine incorporation into liver DNA. This inhibition was followed by a gradual, but incomplete recovery. The second objective of the study was to determine the effects of DNA damage on the size distribution and elongation of nascent hepatocyte DNA. Hepatocytes, which have been shown to demonstrate a pattern of inhibition and subsequent recovery of DNA synthesis following UV irradiation similar to that seen in vivo upon treatment with N-OH-AAF were cultured. The size distribution of nascent DNA after UV irradiation was determined by pH step gradient alkaline elution analysis. The results show that UV irradiation caused a dose-dependent decrease in the size distribution of nascent DNA suggesting an inhibition of elongation. The results show that resumption of DNA synthesis and nascent strand elongation occur on damaged templates. These observations along with previous studies support the idea that DNA damage leading to inhibition of DNA synthesis may induce SOS-type processes which if mutagenic may play a role in the initiation of carcinogenesis. (Auth.)

  8. Rad51 recombinase prevents Mre11 nuclease-dependent degradation and excessive PrimPol-mediated elongation of nascent DNA after UV irradiation

    Science.gov (United States)

    Vallerga, María Belén; Mansilla, Sabrina F.; Federico, María Belén; Bertolin, Agustina P.; Gottifredi, Vanesa

    2015-01-01

    After UV irradiation, DNA polymerases specialized in translesion DNA synthesis (TLS) aid DNA replication. However, it is unclear whether other mechanisms also facilitate the elongation of UV-damaged DNA. We wondered if Rad51 recombinase (Rad51), a factor that escorts replication forks, aids replication across UV lesions. We found that depletion of Rad51 impairs S-phase progression and increases cell death after UV irradiation. Interestingly, Rad51 and the TLS polymerase polη modulate the elongation of nascent DNA in different ways, suggesting that DNA elongation after UV irradiation does not exclusively rely on TLS events. In particular, Rad51 protects the DNA synthesized immediately before UV irradiation from degradation and avoids excessive elongation of nascent DNA after UV irradiation. In Rad51-depleted samples, the degradation of DNA was limited to the first minutes after UV irradiation and required the exonuclease activity of the double strand break repair nuclease (Mre11). The persistent dysregulation of nascent DNA elongation after Rad51 knockdown required Mre11, but not its exonuclease activity, and PrimPol, a DNA polymerase with primase activity. By showing a crucial contribution of Rad51 to the synthesis of nascent DNA, our results reveal an unanticipated complexity in the regulation of DNA elongation across UV-damaged templates. PMID:26627254

  9. Eggplant and related species are promising genetic resources to dissect the plant immune response to Pseudomonas syringae and Xanthomonas euvesicatoria and to identify new resistance determinants.

    Science.gov (United States)

    Clarke, Christopher R; Hayes, Byron W; Runde, Brendan J; Wicker, Emmanuel; Vinatzer, Boris A

    2014-10-01

    The apparent lack of durability of many resistance (R) genes highlights the need for the constant identification of new genetic sources of resistance for the breeding of new disease-resistant crop cultivars. To this end, we screened a collection of accessions of eggplant and close relatives for resistance against Pseudomonas syringae pv. tomato (Pto) and Xanthomonas euvesicatoria (Xeu), foliar plant pathogens of many solanaceous crops. Both pathogens caused substantial disease on most genotypes of eggplant and its relatives. Promisingly, however, some of the genotypes were fully or partially resistant to either of the pathogens, suggesting the presence of effective resistance determinants in these genotypes. Segregation of resistance to the growth of Xeu following infiltration in F2 progeny from a cross of a resistant and susceptible genotype suggests that resistance to Xeu is inherited as a multigenic trait. With regard to Pto, a mutant strain lacking all 28 functional type III secreted effectors, and a Pseudomonas fluorescens strain expressing a P. syringae type III secretion system (T3SS), both elicit a strong cell death response on most eggplant lines. Several genotypes thus appear to harbour a mechanism for the direct recognition of a component of the T3SS. Therefore, eggplant and its close relatives are promising resources to unravel novel aspects of plant immunity and to identify new candidate R genes that could be employed in other Solanaceae in which Xeu and Pto cause agriculturally relevant diseases. © 2014 BSPP AND JOHN WILEY & SONS LTD.

  10. Characterization of a metal resistant Pseudomonas sp. isolated from uranium mine for its potential in heavy metal (Ni2+, Co2+, Cu2+, and Cd2+) sequestration.

    Science.gov (United States)

    Choudhary, Sangeeta; Sar, Pinaki

    2009-05-01

    Heavy metal sequestration by a multimetal resistant Pseudomonas strain isolated from a uranium mine was characterized for its potential application in metal bioremediation. 16S rRNA gene analysis revealed phylogenetic relatedness of this isolate to Pseudomonas fluorescens. Metal uptake by this bacterium was monophasic, fast saturating, concentration and pH dependent with maximum loading of 1048 nmol Ni(2+) followed by 845 nmol Co(2+), 828 nmol Cu(2+) and 700 nmol Cd(2+)mg(-1) dry wt. Preferential metal deposition in cell envelope was confirmed by TEM and cell fractionation. FTIR spectroscopy and EDX analysis revealed a major role of carboxyl and phosphoryl groups along with a possible ion exchange mechanism in cation binding. Binary system demonstrated selective metal binding affinity in the order of Cu(2+)>Ni(2+)>Co(2+)>Cd(2+). A comparison with similar metal uptake reports considering live bacteria strongly indicated the superiority of this strain in metal sequestration, which could be useful for developing efficient metal removal system.

  11. Live cell imaging of the nascent inactive X chromosome during the early differentiation process of naive ES cells towards epiblast stem cells.

    Directory of Open Access Journals (Sweden)

    Aurélia Guyochin

    Full Text Available Random X-chromosome inactivation ensures dosage compensation in mammals through the transcriptional silencing of one of the two X chromosomes present in each female cell. Silencing is initiated in the differentiating epiblast of the mouse female embryos through coating of the nascent inactive X chromosome by the non-coding RNA Xist, which subsequently recruits the Polycomb Complex PRC2 leading to histone H3-K27 methylation. Here we examined in mouse ES cells the early steps of the transition from naive ES cells towards epiblast stem cells as a model for inducing X chromosome inactivation in vitro. We show that these conditions efficiently induce random XCI. Importantly, in a transient phase of this differentiation pathway, both X chromosomes are coated with Xist RNA in up to 15% of the XX cells. In an attempt to determine the dynamics of this process, we designed a strategy aimed at visualizing the nascent inactive X-chromosome in live cells. We generated transgenic female XX ES cells expressing the PRC2 component Ezh2 fused to the fluorescent protein Venus. The fluorescent fusion protein was expressed at sub-physiological levels and located in nuclei of ES cells. Upon differentiation of ES cell towards epiblast stem cell fate, Venus-fluorescent territories appearing in interphase nuclei were identified as nascent inactive X chromosomes by their association with Xist RNA. Imaging of Ezh2-Venus for up to 24 hours during the differentiation process showed survival of some cells with two fluorescent domains and a surprising dynamics of the fluorescent territories across cell division and in the course of the differentiation process. Our data reveal a strategy for visualizing the nascent inactive X chromosome and suggests the possibility for a large plasticity of the nascent inactive X chromosome.

  12. Live cell imaging of the nascent inactive X chromosome during the early differentiation process of naive ES cells towards epiblast stem cells.

    Science.gov (United States)

    Guyochin, Aurélia; Maenner, Sylvain; Chu, Erin Tsi-Jia; Hentati, Asma; Attia, Mikael; Avner, Philip; Clerc, Philippe

    2014-01-01

    Random X-chromosome inactivation ensures dosage compensation in mammals through the transcriptional silencing of one of the two X chromosomes present in each female cell. Silencing is initiated in the differentiating epiblast of the mouse female embryos through coating of the nascent inactive X chromosome by the non-coding RNA Xist, which subsequently recruits the Polycomb Complex PRC2 leading to histone H3-K27 methylation. Here we examined in mouse ES cells the early steps of the transition from naive ES cells towards epiblast stem cells as a model for inducing X chromosome inactivation in vitro. We show that these conditions efficiently induce random XCI. Importantly, in a transient phase of this differentiation pathway, both X chromosomes are coated with Xist RNA in up to 15% of the XX cells. In an attempt to determine the dynamics of this process, we designed a strategy aimed at visualizing the nascent inactive X-chromosome in live cells. We generated transgenic female XX ES cells expressing the PRC2 component Ezh2 fused to the fluorescent protein Venus. The fluorescent fusion protein was expressed at sub-physiological levels and located in nuclei of ES cells. Upon differentiation of ES cell towards epiblast stem cell fate, Venus-fluorescent territories appearing in interphase nuclei were identified as nascent inactive X chromosomes by their association with Xist RNA. Imaging of Ezh2-Venus for up to 24 hours during the differentiation process showed survival of some cells with two fluorescent domains and a surprising dynamics of the fluorescent territories across cell division and in the course of the differentiation process. Our data reveal a strategy for visualizing the nascent inactive X chromosome and suggests the possibility for a large plasticity of the nascent inactive X chromosome.

  13. Growth of Pseudomonas spp. in cottage cheese

    DEFF Research Database (Denmark)

    Østergaard, Nina Bjerre; Dalgaard, Paw

    of spoilage microorganisms in cottage cheese can cause undesirable alterations in flavour, odour, appearance and texture. Contamination and growth of psychrotolerant pseudomonads including Pseudomonas fragi and Pseudomonas putida has been reported for cottage cheese but the influence of these bacteria...... (pH 7.0) showed interesting results. Despite a lower pH value in the cottage cheese, compared to the dressing, more rapid growth was observed. This may be caused by insufficient amounts of oxygen in the cream dressing having a negative effect on growth of Pseudomonas spp. At 15˚C growth...

  14. Pseudomona pseudomallei community acquired pneumonia

    International Nuclear Information System (INIS)

    Severiche, Diego

    1998-01-01

    This is the first published case report en Colombia about pseudomona pseudomallei community acquired pneumonia. This uncommon pathogen is from the epidemiological standpoint a very important one and medical community should be aware to look after it in those patients where no other etiological pathogen is recovered. A brief summary about epidemiology is showed, emphasizing those regions where it can be found. Likewise, comments about the differential diagnosis are important since it should be considered in those patients where tuberculosis is suspected. This is particularly representative for countries with high tuberculosis rates. Furthermore, a microbiological review is shown, emphasizing on isolation techniques, descriptions about therapeutics and other regarding treatment issues according international standards. Finally; a description about the clinical picture, laboratory findings, treatment and evolution of the case reported are shown for discussion

  15. Silver against Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Kirketerp-Møller, K.; Kristiansen, S.

    2007-01-01

    Silver has been recognized for its antimicrobial properties for centuries. Most studies on the antibacterial efficacy of silver, with particular emphasis on wound healing, have been performed on planktonic bacteria. Our recent studies, however, strongly suggest that colonization of wounds involves...... bacteria in both the planktonic and biofilm modes of growth. The action of silver on mature in vitro biofilms of Pseudomonas aeruginosa, a primary pathogen of chronic infected wounds, was investigated. The results show that silver is very effective against mature biofilms of P. aeruginosa......, but that the silver concentration is important. A concentration of 5-10 ig/mL silver sulfadiazine eradicated the biofilm whereas a lower concentration (1 ig/mL) had no effect. The bactericidal concentration of silver required to eradicate the bacterial biofilm was 10-100 times higher than that used to eradicate...

  16. Antibiotic Conditioned Growth Medium of Pseudomonas Aeruginosa

    Science.gov (United States)

    Benathen, Isaiah A.; Cazeau, Barbara; Joseph, Njeri

    2004-01-01

    A simple method to study the consequences of bacterial antibiosis after interspecific competition between microorganisms is presented. Common microorganisms are used as the test organisms and Pseudomonas aeruginosa are used as the source of the inhibitor agents.

  17. Production and characterization of biosurfactant from Pseudomonas ...

    African Journals Online (AJOL)

    Further characterization of biosurfactant using Fourier transform infrared spectroscopy (FTIR) revealed it as a rhamnolipid. Keywords: Mangrove ecosystems, Pseudomonas aeruginosa, biosurfactant, critical micelle concentration (CMC), FT-IR fourier transform infrared spectroscopy (FTIR). African Journal of Biotechnology, ...

  18. Cell death in Pseudomonas aeruginosa biofilm development

    DEFF Research Database (Denmark)

    Webb, J.S.; Thompson, L.S.; James, S.

    2003-01-01

    Bacteria growing in biofilms often develop multicellular, three-dimensional structures known as microcolonies. Complex differentiation within biofilms of Pseudomonas aeruginosa occurs, leading to the creation of voids inside microcolonies and to the dispersal of cells from within these voids...

  19. Pseudomonas aeruginosa (Family Pseudomonadaceae) is an ...

    African Journals Online (AJOL)

    Pseudomonas aeruginosa (Family Pseudomonadaceae) is an obligate aerobic, motile, gram negative bacillus.which is able to grow and survive in almost any environment and resistant to temperature extremes. It is involved in the etiology of several diseases i.

  20. Stratified growth in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Werner, E.; Roe, F.; Bugnicourt, A.

    2004-01-01

    In this study, stratified patterns of protein synthesis and growth were demonstrated in Pseudomonas aeruginosa biofilms. Spatial patterns of protein synthetic activity inside biofilms were characterized by the use of two green fluorescent protein (GFP) reporter gene constructs. One construct...