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  • 1
    Electronic Resource
    Electronic Resource
    Physical and genetic map of the Pseudomonas fluorescens SBW25 chromosome (1996)
    Oxford BSL : Blackwell Science Ltd
    Molecular microbiology 19 (1996), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Pseudomonas fluorescens is a saprophytic bacterium commonly isolated from soil, water, and the surfaces and tissues of plants and animals. The species has important applications in biotechnology because it can enhance plant growth and protect crops against disease. A complete physical map of the 6.63 Mbp P. fluorescens SBW25 chromosome was constructed using data obtained from combinations of one- and two-dimensional electrophoresis of completely or partially digested chromosomal DNA with end labelling. In total, 139 restriction sites (15 PacI, 53 SpeI, 71 XbaI) were placed on the physical map and complete maps of the circular chromosome were obtained for both PacI and SpeI; only XbaI fragments linking SpeI fragments were positioned. The average resolution of restriction sites was 48 kbp. A genetic map was derived from the physical map by Southern hybridization and 31 genes were positioned including oriC, rDNA operons (rnnA–E), recAgacA and pyvD
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1996.391926.x
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  • 2
    Electronic Resource
    Electronic Resource
    Survival, Colonization and Dispersal of Genetically Modified Pseudomonas fluorescens SBW25 in the Phytosphere of Field Grown Sugar Beet (1995)
    [s.l.] : Nature Publishing Company
    Nature biotechnology 13 (1995), S. 1493-1497 
    Details
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] A genetically modified microorganism (GMM), Pseudomonas fluorescens SBW25 (lacZY and kanr-xylE), derived from a strain taken from the leaves of mature sugar beet (Beta vulgaris var. Amethyst), was released as a seed inoculum (ca 1×107 cfu/seed) to field-grown sugar beet in 1993 and 1994. The ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nbt1295-1493
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  • 3
    Electronic Resource
    Electronic Resource
    Description of a novel plasmid replicative origin from a genetically distinct family of conjugative plasmids associated with phytosphere microflora (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 149 (1997), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A novel replicative origin (oriV) from a conjugative, mercury resistance plasmid (pQBR11, 304 kbp) has been cloned and sequenced. Homology to the pQBR11 oriV-containing 3.55 kbp BamHI fragment (pCV1200) was restricted to one of five genetically distinct classes (group I) of narrow host range, mega-plasmids that persist as a genetic component of the pseudomonad community indigenous to the microflora of sugar beet. The oriV of pQBR11 was located within a unique sequence of 300 bp which initiated the replication of pUC derived suicide vectors in Pseudomonas putida UWC1. The limited size of the DNA sequence required to initiate replication, and the presence of two 15/16 bp directly repeated motifs, indicate that this group of mega-plasmids contain a single origin of replication, which initiates replication via a host-polymerase dependent rolling circle mechanism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1997.tb10318.x
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  • 4
    Electronic Resource
    Electronic Resource
    Demonstration of tra+ plasmid activity in bacteria indigenous to the phyllosphere of sugar beet; gene transfer to a recombinant pseudomonad (1993)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 12 (1993), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The presence of transfer proficient plasmids in bacteria isolated from the leaves of sugar beet (Beta vulgaris L.) was studied. Of 435 bacteria sampled 79 (18%) contained plasmids. Pseudomonads (30%), Erwinia (12%) and Klebsiella (9%) were the largest populations sampled of which 22%, 33% and 29%, respectively, contained plasmids. The ability of these plasmids to self-transfer or mediate the mobilization of the tra− mob+ broad host range IncQ plasmid R300B was determined. R300B was maintained in 61/79 natural plasmid containing isolates, the Gram positive isolates could not support R300B. Pseudomonas aureofaciens SBW25, isolated from sugar beet leaves, was chromosomally marked with a tetracycline resistance gene and used as a recipient (SBW25ETc). Five isolates of Erwinia herbicola and one of Erwinia salicis containing natural plasmids were able to mobilize R300B into the recombinant, SBW25ETc. These mobolizing (tra+) plasmids were not maintained in transconjugant SBW25 cells. Analysis of the fragment patterns of Pst I digested plasmid DNA demonstrated that four (pSB139, pSB140, pSB142, pSB146; 110 kb) were identical, one (pSB153; 65 kb) was common to a subset of fragments in these four and another (pSB169; 100 kb) was unique. Other natural isolates were able to transfer copper resistance (Erwinia rhapontici, 2 strains) or mercury resistance (Pseudomonas fluorescens SBW340) to a rifampicin resistant recipient Pseudomonas putida UWC1 but not to SBW25ETc. These self-transferable plasmids were not able to mobilize R300B. These data demonstrate that the phyllosphere supports indigenous microbial populations which have the capacity to transfer genetic material between bacteria of different genera.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1993.tb00032.x
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  • 5
    Electronic Resource
    Electronic Resource
    Temporal fluctuations in the pseudomonad population associated with sugar beet leaves (1999)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 28 (1999), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A total of 473 fluorescent pseudomonads were isolated from sugar beet leaves, sampled approximately every 30–40 days, over three consecutive growing seasons. Total cellular fatty acid composition and restriction fragment length polymorphism data for the flanking regions of the ribosomal RNA operon were obtained for each isolate to assess the temporal variability in diversity of leaf-associated fluorescent pseudomonad populations. A total of 115 distinct ribotypes (genotypes) were detected within the cluster identified by principal component analysis of cellular fatty acid content. Although the abundance of isolates was lognormally distributed, individual ribotypes were detected more often than would be predicted by such a distribution. The number of ribotypes identified in each sample was directly related to the size of that sample. Despite the implication that the sample size was too small, the data suggested that some individual ribotypes were capable of long-term survival and that there was a dynamic, non-random and continuous turnover within the population. A seasonal pattern of isolate reoccurrence was observed. As one group suffered decline, the next group, better adapted to the prevailing conditions, proliferated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1999.tb00589.x
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  • 6
    Electronic Resource
    Electronic Resource
    Two dnaB genes are associated with the origin of replication of pQBR55, an exogenously isolated plasmid from the rhizosphere of sugar beet (2002)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 42 (2002), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Plasmid pQBR55 (∼149 kb) represents one of five (Groups I–V) genetically distinct transfer proficient, mercury resistance plasmid groups that have been observed in the phytosphere pseudomonad community at a single geographic location in Oxford, UK. A 4.9-kb HindIII fragment was cloned from pQBR55 (a Group III plasmid) that facilitates autonomous replication of a narrow host range cloning vector, pKIL29, in the non-permissive host Pseudomonas putida UWC1. Sequencing revealed that the fragment contains a unique tandem array of dnaB genes (one partial and one complete), a homologue of the traA gene of plasmid RP4 and several potential open reading frames with little homology to any known sequences. The fragment also contains: a short region of direct and indirect repeats, two sequences that resemble the consensus Escherichia coli DnaA box motif, an A+T rich region and evidence of a GC-skew inversion. All features associated with origins of replication. Two specific oligonucleotide primer pairs, one targeted at the tandem dnaB genes and the other at the dnaB–traA arrangement were used for PCR analysis of other plasmids isolated from the same field site. PCR generated amplification products were only amplified from Group III plasmids (defined by RFLP typing) but not from plasmids belonging to Group I, II, IV or V. The nucleotide sequences of the amplified fragments were identical to those of pQBR55, even though the other Group III plasmids had distinct RFLP patterns. The results of a more general PCR screen, using primers to amplify a 389-bp fragment of all described pseudomonad dnaB genes, suggests that closely related dnaB sequences are only associated with Group III plasmids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.2002.tb01010.x
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  • 7
    Electronic Resource
    Electronic Resource
    The dispersal and establishment of pseudomonad populations in the phyllosphere of sugar beet by phytophagous caterpillars (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 24 (1997), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The role of phytophagous insects in the dispersal of phyllosphere bacterial populations has been investigated. Following seed inoculation, a leaf colonising population of Pseudomonas fluorescens SBW25EeZY6KX (lacZY and aph-xylE) was established in field grown sugar beet (Beta vulgaris var. amethyst). Third instar, Mamestra brassicae (Lepidoptera: Noctuidae) larvae were placed onto the emerging leaves of mature plants and the dispersal of the marked strain (ca. 5×105 cfu/g leaf) to untreated plants monitored. Forty-two days after caterpillar introduction, approximately twenty days after pupation of the larvae, an established population of transferred bacteria was detected at densities similar to those determined for seed inoculated plants of the same age. Under appropriate conditions phylloplane colonising pseudomonads can be dispersed by invertebrates and establish viable populations on the leaves of recipient plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1997.tb00431.x
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  • 8
    Electronic Resource
    Electronic Resource
    Diversity of mercury resistance plasmids obtained by exogenous isolation from the bacteria of sugar beet in three successive years (1996)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 20 (1996), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract: Self-transmissible plasmids conferring mercury resistance were exogenously isolated from the bacterial populations of sugar beet roots (rhizoplane) and leaves (phyllosphere) into a Pseudomonas putida recipient. Fifty rhizoplane plasmids and 29 phyllosphere plasmids (60–383 kb) were purified. Numerical analysis of plasmid DNA restriction enzyme digest patterns identified five distinct groups. Three of these plasmid groups were isolated from sugar beet crops grown at the same site over three consecutive years, demonstrating their established presence. Each group of plasmids comprised individual isolates with structural additions or deletions. The frequency of exogenous isolation correlated with factors likely to influence plant growth, bacterial activity and the physiological state of donors prior to sampling. All plasmids investigated conferred narrow spectrum mercury resistance with a reductase detoxification mechanism. None of the plasmids conferred resistance to a range of antibiotics, other heavy metals, or to UV, and following transfer to recipient bacteria the range of carbon source utilisation was not altered. This is the first report of the persistence of Pseudomonas spp. plasmid structural types isolated over several years from a terrestrial habitat.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1996.tb00320.x
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  • 9
    Electronic Resource
    Electronic Resource
    Short-term community dynamics in the phyllosphere microbiology of field-grown sugar beet (1995)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 16 (1995), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Short-term fluctuations in the number of culturable microorganisms and the composition of the bacterial community colonising the phyllosphere of field grown sugar beet were determined at 6 h intervals over a 42 h period. The number of culturable bacteria and Erwinia were found to fluctuate diurnally, increasing in size overnight and declining during daylight hours. In contrast, the number of Arthrobacter and yeasts were constant throughout the study, even after rainfall. Over most of the study the number of filamentous fungi and pseudomonads decreased. The short-term fluctuations in bacterial numbers were similar in magnitude to those detected over most of the previous season. No significant variation was observed in microbial numbers between samples taken from three locations within the field. Fatty acid analysis of isolated bacteria revealed the community to be similar in composition to that detected during the same period the previous year and indicated no significant short-term temporal or spatial variation. To limit the effects of diurnal fluctuations on the number of culturable bacteria detected, it is recommended that in long-term studies, samples be collected at the same time of day on each occasion.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1995.tb00284.x
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  • 10
    Electronic Resource
    Electronic Resource
    Autecology of a genetically modified fluorescent pseudomonad on sugar beet (1995)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 17 (1995), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract: The survival, dispersal and impact of a genetically modified microorganism (GMM) in the phyllosphere of glasshouse grown sugar beet was investigated. The GMM, Pseudomonas fluorescens SBW25EeZY6KX (lacZY and kanr-xylE) derived from a bacterium originally isolated from field grown sugar beet, was introduced as a seed inoculum. It survived in the phyllosphere throughout the 531-day study on plants growing in field soil. The bacterium was highly competitive, in some samples representing the largest bacterial population isolated. The GMM also established on a broad range of other plant species when introduced on the seed. Fluctuations in the numbers of GMM correlated with those of the indigenous pseudomonad population, indicating the organism had not been adversely affected by the addition of novel genes. Inoculum density had no significant effect on the number of GMMs detected in the phyllosphere, although it affected the population size in the rhizosphere. The GMM persisted in fallow soil for at least 10 days and subsequently colonised germinating untreated plants. Limited potential for GMM dispersal from the phyllosphere was detected using physical means of transfer, although moisture assisted dispersal from colonised plants. GMM transfer to other plant species growing between treated plants was detected. Insects captured in the glasshouse carried the GMM, indicating a potential for long distance dispersal. Introduction of the wild-type and GMM on the seed had a transient impact on the number and composition of the bacterial community colonising treated plants. Assessment of community diversity by fatty acid methyl ester (FAME) content of bacterial isolates revealed that the GMM had less impact on the bacterial community than the wild-type. Both organisms competitively excluded Enterobacteriaceae taxa, although this impact was attributed in part to the low taxa diversity that developed in the artificial conditions of the glasshouse. Instances of the GMM undergoing phenotypic change after introduction on the seed were detected by FAME analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1995.tb00122.x
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