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J I Prosser

Publications and source records attributed to J I Prosser.

9 recordsLinked to original sources

Luminometric measurement of population activity of genetically modified Pseudomonas fluorescens in the soil.

Genetically modified cells of Pseudomonas fluorescens, chromosomally marked with genes for bioluminescence, were inoculated into sterile soil microcosms. During incubation for 90 days, viable cell concentration did not change significantly but light output, measured by luminometry, decreased, indicating reduced metabolic activity due to lack of substrates. Amendment with nutrients resulted in parallel increases in both luminescence and dehydrogenase activity. Luminometry therefore enables rapid monitoring of the activity of populations of luminescence-marked microbial inocula in the soil, with greater sensitivity and selectivity than traditional techniques.

Bacteriological Techniques

Protection of Nitrosomonas europaea colonizing clay minerals from inhibition by nitrapyrin.

Nitrate production by Nitrosomonas europaea in inorganic liquid medium containing ammonium was limited by reduction in pH. In the presence of montmorillonite and vermiculite, expanding clays with high cation-exchange-capacity (CEC), nitrite yield was increased, ammonia oxidation continued at pH values below those which inhibited growth in the absence of clays and growth was biphasic. The first phase was similar to that in the absence of clays, while the second was characterized by a lower rate of nitrite production. Illite, a non-expanding clay with low CEC, had no significant effect on ammonia oxidation, while oxidation of ammonia-treated vermiculite (ATV) occurred with no significant change in the pH of the medium. ATV, montmorillonite and vermiculite, but not illite, protected cells from inhibition by nitrapyrin at concentrations inhibitory to cells growing in suspended culture. This protection was maintained in ATV homo-ionic to Al3+, but montmorillonite made homo-ionic to Al3+ did not provide protection from inhibition. Attachment of cells to clays with high CEC is therefore advantageous in providing exchange at the clay surface of NH+4 and H+ produced by ammonia oxidation, in reducing pH toxicity, and in protecting cells from inhibition.

Aluminum Silicates

Growth mechanisms and growth kinetics of filamentous microorganisms.

Filamentous microorganisms are of major biotechnological importance, being responsible for production of the majority of secondary metabolites, particularly antibiotics. Two main groups are involved, filamentous fungi and filamentous actinomycetes, particularly the streptomycetes. In terms of cellular growth mechanisms, these groups differ greatly. Eukaryotic fungi possess subcellular organelles and cytoskeletal structures directing growth while prokaryotic streptomycetes have no such cellular organization. Despite these fundamental differences, both groups exhibit similar morphologies, growth patterns, growth forms, and hyphal and mycelial growth kinetics on solid media and in liquid culture both grow as dispersed mycelia and pellets. The article therefore discusses the relationship between cellular growth mechanisms and vegetative growth in both filamentous fungi and actinomycetes, the conceptual and theoretical models applicable to both groups, and the significance of such models in industrial fermentation processes.

Actinomycetales

Apical hyphal extension in Streptomyces coelicolor A3(2).

Hyphal extension in the filamentous actinomycete Streptomyces coelicolor A3(2) was shown to occur by addition of newly synthesized wall material in an apical extension zone. Incubation of mycelia with tritiated N-acetyl-D-glucosamine (GlcNAc), a precursor of peptidoglycan, resulted in localized incorporation of label at the apex, as indicated by light microscopic and electron microscopic autoradiography. Within the hyphal extension zone there was a sharp decrease in incorporation with increasing distance from the apex. Hyphal tip shape, examined by low-temperature scanning electron microscopy, approximated to a semi-ellipsoid of revolution and was not hemispherical. Tip shape could be represented accurately by polynomial equations of degree less than seven. The surface stress theory was successfully applied to hyphal tip growth, with tip shape related qualitatively to the inverse of surface tension within the wall of the extension zone. Surface tension was assumed to be inversely proportional to the rate of incorporation of tritiated GlcNAc. Treatment of surface-grown hyphae with beta-lactam antibiotics resulted in localized swelling of hyphal tips. Lysozyme caused swelling of tips and of other regions of hyphae, frequently giving a beaded morphology associated with septa.

Acetylglucosamine

Luminescence-based nonextractive technique for in situ detection of Escherichia coli in soil.

Measurement of light output by luminometry was used to estimate quantitatively the cell concentrations of luminescent strains of Escherichia coli in liquid culture and inoculated into soil. Strains were constructed in which luciferase production was autoinducible or constitutive. In the former, light output per cell varied considerably during growth but was constant in constitutive strains. In liquid culture, the lower detection limit was in the order of 10(2) cells ml-1. Sensitivity was reduced by approximately 1 order of magnitude for cells inoculated into soil, when 2 x 10(2) to 6 x 10(3) cells g of soil-1 could be detected. Light output measurements were obtained within 5 min of sampling, and luminometry therefore potentially offers a rapid and sensitive detection technique for genetically engineered microorganisms.

Bacteriological Techniques

A model for hyphal growth and branching.

A mathematical model for hyphal growth and branching is described which relates cytological events within hyphae to mycelial growth kinetics. Essentially the model quantifies qualitative theories of hyphal growth in which it is proposed that vesicles containing wall precursors and/or enzymes required for wall synthesis are generated at a constant rate throughout a mycelium and travel to the tips of hyphae where they fuse with the plasma membrane, liberating their contents into the wall and increasing the surface area of the hypha to give elongation. The hypothesis that there is a duplication cycle in hyphae which is equivalent to the cell cycle observed in unicellular micro-organisms is also included in the model. Predictions from the model are compared with experimentally observed growth kinetics of mycelia of Geotrichum candidum and Aspergillus nidulans. The finite difference model which was constructed is capable of predicting changes in hyphal length and in the number and positions of branches and septa on the basis of changes in vesicle and nuclear concentration. Predictions were obtained using the model which were in good agreement with experimentally observed data.

Aspergillus nidulans