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Biodiversity: Applied and Systematic Investigations of Cyanobacteria

Objectif



The cyanobacteria are a phylogenetically coherent taxon of oxygenic phototrophic prokaryotes whose range of morphological diversity is greater than that of any other group of microorganisms. They represent a unique source of biodiversity for both fundamental and applied research but previous difficulties encountered in their culture, axenization and systematics have prevented thorough study of their potential. With the first two problems largely overcome, this project proposes a timely investigation of cyanobacterial systematics (employing phenotypic, genotypic, physiological and biochemical characters), a search for compounds of biotechnological interest, and a study of species diversity in the natural environment. The project will be based on the 600 axenic strains available in the Pasteur Culture Collection of Cyanobacteria (PCC) and is composed of three work packages. I. Biosystematics: comparative approaches for definition of taxa. A characterization of the phenotypic and physiological properties of cyanobacteria will involve: cellular morphology and differentiation; photosynthetic pigments; physiology (heterotrophic potential, N2 fixation, temperature maxima, salt tolerance); biochemical constituents (lipids, fatty acids, carotenoids, exopolysaccharides). These studies will be complemented and extended by high resolution molecular techniques applied to total genomic DNA (determination of molar G+C content and genetic complexity, DNA-DNA hybridization, DNA fingerprinting with RFLP and RAPD) or to individual genes (restriction pattern analysis following hybridization to specific probes). Protein fingerprinting will be performed by electrophoretic separation of total or soluble proteins. Phylogenetic analysis will employ the sequences of 16S rRNA and the intergenic transcribed spacer regions (ITS).
II. Bioactive compounds and others of biotechnological interest. Some of the activities in work package I, although designed for biosystematic studies, will reveal compounds of potential interest (lipids, fatty acids, carotenoids and exopolysaccharides). Work package II is devoted more specifically to bioactive compounds and includes an examination of the synthesis and mode of action of toxins and related compounds, some of which, in spite of their undesirable effects on the health of livestock and Man, have already proven commercially valuable. Furthermore, the study of UV-absorbing compounds (sun screens) as one of major biotechnological potential, must be emphasised.
III. Exploration of species diversity in environmental samples. Although work packages I and II will be performed on axenic strains isolated from a wide range of ecosystems, the species presently in culture represent only a small fraction of the cyanobacterial diversity in the natural habitat. Consequently, cyanobacterial-specific 16S rRNA gene probes will be constructed and tested to evaluate the occurrence of these organisms in the environment. If unusual organisms are detected, new approaches for their isolation and cultivation will be sought, as has been the tradition in the PCC during the last 20 years.

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28,Rue du Docteur Roux 28
75724 Paris
France

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