The course aims at presenting and discussing both the criteria and the methods employed in a biotechnological project based on plant cells or whole plants to produce goods of industrial or environmental relevance. An on-site visit to at least one company or research center is included in the course.
Section 1 This section approaches two fundamental questions of metabolic engineering: 1) which factors regulate metabolic fluxes and 2) what are the best strategies to modify the yields in a certain product (alter concentrations o fluxes). In order to tackle the questions we shall make use of the tools provided by the Metabolic Control Analysis theory. We shall discuss the limits due to supply/demand of metabolic intermediates as well as the evidence in favor of the so-called Universal method (parallel activation) to increase fluxes and the relative implications for manipulation strategies. Several case studies (and the relative effectiveness, merits and limits) of biotechnological interventions will be discussed, among which: inactivation of allergens and toxins and the production of material for industrial use (starches, sugars, lipids, vitamins, aminoacids, dyes, drugs, flavourings...etc.). Section 2 This section aims at providing the technical tools and theoretical bases for the genetic improvements of plants and algae to be used in biofuel production or in agriculture for food production, i.e. plants resistant to herbicide and pathogens or plants with improved nutritional features. During the first part, information about the realization of constructs for plant and algae genetic engineering together with the molecular bases of Agrobacterium-mediated transformation will be given. The different constituents of expression vectors, including constitutive and inducible promoters, enhancers, reporter genes, replication origin and marker genes, together with the strategies to create marker-free transgenes will be discussed. In the second part, the organisms and the strategies used to produce biofuels of first, second and third generation will be described. In addition, several examples of transgenic plants and algae with improved biofuel production capabilities will be analyzed. The third part will be devoted to analyze the characteristics of plants modified by transgenesis or mutagenesis which are commercially available, with respect to their tolerance to herbicide and pathogens and to their improved nutritional qualities.