Advanced Plant Cell Biotechnology

Learning objectives
The aim of the course is to discuss the criteria and the methods to design or improve a biotechnological process exploiting plant cells or whole plants to produce biomass or substances of industrial or environmental relevance (e.g. biofuels, oils, detergents, vitamins, starch, sugars, biodegradable polymers). An 'on-site' visit to at least one company or research centre is envisaged to make the students aware about plant biotechnology outside the academic environment.
Upon completing this course, students will acquire substantial knowledge on metabolic fluxes, on strategies that can be used to modify yield and on the tools available in plant biotechnology to achieve the objectives.
The course is ideally linked to those dealing with functional genomics and other "-omic" technologies, plant breeding, molecular enzymology, bioinformatics and data analysis.

Syllabus
The lectures given by Paolo Pesaresi will first focus on the technical tools and theoretical basis of plant/algae biotechnological improvements for biofuel or food production. The different constructs for plant and algae genetic engineering together with the molecular basis of Agrobacterium-mediated transformation are discussed. The different elements 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 are described. Then the organisms and the strategies used to produce biofuels of first, second and third generation are taken into consideration, with several examples of transgenic plants and algae with improved biofuel production capacity. Finally, we will examine plant traits modified by transgenesis or mutagenesis which are already on the market or which are likely to be commercially available in the near future: herbicide tolerance, pest resistance, improved nutritional content...etc.
In the second part, Piero Morandini will approach two fundamental questions of metabolic engineering: 1) which factors regulate metabolic fluxes and intermediate concentrations and 2) which strategy /target works better to modify the yields in a certain product. In order to tackle the questions, we shall make use of the tools provided by the Metabolic Control Analysis theory (elasticity, flux and concentration control coefficients). We shall discuss the interplay of supply and demand of metabolic intermediates as well as the evidence in favour of the so-called 'parallel activation' to increase fluxes and the implications for manipulation strategies. Several case studies (and the relative successes and failures) of plant biotechnology will be discussed, among which the manipulation of starches, sugars, lipids, vitamins, pesticides, amino acids, and the inactivation of allergens and toxins.

REFERENCE MATERIAL
Power-point presentations, lecture notes and articles on which lectures are based will be uploaded and made available on the dedicated web-page. A major reference book about metabolic control theory is the following: Fell, Understanding the control of Metabolism, Portland Press (1997)

RECOMMENDED PREREQUISITES
A substantial background in basic biochemistry, genetics and functional genomics, as normally gained in the bachelor curriculum, is highly recommended. Good mastering of plant biochemistry is helpful but not an absolute requirement.

EXAMINATION PROCEDURES
The examination is oral and consists of questions on each section of the program. In addition, students are offered the possibility to present and discuss a research article selected from a list provided by the teachers on topics relevant to the syllabus. Examples of the examination test/questions are discussed during classes and made available to students. The two parts of the exam can be sustained separately, but the grade will result from the joint evaluation of each candidate by the two instructors.

TEACHING METHODS
Teaching Mode: classroom lectures supported by projected material with open discussions on experimental design, data analysis and specific case studies.
Attendance: highly recommended.

LANGUAGE
English

PROGRAM INFORMATION
Copies of the slides projected in the classroom as well as other materials are made available on the dedicated course website on the ARIEL platform of the University of Milano (http://ariel.ctu.unimi.it). By no means this material replaces the lectures or a textbook. The material is made available only to registered students of the Degree Course in Molecular Biotechnology and Bioinformatics and should not be distributed to others.

WEBsite
http://ppesaresibvia.ariel.ctu.unimi.it/v3/home/Default.aspx