2023/2024

1) Knowledge and understanding: the course aim to provide the student with the basic principles of biotechnologies applicable to crop breeding and to improve crop productivity. 2) Applied knowledge and understanding: basic biotechnological methodologies are introduced with reference to molecular marker development and in vitro culture, including those useful for the study of genetic variability and for obtaining interspecific hybrids. 3) Making judgements: the advantages and disadvantages of each presented technology are illustrated, to develop the student's critical sense. 4) Communication skills: students are asked questions during the classes, both to keep their attention and to teach them to ask questions and give adequate answers. 5) Learning skills: the theoretical bases of the different genetic biotechnologies presented are provided and, for some of them, also the practical bases, through laboratory and in silico exercises.

It is advisable to attend the course after the Genetics exam and, possibly, Biochemistry as well, although not mandatory,

The course starts with the basic principles of recombinant DNA illustrating the mechanism of action of of restriction and modification enzymes, and gene cloning through host-vector systems.
The mechanism of action of standard and quantitative PCR is presented in detail, but particular importance is given to applications of such methodologies, through specific examples.
Among the different methods of DNA sequencing only the Sanger method is reported, both in its manual and automated versions.
The various techniques of analysis of nucleic acids and proteins, including electrophoresis on agarose and polyacrylamide and through blotting (Southern, Northern and Western blotting), and the different fields of application of these techniques are presented.
In regard to libreries, both the construction process, and the analysis of the genomic, cDNA and expression libraries are shown, with an emphasis on information obtained from their use.
The Arabidopsis thaliana and Homo sapiens genome projects are presented, explaining the criteria of choice of model organisms, and information obtained by the complete knowledge of these genomes.
Structural, functional and comparative genomics, as well as the technologies related to each of these branches, are presented. In particular, technologies related to structural genomics, through analysis of the genomic libraries for the sequencing of extended portions of DNA, and to functional genomics, through the main techniques of analysis of transcriptome and proteome, are explained.
Finally, genetically modified plants are presented: how they are realized through the two main systems (biolistic and Agrobacterium), by illustrating advantages and disadvantages of both these methods; how a genetic construct for expression in plants is made, which genetically modified plants are present on the market, with hints related to the issue of their acceptability.

Practical classes:
• Bacterial transformation using a recombinant plasmid vector, which is extracted in a subsequent practical class and characterized by agarose gel electrophoresis
• Standard PCR to determine the presence of a specific transgene in plants of GM wheats
• SDS-PAGE to analyze the protein composition of wheat kernels
• The use of the spectrophotometer to determine the amount of DNA and proteins

If the students are more than 30, practical classes are performed in turns.

Il corso si tiene in modalità mista (aula e in remoto) e le esercitazioni nell'apposita aula

The course is held in classroom and online whereas practical classes are held in the dedicated classroom

Brown T. Molecular Biotechnology
Rao R, Leone A (Idelson-Gnocchi) Biotecnologie e Genomica delle Piante
Pasqua G, Forni C (Piccin) Biotecnologie vegetali

Slides and other materials will be made available to students on Moodle platform

The lesson is held in hybrid mode (in-person and online mode) whereas practical classes are held in the dedicated laboratory

Attendance is not mandatory.

The oral exam is taken exclusively on the entire program. Three questions are posed: two concern the first part of the course (up to the libraries included) and the third concerns the final part. The final score is the average of the three scores of the respective oral questions.

    Samuela Palombieri