Methods in Plant Genetics and Biotechnology

The course programme includes the presentation and discussion of the following topics:
- Introduction to plant biotechnology
- Origins of agriculture, domestication and the Green Revolution
- Plant breeding: selection, crossing, mutagenesis, the importance of biodiversity conservation
- In vitro cultures of plant cells, tissues and organs
- Genetically modified plants: differences between breeding and genetic engineering
- Transformation methods (Agrobacterium tumefaciens, in vitro transformation, shotgun)
-Design of the transgenic construct
- Methodologies used for plant transformation and transgene identification, forward genetics and reverse genetics.
- Transformation of plants of agronomic interest
- Regulation of gene expression: identification and use of constitutive, inducible, tissue- and cell-specific promoters
- Selection markers and reporters genes (use and implications)
- Gene overexpression and silencing: constructs for gene silencing (antisense RNA and interference)
- Definition of microRNAs and use of artificial microRNAs
- Genome editing techniques and their applications in plant biotechnology
Zinc Finger
Talen
CRISPR
- Programmes for the design of Genome Editing constructs, sequence analysis for the evaluation of mutation induction

- Transgenic plants for herbicide resistance
Roundup Ready soya
. Golden rice
. Phytic acid and mineral biofortification

Practice (8 hours)
Using Arabidopsis thaliana as model species:
1. Floral Dip technique using Agrobacterium tumefaciens to transform Arabidopsis plants;
2. Expression analysis using a GUS promoter/assay in different tissues of the plants (vegetative and reproductive tissues);
3. Sowing in plates of Arabidopsis seeds and identification of transgenic plants (phenotyping).