Plant Biotechnology

The course "Plant Biotechnology" provides an in-depth exploration of the molecular and phenotypic foundations of plant domestication and the Green Revolution, followed by a comprehensive overview of first-generation genetically modified (GM) crops and the engineering methods that enabled their creation. After a brief introduction to the origins of agriculture and the domestication syndrome, the course examines key molecular pathways underlying major domestication traits such as seed shattering, flowering time, and stress responses, as well as the genetic basis of semi-dwarfism and yield improvements characteristic of Green Revolution varieties. The course then describes the molecular markers utilized to genotype spontaneous mutation with a particular focus on PCR and sequence-based techniques. It then covers the principles of plant transformation, with a detailed focus on Agrobacterium tumefaciens biology, Ti plasmid and T-DNA transfer, and binary vector systems, leading to the engineering of first-generation GM crops. Together with that, traditional and innovative cloning techniques and delivery of non-invasive reporter genes (e.g GFP, Ruby, Luciferase) will be illustrated. Additionally, alternative methods of plant transformation, including biolistics (gene gun), protoplast-mediated transformation and Virus-Induced Gene Silencing (VIGS) will also be discussed with an introduction on in vitro cultivation and cell culture. Case studies include glyphosate-tolerant crops through EPSPS gene engineering and insect-resistant plants expressing Bacillus thuringiensis (Bt) toxins, with discussion of molecular mechanisms, performance in the field, and resistance management. The course then addresses the transition to modern genetic engineering and plant genome editing technologies, including advanced delivery strategies, CRISPR-Cas systems (Cas9, Cas12a), base and prime editing, and approaches that enable targeted mutagenesis, precise gene insertion, gene stacking, and the engineering of complex agronomic traits such as disease resistance, abiotic stress tolerance, and nutrient use efficiency. The generation of novel epi-alleles by innovative Cas-based genetic engineering systems (ZFN, TALEN and SunTag) will also be described with their impact on plant growth and development. Regulatory aspects for transgenic and genome-edited crops are also discussed.