66632 - BIOTECNOLOGIE GENETICHE VEGETALI

Learning outcomes

By the end of this course, students will have a foundational understanding of plant morphology and molecular physiology, comprehending the structural and functional organization of plants and how they interact with their environment. These fundamentals are crucial for a solid grasp of the core principles of genetic improvement and plant biotechnologies. This knowledge will enable them to carry out biotechnological interventions on plants of agricultural importance, supporting both the qualitative and quantitative genetic improvement of crop production and its sustainability.

Given the significance of quantitative traits for the primary production of agricultural species, especially cereals, students will gain expertise in both classical and molecular quantitative genetics. This includes the ability to map, clone, and edit genes and Quantitative Trait Loci (QTL), along with a familiarity with traditional methods for inducing genetic variability in plants.

Students will acquire the necessary knowledge to implement marker-assisted selection, gene/QTL cloning, plant transformation, and genetic engineering and gene editing. Additionally, topics in genomics and transcriptomics will be covered, providing students with all the essential tools to utilize biotechnologies for plant breeding.

A particular emphasis will be placed on developing the ability to solve problems related to the practical applications of the techniques presented in the course.

Course contents

1. Course Introduction

A quick overview of the main topics of the course, with targeted connections to current and future applied aspects in the global socio-economic context.

2. Elements of Botany and Plant Physiology

Description of the peculiarities of the plant cell, differentiation into tissues and organs, with a particular focus on meristematic tissues and the anatomy and morphology of stem, root, and leaf. Phytohormones: synthesis, transport, signaling mechanisms, and biological functions, with examples of the role of cytokinins, ethylene, abscisic acid, gibberellins, and auxins.

3. Structure of Plant Genomes

Description of plant genomes, size, transposable elements and repetitive regions, relationships between euchromatin and heterochromatin, number of genes. Genome evolution and polyploidy, synteny and collinearity. Transcription factors and multigene families. Introduction to genome sequencing.

4. Analysis of Plant Transcriptome

Description of the main mechanisms of gene expression regulation and methodologies for studying expression, from microarrays to RNA-Seq. Differential expression and gene networks.

5. Fundamentals of Developmental Genetics and Epigenetics: Embryonic and Post-Embryonic Development, Plant/Environment Interaction

Embryogenesis and meristem differentiation, cell fate control. Response to external stimuli (temperature and light), regulation of flowering and floral organ differentiation. Use of mutants for studying gene functions. Examples of response to biotic and abiotic stresses. Transcription factors involved in the regulation of development and in the response to environmental stimuli.

6. Genetic Control of Reproductive Systems

Description of reproductive organs: flower, fruit, and seed, and modes of plant reproduction. Reproductive systems: Genetic incompatibility; male sterility; apomixis. Polyploid plants. Hybrid vigor and inbreeding depression.

7. Molecular Genetics, Molecular Markers: SNPs identification and use (analysis of genetic variability/domestication of agricultural plant species)

Identification of variations in DNA nucleotide sequence; study of genetic variability and the effect of domestication and selection in major agricultural species.

8. Applications of Molecular Genetics to Genetic Improvement, Mapping of Genes Controlling Mendelian and Quantitative Traits, QTL Mapping and Association Analysis

Definition of QTLs and mapping methods. Association mapping. Experimental populations for mapping Mendelian and quantitative loci, natural populations for association analysis. Marker-Assisted Selection (MAS).

9. Mutagenesis and TILLING

Mutations induced by mutagenic agents. Forward and reverse genetics. Applications for the genetic improvement of agricultural species.

10. Recombinant DNA Technologies

Manipulation of nucleic acids, cloning methodologies, vector production, and bacterial transformation. Applications for DNA library production, heterologous protein expression, and in vitro and in vivo mutagenesis.

11. Introduction to In Vitro Culture Techniques

General principles. Embryogenesis, organogenesis, haploid cell cultures, protoplast culture, somatic fusion, embryo rescue, micropropagation. Preparation of culture media and use of growth regulators.

12. Plant Genetic Transformation

Transformation methods: biolistics, Agrobacterium-mediated transformation, polyethylene glycol gene transfer, microspore transfection, in planta transformation, viral-mediated transformation, floral dip. Transgenesis, cisgenesis, intragenesis. Gene silencing (RNAi, microRNA, VIGS). Selection markers and reporter genes.

13. New Genomic Techniques (NGT)

Oligonucleotide directed mutagenesis (site-directed mutagenesis). Genome editing: Meganucleases, Zinc Finger Nucleases (ZFN), Transcription Activator-Like Effector Nucleases (TALEN), Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9). Applications for plant genetic improvement.

14. Current Global Status of Genetically Modified (GM) Organisms

Historical overview, applications and objectives of genetic transformation, diffusion of GM crops worldwide.

Readings/Bibliography

• Peter J. Russell. "GENETICS A Molecular Approach" Pearson Edit.
• Russell PJ et al. AGRICULTURAL GENETICS. EdiSES, 2016.
• GENETIC IMPROVEMENT OF AGRICULTURAL PLANTS (F. Lorenzetti et al. 2017, Edagricole, Milan).
• MOLECULAR BIOTECHNOLOGY – PRINCIPLES AND TECHNIQUES. (Brown TA, 2017, Zanichelli).
• Plant genes, genomes and genetics. Erich Grotewold, Joseph Chappell, Elizabeth A. Kellogg. Wiley-Blackwell ed.

Teaching methods

The course includes frontal lectures and laboratory exercises. During the frontal lectures (48 hours), using slides, the main aspects of the discipline as reported in the program above are presented. Laboratory activities include 30 hours of practical exercises in biotechnology and genomics/transcriptomics applied to plants of agricultural interest. On the Virtual platform, students will have access to lecture slides and other supplementary teaching materials such as scientific journal articles, videos, and links to web pages.

Assessment methods

The assessment of course learning takes into account the level of knowledge and skills acquired by the student regarding the course content. Learning will be verified through a written exam at the end of the lessons, consisting of a combination of multiple-choice and open-ended questions. There are 20 multiple-choice questions (1 point for each correct answer, 0 if incorrect or missing), and the 4 open-ended questions allow up to 3 points each, evaluating the correctness of the answer and the appropriateness of language. The maximum score obtainable is 30 cum laude. The exam is considered passed with a minimum score of 18/30.

It will also be possible to take the oral assessment through a 30-minute interview where preparation on the course program will be evaluated.

Students with Specific Learning Disorders (SLD) or temporary/permanent disabilities: it is recommended to contact the competent University Office in advance (https://site.unibo.it/studenti-con-disabilita-e-dsa/it ). This office will propose any necessary adaptations to interested students, which must be submitted for the teacher's approval at least 15 days in advance. The teacher will evaluate their appropriateness also in relation to the learning objectives of the course.

Teaching tools

Video projector for frontal lectures, molecular biology laboratory, and computer labs for practical exercises.

Office hours

See the website of Francesco Camerlengo

See the website of Cristian Forestan