72575 - Stress Physiology and Redox Dynamics in Plants

Learning outcomes

At the end of the course, the student will have in-depth knowledge of modern developments in specific areas of plant physiology and the main methodologies targeting the expression of recombinant proteins in plants and their characterization. In particular, the student is able to: - analyze and discuss topics of genomics, redox biology, physiology of abiotic and biotic stress in higher plants; - critically evaluate the scientific literature on the topics covered; - analyze fragments of DNA and transform bacterial cells; - purify recombinant proteins by chromatographic techniques; - conduct analysis of biochemical and molecular characterization of the protein product.

Course contents

The course consists of Module 1 (lectures, 4 credits, Prof. Trost) and Module 2 (individual experimental laboratory, 3 credits, Prof. Sparla).

Module 1 (Prof. Trost)

Elements of plant genomics. Fundamental characteristics of the genome of Arabidopsis thaliana and other model species in plant physiology. Genomic duplications in the evolution of angiosperms. Polyploidy. Synteny. Gene expression databases. Genetic transformation and regeneration in vitro. Reverse genetics techniques applied to plants: insertional mutagenesis, RNA interference, genome editing.

Oxidative stress and redox signalling. Reactive oxygen species: chemistry, sources of ROS in plant cells, antioxidant systems. The thioredoxin system and the redox regulation of metabolism. Glutaredoxins and glutathionylation of proteins. Reactive nitrogen species and cysteine nitrosylation. Measurement of the redox state of glutathione in vivo using genetically encoded fluorescent probes (roGFP and derivatives). Redox proteomics. Post-translational redox modifications and their identification. Reactive sulfur species and cysteine sulfhydration.

Abiotic stress: photoinhibition and photoprotection. Photosynthetic response to light intensity. Photosynthetic production of ROS. Responses to light stress: leaf heliotropism, chloroplast movement. Molecular mechanisms of photoprotection: dissipation of excess energy by non-photochemical quenching, water-water cycle, state transition, cyclic electron transport, chlororespiration, repair cycle of protein D1.

Biotic stress: plant-pathogen interactions. Plant pathogens and pathogenesis. Constitutive defense. Innate immunity in plants: PAMP, DAMP and basal immunity; effectors, R genes and vertical resistance. Phytoalexins. Crosslinking reactions at the cell wall. Oxidative burst. Hypersensitive response. Acquired systemic resistance (SAR); salicylic acid, NPR1 and redox regulation. Intracellular and long-distance signaling.

Module 2 (Prof. Sparla):

During the practical activity, in vitro and in vivo approaches will be used. The in vitro activity will focus on cloning, expression, purification and biochemical characterization of the NAD(P)H:quinone oxidoreductase of Arabidopsis thaliana (AtNQR). The in vivo activity will allow to highlight the onset of the oxidative burst in response to wounding in Arabidopsis leaves and to identify, in spinach, the presence of an immunocorrelated form of the recombinant AtNQR.

Readings/Bibliography

Module 1

- Rascio et al., Elementi di Fisiologia vegetale, seconda edizione, EdiSES 2017

- A.M. Smith et al., Biologia delle piante, Zanichelli 2011

Vol. 1 – Evoluzione, sviluppo, metabolismo

Vol. 2 – Interazioni con l’ambiente, domesticazione

- Updated reviews and scientific articles indicated by the teacher

- Power point files (IOL Unibo)

 

Module 2

- The teaching material (lesson slides; links to publications and links to online databases) will be available on the IOL- University of Bologna platform via username and password

 

Teaching methods

Module 1: Lectures  

Module 2 (laboratory): each student is required to carry out the experimental laboratory activities individually. The teacher will give a theoretical introduction to the lab and continuous technical assistance. The obtained results will be critically discussed.

Assessment methods

The final exam consists in three parts that concur to the final evaluation:

- assessment of laboratory reports

- written test (on line)

- oral exam (on line)

Teaching tools

All lectures will be given with power point presentations. The files of power point presentations are made available to the students at the end of the course (IOL Unibo). 

The lab course will take place in a didactic laboratory with individual workstations.

 

Office hours

See the website of Paolo Bernardo Trost

See the website of Francesca Sparla