1. Methodology / Techniques Open Issues and Opportunities
Understanding desiccation tolerance mechanisms in plants
What it is about
Crop plants are the primary source of food for humans. One of the major problem affecting the crop production is represented by drought. Improve drought tolerance of crop plants is an urgent task as on one side global warming is decreasing the amount of water available for cultivation and on the other side the world population and consequently the food demand is increasing
Lindernia brevidens
Oropetium thomaeum
Resurrection plants such as Craterostigma plantagineum, Oropetium thomaeum and Lindernia brevidens are able to survive desiccation by activating mechanisms to avoid the damage generated by water scarcity
Our research aim to understand the molecular basis of protection mechanisms used by resurrection plants to survive desiccation in order to use this knowledge to improve drought tolerance in crop plants
Craterostigma plantagineum
Methodology / Techniques
Gene and protein
expression
Protein-protein interactions
Several techniques are used to identify genes responsible for desiccation tolerance in resurrection plants and study their function. RNA sequencing is used to get the whole set of genes activated in the plant during desiccation. Candidate desiccation tolerance genes are identified from this set and then functional characterised
RNA sequencing
Yeast two-hybrid screenings
Pull-down assays
Bimolecular fluorescence complementation assays
Gene characterisation is performed on different levels. Using RT-qPCR and protein blots we first confirm that the gene transcript and protein are accumulated during desiccation. We also study the subcellular localization of desiccation-related proteins by using fluorescence tags coupled with fluorescence microscopy
Fluorescence
microscopy
Biolistic transformation
Gene regulation and protein localization
Agro-mediated
transformation
(e.g. Agroinfiltration)
RT-qPCR
We also study regulatory regions of desiccation-related genes to identify factors required for their activation. Part of our research aims also to identify other proteins interacting with desiccation-related proteins by using different techniques such as yeast two-hybrid screenings, pull-down and bimolecular fluorescence complementation assays
Protein
blots
Open Issues and Opportunities
Genome editing
Our comprehension of desiccation tolerance is still limited. This is partially due to the complexity of this trait. Moreover research with resurrection plants is still hampered by the lack of genome sequence and efficient plant transformation protocols
Genome sequence?
Plant transformation
Recently developed genome editing technologies such as CRISPR/Cas9 could greatly contribute to boost our understanding of desiccation tolerance mechanisms but to date there is still no report about their application in resurrection plants
Accumulating data support the hypothesis that protection genes are already present in plants including crops but such genes are not properly activated in plants sensitive to drought. Research on the regulation of desiccation-induced genes in resurrection plants is required to identify key regulatory elements
Contact Person: Valentino Giarola, Institute of Molecular Physiology and Biotechnology of Plants (IMBIO)
Kirschallee 1, 53115, Bonn, DE
Tel: