Objective
Plant cells are surrounded by a strong wall composed mainly of polysaccharides. This wall is essential for the form of the plant and allows the plant to stand upright, and the wall is the first line of defence against invading pathogens. Very little is known about how the wall polysaccharides are formed and integrated into the wall. Many enzymes are necessary for biosynthesis of the polysaccharides; the majority are glycosyltransferases (GTs) but also transglycosidases are involved.
The overall objective of the project is to assign function to genes involved in the biogenesis of plant cell wall polysaccharides. A functional genomics strategy will be pursued using Arabidopsis thaliana as the model:
1) Development and use of bioinformatics to predict which open reading frames are involved in cell wall biosynthesis.
2) Reverse genetics. A high throughput approach will allow investigation of at least 100 GTs. Clones will be produced in GateWay entry vectors for the easy sub-cloning in vectors for expression, epitope tagging etc. The activities of heterologously expressed enzymes will be determined using assays developed in the project.
3) Novel analytical tools for characterization of polysaccharides in the mutants will be developed and applied including antibodies and carbohydrate binding modules for in situ polysaccharide analysis, mass spectrometry, polysaccharide sequencing, substrate-chips for surface plasmon resonance, glyco-chips for activity screening etc.
4) Protein-protein interactions between the identified enzymes and other proteins will be investigated by proteomics, epitope tagging and bioimaging.
The network consists of 10 partners from 7 countries and will provide an outstanding environment for training of young researchers. In addition to the training implied in carrying out a challenging research project with frequent exchange between the partner laboratories, the network will implement a formal training program consisting of courses and workshops.
Fields of science
- natural sciencesbiological sciencesgenetics
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomics
- natural sciencesbiological sciencesbiochemistrybiomoleculescarbohydrates
- natural scienceschemical sciencesanalytical chemistrymass spectrometry
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
Call for proposal
FP6-2002-MOBILITY-1
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