Plants must survive the vagaries of the natural environment and have developed an intricate web of defences to combat stresses to which they are exposed. Unravelling the mechanisms by which plants sense and respond would be a major breakthrough in fundamental scientific understanding and highlight key pathways for targeted manipulation to improve crop yields. A common denominator in the reaction of plants to stress is the production of a suite of hazardous reactive oxygen species (ROS). This causes shifts i n the amount and redox status of key antioxidants, in particular ascorbate (ASC) and glutathione (GSH), a signal which can be sensed and transduced by the plant. To date, attention was focused on defence-related gene expression caused by cellular ASC and G SSH, yet the primary source of ROS is the cell wall. It would thus make sense for key redox sensors and transduction elements to be situated in this poorly understood sub-cellular compartment. Recent work published by the host laboratory has employed novel transgenic plants to illustrate the importance of extracellular redox status of ASC in modulating defence-related gene expression.
The proposed project will draw upon the accumulated knowledge and provide access to characterized transgenic plants plus a u nique mix of state-of-the-art facilities (cat2 O3-fumigation suite co-located with MALDI TOF-TOF MS, SELDI-TOF MS and MUDPIT technology) in a bid to identify changes in the leaf proteome instigated by shifts in the redox status of extra-cellular ASC. Leaf and cell wall fluid will be sampled at regular intervals from plants exposed to O3. Differentially expressed proteins will be sequenced and identified with support from the hosts Bioinformatics Support Unit. The work proposed will clarify the role played by extra-cellular ASC redox status in signal transduction and provide fundamental information about how plants respond to, and discriminate between, stresses to which they are exposed in the field.
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