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Investigation of the transcriptional regulation of the antioxidant network in bundle sheath cells of Arabidopsis using a novel immunocapture technique for bundle sheath specific polysomes

Objetivo

The objective of this proposal is to investigate the transcriptional regulation of genes encoding the antioxidant network and their cognate transcriptional factors (TFs) in bundle sheath cells (BSC) of the Arabidopsis leaf.

Based on previous data concerning the BSC-specific stress-responsive expression of the ascorbate peroxidase 2 gene (APX2), it is postulated that the expression of the antioxidant defence gene network and its controlling TFs in BSC of the leaf subject to excess light stress, wounding or pathogen infection is constituted differently from that in cells of the leaf lamina.

Therefore, it is postulated that a regulatory network model, and by implication its description of cellular ROS metabolism, will have different properties to those derived for the whole leaf. I propose to establish a novel technique based on epitope-tagging of ribosomes only in a particular cell type and then to purify RNA specific to BSC and lamina cells separately from selectively immunoprecipitated polyribosomes. Next, the expression of antioxidant genes and TF genes known to influence their function will be examined by qRT-PCR.

Finally, computational models will be developed to determine the likely impact of the stresses on antioxidant and ROS metabolism in these cells compared with that from lamina tissue. These results will contribute to the development of sustainable agricultural systems since the knowledge of antioxidant networks in specific tissues will allow to determinate the most favourable conditions for the most efficient utilisation of environmental resources and therefore reduce chemical pesticides in crop production.

It will also contribute to other fields, even biomedical, since the technology of epitope-tagging of ribosomes to purify RNA from a specific cell type potentially is universal and, it could be used to decipher the function of genes and gene products relevant to human health and to explore their interactions with each other and with their environment.

Ámbito científico (EuroSciVoc)

Palabras clave

Tema(s)

Convocatoria de propuestas

FP6-2005-MOBILITY-5
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Régimen de financiación

EIF - Marie Curie actions-Intra-European Fellowships

Coordinador

UNIVERSITY OF ESSEX
Aportación de la UE
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Coste total
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