Quercus Ilex proteomic studies on the germination and early seedling growth under drought stress. (QUIPROD)
Proteomics is a high-throughput approach which could provide comprehensive understanding of complex developmental processes. Holm oak (Quercus ilex L.) is a dominant tree in Mediterranean natural communities. Oak seeds are recalcitrant. Seed proteome and germination under stress are studied predominantly in cereals and model plants, with scant information about forest species.
This project aims at studying the germination and early seedling growth in Holm oak at the proteome level, especially how these processes are affected by drought. Emphasis will be made on phosphorylation and ubiquitination as major posttranslational modifications (PTM). Proteome analysis will be complemented with data at metabolite and transcript level and with cytochemical observations.
METHODOLOGY: Holistic and targeted approaches will be combined.
Holistic approach - by using 2-DE coupled to MS, changes in the protein and some PTM profiles will be analyzed. The workflow includes: sampling, protein extraction, separation (1-D/2-DE), gel staining, image capture and analysis, MS of spots, protein identification. PTM will be detected by specific staining or western blot. Appropriate statistical analysis, algorithms, databases and repositories will be used.
Targeted approach -the oxidative status of the tissues will be monitored and more detailed analysis of some proteins of interest will be performed at transcript level. Analyses at metabolite level will include carbohydrates (ethanol extraction, GC-MS), aminoacid derivates (ethanol extraction, HPLC quantification), some secondary metabolites (GC or HPLC). Histochemical staining for carbohydrates, phenolic compounds, and H2O2 will also be performed.
EXPECTED RESULTS: To gain deeper knowledge of the response to drought in a recalcitrant seed producing tree species at the earliest developmental stages. The relevance of specific PTMs will be evaluated. Proteomic data will be validated by complementary targeted studies.
Departamento de Bioquímica y Biología Molecular
Code PAIDI: AGR-164
Prof. J.V. Jorrin Novo. coordinator.
Universidad de Córdoba
Budget of Andalusian group: € 174, 610
- Universidad de Córdoba