Sub-cellular fractionation and gel-based proteomics of Haberlea rhodopensis: a promising approach to open the black box of resurrection plants

Haberlea rhodopensis (Gesneriacea) belongs to the group of the resurrection plants. Their ability to survive extreme desiccation ren ders them excellent models to study plant response to dehydration. Plant metabolic networks are very complex and subcellular proteomics has proven to be a powerful approach to gain functional knowledge about the subcellular compartments. Here we present a protocol for the simultaneous isolation of pure and intact chloroplasts and mitochondria from Haberlea rhodopensis with subsequent gel-based proteomics including BN PAGE and 2-D gel electrophoresis. Differential detergent – based solubilization of chloroplast membranes allows separation of distinct activities in sub-organellar compartments and contributes for drastic reduction in overall sample complexity improving resolution and sensitivity of further proteomic analyses.