An optimized apoplastic fluid extraction method for the simultaneous characterization of the proteome and metabolome of grapevine leaves.

The study of different cellular compartments also allows a broader conception of cell dynamics. Plant apoplast is the cellular compartment external to the plasma membrane including the cell wall. Despite our knowledge on apoplast involvement on several processes from cell growth to biotic and abiotic stress response, its dynamics is yet still poorly studied due to the need for adaptation of the extraction process according to the plant system and consequently, the difficulty to perform a comprehensive profiling of proteins, metabolites and lipids within a single experiment. Moreover, when looking at woody plants as grapevine, more challenges are raised leading to only few studies published so far focusing on apoplast characterization. Being grapevine one of the most important fruit crops worldwide, a wider characterization of this compartment may unveil the importance of grapevine’s secretome. Here, we describe an optimized vacuum-infiltration-centrifugation method for the simultaneous extraction of apoplastic proteins and metabolites from the grapevine leaf on a single experiment, compatible with high-throughput mass spectrometry analysis.