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Banana (Musa spp.) multiple shoot meristems are an excellent model to study the meristem proteome. Using a 2‐DE protocol developed for small amounts of tissue and MS‐based cross species polypeptide identification, we have revealed the meristem proteome and investigated the influence of sucrose‐mediated osmotic stress in a dehydration‐tolerant variety. Proteins that were significantly up‐ or down‐regulated due to the high‐sucrose treatment were classified using non‐parametric univariate statistics. Our results suggest that the maintenance of an osmoprotective intracellular sucrose concentration, the enhanced expression of particular genes of the energy‐conserving glycolysis and the conservation of the cell wall integrity are essential to maintain homeostasis, to acclimate and to survive dehydration. By comparing the dehydration‐tolerant variety with a dehydration‐sensitive variety, we were able to …