Main conclusionTomato plant acclimation to a mild water stress implied tissue-specific hormonal and nutrient adjustments, being the root one of the main modulators of this response.Phytohormones are key regulators of plant acclimation to water stress. However, it is not yet clear if these hormonal responses follow specific patterns depending on the plant tissue. In this study, we evaluated the organ-specific physiological and hormonal responses to a 14 day-long mild water stress in tomato plants (Solanum lycopersicum cv. Moneymaker) in the presence or absence of the arbuscular mycorrhizal fungus Rhizoglomus irregulare, a frequently used microorganism in agriculture. Several physiological, production, and nutritional parameters were evaluated throughout the experiments. Additionally, endogenous hormone levels in roots, leaves, and fruits at different developmental stages were quantified by ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS). Water deficit drastically reduced shoot growth, while it did not affect fruit production. In contrast, fruit production was enhanced by mycorrhization regardless of the water treatment. The main tissue affected by water stress was the root system, where huge rearrangements in different nutrients and stress-related and growth hormones took place. Abscisic acid content increased in every tissue and fruit developmental stage, suggesting a systemic response to drought. On the other hand, jasmonate and cytokinin levels were generally reduced upon water stress, although this response was dependent on the tissue and the hormonal form. Finally, mycorrhization improved plant nutritional status content of certain macro and microelements, specially at the roots and ripe fruits, while it affected jasmonate response in the roots. Altogether, our results suggest a complex response to drought that consists in systemic and local combined hormonal and nutrient responses.