In Mediterranean regions, water scarcity associated with climate change particularly threatens the sustainability of viticulture. Breeding grapevine for reduced water use and maintained production is therefore of major interest. This requires a comprehensive knowledge of the plant physiological responses to drought. In this study we focused on the determinism of transpiration rate as a key trait regulating water status in plant tissues, and on its relationship with water-use efficiency (WUE). We used a F1 progeny from a cross between cultivars Syrah and Grenache and combined powerful phenotyping tools on potted plants submitted to either well-watered or mild deficit conditions with quantitative genetics (for QTL detection) and physiological experiments. Analysis of the genetic control of water status maintenance in the plant, more or less efficient under soil water deficit (i.e. iso- or anisohydric), revealed a dual physiological determinism with a key role for plant hydraulic conductance beside that of stomatal control of transpiration. An indirect role of abscisic acid on stomatal conductance was also evidenced, mediated by the downregulation of leaf hydraulic conductance, with a genetic variability which correlated with genetic variation in iso- or aniso-hydric behaviour. We then revealed wide genetic variations in nocturnal transpiration, which correlated with variations in water use efficiency, and identified corresponding genetic and physiological determinants. In a final switch to the field, we showed consistency between QTLs detected for daytime WUE in pots and in the vineyard. Beyond the potential interest of the QTLs detected in this study for breeding prospects, this work demonstrated the benefits of quantitative genetics to shed light on ecophysiological and physiological processes.