Achieving more environmentally sustainable vineyards, particularly regarding efficient water use, is paramount in semi-arid grape-growing regions. Rootstocks may be a possible strategy to address these challenges, but require a comprehensive evaluation of their effect on the scion, including ecophysiological traits. The objectives of this study were 1) to characterize the physiological response of Tempranillo cultivar grafted onto five commercial (1103 P, 110 R, 140Ru, 420 A, and SO4), and seven recently bred (RG2, RG3, RG4, RG6, RG7, RG8 and RG9) rootstocks and 2) to elucidate the relationships between agronomic and physiological traits conferred by grapevine rootstocks. This was carried out over three seasons (2018–2020) in a typical Mediterranean vineyard by determining water relations, leaf gas exchange, carbon isotope ratios and vegetative development and yield components. The results highlighted the different behaviour of ‘Tempranillo’ vines due to the rootstock effects on vine water status, photosynthetic performance, hydraulic conductance, vegetative growth and yield parameters. Overall, rootstocks inducing vigour and yield in the scion, such as 140Ru and RG8, showed higher leaf gas exchange rates and hydraulic conductance at the whole-plant level due to less negative water potentials, suggesting a higher water uptake and transport capacity than RG2, RG7 and RG9. The RG rootstocks showed a very wide range of ecophysiological responses, but only RG8 outperformed compared to the most widely used commercial rootstocks. Moreover, this response was modulated by the season and the block soil type, suggesting the importance of rootstock selection according to the edaphoclimatic conditions. Therefore, this study highlights the high potential of rootstocks to adapt to water scarcity by improving crop water productivity in vineyards and provides physiological insights for future studies and breeding programmes.