Tobacco (Nicotiana tabacum; C3) plants increase their water use efficiency (WUE) under abiotic stress and are suggested to show characteristics of C4 photosynthesis in stems, petioles, and transmitting tract cells. The tobacco stress-induced Aquaporin1 (NtAQP1) functions as both water and CO2 channel. In tobacco plants, overexpression of NtAQP1 increases leaf net photosynthesis (AN), mesophyll CO2 conductance, and stomatal conductance, whereas its silencing reduces root hydraulic conductivity (Lp). Nevertheless, interaction between NtAQP1 leaf and root activities and its impact on plant WUE and productivity under normal and stress conditions have never been suggested. Thus, the aim of this study was to suggest a role for NtAQP1 in plant WUE, stress resistance, and productivity. Expressing NtAQP1 in tomato (Solanum lycopersicum) plants (TOM-NtAQP1) resulted in higher stomatal conductance, whole-plant transpiration, and AN under all conditions tested. In contrast to controls, where, under salt stress, Lp decreased more than 3-fold, TOM-NtAQP1 plants, similar to maize (Zea mays; C4) plants, did not reduce Lp dramatically (only by approximately 40%). Reciprocal grafting provided novel evidence for NtAQP1's role in preventing hydraulic failure and maintaining the whole-plant transpiration rate. Our results revealed independent, albeit closely related, NtAQP1 activities in roots and leaves. This dual activity, which increases the plant's water use and AN under optimal and stress conditions, resulted in improved WUE. Consequently, it contributed to the plant's stress resistance in terms of yield production under all tested conditions, as demonstrated in both tomato and Arabidopsis (Arabidopsis thaliana) plants constitutively expressing NtAQP1. The putative involvement of NtAQP1 in tobacco's C4-like photosynthesis characteristics is discussed.