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The effects of water availability were evaluated on the photosynthetic tissue anatomy in Aloe vera (L) Burm f. and its relationship with morphological, physiological parameters, and water use efficiency as a function of aerial biomass and gel production. Plants were subjected to four levels of water availability equivalent to 20% (T1), 15% (T2), 10% (T3), and 5% (T4) of the atmospheric evaporative demand. The plants exhibited anatomical, morphological, and physiological responses to the different watering treatments. The extreme treatments produced negative responses due to excess water in T1 and water deficit in T4. Treatments T2 and T3 elicited positive responses in cell characteristics and productivity. Anatomical and structural characteristics were closely linked to physiology. Increased stomata number was negatively related to leaf length, width, and thickness (r = −0.85, −0.81, and −0.59, respectively) and to biomass production (r = −0.84), and positively related to the increase of cuticle thickness (r = 0.78). Treatment T2 showed the maximum efficiency of water use for biomass production (24.6 g L −1), which was closely related to cell size (r = 0.68) and number of stomata (r = −0.70).