In South-western Morocco, water scarcity and high temperature are the main factors determining species survival. Argania spinosa (L.) Skeels is a tree species, endemic to Morocco, which is suffering from ongoing habitat shrinkage. Argan trees play essential local ecological and economic roles: protecting soils from erosion, shading different types of crops, helping maintain soil fertility and, even more importantly, its seeds are used by the local population for oil production, with valuable nutritional, medicinal and cosmetic purposes. The main objective of this study was to identify the sources of water used by this species and to assess the effect of water availability on the photosynthetic rate and stem water potential in two populations: one growing on the coast and a second one 10 km inland. Stem water potential, photosynthetic rate and xylem water isotopic composition (δ18O) were seasonally monitored during 2 years. Trees from both populations showed a similar strategy in the use of the available water sources, which was strongly dependent on deep soil water throughout the year. Nevertheless, during the wet season or under low precipitation a more complex water uptake pattern was found with a mixture of water sources, including precipitation and soil at different depths. No evidence was found of the use of either groundwater or atmospheric water in this species. Despite the similar water-use strategy, the results indicate that Argania trees from the inland population explored deeper layers than coastal ones as suggested by more depleted δ18O values recorded in the inland trees and better photosynthetic performance, hence suggesting that the coastal population of A. spinosa could be subjected to higher stress.