Predicting the future impact of climate change on Stipa tenacissima steppes is of paramount importance to prevent desertification. From this perspective, we selected seven populations, covering a rainfall range from 100 to 600 mm/year at the sites of seed collection, and we grew the seedlings at the same site. Gas exchange measurements of CO2, light response curves, chlorophyll content and isotopic analysis were investigated. Thala and Valencia populations displayed a contrasted gas exchange response. Both populations were subjected to A-Ci curve analysis. No significant difference was detected between these two different populations in the maximum rate of carboxylation (Vcmax) and the maximum rate of electron transport (Jmax). Stipa tenacissima exhibited a variable response in relation to different light intensities. Photosynthesis and chlorophyll fluorescence parameters proved to be significantly different among populations in response to light intensities. All populations exhibited similar chlorophyll content. The isotopic analysis highlighted a different carbon and nitrogen isotopic composition (δ13C and δ15N) and carbon content © among populations. Mesic populations presented a higher gas exchange activity and light response. The xeric populations performed a photosynthesis adjustment along with minimizing chlorophyll content. Under climate change, S. tenacissima limits productivity, which may lead to desertification.