Drought is one of the critical factors limiting tree growth and survival. Clarifying the adaptation to drought will facilitate the cultivation of drought-tolerant varieties. Metabolites, as direct signatures of biochemical functions, can uncover the biochemical pathways involved in drought responses. Here, we investigated the physiological and metabolic responses of drought-tolerant Populus simonii and drought-susceptible P. deltoides cv. 'Danhong' to drought. Under drought conditions, P. simonii grew better and had a higher photosynthetic rate than P. deltoides cv. 'Danhong'. Global untargeted metabolite profiling was analyzed using gas chromatography time-of-flight mass spectrometry system. A total of 69 and 53 differentially accumulated metabolites were identified in drought-stressed P. simonii and P. deltoides cv. 'Danhong', respectively. The metabolisms of carbohydrate, amino acid, lipid and energy were involved in the drought responses common to both poplar species. The citric acid cycle was significantly inhibited to conserve energy, whereas multiple carbohydrates acting as osmolytes and osmoprotectants were induced to alleviate the adverse effects of drought stress. Unlike P. deltoides cv. 'Danhong', P. simonii underwent a specific metabolic reprogramming that enhanced non-enzymatic antioxidants, coordinated the cellular carbon/nitrogen balance, and regulated wax biosynthesis. These results provide a reference for characterizing the mechanisms involved in poplar response to drought and for enhancing the drought tolerance of forest trees. This article is protected by copyright. All rights reserved.