ABSTRACT Due to the predictions of climate change, there is a need to identify forage plants that can keep their productivity and nutritive value under hydric stress. The objective of this study was to evaluate the biomass and chemical responses of three Desmanthus spp. accessions under two water deprivation regimens (7 and 21-day). The experimental design used was randomized blocks in a 3 × 2 factorial arrangement (access and water deprivation) with four replications. There were reductions in the biomasses of leaf and stem fractions of 64% and 51%, respectively. The 43F accession showed greater leaf (1.86 g.plant-1) and stem (1.97 g.plant-1) biomasses under a total water restriction of 21 days, compared to the 89F and AusT accessions. In accessions 43F and 89F, there were carbohydrate accumulations in the leaves of 28% and 51%, respectively, under the 21-day water restriction, while AusT decreased 38% within a 7-day interval. Water deprivation affected the chemical characteristics of the accessions. Free proline was similar among accessions and accumulated 463% more in the leaves of plants submitted to 21-day water deprivation (90.22 mg.kg-1) compared to those subjected to water deprivation for 7 days (16.03 mg.kg-1). Proline and total soluble carbohydrate accumulation in 43F and 89F were insufficient to regulate crude protein, C content, and C:N ratios. These results demonstrate the variability in drought tolerance among accessions. Accessions 43F and 89F were more susceptible to 21-day water deprivation, while AusT showed greater drought tolerance.