Although the photosynthetic responses of sugarcane plants to environmental stresses are well documented, the biomass accumulation and the dynamic of carbohydrate reserves under simultaneous exposure of roots to low temperature and drought are not known. This work aims to investigate the effect of water deficit and low substrate temperature stresses, occurring alone or in combination, on the sugarcane IACSP94-2094, a drought-tolerant genotype. As our hypothesis, we assume that this genotype is also tolerant to low substrate temperature, since low temperatures and water deficit occur simultaneously under field conditions. The water deficit alone or in combination with low substrate temperature caused reductions in leaf water potential and CO2 assimilation, which was not observed in plants subjected only to low substrate temperature. The leaf concentration of non-structural carbohydrates, sucrose and starch increased in plants under root chilling. 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