The use of drought-tolerant rootstocks is an important strategy in maintaining orchard productivity while meeting the increasing need to conserve water resources. The drought tolerance of two new genotypes, Vineland 1 (V.1) and Vineland 3 (V.3), was assessed along with industry standards to test the hypothesis that differences in water-use efficiency exist among these apple rootstocks. One-year-old, non-grafted nursery liners of M.9, MM.111, V.1, and V.3 were grown in a controlled-environment experiment. Plants of each genotype were maintained water-replete or were subjected to a 9-d controlled dry down and then maintained under water stress conditions for 55 d. Water stress reduced biomass accumulation and trunk cross-sectional area for all four genotypes. The two vigorous genotypes, MM.111 and V.1, increased their root-to-shoot ratios in response to water stress, whereas the root-to-shoot ratios of M.9 and V.3 remained unchanged in their water-replete controls. Genotype V.3 maintained its transpiration at a significantly lower soil water content compared to M.9, MM.111, and V.1. Of the four genotypes, V.3 demonstrated a high tolerance to water stress conditions, and therefore deserves further investigation using grafted apple trees in an orchard study.