Tomato (Lycopersicon esculentum Mill.) plants were grown in either a glasshouse (GH) or a controlled environment cabinet (CEC) to assess the effects of partial rootzone drying (PRD) on biomass allocation. Control and PRD plants received the same amounts of water. In control plants, water was equally distributed between two compartments of a split-root system. In PRD plants, only one compartment was watered while the other was allowed to dry. At the end of each drying cycle, wet and dry compartments were alternated. In the GH, total biomass did not differ between PRD and control plants after four cycles of PRD, but PRD increased root biomass by 55% as resources were partitioned away from shoot organs. In the CEC, leaf water potential did not differ between treatments at the end of either of two cycles of PRD, but stomatal conductance of PRD plants was 20% less at the end of the first cycle than at the beginning. After two cycles of PRD in the CEC, biomass did not differ between PRD and control plants, but PRD increased root biomass by 19% over the control plants. The promotion of root biomass in PRD plants was associated with the alternation of wet and dry compartments, with increased root biomass occurring in the re-watered compartment after previous exposure to soil drying. Promotion of root biomass in field-grown PRD plants may allow the root system to access resources (water and nutrients) that would otherwise be unavailable to control plants. This may contribute to the ability of PRD plants to maintain similar leaf water potentials to conventionally irrigated plants, even when smaller irrigation volumes are supplied.