Throughout the world, huge areas have been occupied by monospecific tree plantations in order to fulfil the increasing demand for industrial wood products. The use of mixed-species plantations has recently been advocated as a potential compromise between maintaining high-volume wood production and conserving other ecosystem services. Yet little is known about the impact of tree species mixture on species- and ecosystem-level carbon accumulation and water use in mixed plantations. We combined data on above-ground biomass, xylem sap flux density, and the carbon and oxygen isotope composition of leaves and needles in a boreal plantation to test whether different levels of species mixture would impact tree- and plot-level biomass production, transpiration and water use efficiency. Data were recorded in 22 20-x-20-m plots (7 for transpiration) randomly allocated to either monocultures or to two-, three-, and five-species mixtures of Scots pine (Pinus sylvestris), Norway spruce (Picea abies), Silver birch (Betula pendula), Common alder (Alms glutinosa) and Siberian larch (Larix sibirica). At tree level, for a given species, we found significant differences among mixtures in transpiration and water use efficiency, though species mixture had no impact on above-ground biomass. At plot level, a large variability in functioning among mixtures was observed, but increasing the number of species in the mixture did not enhance productivity, transpiration or water use efficiency. The presence in mixtures of a high performing species like birch brought about changes in the canopy structure which in turn may have led to changes in micro-environmental conditions. Such changes could have contributed to explain differences in transpiration and water use efficiency among mixtures. We concluded that mixing locally-adapted species under non-limiting soil water conditions did not provide any benefit for ecosystem-level carbon accumulation and water use at this young ontogenic stage, but our study does not preclude that some benefits might arise at older stages. (C) 2013 Elsevier B.V. All rights reserved.