Key messageOver-yielding of stand biomass did not occur in a tree polyculture comprised of Betula pendula, Alnus glutinosa and Fagus sylvatica selected for contrasting traits. This was due to antagonistic interactions between the component species. Fine root dynamics and soil C stocks were unaffected by species mixture.ContextIncreasing CO2 fixation in tree biomass through afforestation and forest management actions has potential for cost-effective climate mitigation. The influences of tree mixture on biomass production and subsequent soil C accumulation in polyculture still remain uncertain.AimsWe studied the polyculture of Alnus glutinosa (L.) Gaertn, Betula pendula Roth and Fagus sylvatica L. in a plantation forest to examine the effectiveness of species mixtures as a tool for increased biomass production and soil C accumulation.MethodsTree biomass was estimated by developing species-specific allometric models and 3 years tree measurement. Fine root biomass and production were estimated by root coring and root-mesh methods. The ‘relative yield of mixture’ approach was used to examine the mixture effect.ResultsIn mixture, an additive effect was observed in A. glutinosa (13% increase in basal diameter relative to the monoculture); however, there was no overall effect of mixture on total standing biomass due to the suppression of F. sylvatica (2.75 g m−2 reduction in woody biomass). Fine root biomass production showed no mixture effect. The quantity and quality of soil C (top 0.5 m) was not affected by tree mixture.ConclusionWe conclude that the contrasting growth responses of the A. glutinosa, B. pendula and F. sylvatica in polyculture resulted in no over-yielding of standing biomass despite the complementary traits of the component species.