•Sap flow and stem radial variations were evaluated in mixed floodplain forests.•Stem radial variations and sap flow was reflected in species’ sensitivity to drought.•Tree water deficit was coupled to drying soil and sap flow to evaporation demand.•Narrow-leaved ash showed the most conservative water use but the highest stem growth.•Groundwater decline and frequent droughts may alter tree species composition. Recent changes in the floodplain forests of Central Europe, caused mainly by changes in hydrological management and the increased frequency of droughts due to climate change, have led to severe degradation of floodplain ecosystems. Our main objective was to determine the sensitivity of trees to drought by observing the response of the tree phenology, stem radial growth, and physiology (sap flow) of three predominant tree species, namely English oak, narrow-leaved ash, and common hornbeam, to the environmental variables (climate). Stem radial growth began before bud break in ring-porous oak and ash, whereas in diffuse-porous hornbeam, growth onset occurred after leaf formation. The early onset with intense growth during favorable months (April–May) observed in ring-porous species was a major prerequisite for the successful growth of oak and ash at this site. Tree water deficit (TWD), an indicator of stem hydration, was triggered by decreasing soil moisture in all species, and was most prominent in ash, followed by oak. Intriguingly, sap flow was decoupled from TWD in all species and was driven primarily by evaporative demand from the atmosphere. Oak was the least conservative in regulating sap flow under atmospheric drought followed by hornbeam, whereas ash was most restricted and reduced its transpiration during dry periods. In contrast, ash was characterized by the highest radial growth and growth-based water-use efficiency. The lower water storage capacity of oak and ash is likely compensated by deep rooting and drought avoidance strategies, respectively. Tree species that tend to use surface soil water could be severely limited by more extractive species such as hornbeam. Despite the contrasting leaf and wood phenology, stomatal control, and rooting depth among the studied floodplain tree species, they exhibited analogous sap flow and water storage dynamics responses to drier conditions that enabled them to co-exist in the South Moravian Region. Nevertheless, our results suggest that the severe droughts and human-induced alterations in groundwater pose serious threats to floodplain forests in Central Europe, with certain tree species being unable to adapt to these altered conditions.