Water resource development and drought have altered river flow regimes, increasing average flood return intervals across floodplains in the Murray-Darling Basin, Australia, causing health declines in riparian river red gum (Eucalyptus camaldulensis) forests and woodlands. Environmental flow allocations helped to alleviate water stress during the recent Millennium Drought (1997–2010), however, quantification of the flood frequency required to support healthy E. camaldulensis communities is still needed. We quantified water requirements of E. camaldulensis for two years across a flood gradient (trees inundated at frequencies of 1:2, 1:5 and 1:10 years) at Yanga National Park, New South Wales to help inform management decision-making and design of environmental flows. Sap flow, evaporative losses and soil moisture measurements were used to determine transpiration, evapotranspiration and plant-available soil water before and after flooding. A formula was developed using plant-available soil water post-flooding and average annual rainfall, to estimate maintenance time of soil water reserves in each flood frequency zone. Results indicated that soil water reserves could sustain 1:2 and 1:5 trees for 15 months and six years, respectively. Trees regulated their transpiration rates, allowing them to persist within their flood frequency zone, and showed reduction in active sapwood area and transpiration rates when flood frequencies exceeded 1:2 years. A leaf area index of 0.5 was identified as a potential threshold indicator of severe drought stress. Our results suggest environmental water managers may have greater flexibility to adaptively manage floodplains in order to sustain E. camaldulensis forests and woodlands than has been appreciated hitherto.