Dissolved phosphorus (DP) can be released from wetlands as a result of flooding or shifts in water column concentrations. Our objectives were to determine the long-term (1460 d) DP retention and release characteristics of an in-stream wetland, and to evaluate how these characteristics respond to flooding, draining, and changes in DP concentrations. The studied in-stream wetland drains an agriculturally intensive subwatershed in the North Carolina Coastal Plain region. The wetland's DP retention and release characteristics were evaluated by measuring inflow and outflow DP concentrations, DP mass balance, and DP movement across the sediment-water column interface. Phosphorus sorption isotherms were measured to determine the sediment's equilibria P concentration (EPCo), and passive samplers were used to measure sediment pore water DP concentrations. Initially, the in-stream wetland was undersized (0.31 ha) and released 1.5 kg of DP. Increasing the in-stream wetland area to 0.67 ha by flooding resulted in more DP retention (28 kg) and low outflow DP concentrations. Draining the in-stream wetland from 0.67 to 0.33 ha caused the release of stored DP (12.1 kg). Shifts both in sediment pore water DP concentrations and sediment EPCo values corroborate the release of stored DP. Reflooding the wetland from 0.33 to 0.85 ha caused additional release of stored DP into the outflowing stream (10.9 kg). We conclude that for a time period, this in-stream wetland did provide DP retention. During other time periods, DP was released due to changes in wetland area, rainfall, and DP concentrations.