Study region: Karst watershed in Lower Flint River Basin (LFRB), southwestern Georgia, USA. Study focus: Baseflow discharges in the LFRB have declined for three decades as regional irrigation has increased; yet, the location and nature of connectivity between groundwater and surface water in this karstic region are poorly understood. Because growing water demands will likely be met by further development of regional aquifers, an important management concern is the nature of interactions between groundwater and surface water components under natural and anthropogenic perturbations. We conducted coarse and fine-scale stream sampling on a major tributary of the Lower Flint River (Ichawaynochaway Creek) in southwestern Georgia, USA, to identify locations and patterns of enhanced hydrologic connectivity between this stream and the Upper Floridan Aquifer. New hydrological insights for the region: Prior water resource studies in the LFRB were based on regional modeling that neglected local heterogeneities in groundwater/surface water connectivity. Our results demonstrated groundwater inputs were concentrated around five of fifty sampled reaches, evidenced by increases in multiple groundwater indicators at these sites. These five reaches contributed up to 42% of the groundwater detected along the entire 50-km sampling section, with ∼24% entering through one groundwater-dominated tributary, Chickasawhatchee Creek. Intermittent flows occurred in two of these upstream reaches during extreme drought and heavy groundwater pumping, suggesting reach-scale behaviors should be considered in resource management and policy. Keywords: Karst hydrogeology, Hydrologic connectivity, Groundwater/surface water interaction, Upper Floridan Aquifer, Groundwater Irrigation