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Phytoplankton nutrient limitation experiments were performed from 1994 to 1996 at three stations in the Cape Fear River Estuary, a riverine system originating in the North Carolina piedmont. Nutrient addition bioassays were conducted by spiking triplicate cubitainers with various nutrient combinations and determining algal response by analyzing chlorophyll a production and 14C uptake daily for 3 d. Ambient chlorophyll a, nutrient concentration, and associated physical data were collected throughout the estuary as well. At a turbid, nutrient-rich oligohaline station, significant responses to nutrient additions were rare, with light the likely principal factor limiting phytoplankton production. During summer at a mesohaline station, phytoplankton community displayed significant nitrogen (N) limitation, while both phosphorus (P) and N were occasionally limiting in spring with some N+P co-limitation. Light was apparently limiting during fall and winter when the water was turid and nutrient-rich, as well as during other months of heavy rainfall and runoff. A polyhaline station in the lower estuary had clearer water and displayed significant responses to nutrient additions during all enrichment experiments. At this site N limitation occurred in summer and fall, and P limitation (with strong N+P co-limitation) occurred in winter and spring. The data suggest there are two patterns controlling phytoplankton productivity in the Cape Fear system: 1) a longitudinal pattern of decreasing light limitation and increasing nutrient sensitivity along the salinity gradient, and 2) a seasonal alternation of N limitation, light limitation, and P limitation in the middle-to-lower estuary. Statistical analyses indicated upper watershed precipitation events led to increased flow, turbidity, light attenuation, and nutrient loading, and decreased chlorophyll a and nutrient limitation potential in the estuary. Periods of low rainfall and river flow led to reduced estuarine turbidity, higher chlorophyll a, lower ambient nutrients, and more pronounced nutrient limitation.