Environmental degradation related to uncontrolled development resulted in the passage of the United States Clean Water Act (CWA) in 1972, with the stated purpose “to restore and maintain the integrity of the nation’s waters”. Implementation of the CWA lead to increased research to develop multimetric indicators to better measure and understand the complex patterns of ecological responses to stress occurring across levels of biological, spatial and temporal organization. One area of research is the use of integrated indices of chemical risk, ecotoxicological risk and ecological risk to assess the impact of human activity across disturbance gradients of urbanization. Selecting relevant metrics for constructing multimetric index requires identifying bioindicator organisms with capacities to detect signals from anthropogenic disturbances. This study explored the potential efficacy of a suite of higher plant ecotoxicological assays for use as bioindicators in ecological risk assessment along a gradient of urbanization in a wetland ecosystem. The study was conducted in the Pike River watershed (Racine, Wisconsin, USA) in six wetlands selected across a gradient of dominant land use types (agricultural, commercial, residential, undeveloped and industrial). MicroBioTest Phytotoxkit TM ecotoxicological assays, based on growth inhibition of three plants (Sinapis, Sorghum and Lepidium) were used to assess sediment toxicity. The relationships between Phytotoxkit™ responses and predicted pollutant loadings calculated from surrounding landuse provided clear signals of stress from watershed pollutants draining into the wetland sites. The potential for these ecotoxicological indicators to serve as biological response signatures is strong, and further research and calibration in field and microcosms studies will assist in calibrating responses for use in integrated monitoring efforts.