The growing production and commercial application of engineered nanoparticles (ENPs), such as Ag, CeO2, and TiO2 nanoparticles, induce a risk to the environment as ENPs are released during their use. The comprehensive assessment of the environmental risk that the ENPs pose involves understanding their fate and behavior in wastewater treatment systems. Therefore, in this study, we investigate the effect of plants and different substrates on the retention and distribution of citrate-coated silver nanoparticles (Ag-NPs) in batch experimental setups simulating constructed wetlands (CWs). Sand, zeolite, and biofilm-coated gravel induce efficient removal (85, 55, and 67 %, respectively) of Ag from the water phase indicating that citrate-coated Ag-NPs are efficiently retained in CWs. Plants are a minor factor in retaining Ag as a large fraction of the recovered Ag remains in the water phase (0.42–0.58). Most Ag associated with the plant tissues is attached to or taken up by the roots, and only negligible amounts (maximum 3 %) of Ag are translocated to the leaves under the applied experimental conditions.