Aquatic ecosystems are expected to receive Ag0 and Ag2S nanoparticles (NPs) through anthropogenic waste streams. The speciation of silver in Ag-NPs affects their fate in ecosystems, but its influence on interactions with aquatic plants is still unclear. Here, the Ag speciation and distribution was measured in an aquatic plant, duckweed (Landoltia punctata), exposed to Ag0 or Ag2S NPs, or to AgNO3. The silver distribution in duckweed roots was visualized using synchrotron-based micro X-ray fluorescence (XRF) mapping and Ag speciation was determined using extended X-ray absorption fine structure (EXAFS) spectroscopy. Duckweed exposed to Ag2S-NPs or Ag0-NPs accumulated similar Ag concentrations despite an order of magnitude smaller dissolved Ag fraction measured in the exposure medium for Ag2S-NPs compared to Ag0-NPs. By 24 h after exposure, all three forms of silver had accumulated on and partially in the roots regardless of the form of Ag exposed to the plants. Once associated with duckweed tissue, Ag0-NPs had transformed primarily into silver sulfide and silver thiol species. This suggests that plant defenses were active within or at the root surface. The Ag2S-NPs remained as Ag2S, while AgNO3 exposure led to Ag0 and sulfur-associated Ag species in plant tissue. Thus, regardless of initial speciation, Ag was readily available to duckweed.