AbstractInterstitial invertebrate assemblages of the Rhǒne River were studied at the scale of the entire alluvial plain. Ten stations, located in five former channels, were sampled with a Bou‐Rouch pump at 50 and 100 cm depth. The chemical characteristics of the interstitial water allowed the differentiation of stations without hydrological connections with the subterranean habitats, stations fed by surface water coming through the bank sediments and stations fed by true groundwater. The faunal assemblages reflect the difference between connected and isolated stations but did not demonstrate any differences related to the origin of the water. In fact the interstitial assemblages are distributed along a gradient from the margin of the floodplain (stations with a high physical stability associated with the absence of flood periods) to the area close to the main channel regularly influenced by floods. These results demonstrate that both groundwater characteristics and the position of the station in the floodplain influence the spatial distribution of interstitial fauna in the alluvial valley of the Rhǒne Ri