Spatio-temporal variations in the composition of the animal interactive assemblages may result in variations in selective pressures on the plants. In ant–seed dispersal mutualisms, the study of the magnitude of spatial and temporal variation of ant assemblages is rarely studied, limiting inferences and generalizations on the evolution of this mutualism. Here, we describe the ant–disperser assemblage of the myrmecochorous herb Helleborus foetidus in 14 populations across the Iberian Peninsula, and dissect the variation in the assemblage into spatial and temporal components as a first step to evaluate the evolutionary potential of this interaction. The ant–visitor assemblage of H. foetidus was mainly represented by species of Formicinae and it was highly diverse and variable in composition and function. Ants behaving as legitimate dispersers and those with mixed behaviour numerically dominated the assemblage compared with elaiosome consumers. The magnitude of the spatial variation was higher than the temporal variation, suggesting that the relative frequency of each functional group will be more foreseeable among years in each population than among populations. At the expense of further analysis of the effects of such variation on dispersal success, we can envisage a selection mosaic scenario, where local adaptive responses of plants might arise as a result of local variations in the specific composition and function of the assemblage. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society, 2007, 92, 135–150.