Palaeoecological reconstructions in the Netherlands are commonly based on pollen and macrofossil analysis, but can be limited if the preservation of organic material is poor. Phytoliths, biogenic silica, do not have this limitation and preserve in settings where other macro- and microfossils do not. Little is known about how phytolith assemblages preserved in soils and sediments reflect the parent vegetation in north-western European systems, so it is currently difficult to contextualise past environments. Here, we characterise phytolith assemblages for soil samples recovered from three major vegetation types in the Netherlands to provide reference data for future reconstructions of past vegetation change. We collected 42 soil surface samples from forests, wetlands and agricultural fields across the Netherlands and characterised the phytolith assemblages they contained. We identified the different phytolith morphotypes and quantified the percentages and concentrations (#phytoliths/cm3 soil) in each sample. We used non-metric multidimensional scaling to assess the variation in phytolith assemblage composition within, and between, the three vegetation types. The phytolith assemblages analysed from the forests, wetlands and agricultural fields were clearly distinguishable from each other. Agricultural fields were dominated by four phytolith morphotypes of grass silica short cells (GSSCs): rondel (tabular), cross type 1 (>15 µm), rondel (elongated) and disturbance or crop phytoliths. Forests settings had significantly higher amounts of different arboreal phytoliths (large and small spheroid rugose) compared with other vegetation types. Wetlands could be identified by significantly higher amounts of Cyperaceae phytoliths (papillate) and other GSSCs (saddle and bilobates with thick castula). Phytolith assemblages could distinguish different subtypes of vegetation within forest and wetland areas, while differences between agricultural systems could not be identified. Our study demonstrates that phytoliths preserved in soils or sediments can be used to separate major vegetation types across the Netherlands. Thus, these results support the hypothesis that phytoliths can be used to infer past environmental conditions in palaeoecological reconstructions. We suggest that future work should: (1) focus on characterising which phytolith types are produced by the commonest tree, wetland, shrub and herb species in the Netherlands and (2) characterise phytolith assemblages across a wider array of vegetation types in north-western European systems to increase the capability for quantitative reconstructions using phytolith assemblages.