Urban grasslands often exist as fragmented patches with varying threat levels. These patches are generally inappropriately managed due to limited data on their responses to disturbance and seed bank regenerative potential. This study compared seed banks between non-naturally disturbed and undisturbed patches of South African subtropical grasslands in an urban environment. Standing vegetation was characterised via year-round quadrat and monthly transect sampling. The seed banks were sampled after the two main dispersal periods and assessed for species composition and density. The seed bank species (24) represented a subset (9%) of the standing vegetation, with forbs being the most speciose life form and grasses the most abundant. Disturbance had a moderate impact on seed bank density but significantly affected species composition and ordering. Clustering and ordination analyses separated seed banks based on disturbance history. However, seed banks in disturbed patches maintained the general character of the grassland type. Diagnostic, rare and threatened taxa were absent from seed banks, irrespective of disturbance history. Consequently, restoration will possibly require seed enrichment and species reintroduction rather than exploitation of natural seed banks. Identifying disturbance-sensitive species through seed bank surveys can inform grassland conservation and restoration strategies.