Extensive changes in land cover during the 20th century are known to have had detrimental effects on biodiversity in rural landscapes, but the magnitude of change and their ecological effects are not well known on regional scales. We digitized historical maps from the beginning of the 20th century over a 1652 km² study area in southeastern Sweden, comparing it to modern-day land cover with a focus on valuable habitat types. Semi-natural grassland cover decreased by over 96 % in the study area, being largely lost to afforestation and silviculture. Grasslands on finer soils were more likely to be converted into modern grassland or arable fields. However, in addition to remaining semi-natural grassland, today’s valuable deciduous forest and wetland habitats were mostly grazed grassland in 1900. An analysis of the landscape-level biodiversity revealed that plant species richness was generally more related to the modern landscape, with grazing management being a positive influence on species richness.