Tetrad vascular plant data from Staffordshire (VC 39), accumulated 1995 - 2009, were analysed using Two Way Indicator Species Analysis and Principal Components Analysis, explored using GIS, to examine major floristic distribution patterns. Environmental data were also examined using Redundancy Analysis. The strongest floristic trend was a human influence axis, arranged south—north, with the south being most anthropocentric and the north having a high proportion of semi-natural habitats. Indicator species for different parts of the County showed a corresponding strong biogeographic element, with ‘Southern-temperate’ and ‘Boreo-temperate’ characteristics in south and north respectively. The next most important factor in the distribution of plant species in Staffordshire was habitat quality and richness, which separated intensively-managed farmland from areas with seminatural habitats and from human influenced urban areas. Environmental variables associated with the main division in the data were mainly climate and altitude and the extent of developed land. Historic field patterns, identified by Staffordshire County Council, were associated with semi-natural habitats, linking an historical landscape characterisation with vascular plant analysis for the first time in the UK. The investigation set out to characterise and classify spatial units in terms of biodiversity importance, and to produce an objective ecological classification of the Staffordshire region based on botanical and physical features. The resulting classification characterises different areas of the County in terms of plant species composition, and has many potential applications in terms of nature conservation strategy development, targeting of resources and monitoring. A further aim was to facilitate the selection of indicator species for high quality habitats – the study has enabled revision of Staffordshire’s axiophyte plant list. Both the ecological classification and indicator species will be of strategic value because the information provided is authenticated by being based on objective analysis of a wide range of data. In Staffordshire, further work is required to develop environmental data, especially an effective dataset for hedges, while comparison of the data with historical botanical data could provide further information about the County’s previous environment. The techniques developed in the study could be used more widely to: produce of lists of key species, provide information for environmental modeling, climate change monitoring, target nature conservation activity, and to develop landscape strategies.