Woodland ecosystems within the Duffield landscape, Derbyshire, represent an important component of the southern Peak District ecological network, providing essential ecosystem services while supporting a diverse assemblage of native flora. This study investigated woodland distribution and floral diversity by integrating remote sensing analysis with a field-based botanical survey. Tree cover patterns were assessed using the Global Forest Change dataset across multiple canopy cover thresholds (>10–75%) to evaluate woodland structure, connectivity, and recent woodland gain between 2000 and 2020. A total of 20 vascular plant species were recorded, representing woodland, grassland, wetland, and ecotonal habitats. The flora was dominated by native species, including Prunella vulgaris, Achillea millefolium, Lotus corniculatus, Digitalis purpurea, and two orchid species (Anacamptis pyramidalis and Dactylorhiza fuchsii), indicating relatively diverse semi-natural habitats. Woodland mapping demonstrated that lower canopy thresholds captured fragmented woodland, hedgerows, and riparian vegetation, whereas higher thresholds identified mature woodland concentrated along valley slopes. Woodland gain adjacent to existing forests suggested ongoing natural regeneration and improved habitat connectivity. The occurrence of numerous nectar-rich and nitrogen-fixing species highlights the ecological importance of the landscape for pollinators, nutrient cycling, and ecosystem resilience, while the presence of the invasive Impatiens glandulifera emphasises the need for continued management. Overall, the study demonstrates that Duffield supports a heterogeneous woodland ecosystem of considerable biodiversity value, underscoring the importance of protecting mature woodland, maintaining ecological connectivity, controlling invasive species, and promoting long-term biodiversity monitoring and habitat conservation.