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Land-use and land-cover (LULC) change is a primary driver of terrestrial carbon release, often through the conversion of forest into agriculture or expansion of urban areas. Classification schemes are a key component of landscape analyses. This study creates a novel LULC classification scheme by incorporating ecological data to redefine classes of an existing LULC classification based on variation in above-ground tree carbon. A tree inventory was conducted for 531 plots within a subbasin of the Tampa Bay Watershed, Florida, USA. Above-ground tree carbon was estimated using the i-Tree model. Plots were classified using the Florida Land Use Cover Classification System. Mean quantities of above-ground tree carbon, by class, were tested for statistical differences. A reclassification was conducted based on these differences. Sub-classes within a given “land cover” class were similar for six of the seven classes. Significant differences were found within the “Wetlands” class based on vegetation cover, forming two distinct groups: “Forested Wetlands” and “Non-forested and Mangrove Wetlands”. The urban “land use” class showed differences between “Residential” and “Non-residential” sub-classes, forming two new classes. LULC classifications can sometimes aggregate areas perceived as similar that are in fact distinct regarding ecological variables. These aggregations can obscure the true variation in a parameter at the landscape scale. Therefore, a study’s classification system should be designed to reflect landscape variation in the parameter(s) of interest.