Wetlands are among the most fragile habitats on Earth and have often undergone major environmental changes. As a study case in this context, the present work aims at increasing the floristic knowledge of a reclaimed land now turned into an agricultural lowland with scarce patches of natural habitats. The study area is named Piana di Rosia, and it is located in southern Tuscany (Italy). The compiled checklist consists of 451 specific and subspecific taxa of vascular plants. The life-form spectrum shows a predominance of hemicryptophytes, followed by therophytes. The chorological spectrum highlights a co-dominance of Euri-Mediterranean and Eurasian species along with many widely distributed species. The checklist includes seven species of conservation concern, three Italian endemics (Crocus etruscus Parl., Polygala vulgaris L. subsp. valdarnensis (Fiori) Arrigoni, and Scabiosa uniseta Savi), 41 alien species, 21 segetal species, and 11 aquatic macrophytes of which five helophytes and six hydrophytes. This study suggests that irreversible land-use changes in wetlands can lead towards a simplification of the flora. However, despite the deep transformations that the former wetland has undergone, the presence of some aquatic and protected taxa is interesting. From a conservation point of view, the natural value of this agricultural area could be enhanced and its current management partly reconsidered, thus preserving the remnants of naturalness present.