According to European Union guidelines, the assessment of the ecological status of Transitional Water Systems (TWSs) should be based on the monitoring of biological communities rather than physico-chemical parameters and pollutants. Macrophytes, including aquatic angiosperms and macroalgae, are organisms that respond more quickly to environmental changes by varying the structure and biomass of their assemblages. There are several ecological indices based on macrophytes, among them the Macrophyte Quality Index (MaQI), which has been intercalibrated with water and sediment parameters, nutrient concentrations, and pollutants and is used to determine the ecological status of Italian TWSs. In the Venice Lagoon, it was applied to 87 stations, showing a significant score increase over the last ten years of monitoring (2011–2021) due to progressive lagoon environmental recovery. The dominant taxa assemblages, previously dominated by Ulvaceae, were replaced by species of higher ecological value, with an increase in the number and distribution of sensitive species, as well as the spread and cover of aquatic angiosperms. The rise in the Ecological Quality Ratio (EQR) determined by the MaQI confirms the key role of macrophyte monitoring in detecting environmental changes in TWSs. In fact, a simple check of the presence or absence of aquatic angiosperms and sensitive species is sufficient for an initial rapid assessment of the ecological status of these environments.