Riverine floodplains are recognized as centers of biodiversity, but due to intense anthropogenic pressures, many active floodplains have disappeared during the last century. This research focuses on the long-term changes in phytoplankton diversity in the floodplain lake situated in the Kopački Rit (Croatia), one of the largest conserved floodplains in the Middle Danube. The recent dataset from 2003 to 2016 and historical data from the 1970s and 1980s indicate high phytoplankton diversity, summarising 680 taxa for nearly half a century. The variability of species richness is driven by specific in-lake variables, particularly water temperature, water depth, total nitrogen, pH, and transparency, determined by a redundancy analysis of the current data. The high phytoplankton diversity levels are sustained regardless of intense pressures on the lake environment, including exposure to strong anthropogenic pollution in the past and extreme hydrological events, both droughts and floods, which have increasingly affected this part of the Danube in the last decades. The conserved hydrological connection between various biotopes along the river-floodplain gradient seems crucial in maintaining high phytoplankton diversity. Accordingly, conserving natural flooding is mandatory to maintain high biodiversity in complex and dynamic river-floodplain systems.