Because estuaries are dynamic on many temporal scales, it is difficult to differentiate long-term shifts from those resulting from erratic pulse events like severe storms. Diatoms are abundant and diverse in estuaries, and may characterize these dynamics across relevant time scales. The climate of south Florida is subtropical, with dry winters and wet summers that may also harbor tropical storms and cyclones. We investigated whether these repeated seasonal drivers of change in estuaries differ from the influence of tropical cyclones on planktonic and benthic diatom assemblages. Diatom assemblages and environmental parameters were measured in the Charlotte Harbor watershed of southwest Florida to identify differences between the wet and dry seasons and changes that occurred following Tropical Storm Debby in 2012. Indicator taxa that were significantly affiliated with each season and post-storm conditions were identified. Diatom assemblages across the watershed were similar in wet and dry seasons, but differences increased following the tropical storm. The reduction in community dispersion following the storm was pronounced in each of the major drainages of the estuary. There were spatially-distinct responses of diatoms to environmental changes driven by the storm. These results suggest that past storm activity could be detected using diatoms preserved in estuarine sediments using both single indicator-species as well as community approaches