Population genetic studies provide insights into intraspecific diversity and dispersal patterns of microorganisms such as protists, which help understanding invasions, harmful algal bloom development and occurrence of seafood poisoning. Genetic differentiation across geography has been reported in many microbial species indicating significant dispersal barriers among different habitats. Temporal differentiation has been less studied and its frequency, drivers and magnitude are poorly understood due to a lack of integral studies. The toxic dinoflagellate species /Gambierdiscus caribaeus/ was sampled during two years in the Florida Keys, and repeatedly from 2006 to 2016 at St. Thomas, US Virgin Islands (USVI), including a three-year period with monthly sampling, to compare spatial and temporal genetic differentiation. Samples from the USVI site showed high temporal variability in local population structure, which correlated with changes in salinity and benthic habitat cover. In some cases, temporal variability exceeded spatial differentiation, despite apparent lack of connectivity and dispersal across the Greater Caribbean Region based on the spatial genetic data. Thus, local processes such as selection might have a stronger influence on population structure in microorganisms than geographic distance. The observed high temporal genetic diversity challenges the prediction of harmful algal blooms and toxin concentrations, but illustrates also the evolutionary potential of microalgae to respond to environmental change.