Seasonal changes in taxonomic and functional diversity of microbial communities in polar regions are commonly observed, requiring strategies of microbes to adapt to the corresponding changes in environmental conditions. These natural fluctuations form the backdrop for changes induced by anthropogenic impacts. The main goal of this study was to assess the seasonal and temporal changes in bacterial and archaeal diversity and community structure off the northern Antarctic Peninsula over several seasons (spring, summer, autumn) from 2013 to 2015. Ten monitoring stations were selected across the Gerlache and Bransfield Straits and nearby Elephant Island, and archaeal and bacterial communities examined by amplicon sequencing of 16S rRNA genes. Alpha-diversity indices were higher in spring and correlated significantly with temperature. Spring was characterized by the presence of SAR11, and microbial communities remaining from winter, including representatives of Thaumarchaeota (Nimosopurnilus), Euryarchaeota, members of Oceanospirillales, SAR324. Summer and autumn were characterized by a high prevalence of Flavobacteria (NS5 marine group and Polaribacter), Alphaproizobacteria (Rhodobacterales and SAR11 Glade) and Gammaproteobacteria (Oceanospirillales/Balneatrix and Celivibrionales), generally known to be associated with organic matter degradation. Relatively higher abundance of phytoplankton groups occurred in spring, mainly characterized by the presence of the haptophyte Phaeocystis and the diatom Corethron, influencing the succession of heterotrophic bacterial communities. Microbial diversity and community structure varied significantly over time, but not over space, i.e., were similar between monitoring stations for the same time. In addition, the observed interannual variability in microbial community structure might be related to an increase in sea surface temperature. Environmental conditions related to seasonal variation, including temperature and most likely phytoplankton derived organic matter, appear to have triggered the observed shifts in microbial communities in the waters off the northern Antarctic Peninsula.