Seasonal variations in the picophytoplankton community structure (Synechococcus spp. and picoeukaryotes) were studied by flow cytometry in the coastal ecosystem of the subtropical western Pacific from October 2019 to September 2020. Synechococcus spp. was dominant in abundance during the study period, with its density ranging from 0.05 to 5.6 × 104 cells mL−1; its maximum occurred in July 2020. Picoeukaryotes were less abundant, with their density ranging from 0.2 to 13.6 × 103 cells mL−1. Their highest abundance was recorded in January 2020. The growth rates of Synechococcus spp. and picoeukaryotes ranged from −0.39 to 1.42 d−1 and 0.38 to 2.46 d−1, respectively, throughout the study period. Overall, the growth rate of the picoeukaryotes was significantly higher than that of Synechococcus spp. It is interesting to note that the grazing mortality of Synechococcus spp. and picoeukaryotes during the warmer period (April to September) was relatively low. Based on this study, we suggest that mixotrophic nanoflagellates lowered their feeding activity that obtained nutrients from prey and instead used additional nutrients during the incubation experiments. Our study demonstrated that a shift in the picophytoplankton community composition and grazing activity of predacious nanoflagellates in cold and warm periods can impact on the seasonal dynamics of the microbial food web.