To investigate the temporal-spatial distribution of picophytoplankton in relation to different water masses in the northern East China Sea (ECS), picophytoplankton abundance were investigated using flow cytometry with environmental factors in 2016–2017. The results from the analysis of flow cytometer data showed that Synechococcus appeared across all seasons, exhibiting its minimum abundance in winter and maximum abundance in summer. Furthermore, high abundance was detected in the surface mixed layer during spring and summer when vertical stratification occurs; in particular, Synechococcus exhibited maximum abundance in thermocline layer, indicating a close correlation to water temperature and thermocline formation. In addition, the abundance of Synechococcus indicated a decrease in the western seas in 2017 compared to 2016 under the strong influence of the Changjiang Diluted Water (CDW). This was determined by the significant influence of the CDW on the abundance of Synechococcus during summer in the northern waters of the ECS. In contrast, Prochlorococcus did not appear during winter and spring, and its distribution was limited during summer and autumn in the eastern seas under the influence of the Kuroshio current. The largest range of Prochlorococcus distribution was confirmed during autumn without the influence of the CDW. Thus, the distribution pattern of each picophytoplankton genus was found to be changing in accordance to the extension and reduction of sea current in different seasons and periods of time. This is anticipated to be a useful biological marker in understanding the distribution of sea currents and their influence in the northern waters of the ECS.