Thraustochytrids are unicellular fungi-like (heterotrophic) marine protists that have long been considered to play an important role in the biogeochemical cycles of the coastal oceans. However, the significance of their ecological functions and their diversity in marine ecosystems remain largely unknown. In this report, we examined the spatial and temporal variations of planktonic thraustochytrids, their relationship with other environmental factors, and their diversity in the subtropical coastal waters of China. The abundance of planktonic thraustochytrids ranged from 2.56 × 105 to 17.57 × 105 cells L-1 with highest abundance detected in polluted coastal water in the Spring (March) season. The thraustochytrid biomass was greater than the bacterial biomass in most seawater samples, ranging from 32.29 to 359.51% that of bacterioplankton. The abundance of thraustochytrids appeared to be largely related to that of bacterioplankton and to chemical oxygen demand in water columns. High-throughput sequencing analyses revealed a total of 105 OTUs (97% similarity), which were members of genera Thraustochytrium, Aplanochytrium, Oblongichytrium, Ulkenia, Labyrinthula and undescribed novel phylotypes. Results of this study indicated unprecedented high diversity of labyrinthulomycetes as well as the presence of novel labyrinthulomycete and thraustochytrid lineages, and also provided new information on the significant role of thraustochytrids in microbial food webs in a coastal marine ecosystem.