Abstract. Trophic preference (i.e., food resources and trophic levels) of different copepod groups was assessed along a salinity gradient in the temperate estuarine Gwangyang Bay of Korea, based on seasonal investigation of taxonomic results in 2015 and stable isotope analysis incorporating multiple linear regression models. The δ13C and δ15N values of copepods in the bay displayed significant spatial heterogeneity as well as seasonal variations, which were indicated by their significant relationships with salinity and temperature, respectively. Both spatial and temporal variations reflected those in isotopic values of food sources. The major calanoid groups (marine calanoids and brackish water calanoids) had a mean trophic level of 2.2 relative to nanoplankton as the basal food source, similar to the bulk copepod assemblage; however, they had dissimilar food sources based on the different δ13C values. Calanoid isotopic values indicated a mixture of different genera including species with high δ15N values (e.g., Labidocera, Sinocalanus, and Tortanus), moderate values (Calanus sinicus, Centropages, Paracalanus, and Acartia), and relatively low δ15N values (Eurytemora pacifica and Pseudodiaptomus). Feeding preferences of different copepods probably explain these seasonal and spatial patterns of the community trophic niche. Bayesian mixing model calculations based on source materials of two size fractions of particulate organic matter (nanoplankton at < 20 µm vs. microplankton at 20–200 µm) indicated that Acartia and Centropages preferred large particles; Paracalanus, Calanus, Eurytemora, and Pseudodiaptomus apparently preferred small particles. Tortanus was typically carnivorous with low selectivity on different copepods. Labidocera preferred marine calanoids Acartia, Centropages, and harpacticoids; on the other hand, Sinocalanus and Corycaeus preferred brackish calanoids Paracalanus and Pseudodiaptomus. Overall, our results depict a simple energy flow of the planktonic food web of Gwangyang Bay: from primary producers (nanoplankton) and a mixture of primary producers and herbivores (microplankton) through omnivores (Acartia, Calanus, Centropages, and Paracalanus) and detritivores (Pseudodiaptomus, Eurytemora, and harpacticoids) to carnivores (Corycaeus, Tortanus, Labidocera, and Sinocalanus).