The black southern cod, Patagonotothen tessellata, is the most important notothenioid fish species in terms of abundance in southern Chilean Patagonia. However, studies on its trophic ecology are scarce. Here we assessed the spatial variation in the diet of P. tessellata between two localities, one with oceanic influence (Staples Strait) and another with continental influence (Puerto Bories). We used permutation analysis combined with non-metric multi-dimensional scaling to evaluate spatial differences in diet. In addition, generalized additive models were used to identify the most significant environmental, biological and spatial predictors of variability in diet. The black southern cod presents spatial differences in diet composition among contrasting environmental localities. The diet in Staples Strait was characterized by the dominance of the polychaete Platynereis australis, whereas diet in Puerto Bories was characterized by crustaceans, mainly ostracods, gammarids and algae, mainly the filamentous green alga Rhizoclonium sp. and the red algae Polysiphonia sp. Diet composition did not show significant difference between sexes, whereas diet of small, medium and large fish differed to some degree. Smaller-sized P. tessellata were most likely to contain food. Salinity and temperature had significant influence on diet variability, suggesting that P. tessellata showed a greater diversity of prey items in environmental conditions with greater temperature and lower salinity. The results provide evidence of two dietary patterns depending on the type of environment in which they are distributed, highlighting the potential role of the environmental variables on the availability and abundance of potential prey and in structuring diet.