Spawning substrate is a critical factor in reproduction of commercially valuable percids. Substrate preferences of pikeperch Sander lucioperca were investigated using three types of artificial spawning substrates: long fibred brush, artificial turf and smooth plastic. There was a significantly higher preference for the brush nest, with thick rigid structures, than for artificial turf, while plastic nests remained unoccupied.The majority of pikeperch pairs (94.4%) spawned within 14 days of the beginning of the experiment. No difference was observed between substrates in time from stocking of fish to nest occupation and spawning. After egg laying, nests were moved to controlled conditions of a recirculation aquaculture system for incubation. Hatching rate and larva production did not differ significantly among tested substrates. Either used substrate was suitable for spawning and incubation in a recirculation aquaculture system. Obtained results can be used for semi-artificial reproduction as well as for the support and control of wild pikeperch stocks in natural habitats. K E Y W O R D S artificial nest, broodstock, egg incubation, reproduction, Sander lucioperca ). Information about substrate selectivity and the minimum quantity of suitable spawning grounds How to cite this article: Malinovskyi O, Veselý L, Blecha M, Křišťan J, Policar T. The substrate selection and spawning behaviour of pikeperch Sander lucioperca L. broodstock under pond conditions. Aquac Res. 2018;49:3541-3547.