As coastal lagoons serve as nursery areas for some marine and estuarine fish, selective pressures of these brackish or hypersaline lagoons may influence the genetic structure of species and populations. We examined spatial and temporal genetic patterns at eight microsatellite loci in white seabream [Diplodus sargus (Linnaeus 1758)] recruits from the Mar Menor (Southeast Spain) and compared these loci with those in coastal populations from the open sea, observing a high degree of genetic diversity and spatio-temporal genetic stability. However, the results suggest the presence of subpopulations or genetic substructures in the Mar Menor D. sargus population that could be interpreted as a homogeneous mixture of individuals from three differentiated subpopulations in the Mediterranean and evidence of the Wahlund effect. It also suggests that D. sargus adults return to their original spawning habitat, thus conserving the genetic differences among the respective populations over time. Overall, this study demonstrates the importance of the Mar Menor coastal lagoon as a nursery area for the conservation of genetic diversity of D. sargus populations.