We studied the contribution of stocking with Mediterranean autochthonous brown trout (Salmo trutta) to the lake-migratory spawners in French tributaries of Lake Geneva. This singular lake system is inhabited by both the resident and the migratory life-history form, and by trout of Atlantic and Mediterranean origins. Using two discriminatory microsatellite loci between the Atlantic and Mediterranean alleles, we compared the genetic composition of the migratory Atlantic populations of the four main tributaries before (1999; N = 113) and after (2004, 2005 and 2006; N = 137) recent introductions of native Mediterranean trout. No significant modification of the genetic composition was detected in all four populations over time and only two individuals were statistically assigned to the Mediterranean strain by the Bayesian method implemented in the structure program. These results suggest a negligible contribution of the stockings with autochthonous Mediterranean trout to the lake-migratory spawners migrating into the four tributaries.