Female Northern Pintails (Anas acuta) were sampled in California's three main Central Valley wintering regions (Sacramento Valley, Suisun Marsh, San Joaquin Valley) during September–October before most regional movements occur and microsatellite and mitochondrial DNA were analyzed to examine population structure and relatedness. Despite reportedly high rates of early-fall pairing and regional fidelity, both sets of markers indicated that there was little overall genetic structuring by region. Pintails from Suisun Marsh did exhibit higher relatedness among individuals and capture groups than in the Sacramento or San Joaquin Valleys, likely reflecting a sample comprised of a greater proportion of local breeders. The lack of genetic structuring among regions indicates that a high degree of movement and interchange occurs among pintails wintering in the Central Valley. Thus, although maintaining the existing distribution of pintails among Central Valley regions is important for other reasons, it does not appear to be critical to retain current patterns of population genetic variation. Because of potential lack of independence among highly related study individuals, researchers should consider regional differences in relatedness when designing sampling schemes and interpreting research findings.