The northwestern Pacific, characterized by unique tectonic and hydrological settings, has greatly intrigued marine phylogeographers. However, current studies mostly focus on the influence of Pleistocene isolation of sea basins in population structure of species in the region, leaving the contribution of other factors (such as freshwater outflow and environmental gradients) largely unexploited. Here we shed light on the question by investigating phylogeography of the surf clam Mactra chinensis in the East China Sea (ECS). Genetic information was acquired from 501 specimens collected from its main distribution in the region, represented by mitochondrial cytochrome oxidase I (COI) and nine polymorphic microsatellite loci. A shallow and star-like phylogeny was revealed for all COI haplotypes, indicating the origin of populations from a single refugium. Although no divergent lineages existed, population subdivision was detected in both data sets. The most striking pattern was the significant differentiation between populations north and south of a biogeographic boundary-the Changjiang Estuary, suggesting a barrier effect of the freshwater outflow to gene flow. For the northern group, substructure was revealed by COI result as one southernmost population was significant different from other ones. Clear latitude gradations in allele frequencies were revealed by microsatellite analyses, likely influenced by environmental gradient factors such as temperature. Our results demonstrate that genetic subdivision can arise for populations within the ECS despite they have a single origin, and multiple mechanisms including Changjiang River outflow, environmental gradient factors and life-history traits may act in combination in the process.