Although polyploidization is rare among bivalve mollusks, recent cytogenetic studies have revealed a remarkable degree of genome amplification (up to 13n) in the freshwater bivalve family sphaeriidae. We generated single-copy nuclear gene trees in order to test hypotheses addressing the evolutionary origins of sphaeriid genome duplication. Polyploid North American members of three cosmopolitan sphaeriid genera (Sphaerium, Musculium, and Pisidium) were characterized for their expressed allelic repertoire of a 526 nt c-DNA fragment of 6-phosphogluconate dehydrogenase (PGD). Pronounced levels of intra-individual genetic variation were uncovered in most of the polyploid taxa and a minority of alleles showed strong evidence of recombination. Phylogenetic analyses resolved polyploid sphaeriid PGD alleles into two clades (A, B), each of which contained a subsample of intra-individual allelic diversity of the genus Sphaerium. These two clades were also recovered in Musculium, however one (B) is represented here by a single recombinanat allele. With the exception of a divergent segment in one putatively recombinanat allele, the expressed PGD repertoire of the three Pisidium species investigated was restricted to one of the two clades (A). Major within-clade PGD gene tree branching patterns were congruent with mitochondrial gene tree topologies for these taxa. These results are inconsistent with a pattern of recent independent attainment of a polyploid status by our Sphaerium/Musculium study taxa and indicate that they may share a common genome duplication event predating the Miocene appearance of these two genera in the fossil record.