To expand current knowledge on the molecular aspects of alkaline phosphatase PhoX in shallow eutrophic freshwaters, we investigated the genetic diversity and abundance of the PhoX-encoding gene (phoX) in 4 ecological regions in Lake Taihu, China, following a gradient in total phosphorus concentrations ranging from hypereutrophic to mesotrophic. Bacterial phoX was heterogeneously distributed with the highest diversity in the eutrophic regions and the highest abundance in the mesotrophic Xukou Bay. The concentrations of total phosphorus and enzymatically hydrolyzable phosphorus determined the distribution of bacterial phoX in Lake Taihu. Most (70.8%) of the phoX-translated proteins had <90% similarity to the PhoX proteins in the GenBank database, suggesting the presence of novel phoX genotypes in Lake Taihu. The low overlap in phoX genotypes (15.8%) between Lake Taihu and some marine ecosystems, and the dominance of the translated proteins most similar to the Alphaproteobacteria-affiliated PhoX, demonstrate the uniqueness of PhoX in eutrophic freshwaters.