Dolichospermum flos-aquae (formerly Anabaena flos-aquae) is a diazotrophic cyanobacterium causing harmful blooms worldwide, which is partly attributed to its capacity to compete for nitrogen (N) and phosphorus (P). Preventing the blooms by reducing P alone or both N and P has caused debate. To test the effects alone and together on the growth of cyanobacteria, we performed culture experiments in different eutrophication scenarios. N2 fixation in terms of heterocyst density, nitrogenase activity and nifH expression increased significantly in P-replete cultures, suggesting that P enrichment facilitates N2 fixation. Correspondingly, the expression of genes involved in P uptake, e.g., those involved in P-transport ( pstS) and the hydrolysis of phosphomonoesters ( phoD), was upregulated in P-deficient cultures. Interestingly, N addition enhanced not only the expression of these genes but also polyphosphate formation and alkaline phosphatase activity in P-deficient cultures relative to the P-replete cultures, as evidenced by qualitative (enzyme-labeled fluorescence) and quantitative (fluorogenic spectrophotometry) measurements. Furthermore, after N addition, cell activity and growth increased in the P-deficient cultures, underscoring the risk of N enrichment in P-limited systems. The eco-physiological responses shown here help further our understanding of the mechanism of N and P colimitation and underscore the importance of dual N and P reduction in controlling cyanobacterial blooms.