Poyang Lake is a unique wetland system that has evolved in response to natural seasonal fluctuations in water levels. To better characterize the response of water quality to hydrological variation, historical data were analyzed in combination with dissolved organic matter (DOM) fluorescence samplings conducted in situ. Historical data showed that long-term changes in water quality are mainly controlled by the sewage inputs to Poyang Lake. Monthly changes in water quality recorded during 2008 and 2012 suggest that water level may be the most important factor for water quality during a hydrological year. DOM fluorescence samples were identified as three humic-like components (C1, C2, and C3) and a protein-like component (C4). These obvious compositional changes in DOM fluorescence were considered to be related to the hydrodynamic differences controlled by water regimen. Principal component analysis (PCA) showed higher C1 and C2 signals during a normal season than the wet season, whereas C3 was lower, and C4 was higher in the dry season than in the wet or normal seasons. From the open lake to the Yangtze River mouth, increased C3 component carried by backflows of the Yangtze River to the lake resulted in these unique variations of PCA factor 2 scores during September. These obvious compositional changes in DOM fluorescence were considered to be related to the hydrodynamic differences controlled by water regimen. DOM fluorescence could be a proxy for capturing rapid changes in water quality and thereby provide an early warning signal for the quality of water supply.