Two water sampling campaigns were carried out in April and August 2012 to investigate the spatial and temporal characteristics of 2 H and 18 O in the basin of Dongting Lake and to further understand the interactions between precipitation, river water, lake water, and groundwater. Influenced by different monsoons, precipitation, river water, and lake water are enriched in isotopes in April but depleted in August. Stable isotopes of river water and lake water also have great spatial variations. Unlike river and lake water, groundwater has little seasonal isotopic variation, indicating that the transformation between surface water and groundwater is very slow.