Organic acids are critical as secondary metabolites for plant adaption in a stressful situation. Oxalic acid, tartaric acid, and malic acid can improve plant tolerance under waterlogged conditions. Two prominent woody species (Taxodium distichum-Swamp cypress and Salix matsudana-Chinese willow) have been experiencing long-term winter submergence and summer drought in the Three Gorges Reservoir. The objectives of the present study were to explore the responses of the roots of two woody species during flooding as reflected by root tissue concentrations of organic acids. Potted sample plants were randomly divided into three treatment groups: control, moderate submergence, and deep submergence. The concentrations of oxalic acid, tartaric acid, and malic acid in the main root and lateral roots of the two species were determined at four stages. The results showed that T. distichum and S. matsudana adapted well to the water regimes of the reservoir, with a survival rate of 100% during the experiment period. After experiencing a cycle of submergence and emergence, the height and base diameter of the two species showed increasing trends. Changes in base diameter showed insignificant differences between submergence treatments, and only height was significant under deep submergence. The concentrations of three organic acids in the roots of two species were influenced by winter submergence. After emergence in spring, two species could adjust their organic acid metabolisms to the normal level. Among three organic acids, tartaric acid showed the most sensitive response to water submergence, which deserved more studies in the future. The exotic species, T. distichum, had a more stable metabolism of organic acids to winter flooding. However, the native species, S. matsudana, responded more actively to long-term winter flooding. Both species can be considered in vegetation restoration, but it needs more observations for planting around 165 m above sea level, where winter submergence is more than 200 days.