The hybrid 'Zhongshanshan 406' (. hybrid 'Zhongshanshan 406') [ Tenore × (L.). Rich] has an outstanding advantage in flooding tolerance and thus has been widely used in wetland afforestation in China. () played key roles in ethanol metabolism to maintain energy supply for plants in low-oxygen conditions. Two genes were isolated and characterized- and (GenBank ID: AWL83216 and AWL83217-basing on the transcriptome data of . hybrid 'Zhongshanshan 406' grown under waterlogging stress. Then the functions of these two genes were investigated through transient expression and overexpression. The results showed that the ThADH1 and ThADH4 proteins both fall under ADH III subfamily. ThADH1 was localized in the cytoplasm and nucleus, whereas ThADH4 was only localized in the cytoplasm. The expression of the two genes was stimulated by waterlogging and the expression level in roots was significantly higher than those in stems and leaves. The respective overexpression of and in caused the opposite phenotype, while waterlogging tolerance of the two transgenic significantly improved. Collectively, these results indicated that genes and were involved in the tolerance and adaptation to anaerobic conditions in . hybrid 'Zhongshanshan 406'.