This paper adopted high-performance liquid chromatography-mass spectrograph (HPLC-MS) to investigate the occurrence and seasonal changes of tetracycline antibiotics in the Xiang River and its sediments. Five representative antibiotic resistance genes (tet A, tet C, tet G, tet M, tet X) in water and sediment samples were quantified with real-time PCR. The concentrations of tetracycline showed sensitive seasonal changes, ranging from 16.23 ng·L-1 to 496.73 ng·L-1 in the water samples, and from 347.77 ng·kg-1 to 3 829.75 n·kg-1 in the sediment samples, respectively. The target resistance genes were detected in all samples. Compared with other resistance genes, tet A and tet C were higher in absolute copy number and relative abundance, suggesting that the tet A and tet C were dominant in the Xiang River. Statistical analysis indicated a positive correlation between the relative abundance of tet A and tet C in the water samples and the sediment samples in winter. There was no linearly relationship between antibiotics and resistance genes.