Abstract: Antibiotic resistance genes (ARGs) are emerging pollutants, which is widely present in a variety of environmental media. The lake water can sustain ARGs for a long time due to the low circulation rate of pollutants. Hence, researchers in the ARGs field pay much attention to lake water studies. However, previous studies mainly focused on the pollution level of ARGs in lakes, and the sources of pollution remain uncertain. East Dongting Lake is a National Nature Reserve, which plays an important role in protecting the ecological environment and mitigating the floods in Yangtze River. Samples of surface water and land pollution sources (aquaculture areas and sewage treatment plants) of East Dongting Lake were collected in November 2019 to explore the occurrence characteristics of antibiotics and ARGs in the surface water of East Dongting Lake, analyze the correlation between ARGs and environmental parameters (e.g., antibiotics), and understand the contribution of land pollution sources to ARGs in the lake. Antibiotics and ARGs were detected by using ultra performance liquid chromatography tandem mass spectrometry and fluorescence quantitative PCR, respectively. Meanwhile, water quality parameters were calibrated and redundancy analysis was carried out. The results showed that: (1) The concentration of antibiotics in the surface water of East Dongting Lake was in the range of ND (Non-Detected)-486.59 ng/L, and ofloxacin concentration was the highest. The spatial distribution analysis showed that the concentration of antibiotics was closely related to the surrounding pollution sources. (2) The detection rate of ARGs was 100%, and the concentration of sul2 was higher than others, with an average concentration of 1.3×10³ copies/mL. In addition, the spatial distribution also showed that the pollution levels of ARGs was affected by pollution sources along the lake. (3) Sewage treatment plants can reduce ARGs to a certain extent, but cannot completely remove ARGs, and may even increase the possibility of ARGs transmission. Furthermore, the contribution of wastewater treatment plants to the ARGs of the lake is much higher than that of aquaculture areas. (4) The results of redundancy analysis between ARGs and antibiotics were consistent with the characteristics of ARGs under the selective pressure of their corresponding antibiotics. Total phosphorus (contribution rate was 28.9%), conductivity (contribution rate was 15.4%), nitrate nitrogen (contribution rate was 13.3%) and temperature (contribution rate was 12.7%) were the four most important water quality parameters affecting ARGs abundance. The results showed that ofloxacin and roxithromycin were the main antibiotics in terms of concentration, and the concentration of sulfonamides and tetracyclines resistance genes was high. The abundance of ARGs was not only related to the selective pressure of the corresponding antibiotics, but also to the pressure of some environmental factors.