The threat of antibiotic resistance genes (ARGs) caused by animal manure application to human health has been the focus of attention in agriculture. Applying lime to acid soil for the amelioration of soil acidity is a prevailing agricultural practice. However, the role of lime on the spread of antibiotic resistome from soil to plant is unknown. In this study, a pot experiment of lettuce was established in the acid black soil with lime addition at the rate (w/w) of 0%, 0.08%, 0.16%, and 0.32% of the total soil mass to explore the transmission of ARGs introduced by the fresh poultry manure in the soil-plant system. The bulk and rhizosphere soils as well as the leaf samples were collected after lettuce was cultivated for 60 days, the bacterial community and antibiotic resistome in these samples were determined by using Illumina sequencing and high-throughput quantitative PCR (HT-qPCR) methods, respectively. Results showed that lime application decreased the number and abundance of ARGs and slowed down the spread of manure-derived ARGs in the soil-plant system. The ARGs and bacterial community composition were significantly varied among bulk soils, rhizosphere soils and leaf endophyte, and also influenced by lime within the same sampling types. The structural equation model further demonstrated that the lime addition had a negative effect on ARG diversity, which was also indirectly regulated by bacterial community diversity. These findings suggest that lime addition can alleviate the level and dissemination of ARGs in soils and provide a potential measure to control the spread of ARGs derived from animal manure.