Black soils (Mollisols) are important soil resources for crop production and maintain food safety in China. For keeping soil fertility, the application of animal manure is commonly practiced in black soils. However, the impact of this application on abundance and diversity of antibiotic resistance genes (ARGs) in black soils of China remains unclear. Here, we surveyed the profiles of ARGs in 72 soil samples collected from four long-term experimental stations with different fertilization regimes and from open farmlands in two sites across northeast China using high-throughput quantitative PCR. Results showed that a total of 178 ARGs including mobile genetic elements (MGEs) were detected, and the diversity and abundance of ARGs were significantly increased with manure application. Additionally, the finding of a significant positive correlation between relative abundance of ARGs and MGEs (P < 0.0001), suggesting that horizontal gene transfer may potentially impact the transmission of ARGs. Furthermore, two genes aadA-1-01 and mexF, encoding resistance to aminoglycoside and multidrug, respectively, were recognized as indicators to estimate the abundance of other co-occurring ARGs. These findings provided insights into the soil resistome in black soils of northeast China and also highlighted the environmental risks caused by manure application should not be ignored.