Agricultural intensification has strongly impacted ecosystems and accelerated the process of global change. Consequently, agroecological practices are being increasingly adopted. Agroecological practices are biodiversity-based solutions that aim to generate sustainable and resilient agroecosystems, which could enhance the supply of ecosystem services. This study compared agroecological and conventional horticultural farms in terms of agroecological practices and ecosystem services supply. We conducted biophysical samplings and interviews on 24 agroecological and conventional farms over two summers in the Madrid Region (Spain). We used multiple indicators as proxies of the supply of 12 ecosystem services, and we identified the agricultural practices applied at each farm. We found that agroecological farmers applied more agroecological practices compared to conventional farmers, and agroecological farms had a higher potential to supply regulating, provisioning, and cultural services. Some agroecological practices, such as crop diversification, light tillage, and the use of organic pesticides, were associated with enhancing soil fertility, pest control, and pollination services. Our study provided empirical evidence that agroecological practices enhance ecosystem services at horticultural farms, which is extremely relevant to upscaling agroecology in the current context of ongoing European policy reforms.