Subsoil carbon (C) stocks are a prime target for efforts to increase soil C storage for climate change mitigation. However, subsoil C dynamics are not well understood, especially in soils under long-term intensive agricultural management. We compared subsoil C storage and soil organic matter (SOM) composition in tomato–corn rotations after 25 years of differing C and nutrient management in the California Central Valley: CONV (mineral fertilizer), CONV+WCC (mineral fertilizer and cover crops), and ORG (composted poultry manure and cover crops). The cover crop mix used in these systems is a mix of oat (Avena sativa L.), faba bean (Vicia faba L.), and hairy vetch (Vicia villosa Roth). Our results showed a ∼19 Mg ha−1 increase in soil organic C (SOC) stocks down to 1 m under ORG systems, no significant SOC increases under CONV+WCC or CONV systems, and an increased abundance of carboxyl-rich C in the subsoil (60–100 cm) horizons of ORG and CONV+WCC systems. Our results show the potential for increased subsoil C storage with compost and cover crop amendments in tilled agricultural systems and identify potential pathways for increasing C transport and storage in subsoil layers.