1. Soil organic carbon (SOC) dynamics is regulated by a complex interplay of factors such as climate and potential anthropogenic activities. Livestocks play a key role in regulating the C cycle in grasslands. However, the interrelationship between SOC and these drivers remains unclear at different soil layers, and their potential relationships network have rarely been quantitatively assessed.

2. Here, we completed a six-year manipulation experiment of grazing exclusion (no grazing: NG) and increasing grazing intensity (light grazing: LG, medium grazing: MG, heavy grazing: HG). We measurements of light fraction organic carbon (LFOC) and heavy fraction organic carbon (HFOC) in 12 plots along grazing intensity in three soil layers (topsoil: 0-10 cm, mid-soil: 10-30 cm, subsoil: 30-50 cm) to assess their underlying controls.

3. Grazing significantly reduced SOC of the soil profile, but with significant depth and time dependencies. (1) SOC and SOC stability of the topsoil is primarily regulated by grazing duration (years). Specifically, grazing duration and grazing intensity increased the SOC lability of topsoil due to an increase in LFOC. (2) Grazing intensity was the major factor affecting the mid-soil SOC dynamics, among which MG had significantly lower SOC than did NG. (3) Subsoil organic carbon dynamics were mainly regulated by climatic factors. The increase in mean annual temperature (MAT) may have promoted the turnover of LFOC to HFOC in the subsoil.

4. Synthesis and applications. When evaluating the impacts of grazing on soil organic fraction, we need to consider the differences in sampling depth and the duration of grazing years. Our results highlight that the key factors influencing SOC dynamics differ among soil layers. Climatic and grazing factors have different roles in determining SOC in each soil layer.