【Objective】 With the fluctuation in crayfish market, some integrated rice-crayfish co-culture paddies are restored to traditional rice-upland rotations. However, the changes in soil phosphorus (P) availability following the restoration remain underexplored. 【Method】In this study, soil samples from 0 to 50 cm depth were collected in May 2022 in Yanwo Town, Honghu City, Hubei Province. Three field types were selected: rice-crayfish co-culture with restored rice-upland rotation (IRCC-PR), continuous rice-crayfish co-culture (IRCC), and traditional rice-upland rotation (PR). Soil available P content and alkaline phosphatase activity (APA) were measured. The modified Hedley P fractionation method was used to determine soil P forms. 【Result】Results indicated that soils showed the highest proportion of moderate P (Moderate-P: 64.8%–66.9%), followed by stable P (Residual-P: 25.5%–40.0%), with labile P having the lowest proportion (Labile-P: 3.3%–4.0%). Compared to IRCC and PR soils, IRCC-PR soils showed increased proportions of Labile-P and Residual-P, while the proportion of Moderate-P decreased. Similar to PR soils, IRCC-PR soils exhibited a clear surface accumulation pattern for available P and APA. The concentrations in surface soils (0–5 cm) were 5.1 and 3.0 times higher, respectively, than those in subsurface soils (30–50 cm). In contrast, these parameters slightly increased with depth in IRCC soils.【Conclusion】Our results reveal that the restoration of rice-upland rotation altered the vertical distribution of soil available P, emphasizing the need to shift the risk control strategy for P loss in paddy fields from managing leaching losses to controlling P loss with soil erosion and surface runoff.