Abstract: Field in situ infiltration experiments at different depths can be used to express the state of soil stratification, and the physical differentiation of soil layers, in order to quantify the changes in the water of the soil profile. This study aims to obtain the infiltration capacity and water retention of paddy soils. The field in situ soil infiltration experiments were also carried out at different depths to identify the soil stratification in the representative plot under long-term mechanized tillage of smallholder farmers in the rice-wheat rotation region of eastern China. Seven infiltration pits of different depths were excavated in the experimental plots. After that, the infiltration experiments were conducted at the bottom of the pits. Soil water content was then measured in the layers for 48 hours of infiltration. A systematic investigation was made to explore the soil infiltration capacity at the bottom of the pits and the soil water content of each layer. The results showed that an accurate description was achieved in the water infiltration and water holding capacity of the soil at different pits after the infiltration experiments at different depths. The location and thickness of the plow pan were also clearly identified during this time. The plow pan was started at a depth of about 15 cm, indicating the outstanding difference between the cultivated layer and the plow pan. The average soil penetration resistances of the cultivated layer and the plow pan were 1 005.79 kPa, and 1 910.73 kPa, respectively. The soil profile showed that the soil in the cultivated layer shared a loose morphology and dense root distribution, whereas, the soil in the plow pan presented a high bulk density, low porosity, and poor permeability, while the soil in the subsoil layer was in more iron-manganese spots and poor structure. Furthermore, the soil infiltration parameters decreased with the increasing pit depth. The average infiltration rate and cumulative infiltration in the 0-15 cm pit depth range were 17.04 and 18.06 times higher, respectively, than those in the >20-30 cm pit depth range. The three infiltration models were fitted using Horton, Kostiakov and Philip. Specifically, the Kostiakov model had the highest R2 (0.98-0.99) and the smallest RMSE (0.01-0.77 mm/min), indicating the consistency of the fitted parameters. Infiltration parameters were extremely significant correlated with the soil bulk density, water content, total porosity, and field capacity (P<0.01), but not with the soil penetration resistance (P>0.05). Field in situ infiltration experiment at different depths was an important tool to identify the soil stratification and quantify the water function differentiation in the soil profiles, according to the soil profile, cone penetration and sampling. Long-term mechanized tillage under the smallholder production model in the rice-wheat rotation region of eastern China can be expected to result in the apparent stratification and vertical differentiation of soil water functions in paddy soils. In turn, there are also some significant differences in the infiltration capacity between the cultivated layer and soil layers below the cultivated layer. This study can provide a reference for mechanical cultivation and irrigation in rice-wheat rotation regions.