To explore the isotopic distribution and differentiation of water along the hydraulic flow gradients and plant-bed/ditch systems in constructed root-channel wetlands, surface and subsurface water samples were collected from four ecological wetlands, namely Shijiuyang and Guanjinggang in Jiaxing, as well as Changshuitang and Taishangang in Haining. All samples were collected along water flow pathways during the wet and rainy summer season in August 2019, except for those from Taishangang, which were collected within the plant-bed/ditch system during the dry and cold winter season in January 2020. The abundance of deuterium (δD) and δ18 O was determined in each functional area of the wetlands to assess the influence of wetlands on water differentiation. Stable isotope technology and mathematical statistics were used to analyze the distribution of δD and δ18 O in constructed root-channel wetlands and to reveal the influence of plant-bed/ditch systems on stable isotopes of water. A variety of data mining methods were used to examine the differentiation of stable isotopes of water, at various dimensions and scales, including nonparametric Kendall's tau-b correlation, stepwise regression, gray relational analysis, and machine learning (random forest) combined with scatter diagrams and model hypothesis diagnosis analysis. The main results were as follows:① The spatiotemporal variations in water isotopes of stream networks were largely affected by different water supply and evaporation enrichment effects. The slope and intercept of the wetland water line in Jiaxing were both significantly lower than the regional precipitation line of the adjacent Changshu Station (CHNIP). This showed that the wetlands area had undergone hydrogen and oxygen isotope enrichment. The δD values in Shijiuyang wetland water ranged from -52.2‰ to -49.4‰, and δ18 O values ranged from -7.6‰ to -6.9‰. In Guanjinggang wetland water samples, δD ranged from -48.1‰ to -45.1‰, and δ18 O ranged from -6.8‰ to -5.8‰. The δD values in Changshuitang wetland water ranged from -49.8‰ to -48.4‰, and δ18 O ranged from -7.2‰ to -6.6‰. The δD values in Taishangang wetland water ranged from -55.3‰ to -51.6‰, and δ18 O ranged from -7.8‰ to -7.2‰. ② Hydrogen and oxygen isotope abundance and composition of water showed complex nonlinear changes in the vertical and horizontal dimensions at different scales. At the regional scale, water level elevation in the vertical dimension had a greater impact on water isotope distribution than the length of the hydraulic flow pathway in the horizontal dimension. Water isotopes tended to be enriched in low-lying areas with low water levels. At the local scale, the influence of hydraulic process often played a greater role in determining water isotope distributions. The spatial variations of water isotopes were comprehensively determined by the evaporation of regional water and meandering hydraulic processes inside the wetland. ③ Compared with other wetland functional areas, the central constructed root-channel area (middle treatment zone) was more enriched in water isotopes. ④ The underground macropore network formed by plants with developed rhizomes or roots (e.g., Phragmites communis Trin. and Typha orientalis Presl), mineral-rich substrate soil, and aquatic plants in the plant bed had a significant influence on the abundance of hydrogen and oxygen isotopes in the plant-bed/ditch system. Therefore, when water passed through the plant-bed/ditch system, the values of δD and δ18 O in the lower ditch (outlet) were lower than those in the higher ditch (inlet). ⑤ The abrupt change in isotopic contents of the plant-bed/ditch system might indicate an inflection point in water quality purification. ⑥ The deuterium excess (d-excess) in subsurface water of the plant-bed/ditch system was significantly higher than that in ditch water, and the coefficient of variation in subsurface water was considerably greater than that in ditch water. The d-excess in the wetland root-channel ecological purification zone showed significant temporal differences and was negative in the summer and positive in the winter, which reflected the seasonal variation in water vapor sources and the spatial variation in isotope fractionation behavior in wetlands. These results provide some understanding of the distribution of water isotopes in constructed wetlands, which will strengthen their operation and management. This study also provides some ideas regarding new technologies for water quality improvement and shows that water isotope technology may be a reliable method for analyzing wetland hydrology.