This study aims to reveal the relationship between farmland soil factors and production of Sphagnum under the conditions of different water levels, then reveals the most primary soil factor which affected the output of Sphagnum. Sphagnum with similar planting ages was collected as research object in Shuitai Village, Longli country, Guizhou Province. Combined with Redundancy analysis, the impacts of interaction between different water levels and soil factors on the characteristics of the Sphagnum physiological and ecological indexes were discussed. The results indicated that after identification, 5 Sphagnum species with the same genus were found, among which Sphagnum palustre L. ssp. Palustre was the dominant species. The physiological and ecological indexes increased with the rise of water level, which indicated that the higher water level would make for the growth of Sphagnum. Soil Water Content (SWC) and soil porosity (STP) increased as the water level rose, indicating that the rise of water level played a certain role in enhancing soil permeability, air permeability, and water storage capacity. Total nitrogen (TN), total phosphorus (TP), total potassium (TK) and available phosphorus (AP) were dropped down as the water level rose, indicating that the higher water level may accelerate the leaching of nutrient elements such as nitrogen, phosphorus and potassium. Urease (URE) and catalase (CAT) were growing with the rise of water level, which can be inferred that soil moisture content can improve soil enzyme activity. The physiological and ecological indexes of S.palustre showed a clearly negative correlation with soil factors such as TN, TP, TK and AP (P<0.05), speculating that nutrient elements like N, P and K may inhibit the growth of Sphagnum. The other soil factors were significantly correlated positively with indexes of S. perustre (P<0.05), among which water level (WAT), URE and SWC also showed a clear positive correlation (P<0.01), indicating that WAT, URE and SWC may contribute to the growth of spitmoss. However, it is necessary to ensure that the soil is acidic and nutri-poor in order to increase the yield of Sphagnum. It is important to increase the water level of the groove to about 0-5cm with increasing the porosity and relaxation of the soil.