Abstract: Climate warming has an obvious asymmetry between day and night, with a greater increment of air temperature at nighttime than at daytime. By adopting passive nighttime warming (PNW) system, a two-year field experiment of nighttime warming was conducted in the main production areas of winter wheat in China (Shijiazhuang of Hebei Province, Xuzhou of Jiangsu Province, Xuchang of Henan Province, and Zhenjiang of Jiangsu Province) in 2009 and 2010, with the responses of soil pH and available nutrient contents during the whole growth periods and of wheat root characteristics at heading stage determined. As compared with the control (no nighttime warming), nighttime warming decreased the soil pH and available nutrient contents significantly, and increased the root dry mass and root/shoot ratio to a certain extent. During the whole growth period of winter wheat, nighttime warming decreased the soil pH in Shijiazhuang, Xuzhou, Xuchang, and Zhenjiang averagely by 0.4%, 0.4%, 0.7%, and 0.9%, the soil alkaline nitrogen content averagely by 8.1%, 8.1%, 7.1%, and 6.0%, the soil available phosphorus content averagely by 15.7%, 12.1%, 19.6%, and 25.8%, and the soil available potassium content averagely by 11.5%, 7.6%, 7.6%, and 10.1%, respectively. However, nighttime warming increased the wheat root dry mass at heading stage in Shijiazhuang, Xuzhou, and Zhenjiang averagely by 31.5%, 27.0%, and 14.5%, and the root/shoot ratio at heading stage in Shijiazhuang, Xuchang, and Zhenjiang averagely by 23.8%, 13.7% and 9.7%, respectively. Our results indicated that nighttime warming could affect the soil nutrient supply and winter wheat growth via affecting the soil chemical properties.