It is of great significance to clarify the effects of cultivars and soil factors on wheat manganese nutrition for optimizing wheat manganese nutrition and achieving high yield and high-quality wheat production. From 2016 to 2020, a field experiment was carried out at 38 test sites in 13 provinces in three wheat production regions: Northwest dry-farming wheat area (DW), Huanghuai wheat-maize rotation area (WM), and Southern rice-wheat rotation area (RW). Wheat yield, yield components, grain Mn concentration, soil available manganese, pH value, and other indicators were tested. Results showed that, wheat yield was in the following order: WM > RW > DW, with the average value of 8.1, 5.9, and 5.9 t hm-2, respectively. Mn concentration in wheat grains was in the following order: RW > DW > WM, with the average value of 46.9, 45.4, and 41.4 mg kg-1, respectively. In different wheat production regions, the relationships of grain Mn concentration with the dry matter accumulation distribution, yield components, Mn uptake, and the utilization of wheat cultivars were different. The correlation of grain Mn concentration was significantly negative with the yield, biomass, and harvest index of wheat cultivars in DW, and was significantly negative with yield and harvest index in WM, but not significant in RW. There was a significant negative correlation between wheat grain Mn content and soil pH value, available Cu, and the total N, but not significant correlation between wheat grain Mn content and soil available Mn. Grain Mn concentration was positively correlated with spike number, but significantly negative with 1000-grain weight in RW. Grain Mn concentration was positively correlated with Mn uptake in shoots and Mn uptake in grains in WM and RW, and only positively correlated with Mn uptake in grains in DW, while its correlation with Mn harvest index was significantly negative in RW but positive in DW and WM. The main soil factors affecting wheat grain Mn concentration included soil total nitrogen, pH value, available Fe, available Mn, and available Cu. In WM, the grain Mn concentration was significantly positively correlated with soil available Fe, available Cu and Mn, but negatively correlated with soil pH. In RW, grain Mn concentration was significantly and negatively correlated with soil pH value, available Cu, and the total nitrogen, but not correlated with soil available Mn. Soil available P and available K were the main factors affecting grain Mn concentration in DW. In conclusion, wheat cultivars in RW of China had higher grain Mn concentration. Lower soil pH value, the total nitrogen, and the higher soil available Fe and Mn were beneficial to the increase of grain Mn concentration, while the effects of soil available Cu on grain Mn concentration varied with wheat regions. The yield had a dilution effect on Mn concentration, and the increase of spike number, grain number per spike, and 1000-grain weight were beneficial to the decrease of Mn content in wheat cultivars.