The study with three wheat cultivars grown in two places of Shandong Province showed that the nitrate reductase (NR) and glutamine synthetase (GS) activities in flag leaf and the GS activity in grain were in the sequence of Jimai 20 > Youmai 3 > PH971942, and higher in Longkou than in Taian. The strong gluten wheat cultivars in Longkou had better grain qualities than those in Taian. There were significant correlations between the environmental factors at grain-filling stage and the grain qualities and enzyme activities of wheat. Higher temperature, moderate drought and less sunshine at grain filling stage were benefit to the grain qualities. The protein content of grain was positively correlated with the NR and GS activities in flag leaf for the medium and strong gluten wheat cultivars in Longkou and for the medium gluten wheat cultivars in Taian. Wheat cultivars for different use needed different environmental conditions, while suitable environmental conditions could promote the enzyme activities in nitrogen metabolism of wheat, and thus, improve the qualities of wheat grain.