The flag leaf can be an important vehicle for high grain yield due to its position and photosynthetic characteristics. To identify the most adaptive and stable yielding genotype, three winter wheat genotypes were studied during two experimental years under field conditions to quantify the influence of flag leaf removal in different phenological stages on the grain yield and grain quality. To quantify the influence of the flag leaf on the main yield components, the flag leaf was removed every 7 days, starting from the booting stage to ripening. Chlorophyll a, chlorophyll b, and carotenoids were determined from the removed leaves. As a complex trait, the number of grains/spikes and the weight of the grains/spikes were highly influenced by the flag leaf removal during the early generative stages, causing a yield loss from 9% to more than 40%, depending on the variety. It was established that the photosynthetic pigments were highly influenced by the plant phenology stages and the environmental conditions, especially carotenoids, which act as photoprotective and antioxidant agents under stress conditions. Regarding grain quality, flag leaf removal had a significant influence on the accumulation of grain protein and the wet gluten content, the variability of these quantitative traits being also influenced by the climatic conditions.