Three winter wheat varieties with differing breadmaking quality were grown at two locations in two years at 0 or 3 × 60 kg of nitrogen application. The effect of nitrogen on amount of different components of gluten proteins was determined by reverse‐phase HPLC. A high amount of nitrogen led generally to a significant increase of total protein content. However, this increase was obvious only for the gluten proteins; albumins and globulins remained nearly unaffected. The effect of increased protein content on gliadin to glutenin (gli‐glu) ratio was inconsistent. While increased protein content increased the gli‐glu ratio in the variety Capo, the opposite was true for the variety Renan. Gli‐glu ratio of the variety Lindos showed no discernible tendency. As total protein content increased, the ratio of low molecular weight (LMW) to high molecular weight (HMW) glutenins decreased consistently, i.e., in all varieties, in both years and locations. Change of LMW to HMW ratio showed a significant negative correlation to sedimentation value and bread volume. There was no consistent change in the ratio between x‐ and y‐type HMW subunits due to fertilization, as could be shown by densitometric measurements on SDS‐PAGE gels. This ratio appeared to be dependent on the genotype and has decreased with decreasing quality. The amount of x‐type subunits correlated closely with sedimentation value and bread volume. These results suggest that ratio of HMW glutenins, especially x‐type subunits, to total protein content could be the best early detectable parameter with high predictive value for breadmaking quality.

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