The current study aimed to determine and compare the grain yield, some quality characteristics and high-molecular-weight glutenin subunits (HMW-GS) of bread wheat cultivars cultivated in several agro-ecological regions using a total of 46 registered bread wheat cultivars in the 2016–2017 and 2017–2018 growing seasons. The results determined that the environment had the largest share in the total variation (Genotype + Environment + Genotype×Environment Interaction). The Mediterranean region had the highest average grain yield with 8137 kg ha− 1, while the Central Anatolia region (under the rainfed conditions) had the lowest average with 4260 kg ha− 1. The average thousand kernel weight of the cultivars was 35.3–39.9 g, test weight 77.2–79.2 kg hL− 1, protein content 13.4–14.7%, Zeleny sedimentation 39.2–53.3 mL, and alveograph energy value varied between 191.2-276.4 10− 4 J. Regarding the HMW-GS, 18 of the 46 cultivars scored 10, and one scored 5. The highest mean protein content and alveograph energy value was determined in cultivars with 9 Glu-1 score. In Zeleny sedimentation, cultivars with 10 Glu-1 score showed the highest mean value. The most common subunits in loci; it is 2* in Glu-A1, 7 + 8 and 7 + 9 in Glu-B1, and 5 + 10 in Glu-D1. The fact that registered cultivars predominantly carry these subunits at Glu-1 loci, which could be the result of yield and quality-oriented selection in the breeding process. It was concluded that high quality new varieties could be developed by HMW-GS oriented crosses and selections in wheat breeding programs.