The results of a comparative analysis of the breeding value of winter wheat varieties based on the number of spikelets per main spike during 2021–2023 are presented. Three ecotype groups were studied: forest-steppe, steppe, and Western European. It was established that the highest mean spikelet number (x̄) was formed by the Western European ecotype varieties, particularly Acratos (19.6 spikelets). Forest-steppe ecotype varieties exhibited stable trait expression within the range of 15.5–16.9 spikelets, whereas the steppe ecotype was characterized by somewhat lower values. Variability limits (Lim) and the breeding value index (Sc) were analyzed, enabling the identification of the most promising genotypes for further breeding work aimed at improving crop productivity. Breeding value (Sc) was calculated according to V. V. Khangildyn and M. A. Lytvynenko (1981): Sc = X × (Xlim / Xopt), where: Sc – breeding value; X – the mean value of the studied trait; Xlim – the minimum value of the trait; Xopt – the maximum value of the trait.