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Proline is one of the organic osmolytes that accumulates in plants in response to stressful environmental conditions. The aim of this study was to determine the concentration of proline in the grain filling phase in ten winter wheat cultivars under conditions of prolonged periods of air temperatures above 30°C and to assess the variability of wheat variety responses. The correlation between proline accumulation and photosynthetic pigments, accumulated starch and yield was also determined. In the experimental field, flag leaves were sampled for each variety in the grain filling phase. Statistical data analysis and parameter correlation were performed using the SPSS program. The results showed that there is a statistically significant difference between proline values in moderate and high air temperature conditions when proline accumulation occurs. The significant correlation of accumulated starch content and yield with proline concentration showed that in addition to higher proline values under heat stress, wheat varieties Apac, Talas and Futura also have higher yield and starch content. A correlation was also found between proline concentration and photosynthetic pigment contents, where the Apac wheat variety showed the best adaptive response to the investigated traits and was characterized by a smaller reduction in photosynthetic pigment content under heat stress. The interrelation of proline with the photosynthetic pigment content and wheat quality parameters may be important in breeding technologies aimed at improving wheat stress tolerance.