Climatic change affects global agriculture and is a severe threat to global food security due to variability of the three factors measuring climate change (CO2, temperature and precipitation) with temperature being the most crucial one. Wheat is severely affected by high temperatures with reproductive and grain-filling phases being most sensitive, impacting grain number, size and weight. Seed size and weight are the key agronomic traits subjected to artificial selection and involved in the domestication process since the origin of agriculture. Three genotypes Banks, EGA Gregory and Fang-60 with the latter known to be heat tolerant were grown under glass house conditions and subjected to heat stress for 3 days during early- (11–14 dpa – days post anthesis) and late- (27–30 dpa) grain filling stages in a mutually exclusive fashion. The impact of heat stress during early- and late- grain filling on the four major grain characteristics, thousand grain weight (TGW), grain length, grain width and grain thickness was assessed. The tolerant genotype Fang-60 exhibited significantly higher TGW during early-grain filling heat stress than the control possibly due to an ability to exploit the accelerated release of fertilizer under high temperature. Banks and EGA Gregory were moderately tolerant to susceptible to heat stress, respectively, at early- and late-grain filling with Fang-60 being tolerant to both early- and late- grain filling heat stress. This study confirms the availability of significant genetic variation in heat stress response in wheat that might be exploited to adapt wheat to higher growth temperatures.