Meiosis is the least explored stage for thermotolerance in wheat. We evaluated the impact of 5 d of moderate transient daily heat stress during meiosis in the main stem spike on physiological and grain yield traits in 30 diverse wheat cultivars which vary widely in heat tolerance and sensitivity. We found that a moderate heat stress event during meiosis in the main stem spike had lasting impacts on plant growth and reproduction in heat-sensitive, but not heat-tolerant, wheat cultivars. Heat-tolerant cultivars maintained grain yield, grain number and individual grain weight in the main stem spike and also total plant grain yield and biomass in the heat stress treatment relative to the control. Heat-sensitive cultivars responded to heat stress by producing fewer and smaller grains per spikelet on the main stem, fewer tillers, lower biomass and lower total plant grain yield in the high temperature treatment relative to the control. Heat-sensitive cultivars produced higher flag leaf chlorophyll content in the high temperature treatment relative to the control than heat-tolerant cultivars during the first 3 d of heat treatment. There was small reduction in pollen viability from 98% to 96% following heat stress during meiosis which was unrelated to heat tolerance or sensitivity. Moderate transient heat stress during meiosis did not greatly reduce the production of viable male gametes, but had long-lasting negative impacts on fertilization and subsequent seed production in heat-sensitive cultivars.