Limitations in carbohydrate supplies have been implicated as a factor responsible for reproductive failure under heat stress. Heat stress affects two stages of reproductive development in cowpea [Vigna unguiculata (L.) Walp.], and genotypes are available with tolerance and sensitivity to heat during these different stages. The objectives of this study were to determine the responses of these cowpea lines to ambient and elevated [CO2], under heat stress and optimal temperature, and test whether differences in carbohydrate supplies due to genotypes, CO2 enrichment and heat stress are associated with differences in sensitivity to heat during reproductive development. Plants were grown in reach-in growth chambers and subjected to day/night temperatures of either 33/20 or 33/30°C, and [CO2] levels of either 350 or 700 μmol mol-1. Under intermediate night temperature (33/20°C), all lines set substantial numbers of pods. Under high night temperature (33/30°C) with either ambient or elevated [CO2], one heat-sensitive line produced no flowers and the other set no pods, whereas the heat-tolerant line abundantly set pods. High night temperature reduced the overall carbohydrate content of the plants, especially peduncle sugars, and caused decreases in photosynthetic rates. The high pod set of the heat-tolerant line, under high night temperature, was associated with higher levels of sugars in peduncles compared with the heat-sensitive lines. The heat-tolerant line accumulated substantial shoot biomass, exhibited less accumulation of starch in leaves, and possibly had less down-regulation of photosynthesis in response to CO2 enrichment and heat stress than the heat-sensitive lines. Elevated [CO2] resulted in higher overall carbohydrate levels in heat-sensitive lines (starch in leaves, stems and peduncles), but it did not increase their heat tolerance with respect to flower production or pod set. Heat-induced damage to floral buds and anthers in the sensitive lines was associated with low sugars levels in peduncles, indicating that heat had greater effects on assimilate demand than on leaf assimilate supply. The heat-tolerant line was the most responsive genotype to elevated [CO2] with respect to pod production under either high or intermediate temperatures.