Heat tolerance is a physiologically and genetically complex trait regulated by multiple genes. To investigate the genetic basis of heat tolerance, eight parents (five lines and three testers) and their fifteen F1 hybrids were evaluated under normal and high-temperature stress conditions for two consecutive years. Data were recorded for plant height, number of bolls, boll weight, seed cotton yield, ginning out turn (GOT%), H₂O₂, catalase, peroxidase, super-oxidase dismutase, total soluble proteins, carotenoids, chlorophyll a & b contents, short fiber index, fiber strength, UHML, micronaire value, reflectance, and uniformity index. Line × tester analysis suggested that the contribution of lines was higher than testers. Non-additive gene action was observed for all studied traits. The variances of SCA were greater than GCA variances for all studied traits revealed that these traits were governed by a few largely dominant genes. Fb-Shaheen, Eagle-2 and JSQ White Gold were found good general combiner whereas the cross Fb-Shaheen × JSQ White Gold was a good specific combiner and revealed significant better parent heterosis for most of the traits during two years under normal and high temperature stress conditions. The information obtained could be utilized in a breeding program for the development of new synthetic varieties of heat tolerance.