The high carotenoid content in tritordeum (×Tritordeum Ascherson et Graebner) grains is derived from its wild parent, Hordeum chilense Roem. et Schulz. Phytoene synthase 1 (Psy1) is located on chromosome 7H^chS and plays a major role in this trait. This study investigates the impact of the introgression of chromosome 7H^ch into common wheat background on carotenoid composition, including xanthophylls esterified with fatty acids (monoesters and diesters). All of the genetic stocks carrying Psy1 from H. chilense increased their carotenoid content relative to common wheat. In addition, significant changes in the carotenoid profile were detected in different genetic stocks. The most relevant was the increase in content of lutein diesters when both 7H^ch and 7D were present, which indicates the existence of genes involved in the esterification of xanthophylls in both chromosomes. Furthermore, our results suggest that 7H^ch genes preferentially esterify lutein with palmitic acid, whereas 7D is either indifferent to the fatty acid or it prefers linoleic acid for lutein esterification. The involvement and complementarity of 7H^ch and 7D are highly significant considering the scarcity of previous results on lutein esterification in wheat.