The important cyclic translocation 4AL-5AL-7BS is an evolutionary signature of polyploidy in wheat. This study aimed to determine its distribution within the subspecies of Triticum turgidum L., using genomic in situ hybridization and fluorescence in situ hybridization. As it exists in all eight subspecies, this translocation appeared before the differentiation of the subspecies of T. turgidum. This translocation probably first appeared in T. turgidum subsp. dicoccoides and was then transmitted into the other subspecies. Its existence in all of the analyzed subspecies suggests that this translocation may confer an adaptive advantage during the course of evolution.