In situ hybridization of total genomic DNA was used to analyse lines derived from an amphiploid between tetraploid wheat, Triticum durum Desf. (2n l 4x l 28), and the wheatgrasses Thinopyrum distichum (Thunb.) A. Lo$ ve (2n l 4x l 28) and Lophopyrum elongatum (Host) A. Lo$ ve (2n l 2x l 14). A range of chromosome numbers was detected, arising from loss or gain of chromosomes. Total genomic DNA probes from Thinopyrum species, L. elongatum and Triticum monococcum L. were able to discriminate chromosomes from the A and B genomes of tetraploid wheat and those of wheatgrass-origin. The method did not discriminate the two wheatgrass genomes, J and E, indicating their close similarity. Chromosomal aberrations-including telocentric and ring chromosomes-were frequent. Distal inter-genomic translocations of parts of A and B genome chromosome arms, unusual in wheat itself, were more frequent than translocations between T. durum and wheatgrass. In situ hybridization of an rDNA probe most frequently revealed four sites associated with secondary constrictions on T. durum chromosomes and four on Th. distichum or L. elongatum chromosomes, although there was variation in the number of loci between and within plants. Within interphase and prophase nuclei, the three genomes were not intermixed and often lay in distinct sectors.# 1996 Annals of Botany Company