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To determine the composition of chromosome aberrations in a wheat‒<i>Dasypyrum breviaristatum</i> substitution line with seeds treated by a dose of gamma-rays (200 Gy), sequential non-denaturing fluorescence <i>in situ</i> hybridization (ND-FISH) with multiple oligonucleotide probes was used to screen individual plants of the mutagenized progenies. We identified 122 types of chromosome rearrangements, including centromeric, telomeric, and intercalary chromosome translocations from a total of 772 M1 and 872 M2 plants. The frequency of reciprocal translocations between B- and D-chromosomes was higher than that between A- and D-chromosomes. Eight translocations between <i>D. breviaristatum</i> and wheat chromosomes were also detected. The 13 stable plants with multiple chromosome translocations displayed novel agronomic traits. The newly developed materials will enhance wheat breeding programs through wheat‒<i>Dasypyrum</i> introgression and also facilitate future studies on the genetic and epigenetic effects of translocations in wheat genomics.