A set of spring triticale selection forms was studied with the use of microsatellite markers specific for D-genome chromosomes and genomic in situ hybridization. 2R/2D substitutions were found in six of twelve forms. Two lines displayed segregation for the substitution: plants with the substitution and without it, and plants heterozygous for the substitution were revealed. Segregation of the lines and the presence in them of telocentric chromosomes of wheat and rye in the monosomic state suggest that the process of formation is still in progress. The data obtained also demonstrate that triticale is under selection for the 2R/2D substitution as controlling the spectrum of adaptively and selectively valuable traits.