This article aimed to evaluate the efficacy of the iPBS (inter-Primer Binding Site) molecular marker system, leveraging mobile genetic elements, for genotyping diverse triticale accessions sourced from the collection of the Institute of Genetics, Physiology and Plant Protection. The objectives included the isolation and purification of genomic DNA from 7-day-old triticale seedlings of 'Ingen-54' and 'Ingen-93' genotypes, the empirical determination of optimal iPBS primer efficiency through a series of experimental amplifications, and the subsequent selection of highly informative primers for triticale genotyping. Standard polymerase chain reaction (PCR) amplifications were conducted utilizing specific iPBS primers (Pb13, Pb20, Pb21), followed by electrophoretic separation of amplification products on agarose gel. Amplification produced 33 clear, 100% polymorphic loci. POPGENE analysis revealed high informativeness (Nei’s gene diversity h=0.1290, Shannon index I=0.2403) despite low genetic distance (0.0074–0.0169) between the closely related varieties. Primers Pb20 and Pb21 generated the most polymorphic loci. These results confirm the high utility of iPBS markers for genetic diversity assessment and cultivar identification in triticale breeding, even for closely related genotypes.
