Genomes of wheat varieties provide an available and highly efficient polymorphic system of genetic markers. Suitable protein markers in wheat are endosperm storage proteins:gliadins and glutenins.The objective of this study was to determine the composition of high molecular weight glutenin subunits and presence or absence of economically relevant Gli-1B3gliadin block of 41 Slovak varieties of hexaploid wheat (Triticumaestivum L.) included in the List of Registered Varieties from 2010. Gliadin polymorphism was detected by electrophoresis according to the PAGE ISTA methodology. HMW-GS were assessed using the SDS-PAGE method. The allelic block Gli-1B3, the marker of rye translocation 1BL.1RS, also the marker of poor bread-making quality was detected in 6 genotypes (Bonita, Lívia, Malvína, Sana, Verita and Veldava). Fourteen high molecular weight (HMW) - glutenin subunits (GS) were found, three belonged to Glu-1A, seven to Glu-1B and four to Glu-1D locus. The most frequent (37%) HMW-GS at the Glu-1A, Glu-1B and Glu-1D complex loci were 0, 7+9, 5+10, respectively. However, low frequent alleles such as 17+18 and 20 were observed. Bread quality (class) in examined accession varied from minimum value 1 to maximum value 9. The glutenin-based quality score ranged from 4 to10.