Molecular markers were used to identify the allele/gene composition of complex lociGlu-A1 andGlu-B1 of high-molecular-weight (HMW) glutenin subunits in triticale cultivars. Forty-six Polish cultivars of both winter and spring triticale were analysed with 7 PCR-based markers. Amplified DNA fragments of HMW gluteninGlu-1 genes were separated by agarose slab-gel electrophoresis. Differences between all 3 alleles at the locusGlu-A1 [Glu-A1a (encoding Ax1),1b (Ax2*), and1c (AxNull)], 4 alleles atGlu-B1-1 [Glu-B1-1a (Bx7),1b (Bx7*),1d (Bx6),1ac (Bx6.8)], and 5 alleles atGlu-B1-2 [Glu-B1-2a (By8),2b (By9),2o (By8*),2s (By 18*), and2z (By20*)] were revealed. In total, 16 allele combinations were observed. Molecular markers are particularly helpful in distinguishing the wheatGlu-A1a andGlu-A1b alleles from the ryeGlu-R1a andGlu-R1b alleles in triticale genotypes, respectively, as well as subunits Bx7 from Bx7* and By8 from By8*, which could not be distinguished by SDS-PAGE. Novel glutenin subunits By 18* and By20* (unique to triticale) were identified. HMW glutenin subunit combinations of Polish triticale cultivars, earlier identified by SDS-PAGE analyses, were verified by PCR-based DNA markers. Rapid identification of wheatGlu-1 alleles by molecular markers can be an efficient alternative to the standard separation procedure for early selection of useful triticale genotypes with good bread-making quality.