High molecular weight (HMW) glutenin subunits are important seed storage proteins in wheat and its related species. Novel HMWglutenin subunits in Aegilops tauschii accession of TA2484 were detected and characterized. SDS-PAGE analysis revealed the y-type subunit from TA2484 displayed similar electrophoretic mobility compared to that of 1Dy12 subunit. However, the electrophoretic mobility of x-type subunit was faster than that of 1Dx2 subunit. The primary structure of the two cloned subunits from TA2484 was similar to that of the x- and y-type subunits reported before. However, the 148 residues of the x-type subunit, which contained the sequence element GHCPTSLQQ, in the middle of the repetitive domain was quite different from other x-type subunits. Moreover, the 68 residues in this region were identical to those of the y-type subunits from the same accession. Consequently, 1Dx2.3*t (x-type subunit of TA2484) contains an extra cystenin residue located at the repetitive domain, which is novel compared to the x-type subunits reported so far. Phylogenetic analysis indicated that two subunits from accession TA2484 were in the x- and y-type subunit cluster, but bootstrapping value of 100% gave high support for the spilt between two subunits (1Dx2.3*t and 1Dy12.3*t) and their alleles, respectively. A hypothesis on the genetic mechanism generating this novel sequence of 1Dx2.3*t subunit is suggested.