In this work, 9 novel LMW-GS genes (6 LMW-m and 3 LMW-i type) from 4 diploid and 1 tetraploid Aegilops species were amplified and cloned by allelic-specific PCR. Analysis of the deduced amino acid sequences showed that 7 and 2 LMW-GS had 9 and 7 cysteines, respectively. Four LMW-m type subunits genes had an extra cysteine at the C-terminal III, which could form intermolecular disulphide bonds to extend the chains, and therefore would facilitate to form larger gluten polymers. This suggested that these genes are expected to be used as candidate genes for wheat quality improvement. The correlation between specific N-terminal sequences and a decapeptide deletion in the C-terminal II in LMW-GS encoded by D genome was found. Particularly, if LMW-GS possessed a METRCIPG-N-terminal beginning sequences and a decapeptide (LGQCSFQQPQ) deletion in the C-terminal II, they could be encoded by D genome.