Analysis of puroindoline a and b sequences from Triticum aestivum cv. 'Penawawa' and related diploid taxa

Flanking sequences of the puroindoline a (pinA) and puroindoline b (pinB) genes from Triticum monococcum, T. urartu, Aegilops speltoides and A. tauschii were obtained by inverse PCR. Two accessions from each of these related diploid taxa were sequenced, and the sequences compared to those of the soft hexaploid wheat cultivar `Penawawa'. As expected from the origin of the D-genome in hexaploid wheat, the highest sequence identity was observed for the A. tauschii sequences, whereas the sequences for A. speltoides were the most divergent. The promoter sequences were further analyzed for putative regulatory elements, and the possible significance of several highly conserved sequence motifs is discussed. Western blots of Triton X-114 extracted proteins separated by SDS-PAGE confirmed the accumulation of pinA and pinB gene products in the endosperm of all the germplasm lines studied. The deduced amino acid sequences for the related diploid taxa do not imply any major structural changes as compared to the wild-type T. aestivum puroindolines, and is therefore in agreement with the soft endosperm texture of these species.