Molecular Cloning Of ω-gliadin Genes From Wheat Progenitiors And Transformation Of Wheat With α-gliadin Gene

In order to exploit the relationship between a-gliadin gene and its functional properties of flour and evaluate the effect on wheat quality breeding, we transformed one a-gliadin gene from Elymus sibiricus into different wheat cultivars by using Agrobacterium-mediated transformation method. Nineteen positive To transgenic plants were obtained after two sets of primers screening by PCR. Two of SR3 and one of J21 transformed strains were obtained by PCR analysis, protein electrophesis and inherited stable analysis. It will be based upon functional characterization of the gliadin by dough micro-mix test in the latest future.A genomic PCR cloning strategy was applied to isolateω-gliadin sequences from three diploid A genome wheats (Triticum monococcum, Triticum boeoticum and Triticum urartu) and one diploid D genome ancester (Aegilops tauschii). Nineteen amplicon lengths varied from 744-1044 bp, and those of the corresponding deduced mature proteins from 248-348 residues. The primary structure of the deduced polypeptides comprised a short N- and C-terminal conserved domain, and a long, variable repetitive domain. A phylogenetic analysis of 12 full length sequences from A genome diploids combined with the other 6 from common wheat recognized three clades, one containing one T. boeoticum, two T. monococcum, three T. aestivum and four T. urartu sequences; the second grouped one T. boeoticum and four T. monococcum sequences, and the third contained only sequences from T. aestivum. Among the functional (non-pseudogene) ARQ/E-type co-gliadin genes, two were derived from T. boeoticum and three from T. monococcum; one of the latter sequences appeared to be a chimera originating via illegitimate recombination between the other two T. monococcum ones. None of the 13 intact co-gliadin DNA sequences contained any cysteine or methionine residues. The type of epitopes associated with coeliac disease was greater in the T. monococcum or T. boeoticum gliadins than in the T. urartu. We discussed the evolution of A genomeω-gliadin genes in the paper.