Authors:Z.L. Li, H.Y. Li, G. Chen, X.J. Liu, C.L. Kou, S.Z. Ning, Z.W. Yuan, M. Hao, D.C. Liu, and L.Q. Zhang
Seven Glu-A1m allelic variants of the Glu-A1mx genes in Triticum monococcum ssp. monococcum, designated as 1Ax2.1a, 1Ax2.1b, 1Ax2.1c, 1Ax2.1d, 1Ax2.1e, 1Ax2.1f, and 1Ax2.1g were characterized. Their authenticity was confirmed by successful expression of the coding regions in E. coli, and except for the 1Ax2.1a with the presence of internal stop codons at position of 313 aa, all correspond to the subunit in seeds. However, all the active six genes had a same DNA size although their encoding subunits showed different molecular weight. Our study indicated that amino acid residue substitutions rather than previously frequently reported insertions/deletions played an important role on the subunit evolution of these Glu-A1mx alleles. Since variation in the Glu-A1x locus in common wheat is rare, these novel genes at the Glu-A1mx can be used as candidate genes for further wheat quality improvement.
Authors:Z. L. Li, D. D. Wu, H. Y. Li, G. Chen, W. G. Cao, S. Z. Ning, D. C. Liu, and L. Q. Zhang
Gliadin is a main component of gluten proteins that affect functional properties of bread making and contributes to the viscous nature of doughs. In this study, thirteen novel ω-gliadin genes were identified in several Triticum species, which encode the ARH-, ATDand ATN-type proteins. Two novel types of ω-gliadins: ATD- and ATN- have not yet been reported. The lengths of 13 sequences were ranged from 927 to 1269 bp and the deduced mature proteins were varied from 309 to 414 residues. All 13 genes were pseudogenes because of the presence of internal stop codons. The primary structure of these ω-gliadin genes included a signal peptide, a conserved N-terminal domain, a repetitive domain and a conserved C-terminus. In this paper, we first characterize ω-gliadin genes from T. timopheevi ssp. timopheevi and T. timopheevi ssp. araraticum. The ω-gliadin gene variation and the evolutionary relationship of ω-gliadin family genes were also discussed.
Premature termination codons (PTCs) are an important reason for the silence of highmolecular- weight glutenin subunits in Triticum species. Although the Glu-A1y gene is generally silent in common wheat, we here isolated an expressed Glu-A1y gene containing a PTC, named 1Ay8.3, from Triticum monococcum ssp. monococcum (AmAm, 2n = 2x = 14). Despite the presence of a PTC (TAG) at base pair positions 1879–1881 in the C-terminal coding region, this did not obviously affect 1Ay8.3 expression in seeds. This was demonstrated by the fact that when the PTC TAG of 1Ay8.3 was mutated to the CAG codon, the mutant in Escherichia coli bacterial cells expressed the same subunit as in the seeds. However, in E. coli, 1Ay8.3 containing the PTC expressed a truncated protein with faster electrophoretic mobility than that in seeds, suggesting that PTC translation termination suppression probably occurs in vivo (seeds) but not in vitro (E. coli). This may represent one of only a few reports on the PTC termination suppression phenomenon in genes.
High-molecular-weight glutenin subunits (HMW-GSs) are important seed storage proteins associated with bread-making quality in common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD). Variation in the Glu-A1x locus in common wheat is scare. Diploid Triticum monococcum ssp. monococcum (2n = 2x = 14, AmAm) is the first cultivated wheat. In the present study, allelic variations at the Glu-A1mx locus were systematically investigated in 197 T. monococcum ssp. monococcum accessions. Out of the 8 detected Glu-A1mx alleles, 5 were novel, including Glu-A1m-b, Glu-A1m-c, Glu-A1m-d, Glu-A1m-g, and Glu-A1m-h. This diversity is higher than that of common wheat. Compared with 1Ax1 and 1Ax2*, which are present in common wheat, these alleles contained three deletions/insertions as well as some single nucleotide polymorphism variations that might affect the elastic properties of wheat flour. New variations in T. monococcum probably occurred after the divergence between A and Am and are excluded in common wheat populations. These allelic variations could be used as novel resources to further improve wheat quality.