Molecular Characterization of Seven Novel Glu-[A1.sup.m]x Alleles from Triticum monococcum ssp. monococcum

Seven Glu-A[1.sup.m] allelic variants of the Glu-A[1.sup.m]x genes in Triticum monococcum ssp. monococcum, designated as 1Ax2.[1.sup.a], 1Ax2.[1.sup.b], 1AX2.[1.sup.c], 1Ax2.[1.sup.d], 1Ax2.[1.sup.e], 1AX2.[1.sup.f] and 1Ax2.[1.sup.g] were characterized. Their authenticity was confirmed by successful expression of the coding regions in E. coli, and except for the lAx2.[1.sup.a] 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-A[1.sup.m]x alleles. Since variation in the Glu-A1x locus in common wheat is rare, these novel genes at the Glu-A[1.sup.m]x can be used as candidate genes for further wheat quality improvement. Keywords: cultivated einkorn, glutenin, gene expression, quality breeding Abbreviations: aa, amino acid(s); bp, base pair(s); BLAST, basic local alignment search tool; DTT, DL-Dithiothreitol; HMW-GS, high molecular weight glutenin subunit; IPTG, isopropyl P-A-thiogalactopyranoside; NCBI, National Center for Biotechnology Information; ORF, open reading frame; PCR, polymerase chain reaction; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; SNP, single nucleotide polymorphism.