Cloning and functional analysis of a novel x-type high-molecular-weight glutenin subunit with altered cysteine residues from Aegilops umbellulata

Abstract

In common wheat (Triticum aestivum L.) and its relative species, considerable progress has been made in understanding the structure and function of the high-molecular-weight glutenin subunit (HMW-GS). As a species closely related to wheat, Aegilops umbellulata is an important resource for wheat genetic improvement. In this paper, we report a novel HMW-GS 1Ux3.5 in Aegilops umbellulata Y361. The complete open reading frame (ORF) coding for 1Ux3.5 was cloned and sequenced. Analysis of the deduced amino acid sequence revealed that the primary structure of 1Ux3.5 was similar to those of previously published HMW-GSs. The 1Ux3.5 possessed an extra cysteine residue in the repetitive domain, indicating that the subunit may be related to excellent dough quality. Subsequently, the single proteins of 1Ux3.5 and 1Dx5 (used as positive control) were purified at a scale sufficient for incorporation into flour for a dough quality test. Both the SDS sedimentation volume and mixograph parameters demonstrated that 1Ux3.5 showed a greater contribution to the dough quality than 1Dx5. Therefore, the 1Ux3.5 subunit from Aegilops umbellulata may have potential value in improving the processing properties of hexaploid wheat varieties.

Cloning and characterization of high molecular weight glutenin gene from Triticum aestivum cultivar Dacke

“…In the past, many researchers have been focused on HMW glutenin genes of different cultivars of wheat and wheat related species. [20][21][22][23][24] Dacke is reported as a Swedish spring variety. It has a high protein content in the kernel, especially the gluten protein (37%).…”

High-Molecular-Weight Glutenin Subunits: Genetics, Structures, and Relation to End Use Qualities

“…In particular, the Glu-1B and Glu-1D alleles display higher variable frequency while that of the Glu-1A alleles is a much lower. Recently, a number of novel Glu-1 genes from wheat landraces or related species have been identified to be of potential application values to wheat breeding, and efforts have also been made to transfer desirable Glu-1 alleles to cultivated wheat by wide hybridization and chromosomal engineering approaches [ 14 , 31 , 32 , 33 , 34 , 35 , 36 ].…”

“…Besides, HMW-GSs with higher content of β-strands, indicated by secondary structure prediction, may be helpful to form a better gluten structure [ 131 ]. A longer repetitive domain or a higher proportion of repeats units of HMW-GSs may produce more β-turns, which confer elasticity to the polymers and are supposed to be positively related to dough and bread-making qualities [ 33 , 132 , 133 , 134 , 135 , 136 ].…”

1Mx2.1, a novel high-molecular-weight glutenin subunit from Aegilops comosa and its relationship to high-quality dough

“…comosa (accession number PI 551026), durum wheat Langdon and wheat cultivar Chinese spring were extracted, followed by protein separation in a 10% SDS-PAGE (Wan et al ., 2002). Cloning of the coding sequences of HMW-GS, Escherichia coli expression and a mixing test in vitro were performed according to the procedures described in our previous report (Hou et al ., 2017).…”