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

Background: High molecular weight (HMW) glutenin protein plays a crucial role in determining dough viscoelastic properties that determines the quality of wheat flour. The aim of the present study was to isolate, clone and analyze (in silico) the HMW glutenin gene of Triticum aestivum cultivar Dacke.Methods: Primers were designed to amplify a 2445 bp fragment of HMW glutenin gene. Ax type HMW glutenin gene from Triticum aestivum cultivar Dacke was isolated using PCR and it was sequenced by primer walking. Results: Amplified HMW glutenin gene was designated as HMWGAx. Sequence analysis revealed a complete open reading frame encoding 815 amino acid residues with N- and C terminal non-repetitive domain and a central repetitive domain. The calculated molecular weight of the deduced HMW glutenin protein was ~88 kDa and the number of cysteine residues in the HMWGAx was four, in accordance with other x type HMW glutenin proteins. Phylogenetic analysis revealed 100% homology to the previously studied Ax2* type HMW glutenin gene from cultivar Cheyenne. Predicted secondary structure results showed that it was similar to1Ax1 type of common wheat (Triticum aestivum), having superior flour milling quality.Conclusions: Sequence analysis suggests that HMWGAx protein significantly and positively correlates with the properties of elasticity and extensibility of gluten.

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Polymorphism of high-molecular-weight glutenin loci in Ukrainian wheat landraces Triticum aestivum L.

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v29.1416 ◽

2021 ◽

Vol 29 ◽

pp. 111-116

Author(s):

T. O. Sobko ◽

G.M. Lisova ◽

O.M. Blagodarova

Keyword(s):

Molecular Weight ◽

Triticum Aestivum ◽

Winter Wheat ◽

High Molecular Weight ◽

Gene Pool ◽

Allelic Diversity ◽

Triticum Aestivum L ◽

Hmw Glutenin ◽

Wheat Landraces ◽

In The Beginning

Aim. The aim of the study was to investigate allelic variability of high-molecular-weight glutenin loci Glu-A1, Glu-B1, Glu-D1 in Ukrainian winter wheat landraces and obsolete cultivars Triticum aestivum L. Methods. Allelic diversity at the Glu-1 loci were analyzed in 54 collection accessions, including 41 landraces (Krymka, Banatka, Girka, Theyka and others), and 13 first breeding cultivars that were developed in the beginning of the last century by selection from local wheat. Method of SDS-PAG electrophoresis according to Laemmli was used for fractionation of HMW glutenin subunits. Results. A total 11 alleles at the Glu-1 loci were identified, including 3 alleles at the Glu-A1 (a, b, c) and Glu-D1 (a, b, d) loci, and 5 – at the Glu- B1 (c, u, an, aj and subunit 9). Differences in frequencies of glutenin alleles were revealed. Conclusions. In the gene pool of Ukrainian winter bread wheat landraces the most widespread alleles were Glu-A1a (43.3 %), Glu-A1b (40.5 %), Glu-B1c (58 %), Glu-B1u (23 %), Glu-D1d (48.6 %), Glu-D1a (47.2 %). All these alleles (except of the Glu-D1a) are also predominant in the gene pool of modern commercial Ukrainian cultivars. A distinctive feature of Ukrainian landraces are the rare allelic variants of the Glu-B1 locus, which encode the subunits 1By9 and 1By8 (allele Glu-B1aj). Keywords: Triticum aestivum L., winter wheat, landraces, high-molecular-weight glutenin, alleles.

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Conformational differences between two wheat (Triticum aestivum) ‘high-molecular-weight’ glutenin subunits are due to a short region containing six amino acid differences

Biochemical Journal ◽

10.1042/bj2630837 ◽

1989 ◽

Vol 263 (3) ◽

pp. 837-842 ◽

Cited By ~ 32

Author(s):

A P Goldsbrough ◽

N J Bulleid ◽

R B Freedman ◽

R B Flavell

Keyword(s):

Molecular Weight ◽

Triticum Aestivum ◽

Amino Acid ◽

High Molecular Weight ◽

Polyacrylamide Gel Electrophoresis ◽

Amino Acid Sequences ◽

Glutenin Subunits ◽

Sds Page ◽

Hmw Glutenin

‘High-molecular-weight’ (HMW, high-Mr) glutenin subunits are protein constituents of wheat (Triticum aestivum) seeds and are responsible in part for the viscoelasticity of the dough used to make bread. Two subunits, numbered 10 and 12, are the products of allelic genes. Their amino acid sequences have been derived from the nucleic acid sequences of the respective genes. Subunit 10 has fewer amino acids than subunit 12, but migrates more slowly on SDS/PAGE (polyacrylamide-gel electrophoresis). This anomaly is due to between one and six of the amino acid differences between the subunits, localized towards the C-terminal end of the proteins. This has been established by making chimaeric genes between the genes for subunits 10 and 12, transcribing and translating them in vitro and analysing the products by SDS/PAGE. The postulated conformational differences between subunits 10 and 12 are discussed in relation to current hypotheses for the structure of HMW glutenin subunits.

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Characterisation of a novel high-molecular-weight glutenin subunit 1Dy12.3 from hexaploid wheat (Triticum aestivum L.)

Czech Journal of Genetics and Plant Breeding ◽

10.17221/111/2012-cjgpb ◽

2012 ◽

Vol 48 (No. 4) ◽

pp. 157-168 ◽

Cited By ~ 4

Author(s):

D. Mihálik ◽

E. Gregová ◽

P. Galuszka ◽

L. Ohnoutková ◽

T. Klempová ◽

...

Keyword(s):

Molecular Weight ◽

Triticum Aestivum ◽

High Molecular Weight ◽

Hexaploid Wheat ◽

Signal Sequence ◽

Triticum Aestivum L ◽

Glutenin Subunit ◽

Repetitive Domain ◽

Sds Page ◽

Methionine Residues

A novel high-molecular-weight glutenin subunit encoded by the Glu-1D locus was identified in hexaploid wheat (Triticum aestivum L.) cultivar Noe and was designated as 1Dy12.3. This subunit differed in SDS-PAGE mobility from the well-known 1Dy10 and 1Dy12 subunits that are also encoded by this locus. An analysis of the gene sequences confirmed the uniqueness of 1Dy12.3 and revealed that it is most closely related to the 1Dy12 subunit. The size of the deduced protein was calculated to be 67 884 Da, which is different from the 1Dy10 and 1Dy12 subunits (67 475 Da and 68 713 Da, respectively). The 1Dy12.3 protein consists of 652 residues, with a highly conserved signal sequence and N- and C-terminal domains, although the central repetitive domain comprising motifs of hexapeptide (PGQGQQ) and nonapeptide (GYYPTSLQQ) repeats was less conserved. The 1Dy12.3 subunit demonstrates fewer QHPEQG hexapeptide motifs and exhibits an increased number of methionine residues in comparison to the other characterised high-molecular-weight glutenin subunits. The 1Dy12.3 subunit was cloned and expressed in Escherichia coli and was detected with a prolamin-specific antibody. The size of the detected immunocomplex corresponded to the native 1Dy12.3 protein isolated from grains. The existence and characterisation of this novel high-molecular-weight glutenin subunit increases the diversity of the glutenins encoded by the Glu-1D locus.

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