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


  • Kirushanthy Kajendran Department of Chemistry, Faculty of Science, University of Colombo, Colombo
  • Naduviladath Vishvanath Chandrasekharan Department of Chemistry, Faculty of Science, University of Colombo, Colombo
  • Chamari Madhu Hettiarachchi Department of Chemistry, Faculty of Science, University of Colombo, Colombo
  • Wijerupage Sandhya Sulochana Wijesundera Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Colombo



Dough, Dacke, HMW glutenin


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. 


Metrics Loading ...

Author Biographies

Kirushanthy Kajendran, Department of Chemistry, Faculty of Science, University of Colombo, Colombo

Dept. of chemistry university of colombo

world rank - 2158

Naduviladath Vishvanath Chandrasekharan, Department of Chemistry, Faculty of Science, University of Colombo, Colombo

Dept. of Chemistry university of colombo

world rank - 2158

Chamari Madhu Hettiarachchi, Department of Chemistry, Faculty of Science, University of Colombo, Colombo

Dept. of Chemistry university of colombo

world rank - 2158

Wijerupage Sandhya Sulochana Wijesundera, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Colombo

Dept. of Biochemistry and Molecular biology

world rank - 2158


Gianibelli MC, Larroque OR, MacRitchie F, Wrigley CW. Biochemical, genetic, and molecular characterization of wheat endosperm proteins. Cereal Chemistry. 2001;78:635–46.

Belton PS. On the elasticity of wheat gluten. J Cereal Sci. 1999;29:103-7.

Wieser H. Chemistry of gluten proteins. Food Microbiol. 2007;24:115-9.

Larwance GJ, Shepherd KW. Chromsomal location of genes controlling seed protein in species realted to wheat. Theor Appl Genet. 1981;59:25–31

Liu L, Wang AL, Appels R, Ma JH, Xia XC, Lan P. A MALDI-TOF based analysis of high molecular weight glutenin subunits for wheat breeding. J. Cereal Sci. 2009;50:295–301.

Halford NG, Field JM, Blair H, Urwin P, Moore K, Robert L, et al. Analysis of HMW glutenin subunits encoded by chromosome 1A of bread wheat (Triticum aestivum L.) indicates quantitative effects on grain quality. Theor Appl Genet. 1992;83:373-8.

Shewry PR, Halford NG, Tatham AS.The high molecular weight subunits of wheat glutenin. J Cereal Sci. 1992;15:105–20.

MacRitchie F. Physicochemical properties of wheat proteins in relation to functionality. Adv Food Nutr Res. 1992;36:1–87.

Wrigley CW. Giant proteins with flour power. Nature. 1996;381:738–9.

Don C, Lichtendonk WJ, Plijter JJ, Hamer RJ Understanding the link between GMP and dough: from glutenin particles in flour towards developed dough. J Cereal Sci. 2003;38:157–65.

Anderson OD, Greene FC. The characterisation and comparative analysis of high molecular-weight glutenin genes from genomes A and B of hexaploid wheat. Theoret. Appl. Genet. 1989;77:689-700.

Brönneke V, Zimmermann G, Killermann B. Effect of high molecular weight glutenins and D-zone gliadins on bread-making quality in German wheat varieties. Cereal Res Commun. 2000;28:187-94.

Tabiki T, Ikeguchi S, Ikeda TM. Effects of high-molecular-weight and low-molecular-weight glutenin subunit alleles on common wheat flour quality. Breed. Sci. 2006;56:131-6.

Murray MG, Thompson WF) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980;8:4321–5.

Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33:1870-4.

McGuffin LJ, Bryson K, Jones DT. The PSIPRED protein structure prediction server. Bioinformatics. 2000;16:404–5.

Bryson K, McGuffin LJ, Marsden RL, Ward JJ, Sodhi JS, Jones DT Protein structure prediction servers at University College London. Nucleic Acids Res. 2005;33:36–8.

Altpeter F, Vasil VV, Vasil IK. Integration and expression of the high molecular weight glutenin subunit 1Ax1 gene into wheat. Nat. Biotechnol. 1996;14:1155-1159.

Deng ZY, Tian JC, Sun GX. Influence of high molecular weight glutenin subunit substitution on rheological behaviour and bread-baking quality of near-isogenic lines developed from Chinese wheats. Plant Breed. 2005;124:428-31.

Moloi MJ, BiljonAV, Labuschagne MT. Effect of quantity of HMW-GS 1Ax1, 1Bx13, 1By16, 1Dx5 and 1Dy10 on baking quality in different genetic backgrounds and environments. LWT-food science and technology.2017;78:160-4.

Hou W, Feng W, Yu G, Du X, Ren M. Cloning and functional analysis of a novel x-type high-molecular-weight glutenin subunit with altered cysteine residues from Aegilops umbellulata. Crop & Pasture Science. 2017;68:409–14.

Zhang D, He T, Liang H, Huang L, Su Y, Li Y, Li S. The flour quality and related molecular characterization of HMW-GS genes from wild emmer wheat accession TD-256. J Agric Food Chem. 2016;64:5128-36.

Li X, Liu T, Song L, Zhang H, Li L, Gao X. Influence of high-molecular-weight glutenin subunit composition at Glu-A1 and Glu-D1 loci on secondary and micro structures of gluten in wheat (Triticum aestivumL.). Food Chem. 2016;213:728-34.

Schalk K, Lexhaller B, Koehler P, Scherf KA. Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials. PLOS ONE. 2017;12.

Belderok B, Mesdag J, Donner DA. Sweden. In: Donner D.A. (eds) Bread- making quality of wheat. Springer, Dordrecht; 2000: 106-124.

Gilbert SM, Wellner N, Belton PS, Greenfield JA, Siligardi G, Shewry PR, Tatham AS. Expression and characterization of a highly repetitive peptide derived from a wheat seed storage protein. Acta Biochimica et Biophysica Sinica. 2000;1479:135-46.

Pandey R, Mishra A, Garg GK. Plant promoter driven heterologous expression of HMW glutenin gene(s) subunit in E. coli. Mol Biol Rep. 2008;35:153–62.

Georget DMR, Belton PS. Effects of temperature and water content on the secondary structure of wheat gluten studied by FTIR spectroscopy. Biomacromolecules. 2006;7:469-75.

Li QY, Yan YM, Wang AL, An XL, Zhang YZ, Hsam SLK, Zeller FJ. Detection of HMW glutenin subunit variations among 205 cultivated emmer accessions (Triticum turgidum spp. dicoccum). Plant Breeding. 2006;125:120-4.

Jin M, Xie Z, Li J, Jiang S, Ge P, Subburaj S, et al. Identification and molecular characterization of HMW glutenin subunit 1By16* in wild emmer.J. Appl. Genet. 2012;53:249-58.

Guo XH, Hu JL, Wu BH, Wang ZZ, Wang D, Liu DC, Zheng YL. Special HMW-GSs and their genes of Triticum turgidumsubsp. dicoccoidesaccession D141 and the potential utilization in common wheat. Genetic Resources and Crop Evolution. 2016;63:833-44.

Lafiandra D, D’Ovidio R, Porceddu E, Margiotta B, Colaprico G. New data supporting high Mrglutenin subunit 5 as determinant of quality differences among the pairs 5+10 vs 2+12. J Cereal Sci. 1993;18:197–205.

Halford, NG, Forde J, Anderson OD, GreeneE C and Shewry PR. The nucleotide and deduced amino acid sequences of an HMW glutenin subunit gene from chromosome 1B of bread wheat (Triticum aestivum L.), and comparison with those of genes from chromosomes 1A and 1D. Theoret Appl Genet. 1987;75:117-26.

Thompson RD, Bartels D, Harberd NE. Nucleotide sequence of a gene from chromosome 1D of wheat encoding a HMW glutenin subunits. Nucl Acids Res. 1985;13:6833-46.






Short Communication