Skip to main content
SpringerLink
Log in

Molecular cloning and functional characterization of two novel high molecular weight glutenin subunit genes in Aegilops markgrafii

  • Research Article
  • Published:
Journal of Genetics Aims and scope Submit manuscript

Abstract

The high molecular weight glutenin subunits (HMW-GS) in bread wheat are major determinants of the viscoelastic properties of dough and the end-use quality of wheat flour. Two novel HMW-GSs, 1Cx1.1 and 1Cy9.1, from the diploid species Aegilops markgrafii (CC) were identified in the present study. The corresponding open-reading frames of the genes of 1Cx1.1 and 1Cy9.1 were isolated and sequenced using allele-specific polymerase chain reaction. Sequence comparison demonstrated that the HMW-GSs from Ae. markgrafii possess a similar primary structure to the homologous proteins in wheat and related species. A tandem tripeptide exists in the central repetitive domain of 1Cx1.1, and this unique structure is very rare in the HMW-GSs of other genomes. To confirm the authenticity of these isolated endogenous HMW-GS, the heterologous proteins produced by removing the signal peptides expressed by E. coli exhibited the same electrophoretic mobility as the native proteins. Subsequently, the single protein was purified at a sufficient scale for incorporation into flour to perform sodium dodecyl sulphate (SDS) sedimentation testing. Notably, the SDS sedimentation volume was less with the addition of 1Cx1.1 than it was with 1Cy9.1.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Altpeter F., Popelka J. C. and Wieser H. 2004 Stable expression of 1Dx5 and 1Dy10 high-molecular-weight glutenin subunit genes in transgenic rye drastically increases the polymeric glutelin fraction in rye flour. Plant Mol. Biol. 54, 783–792.

    Article  CAS  PubMed  Google Scholar 

  • An X. L., Li X. H., Xiong X. J., Yan Y. M., Zhang Y. Z., Gao L. Y. et al. 2009 Identification and isolation of a new x-type HMW glutenin subunit \({\mathit{1}Dx\mathit{1.6}}^{t}\) gene from Aegilops tauschii. Plant Breed. 128, 41–45.

  • Anderson O. D. and Greene F. C. 1989 The characterization and comparative analysis of high-molecular-weight glutenin genes from genomes A and B of a hexaploid bread wheat. Theor. Appl. Genet. 77, 689–700.

    Article  CAS  PubMed  Google Scholar 

  • Belton P. S., Gil A. M., Grant A., Alberti E. and Tatham A. S. 1998 Proton and carbon NMR measurements of the effects of hydration on the wheat protein \({\upomega }\)-gliadin. Spectrochim. Acta A 54, 955–966.

    Article  Google Scholar 

  • Brark S., Mudgil D. and Khatker B. S. 2013 Relationship of gliadin and glutenin proteins with dough rheology, flour pasting and bread making performance of wheat varieties. LWT - Food Sci. Technol. 51, 211–217.

    Article  Google Scholar 

  • Cao S., Xu H., Li Z., Wang X., Wang D., Zhang A. et al. 2007 Identification and characterization of a novel Ag. intermedium HMW-GS gene from T. aestivum-Ag. intermedium addition lines TAI-I series. J. Cereal Sci. 45, 293–301.

    Article  CAS  Google Scholar 

  • Chen F. G., Liu S. W., Zhao F., Xu C. H. and Xia G. M. 2010 Molecular characterization of the low-molecular weight glutenin subunit genes of tall wheatgrass and functional properties of one clone Ee34. Amino Acids 38, 991–999.

    Article  CAS  PubMed  Google Scholar 

  • Chen F. G., Zhao F., Liu R. K. and Xia G. M. 2011 Functional properties of two low-molecular-weight glutenin subunits carrying additional cysteine residues from hybrid introgression line II-12 derived from Triticum aestivum and Agropyron elongatum. Food Chem. 127, 1773–1776.

    Article  CAS  Google Scholar 

  • Don C., Lichtendonk W. J., Plijter J. J. and Hamer R. J. 2005 The effect of mixing on glutenin particle properties: aggregation factors that affect gluten function in dough. J. Cereal Sci. 41, 69–83.

    Article  CAS  Google Scholar 

  • Don C., Mann G., Bekes F. and Hamer R. J. 2006 HMW-GS affect the properties of glutenin particles in GMP and thus flour quality. J. Cereal Sci. 44, 127–136.

    Article  CAS  Google Scholar 

  • Fang J. Y., Liu Y., Luo J., Wang Y. S., Shewry P. R. and He G. Y. 2009 Allelic variation and genetic diversity of high molecular weight glutenin subunit in Chinese endemic wheats (Triticum aestivum L.). Euphytica 166, 177–182.

    Article  CAS  Google Scholar 

  • Gao X., Zhang Q. S., Newberry M. P., Chalmers K. J. and Mather D. E. 2013 A cysteine in the repetitive domain of a high-molecular-weight glutenin subunit interferes with the mixing properties of wheat dough. Amino Acids 44, 1061–1071.

  • Gitte P., Ole S., Merete Y. and Berthelsen K. 2006 Phylogenetic relationships of Triticum and Aegilops and evidence for the origin of the A, B, and D genomes of common wheat (Triticum aestivum). Mol. Phylogenet. Evol. 39, 70–82.

    Article  Google Scholar 

  • Halford N. G., Field J. M., Blair H., Urwin P., Moore K., Robert L. et al. 1992 Analysis of HMW glutenin subunits encoded by chromosome 1A of bread wheat (Triticum aestivum L.) indicates quantitative effects on grain quality. Theor. Appl. Genet. 83, 373–378.

    Article  CAS  PubMed  Google Scholar 

  • Huang X. Q. and Cloutier S. 2008 Molecular characterization and genomic organization of low molecular weight glutenin subunit genes at the Glu-3 loci in hexaploid wheat (Triticum aestivum L.). Theor. Appl. Genet. 116, 953–966.

    Article  CAS  PubMed  Google Scholar 

  • Ji J., Liu D. C., Wang J., Li J. M. and Zhang A. M. 2008 A method of extraction and separation of wheat gluten. Hereditas 30, 123–126.

    Article  CAS  PubMed  Google Scholar 

  • Kimber G. and Athwal R. S. 1972 A reassessment of the course of evolution of wheat. Proc. Natl. Acad. Sci. USA 69, 912–915.

  • León E., Marín S., Gimínez M. J., Piston F., Rodróguez-Quijano M., Shewry P. R. and Barro F. 2009 Mixing properties and dough functionality of transgenic lines of a commercial wheat cultivar expressing the 1Ax1, 1Dx5 and 1Dy10 HMW glutenin subunit genes. J. Cereal Sci. 49, 148–156.

    Article  Google Scholar 

  • Liu S. W., Zhao F., Gao X., Chen F. G. and Xia G. M. 2010 A novel high molecular weight glutenin subunit from Australopyrum retrofractum. Amino Acids 39, 385–392.

    Article  CAS  PubMed  Google Scholar 

  • Liu Z., Yan Z., Wan Y., Liu K., Zheng Y. and Wang D. 2003 Analysis of HMW glutenin subunits and their coding sequences in two diploid Aegilops species. Theor. Appl. Genet. 106, 1368–1378.

    Article  CAS  PubMed  Google Scholar 

  • Li F., Jiang X., Wei Y. F., Xia G. M. and Liu S. W. 2011 Characterization of a novel type of HMW subunit of glutenin from Australopyrum retrofractum. Gene 492, 65–70.

    Article  PubMed  Google Scholar 

  • Li X., Liu D. C., Sun J. Z., Yang W. L., Guo X. L., Wang D. W. and Zhang A. M. 2015 Characterization of novel high-molecular-weight glutenin subunits and their coding sequences in Aegliops markgrafii. J. Cereal Sci. 65, 9–18.

    Article  CAS  Google Scholar 

  • Luo C., Branlard G., Griffin W. B. and McNeil D. L. 2000 The effect of nitrogen and sulphur fertilisation and their interaction with genotype on wheat glutenins and quality parameters. J. Cereal Sci. 31, 185–194.

    Article  CAS  Google Scholar 

  • Matsuo H., Kohno K. and Morita E. 2005 Molecular cloning, recombinant expression and IgE-binding epitope of \({\upomega }\)-5 gliadin, a major allergen in wheat-dependent exercise induced anaphylaxis. FEBS J. 272, 4431–4438.

  • Matsuo H., Dahlström J., Tanaka A., Kohno K., Takahashi H., Furumura M. and Morita E. 2008 Sensitivity and specificity of recombinant \({\upomega }\)-5 gliadin-specific IgE measurement for the diagnosis of wheat-dependent exercise-induced anaphylaxis. Allergy 63, 233–236.

    Article  CAS  PubMed  Google Scholar 

  • Ma C. Y., Yang Y., Li X. H., Ge P., Guo G. F., Subburaj S. et al. 2013 Molecular cloning and characterization of six novel HMW-GS genes from Aegilops speltoides and Aegilop skotschyi. Plant Breed. 132, 284–289.

    Article  CAS  Google Scholar 

  • Pirozi M. R., Margiottac B., Lafiandrad D. and MacRitchie F. 2008 Composition of polymeric proteins and bread-making quality of wheat lines with allelic HMW-GS differing in number of cysteines. J. Cereal Sci. 48, 117–122.

    Article  CAS  Google Scholar 

  • Payne P. I., Corfield K. G. and Blackman J. A. 1979 Identification of a high molecular weight subunit of glutenin whose presence correlates with bread-making quality in wheats of related pedigree. Theor. Appl. Genet. 55, 153–159.

    Article  CAS  PubMed  Google Scholar 

  • Pei Y. H., Sun H., Song X. Y., Yan Y. M., Ji K. M., Li Q. Y.et al. 2008 In vitro functional testing of high molecular glutenin subunits in wheat. Acta. Agron. Sin. 34, 1910–1915.

    Article  CAS  Google Scholar 

  • Shewry P. R., Halford N. G. and Tatham A. S. 1992 High molecular weight subunits of wheat glutenin. J. Cereal Sci. 15, 105–120.

    Article  CAS  Google Scholar 

  • Shewry P. R., Tatham A. S. and Lazzeri P. 1997 Biotechnology of wheat quality. J. Sci. Food Agric. 73, 379–406.

    Article  Google Scholar 

  • Tang Y. L., Yang W. Y., Tian J. C., Li J. and Cheng F. 2008 Effect of HMW-GS 6+8 and 1.5+10 from synthetic hexaploid wheat on wheat quality traits. J. Integr. Agric. 10, 161–171.

    Google Scholar 

  • Van Herpen T. W. J. M., Goryunova S. V., van der Schoot J., Mitreva M., Salentijn E., Vorst O. et al. 2006 Alpha-gliadin genes from the A, B, and D genomes of wheat contain different sets of celiac disease epitopes. BMC Genomics 7, 1–4.

    Article  PubMed  PubMed Central  Google Scholar 

  • Wan Y., Liu K., Wang D. and Shewry P. R. 2000 High-molecular-weight glutenin subunits in the Cylindropyrum and Vertebrata section of the Aegilops genus and identification of subunits related to those encoded by the Dx alleles of common wheat. Theor. Appl. Genet. 101, 879–884.

    Article  CAS  Google Scholar 

  • Wan Y. F., Shewry P. R. and Hawkesford M. J. 2012 A novel family of \({\upgamma }\)-gliadin are highly regulated by nitrogen supply in developing wheat grain. J. Exp. Bot. 318, 1–8.

    Google Scholar 

  • Wang G. Z., Miyashita N. T. and Tsunewaki K. 1997 Plasmon analyses of Triticum (wheat) and Aegilops: PCR-single-strand conformational polymorphism (PCR-SSCP) analyses of organellar DNAs. Proc. Natl. Acad. Sci. USA 94, 14570–14577.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang J. R., Yan Z. H., Jiang Q. T., Wei Y. M., Banu B. R. and Zheng Y. L. 2007 Sequence variations and molecular phylogenetic analysis of the HMW-GS genes from different genomes in Triticeae. Biochem. S. Ecol. 35, 421–433.

  • Wang S. L., Li X. H., Wang K., Wang X. Z., Li S. S., Zhang Y. Z.et al. 2011 Phylogenetic analysis of C, M, N, and U genomes and their relationships with Triticum and other related genomes as revealed by LMW-GS genes at Glu-3 loci. Genome 54, 273–284.

  • Wang S. L., Yu Z. T., Cao M., Shen X. X., Li N., Li X. H. et al. 2013 Molecular mechanisms of HMW glutenin subunits from \({\rm 1S}^{\rm l}\) genome of Aegilops longissima positively affecting wheat breadmaking quality. PLoS One 8, e58947.

  • Xu L. L., Li W., Wei Y. M. and Zheng Y. L. 2009 Genetic diversity of HMW glutenin subunits in diploid, tetraploid and hexaploid Triticum species. Genet. Resour. Crop Evol. 56, 377–391.

    Article  CAS  Google Scholar 

  • Yan Z. H., Wei Y. M., Wang J. R., Liu D. C., Dai S. F. and Zheng Y. L. 2006 Characterization of two HMW glutenin subunit genes from Taenitherum Nevski. Genetica 127, 267–276.

Download references

Acknowledgements

This work was supported by the financial support from Postdoctoral Foundation of Shandong Agricultural University.

Author information

Authors and Affiliations

  1. College of Life Science, Guizhou Normal University, No. 116, Baoshan North Street, Guiyang , 550001, Guizhou Province, People’s Republic of China

    Xuye Du

  2. College of Food Science and Engineering, Shandong Agricultural University, No. 61, Daizong Street, Taian , 271018, Shandong Province, People’s Republic of China

    Xuye Du & Xiaocun Zhang

Authors
  1. Xuye Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaocun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaocun Zhang.

Additional information

Corresponding editor: U. C. Lavania

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Du, X., Zhang, X. Molecular cloning and functional characterization of two novel high molecular weight glutenin subunit genes in Aegilops markgrafii . J Genet 96, 563–570 (2017). https://doi.org/10.1007/s12041-017-0799-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12041-017-0799-2

Keywords

Search

Navigation