Molecular characterization of high-molecular weight glutenin subunits 1Bx6.1 and 1By22.1 from Triticum spelta К1731 accession

  • O. A. Orlovskaya
  • K. K. Yatsevich
  • S. I. Vakula
  • L. V. Khotyleva
  • A. V. Kilchevsky
DOI: https://doi.org/10.7124/FEEO.v25.1156

Abstract

Aim. Some spelt varieties, along with alleles of gliadins and high-molecular glutenin subunits (HMW-GS), identical to common wheat, contain specific alleles, that are source of Triticum aestivum gene pool enrichment. The aim of this work is the identification, molecular analysis of HMW – GS from T. spelta K1731 and evaluation of their effect on the elastic properties of gluten. Methods. Identification of HMW-GS was carried out by SDS-electrophoresis and PCR analysis. Nucleotide gene sequences were determined by Sanger sequencing. The secondary structure of proteins was predicted on the on-line CFSSP server. Results. Subunits 6.1 + 22.1 of the Glu-B1 locus encoded by the Glu-B1be allele were detected in the T. spelta K1731. The nucleotide sequences of the 1Bx6.1, 1By22.1 genes from spelt were determined, the amino acid sequence and the protein secondary structure of 6.1 + 22.1 subunits were analyzed. Conclusions. Molecular analysis of HMW-GS 1Bх6.1 and 1By22.1 from T. spelta К1731 established a low contribution to the bread-making quality of these subunits.

Keywords: Triticum spelta K1731, HMW-GS, SDS electrophoresis, sequencing, secondary protein structure, gluten quality.

References

Shewry P.R., Halford N.G. Cereal seed storage proteins: structures, properties and role in grain utilization. J. Exp. Botany. 2002. Vol. 53. P. 947–958.

Niu Z.X., Klindworth R., Wang R.R.-C., Jauhar P.P., Larkin P.J., Xu S.S. Characterization of HMW glutenin subunits in Thi-nopyrum intermedium, Th. bessarabicum, Lophopyrum elongatum, Aegilops markgrafii, and their addition lines in wheat. Crop Sci. 2011. Vol. 51, № 2. P. 667 677. doi: 10.2135/cropsci2010.04.0235.

Xu S.S., Khan K., Klindworth D.L., Nygard G. Evaluation and characterization of high-molecular weight 1D glutenin subunits from Aegilops tauschii in synthetic hexaploid wheats. J. Cereal Sci. 2010. Vol. 52, № 2. P. 333 336. doi: 10.1016/j.jcs.2010.05.004.

Singh N.K., Shepherd K.W., Cornish G.B. A simplified SDS-PAGE procedure for separating LMW subunits of glutenin. Journal of Cereal Science. 1991. Vol. 14. P. 203–208.

Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970. No. 227 (5259). P. 680–685.

Payne P.I., Lawrence G.J. Catalogue of alleles for the complex gene loci, Glu-A1, Glu-B1, and Glu-D1 which code for high-molecular-weight subunits of glutenin in hexaploid wheat. Cereal Res. Com. 1983. Vol. 11. P. 29–35.

Ma W., Zhang W., Gale K.R. Multiplex-PCR typing of high molecular weight glutenin alleles in wheat. Euphytica. 2003. Vol. 134, № 1. P. 51–60. doi: 10.1023/A:1026191918704.

Ahmad M. Molecular marker-assisted selection of HMW glutenin alleles related to wheat bread quality by PCR-generated DNA markers. Theor. Appl. Genet. 2000. Vol. 101, № 5–6. P. 892–896. doi: 10.1007/s001220051558.

Lafiandra D., Tucci G.F., Pavoni A., Turchetta T., Margiotta B. PCR analysis of x- and y-type genes present at the complex Glu-A1 locus in durum and bread wheat. Theor. Appl. Genet. 1997. Vol. 94. P. 235–240.

Lei Z.S., Gale K.R., He Z.H., Gianibelli C., Larroque O., Xia X.C., Butow B.J., Ma W. Y-type gene specific markers for enhanced discrimination of high-molecular weight glutenin alleles at the Glu-B1 locus in hexaploid wheat. J. Cereal Sci. 2006. Vol. 43, № 1. Р. 94–101. doi: 10.1016/j.jcs.2005.08.003.

Kozub N.A., Boguslavskkii R.L., Sozinov I.A., Tverdokhleb Ye.V., Xynias I.N., Blume Ya.B., Sozinov A.A. Alleles at storage protein loci in Triticum spelta L. accessions and their occurrence in related wheats. Cytology and genetics. 2014. Vol. 48, № 1. P. 33–41.

Yan Y., Hsam S.L.K., Yu J.Z., Jiang Y., Ohtsuka I., Zeller F.J. HMW and LMW glutenin alleles among tetraploid and hexaploid European spelt wheat (Triticum spelta L.) progenitors. Theor. Appl. Genet. 2003. Vol. 107. P. 1321 1330. doi: 10.1007/s00122-003-1315-z.

Wieser H. Chemistry of gluten proteins. Food Microbiology. 2007. Vol. 24, № 2. P. 115–119. doi: 10.1016/j.fm.2006.07.004.

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

Shewry P.R., Tatham A.S. Disulphide bonds in wheat gluten proteins. J. Cereal Sci. 1997. Vol. 25. P. 207–227.

Tatham A.S, Miflin B.J, Shewry P.R. The beta-turn conformation in wheat gluten proteins: relationship to gluten elasticity. Cereal Chem. 1985. Vol. 62. P. 405–412.

Guo X.H., Hu J.L., Wu B.H., Wang Z.Z., Wang D., Liu D.C., Zheng Y.L. Special HMW-GSs and their genes of Triticum tur-gidum subsp. dicoccoides accession D141 and the potential utilization in common wheat. Genet. Resour. Crop. Evol. 2016. Vol. 63, № 5. P. 833 844. doi: 10.1007/s10722-015-0287-6.

Morgun V.V., Sichkar S.M., Pochinok V.M., Ninieva A.K., Chugunkova T.V. Characterization of spelt collection samples (Triticum spelta L.) by elements of plant productivity structure and baking quality. Plant physiology and genetics. 2016. Vol. 48, № 2. P. 112–119. [in Ukrainian]