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Cloning and characterization of a novel low molecular weight glutenin subunit gene at the Glu-A3 locus from wild emmer wheat (Triticum turgidum L. var. dicoccoides)

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Abstract

Amplification of the coding region, and upstream and downstream sequences of a low-molecular-weight glutenin subunits (LMW-GS) gene from wild emmer wheat (Triticum turgidum L. var. dicoccoides, 2n = 4x = 28, AABB) accession TD22 was carried out using designed allele-specific PCR (AS-PCR) primers. The complete 1,176 bp sequence of a novel LMW-i type subunit gene at the Glu-A3 locus, named LMW-TD22, is described. Analysis of the nucleotide and deduced amino acid sequences showed that this gene possessed striking characteristics although its molecular structure was generally similar to those of previously reported i-type LMW-GS genes that were isolated from common wheat and related species. The deduced amino acid sequence of LMW-TD22 gene contained 390 amino acid residues with the predicted molecular weight being 43,009.3 Da, which appeared to be the longest gene among the cloned LMW-i type genes from bread wheat and related species. The distinct feature of LMW-TD22 was two long polyglutamine stretches of 12 and 17 glutamines occurring in the repetitive and C-terminal domains as well as a cysteine residue present in the seventh amino acid residue of the signal peptide. These polyglutamine repeats are believed to improve the structure of gluten polymer and increase the strength of dough formed from the polymer. In addition, the putative 44 k subunit encoded by LMW-TD22 was verified by N-terminal microsequencing, gel electrophoresis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis. Certain types of post-translational modifications, such as phosphorylation and glycosylation, may be associated with this LMW-i type subunit.

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References

  • An X, Zhang Q, Yan Y, Li Q, Zhang Y, Wang A, Pei Y, Tian J, Wang H, Hsam SLK, Zeller FJ (2006) Cloning and molecular characterization of three novel LMW-i glutenin subunit genes from cultivated einkorn (Triticum monococcum L.). Theor Appl Genet 113:383–395

    Article  PubMed  CAS  Google Scholar 

  • Anderson OD, Greene FC, Yip RE, Halford NG, Shewry PR, Malpica-Romero JM (1989) Nucleotide sequences of the two high-molecular-weight glutenin genes from the D-genome of a hexaploid bread wheat, Triticum aestivum L. cv Cheyenne. Nucleic Acids Res 17:461–462

    Article  PubMed  CAS  Google Scholar 

  • Bartels D, Thompson RD (1986) Synthesis of mRNAs coding for abundant endosperm proteins during wheat grain development. Plant Sci 46:117–125

    Article  CAS  Google Scholar 

  • Cassidy BG, Dvorak J, Anderson OD (1998) The wheat low-molecular-weight glutenin genes: characterization of six new genes and progress in understanding gene family structure. Theor Appl Genet 96:743–750

    Article  CAS  Google Scholar 

  • Ciaffi M, Lee YK, Tamas L, Gupta R, Skerritt J, Appels R (1999) The low-molecular-weight glutenin subunit proteins of primitive wheats III. The genes from D-genome species. Theor Appl Genet 98:135–148

    Article  CAS  Google Scholar 

  • Cloutier S, Rampitsch C, Penner GA, Lukow OM (2001) Cloning and expression of a LMW-i glutenin gene. J Cereal Sci 33:143–154

    Article  CAS  Google Scholar 

  • Colot V, Robert LS, Kavanagh TA, Bevan MW, Thompson RD (1987) Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. EMBO J 6:3559–3564

    PubMed  CAS  Google Scholar 

  • Colot V, Bartels D, Thompson R, Flavell R (1989) Molecular characterization of an active wheat LMW glutenin gene and its relation to other wheat and barley prolamin genes. Mol Gen Genet 216:81–90

    Article  PubMed  CAS  Google Scholar 

  • D’Ovidio R, Marchitelli C, Cardelli EL, Porceddu E (1999) Sequence similarity between allelic Glu-B3 genes related to quality properties of durum wheat. Theor Appl Genet 98:455–461

    Article  CAS  Google Scholar 

  • D’Ovidio R, Masci S (2004) The low-molecular-weight glutenin subunits of wheat gluten. J Cereal Sci 39:321–339

    Article  CAS  Google Scholar 

  • Forde J, Malpica JM, Halford NG, Shewry PR, Anderson OD, Greene FC, Miflin BJ (1985) The nucleotide sequence of a HMW glutenin subunit gene located on chromosome 1A of wheat (Triticum aestivum L.). Nucleic Acids Res 13:6817–6832

    Article  PubMed  CAS  Google Scholar 

  • Ikeda TM, Nagamine T, Fukupka H, Yano H (2002) Identification of new low-molecular-weight glutenin subunit genes in wheat. Theor Appl Genet 104:680–687

    Article  PubMed  CAS  Google Scholar 

  • Ikeda TM, Araki E, Fujita Y, Yano H (2006) Characterization of low-molecular-weight glutenin subunit genes and their protein products in common wheats. Theor Appl Genet 112:327–334

    Article  PubMed  CAS  Google Scholar 

  • Johal J, Gianibelli MC, Rahman S, Morell MK (2004) Characterization of low-molecular-weight glutenin genes in Aegilops tauschii. Theor Appl Genet 109:1028–1040

    Article  PubMed  CAS  Google Scholar 

  • Lauriere M, Bouchez I, Doyen C, Eynard L (1996) Identification of glycosylated forms of wheat storage proteins using two-dimensional electrophoresis and blotting. Electrophoresis 17:497–501

    Article  PubMed  CAS  Google Scholar 

  • Lee YK, Ciaffi M, Appels R, Morell MK (1999) The low-molecular-weight glutenin subunits of primitive wheats II. The genes from A-genome species. Theor Appl Genet 98:126–134

    Article  CAS  Google Scholar 

  • Levy AA, Galili G, Feldman M (1988) Polymorphism and genetic control of high molecular weight glutenin subunits in wild tetraploid wheat Triticum turgidum var. dicoccoides. Heredity 61:63–72

    CAS  Google Scholar 

  • Lew EJL, Kuzmicky DD, Kasarda DD (1992) Characterization of low molecular weight glutenin subunits by reversed-phase high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and N-terminal amino acid sequencing. Cereal Chem 69:508–515

    CAS  Google Scholar 

  • Luo Z, Chen F, Feng D, Xia G (2005) LMW-GS genes in Agropyron elongatum and their potential value in wheat breeding. Theor Appl Genet 111:272–280

    Article  PubMed  CAS  Google Scholar 

  • Maruyama-Funatsuki W, Takata K, Nishio N, Tabiki T, Yabata E, Kato A, Saito K, Funatsuki H, Saruyama H, Yamauchi H (2004) Identification of low-molecular weight glutenin subunits of wheat associated with bread-making quality. Plant Breed 123:355–360

    Article  CAS  Google Scholar 

  • Maruyama-Funatsuki W, Takata K, Funatsuki H, Tabiki T, Ito M, Nishio Z, Kato A, Saito K, Yabata E, Saruyama H, Yamauchi H (2005) Identification and characterization of a novel LMW-s glutenin gene of a Canadian Western Extra-Strong wheat. J Cereal Sci 41:47–57

    Article  CAS  Google Scholar 

  • Masci S, D’Ovidio R, Lafiandra D, Kasarda DD (1998) Characterization of a low-molecular-weight glutenin subunit gene from bread wheat and the corresponding protein that represents a major subunit of the glutenin polymer. Plant Physiol 118:1147–1158

    Article  PubMed  CAS  Google Scholar 

  • Masci S, D’Ovidio R, Lafiandra D, Kasarda DD (2000) A 1B coded low-molecular-weight glutenin subunit associated with quality in durum wheats show strong similarity to subunits present in some bread wheat cultivars. Theor Appl Genet 100:396–400

    Article  CAS  Google Scholar 

  • McDonald MB, Elliot LJ, Sweeney PM (1994) DNA extraction from dry seeds for RAPD analyses in varietal identification studies. Seed Sci Technol 22:171–176

    Google Scholar 

  • Okita TW, Cheesbrough V, Reeves CD (1985) Evolution and heterogeneity of the alpha-/beta-type and gamma-type. J Bio Chem 260:8203–8213

    CAS  Google Scholar 

  • Ozdemir N, Cloutier S (2005) Expression analysis and physical mapping of low-molecular-weight glutenin loci in hexaploid wheat (Triticum aestivum L.). Genome 48:401–410

    Article  PubMed  CAS  Google Scholar 

  • Payne PI (1987) Genetics of wheat storage proteins and the effect of allelic variation on bread-making quality. Ann Rev Plant Physiol 38:141–153

    Article  CAS  Google Scholar 

  • Pflüger LA, Martín LM, Alvarez JB (2001) Variation in the HMW and LMW glutenin subunits from Spanish accessions of emmer wheat (Triticum turgidum ssp. dicoccum Schrank). Theor Appl Genet 102:767–772

    Article  Google Scholar 

  • Pitts EG, Rafalski JA, Hedhcoth C (1988) Nucleotide sequence and encoded amino acid sequence of a genomic gene region for a low molecular weight glutenin. Nucleic Acids Res 16:11376

    Article  PubMed  CAS  Google Scholar 

  • Tanaka H, Shimizu R, Tsujimoto H (2005) Genetical analysis of contribution of low-molecular-weight glutenin subunits to dough strength in common wheat (Triticum aestivum L.). Euphytica 141:157–162

    Article  CAS  Google Scholar 

  • Tatham AS, Field JM, Smith SJ, Shewry PR (1987) The conformations of wheat gluten proteins II. Aggregated gliadins and low molecular weight subunits of glutenin. J Cereal Sci 51:203–214

    Article  Google Scholar 

  • Thompson S, Bishop DHL, Madgwick P, Tatham AS, Shewry PR (1994) High-level expression of a wheat LMW glutenin subunit using a baculovirus system. J Agric Food Chem 42:426–431

    Article  CAS  Google Scholar 

  • Valkoun JJ (2001) Wheat pre-breeding using wild progenitors. Euphytica 119:17–23

    Article  Google Scholar 

  • Xu SS, Khan K, Kindworth DL, Faris JD, Nygard G (2004) Chromosomal location of genes for novel glutenin subunits and gliadins in wild emmer wheat (Triticum turgidum L. var. dicoccoides). Theor Appl Genet 108:1221–1228

    Article  PubMed  CAS  Google Scholar 

  • Yan Y, Prodanovic S, Mladenov N, Milovanovic M (1999) Genetic control of low molecular weight glutenin subunits in wheat by A-PAGE analysis. Cereal Res Comm 27:251–257

    CAS  Google Scholar 

  • Yan Y, Hsam SLK, Yu JZ, Jiang Y, Ohtsuka I, Zeller FJ (2003) HMW and LMW glutenin alleles among putative tetraploid and hexaploid European spelt wheat (Triticum spelta L.) progenitors. Theor Appl Genet 107:1321–1330

    Article  PubMed  CAS  Google Scholar 

  • Yan Y, Zheng J, Xiao Y, Yu J, Hu Y, Cai M, Li Y, Hsam SLK, Zeller FJ (2004) Identification and molecular characterization of a novel y-type Glu-D t 1 glutenin gene of Aegilops tauschii. Theor Appl Genet 108:1349–1358

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This research was supported by grants from the National Natural Science Foundation of China (30571154), and the Ministry of Science and Technology of China (2002CB111300 and 2006AA10Z186).

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Authors and Affiliations

  1. Key Laboratory of Genetics and Biotechnology, College of Life Science, Capital Normal University, 100037, Beijing, China

    Xiaohui Li, Aili Wang, Yinghua Xiao & Yueming Yan

  2. Institute of Crop Science/National Wheat Improvement Centre, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Zhonghu He

  3. CIMMYT-China Office, c/o CAAS, Beijing, 100081, China

    Zhonghu He

  4. Molecular Plant Breeding CRC, Western Australian Department of Agriculture and Food, State Agriculture Biotechnology Centre, Perth, WA, 6150, Australia

    Rudi Appels & Wujun Ma

  5. Plant Breeding, Technical University of Munich, 85350, Freising-Weihenstephan, Germany

    S. L. K. Hsam & F. J. Zeller

Authors
  1. Xiaohui Li
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  2. Aili Wang
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  3. Yinghua Xiao
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  4. Yueming Yan
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  5. Zhonghu He
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  6. Rudi Appels
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  7. Wujun Ma
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  8. S. L. K. Hsam
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  9. F. J. Zeller
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Corresponding authors

Correspondence to Yueming Yan or F. J. Zeller.

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A. Wang and Y. Xiao made equal contribution to the research as the first author.

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Li, X., Wang, A., Xiao, Y. et al. Cloning and characterization of a novel low molecular weight glutenin subunit gene at the Glu-A3 locus from wild emmer wheat (Triticum turgidum L. var. dicoccoides). Euphytica 159, 181–190 (2008). https://doi.org/10.1007/s10681-007-9471-x

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  • DOI: https://doi.org/10.1007/s10681-007-9471-x

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