Skip to main content
SpringerLink
Log in

Molecular characterisation of the low-molecular weight glutenin subunit genes of tall wheatgrass and functional properties of one clone Ee34

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

Wild tall wheatgrass (Lophopyrum elongatum L., 2x = 14) is an important resource for improving bread wheat (Titicum aestivum L.), including HMW-GS and LMW-GS relevant to end-use quality of the wheat flour. A set of 14 distinct sequences were amplified from the genomic DNA of the tall wheatgrass, using degenerate primers targeted at Glu-3, the locus containing the genes encoding the low-molecular weight glutenin subunits (LMW-GS). Three sequences contained an internal stop codon and were classified as pseudogenes. The other 11 all consisted of a single intron-less intact open-reading frame. An alignment of deduced protein sequences showed that the primary structure of all 11 sequences was similar to that of wheat and other wheat-related grass Glu-3 genes. All 11 sequences carried the 14 amino acid residue N-terminal motif MESNIIISFLK/RPWL, and were classified as LMW-m genes, based on the identity of the first amino acid of the mature protein. All but one of the sequences contained seven cysteine residues (the exception had 6). Their repetitive domain differs significantly from that present in Glu-3 genes isolated from the close relative intermediate wheatgrass (Thinopyrum Intermedium, 6x). A phylogenetic analysis showed that the tall wheatgrass sequences were closely related to those of the intermediate wheatgrass, but only distantly so to those from decaploid tall wheatgrass. One of the 11 LMW-GS peptides with a free-cysteine residue was heterologously expressed in E. coli and purified in sufficient scale to perform a flour supplementation test. This showed that the dough strength of bread wheat flour was significantly increased by the presence of the tall wheatgrass LMW-GS.

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

  • An X, Zhang Q, Yan Y, Li Q, Zhang Y, Wang A, Pei Y, Tian J, Wang H, Hsam SL, 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. doi:10.1007/s00122-006-0299-x

    Article  CAS  PubMed  Google Scholar 

  • Bekes F, Anderson OD, Gras PW, Gupta RB, Tam A, Wrigley CW, Appels R (1994) The contribution to mixing properties of 1D HMW glutenin subunits expressed in a bacterial system. In: Henry JR, Ronalds JA (eds) Improvement of cereal quality by genetic engineering. Plenum Press, New York, pp 97–103

    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. doi:10.1007/s001220050797

    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. doi:10.1006/jcrs.2000.0359

    Article  CAS  Google Scholar 

  • D’Ovidio R, Marchitelli C, Ercoil Cardelli L, Porceddu E (1999) Sequence similarity between allelic Glu-B3 genes related to quality properties of durum wheat. Theor Appl Genet 98:455–461. doi:10.1007/s001220051091

    Article  Google Scholar 

  • Dong YS, Zheng DS (2000) Genetic resource of Chinese wheat. China Agriculture Press, Beijing, pp 198–201

    Google Scholar 

  • Gupta RB, Shepherd KW (1990) Two-step one-dimensional SDS-PAGE analysis of LMW subunits of glutenin. Theor Appl Genet 80:65–74

    CAS  Google Scholar 

  • Ikeda TM, Nagamine T, Fukuoka H, Yano H (2002) Identification of new low-molecular-weight glutenin subunit genes in wheat. Theor Appl Genet 104(4):680–687. doi:10.1007/s001220100756

    Article  CAS  PubMed  Google Scholar 

  • Jiang C, Pei Y, Zhang Y, Li X, Yao D, Yan Y, Ma W, Hsam SL, Zeller FJ (2008) Molecular cloning and characterization of four novel LMW glutenin subunit genes from Aegilops longissima, Triticum dicoccoides and T. zhukovsk. Hereditas 145:92–98. doi:10.1111/j.0018-0661.2008.02035.x

    Article  PubMed  Google Scholar 

  • Johal J, Gianibelli MC, Rahman S, Morell MK, Gale KR (2004) Characterization of low-molecular- weight glutenin genes in Aegilops tauschii. Theor Appl Genet 109(5):1028–1040. doi:10.1007/s00122-004-1711-z

    Article  CAS  PubMed  Google Scholar 

  • Kasarda DD, Okita TW, Bemardin JE, Baecker PA, Nimmo CC, Lew EJ, Dietler MD, Greene FC (1984) Nucleic acid (cDNA) and amino acid sequences of alpha-type gliadins from wheat (Triticum aestivum). Proc Natl Acad Sci USA 81(15):712–716. doi:10.1073/pnas.81.15.4712

    Article  Google Scholar 

  • Khatkar BS, Fido RJ, Tatham AS, Schofield JD (2002) Functional properties of wheat gliadins. I. Effects on mixing characteristics and bread making quality. J Cereal Sci 35:307–313. doi:10.1006/jcrs.2001.0430

    Article  CAS  Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5(2):150–163. doi:10.1093/bib/5.2.150

    Article  CAS  PubMed  Google Scholar 

  • Lee YK, Bekes F, Gras PW, Appels R, Morell MK (1999) The low-molecular-weight glutenin subunit proteins of primitive wheats. IV. Functional properties of products from individual genes. Theor Appl Genet 98:149–155. doi:10.1007/s001220051051

    Article  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 sulfatepolyacrylamide gel electrophoresis, and N-terminal amino-acid sequencing. Cereal Chem 69:508–515

    CAS  Google Scholar 

  • Li QL, An XL, Xiao YH (2007) Cloning and molecular characterization of LMW-glutenin subunit genes in Triticum dicoccoides. Scientia Agric Sin 40(3):457–463

    CAS  Google Scholar 

  • Liu H, Shi L, Zhao J, Xia G (2006) Genetic characteristic of high molecular weight glutenin subunits in somatic hybrid wheat lines-potential application to wheat breeding. J Agric Food Chem 54:5007–5013. doi:10.1021/jf060321f

    Article  CAS  Google Scholar 

  • Luo Z, Chen FG, Feng DS, Xia GM (2005) LMW-GS genes in Agropyron elongatum and their potential value in wheat breeding. Theor Appl Genet 111(2):272–280. doi:10.1007/s00122-005-2021-9

    Article  CAS  PubMed  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 glutenin polymer. Plant Physiol 118:1147–1158. doi:10.1104/pp.118.4.1147

    Article  CAS  PubMed  Google Scholar 

  • Masci S, Rovelli L, Kasada DD, Vensel WH, Afiandra DL (2002) Characterization and chromosomal localization of C-type LMW-GS in the bread wheat cultivars Chinese Spring. Theor Appl Genet 104:422–428. doi:10.1007/s001220100761

    Article  CAS  PubMed  Google Scholar 

  • Miiller S, Wieser H (1995) The location of disulphide bonds in α-type gliadins. J Cereal Sci 22:21–27. doi:10.1016/S0733-5210(05)80004-9

    Article  Google Scholar 

  • Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8(19):4321–4325. doi:10.1093/nar/8.19.4321

    Article  CAS  PubMed  Google Scholar 

  • Payne PI (1987) Genetics of wheat storage proteins and the effect of allelic variation on bread-making quality. Annu Rev Plant Physiol 38:141–153. doi:10.1146/annurev.pp.38.060187.001041

    Article  CAS  Google Scholar 

  • Pei YH, Wang AL, An XL, Li XH, Zhang YZ, Huang XQ, Yan YM (2007) Characterization and comparative analysis of three low molecular weight glutenin C-subunit genes isolated from Aegilops tauschii. Can J Plant Sci 87:273–280

    CAS  Google Scholar 

  • Porceddu E, Turchetta T, Masci S, D’Ovidio R, Lafiandra D, Kasarda DD, Impiglia A, Nachit MM (1998) Variation in endosperm protein composition and technological quality properties in durum wheat. Euphytica 100:197–205. doi:10.1023/A:1018338308795

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Shewry PR, Halford NG (2002) Cereal seed storage proteins: structures, properties and role in grain utilization. J Exp Bot 53(370):947–958. doi:10.1093/jexbot/53.370.947

    Article  CAS  PubMed  Google Scholar 

  • Shewry PR, Tatham AS (1997) Disulfide bonds in wheat gluten proteins. J Cereal Sci 25:207–227. doi:10.1006/jcrs.1996.0100

    Article  CAS  Google Scholar 

  • Shewry PR, Halford NG, Lafiandra D (2003) Genetics of wheat gluten proteins. Adv Genet 49:111–184. doi:10.1016/S0065-2660(03)01003-4

    Article  CAS  PubMed  Google Scholar 

  • Walker AE, Walker CE (1992) Documentation and User Instructions for Mixsmart Software, Chapter 10. National Mfg, Lincoln

    Google Scholar 

  • Xu H, Zhang XQ, Wang XP, Guo AG (2004) Molecular cloning of Agropyron intermedium low-molecular-weight glutenin subunit genes from a Triticum aestivum-intermedium addition line TAI-13. Acta Bot Sin 46(5):595–602

    CAS  Google Scholar 

  • Xu H, Wang RJ, Shen X, Zhao YL, Sun GL, Zhao HX, Guo AG (2006) Functional properties of a new low-molecular-weight glutenin subunit gene from a bread wheat cultivar. Theor Appl Genet 113:1295–1303. doi:10.1007/s00122-006-0383-2

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

F. Chen, S. Liu and F. Zhao contributed equally to this work. The authors thank Dr. Zhonghu He (Institute of Crop Science, Chinese of Agricultural Sciences) for mixing test. This research was supported by grants from Ministry of Science and Technology of China (2006AA10Z173).

Author information

Authors and Affiliations

  1. The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education, School of Life Science, Shandong University, 250100, Jinan, People’s Republic of China

    Fanguo Chen, Shuwei Liu, Feng Zhao, Chunhui Xu & Guangmin Xia

Authors
  1. Fanguo Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuwei Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Feng Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chunhui Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Guangmin Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guangmin Xia.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, F., Liu, S., Zhao, F. et al. Molecular characterisation of the low-molecular weight glutenin subunit genes of tall wheatgrass and functional properties of one clone Ee34. Amino Acids 38, 991–999 (2010). https://doi.org/10.1007/s00726-009-0307-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-009-0307-y

Keywords

Search

Navigation