Abstract
A novel chimeric high-molecular-weight (HMW) glutenin subunit gene from a new common wheat line W958 (2n = 6x = 42) was isolated and characterized. SDS–PAGE analysis revealed that this glutenin subunit has similar electrophoretic mobility to 1Dx5, so it was designated 1Dx5′. Genomic DNA from W958 was amplified and a 2,505-bp fragment was obtained. The 1Dx5′ subunit showed a chimeric primary structure of 1Dx5 and 1Dx2, with the 1Dx5 sequence in the 5′ and middle repetitive regions and the 1Dx2 sequence in the repetitive domain and 3′ region. MALDI-TOF-MS analysis demonstrated that 1Dx5′ had a molecular weight of 86815.1 Da, close to that of an x-type glutenin subunit. Secondary structure analysis showed that this subunit had six helixes and one strand, including four helixes in the repetitive domain which could enhance the dough properties. Additionally, the promoter of 1Dx5′ was obtained and showed the same sequence as 1Dx5 or 1Dx2 except for a few base conversions. The promoter analysis indicated that the cis-acting regulatory elements of 1Dx5′ were the same as those of 1Dx5 and/or 1Dx2. Previously, we have demonstrated that this novel glutenin subunit is associated with good bread-making quality and comprises a very large proportion of the F2 segregation population. Consequently, we suggest that the amino acid residue composition and the secondary structure of the subunit may contribute to the bread-making quality. In summary, the novel 1Dx5′ gene could have greater potential in wheat quality improvement.
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References
Anderson OD, Greene FC (1989) The characterization and comparative analysis of high-molecular-weight glutenin genes from genomes A and B of a hexaploid wheat. Theor Appl Genet 77:689–700
Arguello JR, Chen Y, Yang S, Wang W, Long M (2006) Origination of an X-linked testes chimeric gene by illegitimate recombination in Drosophila. PLoS Genet 2:745–754
Belton PS (1999) Mini review: on the elasticity of wheat gluten. J Cereal Sci 29:103–107
Bryson K, McGuffin LJ, Marsden RL, Ward JJ, Sodhi JS, Jones DT (2005) Protein structure prediction servers at University College London. Nucleic Acids Res 33(Web Server issue):W36–W38
Ciaffi M, Lafiandra D, Porceddu E, Benedettelli S (1993) Storage-protein variation in wild emmer (Triticum turgidum ssp. dicoccoides) from Jordan and Turkey. Theor Appl Genet 86:518–525
Cloutier S, Rampitsch C, Penner GA, Lukow OM (2001) Cloning and expression of a LMW-i glutenin gene. J Cereal Sci 33:143–154
FAO (2009) FAO statistical year book 2008–2009. Web Edition
Gupta RB, MacRitchie F (1994) Allelic variation at glutenin subunit and gliadin loci, Glu-1, Glu-3 and Gli-1, of common wheats. II. Biochemical basis of the allelic effects on dough properties. J Cereal Sci 19:19–29
Hedgcoth C, El-Shehawi AM, Wei P, Clarksonand M, Tamalis D (2002) A chimeric open reading frame associated with cytoplasmic male sterility in alloplasmic wheat with Triticum timopheevi mitochondria is present in several Triticum and Aegilops species, barley, and rye. Curr Genet 41:357–365
Hickman DR, Roepstorff P, Shewry PR, Tatham AS (1995) Molecular weights of high molecular weight subunits of glutenin determined by mass spectrometry. J Cereal Sci 22:99–103
Lafiandra D, Ovidio R, Porceddu E, Margiotta B, Colaprico G (1993) New data supporting high Mr glutenin subunit 5 as the determinant of quality differences among the pairs 5 + 10 vs 2 + 12. J Cereal Sci 18:197–205
Li X, Ma W, Gao L, Zhang Y, Wang A, Ji K, Wang K, Appels R, Yan Y (2008) A novel chimeric low-molecular-weight glutenin subunit gene from the wild relatives of wheat Aegilops kotschyi and Ae. juvenalis: evolution at the Glu-3 loci. Genetics 180:93–101
Liu S, Zhao F, Gao X, Chen F, Xia G (2010) A novel high molecular weight glutenin subunit from Australopyrum retrofractum. Amino Acids. doi:10.1007/s00726-009-0450-5
Lumbreras V, Alba MM, Kleinow T, Koncz C, Pagès M (2001) Domain fusion between SNF1-related kinase subunits during plant evolution. EMBO Rep 2(1):55–60
Mackie AM, Lagudah ES, Sharp PJ, Lafiandra D (1996) Molecular and biochemical characterisation of HMW glutenin subunits from T. tauschii and the D genome of hexaploid wheat. J Cereal Sci 23:213–225
Nagy IJ, Takács I, Juhász A, Tamás L, Bedo Z (2005) Identification of a new class of recombinant prolamin genes in wheat. Genome 48:840–847
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
Ren Y, Wang T, Xu ZB, Yang ZJ, Ren ZL (2008) Molecular characterization of a novel HMW-GS 1Dx5′ associated with good bread making quality (Triticum aestivum L.) and the study of its unique inheritance. Genet Resour Crop Evol 55:585–592
Shewry PR, Halford NG (2002) Cereal seed storage proteins: structures, properties and role in grain utilization. J Exp Bot 53:947–958
Shewry PR, Halford NG, Tatham AS (1992) High molecular weight subunits of wheat glutenin. J Cereal Sci 15:105–120
Shewry PR, Halford NG, Tatham AS, Popineau Y, Lafiandra D, Belton PS (2003) The high molecular weight subunits of wheat glutenin and their role in determining wheat processing properties. Adv Food Nutr Res 45:219–302
Sugiyama T, Rafalski A, Peterson D, Sol D (1985) A wheat HMW glutenin subunit gene reveals a highly repeated structure. Nucleic Acid Res 13:8729–8737
Sun M, Yan Y, Jiang Y, Xiao Y, Hu Y, Cai M, Li Y, Hsam SL, Zeller FJ (2004) Molecular cloning and comparative analysis of a y-type inactive HMW glutenin subunit gene from cultivated emmer wheat (Triticum dicoccum L.). Hereditas 141:46–54
Wan Y, Wang D, Shewry R, Halford G (2002) Isolation and characterization of five novel high molecular weight subunit of glutenin genes from Triticum timopheevi and Aegilops cylindrica. Theor Appl Genet 104:828–839
Wrigley CW (1996) Giant proteins with flour power. Nature 381:738–739
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-Dt1 glutenin gene of Aegilops tauschii. Theor Appl Genet 108:1349–1358
Zhang Y, Li Q, Yan Y, Zheng J, An X, Xiao Y, Wang A, Pei Y, Wang H, Hsam SL, Zeller FJ (2006) Molecular characterization and phylogenetic analysis a novel glutenin gene (Dy10.1t) from Aegilops tauschii. Genome 49:735–774
Zhang YZ, Li XH, Wang AL, An XL, Zhang Q, Pei YH, Gao LY, Ma WJ, Appels R, Yan YM (2008) Novel x-type HMW glutenin genes from Aegilops tauschii and their implications on the wheat origin and evolution mechanism of Glu-D1-1 proteins. Genetics 178:23–33
Acknowledgments
This work was supported by the Knowledge Innovation Program of the Chinese Academy of Sciences (Grant No. KSCX2-YW-N-046) and the National Key Project of Transgenic Biologic Varieties Breeding of China (Grant No.2008ZX08009-003).
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Figure S1. Multiple alignments of promoter region of 1Dx5′, 1Dx5 and 1Dx2. The consensus bases were marked by stars (PDF 15 kb)
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Feng, B., An, X., Xu, Z. et al. Molecular cloning of a novel chimeric HMW glutenin subunit gene 1Dx5′ from a common wheat line W958. Mol Breeding 28, 163–170 (2011). https://doi.org/10.1007/s11032-010-9470-9
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DOI: https://doi.org/10.1007/s11032-010-9470-9