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Background  Low-molecular-weight glutenin subunits (LMW-GS) play a crucial role in determining end-use quality of common wheat by influencing the viscoelastic properties of dough. Four different methods - sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE, IEF × SDS-PAGE), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and polymerase chain reaction (PCR), were used to characterize the LMW-GS composition in 103 cultivars from 12 countries.    Results  At the  Glu-A3  locus, all seven alleles could be reliably identified by 2-DE and PCR. However, the alleles  Glu-A3e  and  Glu-A3d  could not be routinely distinguished from  Glu-A3f  and  Glu-A3g , respectively, based on SDS-PAGE, and the allele  Glu-A3a  could not be differentiated from  Glu-A3c  by MALDI-TOF-MS. At the  Glu-B3  locus, alleles  Glu-B3a ,  Glu-B3b ,  Glu-B3c ,  Glu-B3g ,  Glu-B3h  and  Glu-B3j  could be clearly identified by all four methods, whereas  Glu-B3ab ,  Glu-B3ac ,  Glu-B3ad  could only be identified by the 2-DE method. At the  Glu-D3  locus, allelic identification was problematic for the electrophoresis based methods and PCR. MALDI-TOF-MS has the potential to reliably identify the  Glu-D3  alleles.    Conclusions  PCR is the simplest, most accurate, lowest cost, and therefore recommended method for identification of  Glu-A3  and  Glu-B3  alleles in breeding programs. A combination of methods was required to identify certain alleles, and would be especially useful when characterizing new alleles. A standard set of 30 cultivars for use in future studies was chosen to represent all LMW-GS allelic variants in the collection. Among them, Chinese Spring, Opata 85, Seri 82 and Pavon 76 were recommended as a core set for use in SDS-PAGE gels.  Glu-D3c  and  Glu-D3e  are the same allele. Two new alleles, namely,  Glu-D3m  in cultivar Darius, and  Glu-D3n  in Fengmai 27, were identified by 2-DE. Utilization of the suggested standard cultivar set, seed of which is available from the CIMMYT and INRA Clermont-Ferrand germplasm collections, should also promote information sharing in the identification of individual LMW-GS and thus provide useful information for quality improvement in common wheat.

Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat