Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat
<strong>Background</strong> 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...
| Autores: | , , , , , , , , , , , , , , |
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| Formato: | artículo |
| Estado: | Versión enviada para evaluación y publicación |
| Fecha de publicación: | 2010 |
| País: | Argentina |
| Recursos: | Comisión de Investigaciones Científicas de la Provincia de Buenos Aires |
| Repositorio: | CIC Digital (CICBA) |
| Idioma: | inglés |
| OAI Identifier: | oai:digital.cic.gba.gob.ar:11746/7137 |
| Acesso em linha: | https://digital.cic.gba.gob.ar/handle/11746/7137 |
| Access Level: | acceso abierto |
| Palavra-chave: | Agronomía, reproducción y protección de plantas Common Wheat Glutenin Subunit Standard Cultivar Glutenin Protein Glutenin Extract |
| Resumo: | <strong>Background</strong> 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. <strong>Results</strong> At the<em>Glu-A3</em>locus, all seven alleles could be reliably identified by 2-DE and PCR. However, the alleles<em>Glu-A3e</em>and<em>Glu-A3d</em>could not be routinely distinguished from<em>Glu-A3f</em>and<em>Glu-A3g</em>, respectively, based on SDS-PAGE, and the allele<em>Glu-A3a</em>could not be differentiated from<em>Glu-A3c</em>by MALDI-TOF-MS. At the<em>Glu-B3</em>locus, alleles<em>Glu-B3a</em>,<em>Glu-B3b</em>,<em>Glu-B3c</em>,<em>Glu-B3g</em>,<em>Glu-B3h</em>and<em>Glu-B3j</em>could be clearly identified by all four methods, whereas<em>Glu-B3ab</em>,<em>Glu-B3ac</em>,<em>Glu-B3ad</em>could only be identified by the 2-DE method. At the<em>Glu-D3</em>locus, allelic identification was problematic for the electrophoresis based methods and PCR. MALDI-TOF-MS has the potential to reliably identify the<em>Glu-D3</em>alleles. <strong>Conclusions</strong> PCR is the simplest, most accurate, lowest cost, and therefore recommended method for identification of<em>Glu-A3</em>and<em>Glu-B3</em>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.<em>Glu-D3c</em>and<em>Glu-D3e</em>are the same allele. Two new alleles, namely,<em>Glu-D3m</em>in cultivar Darius, and<em>Glu-D3n</em>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. |
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