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Artículo

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

Liu, Li; Ikeda, Tatsuya M.; Branlard, Gerard; Peña, Roberto J.; Rogers, William JohnIcon ; Lerner, Silvia E.; Kolman, Maria de Los AngelesIcon ; Xia, Xianchun; Wang, Linhai; Ma, Wujun; Appels, Rudi; Yoshida, Hisashi; Wang, Aili; Yan, Yueming; He, Zhonghu
Fecha de publicación: 06/2010
Editorial: Biomed Central
Revista: Bmc Plant Biology
ISSN: 1471-2229
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Biotecnología Agrícola y Biotecnología Alimentaria

Resumen

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.
Palabras clave: Bread Wheat , Low Molecular Weight Glutenin , Sds-Page , 2-De , Maldi-Tof-Ms , Pcr
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution 2.5 Unported (CC BY 2.5)
Identificadores
URI: http://hdl.handle.net/11336/15870
URL: http://www.biomedcentral.com/1471-2229/10/124
DOI: http://dx.doi.org/10.1186/1471-2229-10-124
Colecciones
Articulos(CCT - MAR DEL PLATA)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - MAR DEL PLATA
Articulos(INBIOTEC)
Articulos de INSTITUTO DE INV. EN BIODIVERSIDAD Y BIOTECNOLOGIA
Articulos(OCA PQUE. CENTENARIO)
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA PQUE. CENTENARIO
Citación
Liu, Li; Ikeda, Tatsuya M.; Branlard, Gerard; Peña, Roberto J.; Rogers, William John; et al.; Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat; Biomed Central; Bmc Plant Biology; 10; 6-2010; 124-148
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