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Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat.
BMC Plant Biol. 2010 Jun 24; 10:124.BP

Abstract

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 x 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.

Authors+Show Affiliations

Institute of Crop Science, National Wheat Improvement Center/The National Key Facility for Crop Genetic Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences (CAAS), 12 Zhongguancun South Street, Beijing 100081, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20573275

Citation

Liu, Li, et al. "Comparison of Low Molecular Weight Glutenin Subunits Identified By SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in Common Wheat." BMC Plant Biology, vol. 10, 2010, p. 124.
Liu L, Ikeda TM, Branlard G, et al. Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat. BMC Plant Biol. 2010;10:124.
Liu, L., Ikeda, T. M., Branlard, G., Peña, R. J., Rogers, W. J., Lerner, S. E., Kolman, M. A., Xia, X., Wang, L., Ma, W., Appels, R., Yoshida, H., Wang, A., Yan, Y., & He, Z. (2010). Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat. BMC Plant Biology, 10, 124. https://doi.org/10.1186/1471-2229-10-124
Liu L, et al. Comparison of Low Molecular Weight Glutenin Subunits Identified By SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in Common Wheat. BMC Plant Biol. 2010 Jun 24;10:124. PubMed PMID: 20573275.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat. AU - Liu,Li, AU - Ikeda,Tatsuya M, AU - Branlard,Gerard, AU - Peña,Roberto J, AU - Rogers,William J, AU - Lerner,Silvia E, AU - Kolman,María A, AU - Xia,Xianchun, AU - Wang,Linhai, AU - Ma,Wujun, AU - Appels,Rudi, AU - Yoshida,Hisashi, AU - Wang,Aili, AU - Yan,Yueming, AU - He,Zhonghu, Y1 - 2010/06/24/ PY - 2009/12/12/received PY - 2010/06/24/accepted PY - 2010/6/25/entrez PY - 2010/6/25/pubmed PY - 2010/8/13/medline SP - 124 EP - 124 JF - BMC plant biology JO - BMC Plant Biol VL - 10 N2 - 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 x 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. SN - 1471-2229 UR - https://www.unboundmedicine.com/medline/citation/20573275/Comparison_of_low_molecular_weight_glutenin_subunits_identified_by_SDS_PAGE_2_DE_MALDI_TOF_MS_and_PCR_in_common_wheat_ L2 - https://bmcplantbiol.biomedcentral.com/articles/10.1186/1471-2229-10-124 DB - PRIME DP - Unbound Medicine ER -