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Development and validation of a PCR-based marker assay for negative selection of the HMW glutenin allele Glu-B1-1d (Bx-6) in wheat.
Theor Appl Genet. 2004 Sep; 109(5):1064-9.TA

Abstract

Polymorphisms between the coding sequences of high-molecular-weight (HMW) glutenin x-type genes at the Glu-1 locus were used to amplify Glu-1B x-type-specific PCR fragments. PCR analysis in a wheat cultivar subset carrying different Glu-1B x-type alleles resulted in PCR fragments that differed in size for Glu-B1-1d (B-x6) and non -Glu-B1-1d (B-x6) genotypes. Subsequent sequencing analysis revealed a 15-bp in-frame insertion in the coding regions of all Glu-B1-1d (B-x6) genotypes which allowed the development of a B-x6-specific PCR assay for high-throughput allele sizing by ion-pair reversed-phase high-performance liquid chromatography. The assay was validated in a set of 86 German wheat cultivars, and genotyping data unequivocally verified the presence of HMW glutenin subunits GLU-B1-1D (Bx-6) + GLU-B1-2A (By-8) by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. These results demonstrate that the PCR assay can be applied for the detection and negative selection of the 'poor breadmaking quality' Glu-B1-1d (B-x6) alleles in wheat breeding programs.

Authors+Show Affiliations

Schwarz G
EpiGene GmbH, Biotechnology in Plant Protection, Hohenbachernstrasse 19-21, 85354 Freising, Germany. gschwarz@wzw.tum.de
Felsenstein FG
No affiliation info available
Wenzel G
No affiliation info available

MeSH

AllelesBase SequenceBreedingChromatography, High Pressure LiquidChromosome MappingElectrophoresis, Polyacrylamide GelGenetic MarkersGlutensMolecular Sequence DataPolymerase Chain ReactionPolymorphism, GeneticSelection, GeneticSequence AlignmentSequence Analysis, DNATriticum

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

15175854

Citation

Schwarz, G, et al. "Development and Validation of a PCR-based Marker Assay for Negative Selection of the HMW Glutenin Allele Glu-B1-1d (Bx-6) in Wheat." TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik, vol. 109, no. 5, 2004, pp. 1064-9.
Schwarz G, Felsenstein FG, Wenzel G. Development and validation of a PCR-based marker assay for negative selection of the HMW glutenin allele Glu-B1-1d (Bx-6) in wheat. Theor Appl Genet. 2004;109(5):1064-9.
Schwarz, G., Felsenstein, F. G., & Wenzel, G. (2004). Development and validation of a PCR-based marker assay for negative selection of the HMW glutenin allele Glu-B1-1d (Bx-6) in wheat. TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik, 109(5), 1064-9.
Schwarz G, Felsenstein FG, Wenzel G. Development and Validation of a PCR-based Marker Assay for Negative Selection of the HMW Glutenin Allele Glu-B1-1d (Bx-6) in Wheat. Theor Appl Genet. 2004;109(5):1064-9. PubMed PMID: 15175854.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and validation of a PCR-based marker assay for negative selection of the HMW glutenin allele Glu-B1-1d (Bx-6) in wheat. AU - Schwarz,G, AU - Felsenstein,F G, AU - Wenzel,G, Y1 - 2004/06/03/ PY - 2004/01/22/received PY - 2004/05/01/accepted PY - 2004/6/4/pubmed PY - 2004/12/22/medline PY - 2004/6/4/entrez SP - 1064 EP - 9 JF - TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik JO - Theor Appl Genet VL - 109 IS - 5 N2 - Polymorphisms between the coding sequences of high-molecular-weight (HMW) glutenin x-type genes at the Glu-1 locus were used to amplify Glu-1B x-type-specific PCR fragments. PCR analysis in a wheat cultivar subset carrying different Glu-1B x-type alleles resulted in PCR fragments that differed in size for Glu-B1-1d (B-x6) and non -Glu-B1-1d (B-x6) genotypes. Subsequent sequencing analysis revealed a 15-bp in-frame insertion in the coding regions of all Glu-B1-1d (B-x6) genotypes which allowed the development of a B-x6-specific PCR assay for high-throughput allele sizing by ion-pair reversed-phase high-performance liquid chromatography. The assay was validated in a set of 86 German wheat cultivars, and genotyping data unequivocally verified the presence of HMW glutenin subunits GLU-B1-1D (Bx-6) + GLU-B1-2A (By-8) by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. These results demonstrate that the PCR assay can be applied for the detection and negative selection of the 'poor breadmaking quality' Glu-B1-1d (B-x6) alleles in wheat breeding programs. SN - 0040-5752 UR - https://www.unboundmedicine.com/medline/citation/15175854/Development_and_validation_of_a_PCR_based_marker_assay_for_negative_selection_of_the_HMW_glutenin_allele_Glu_B1_1d__Bx_6__in_wheat_ DB - PRIME DP - Unbound Medicine ER -