Theoretical and Applied Genetics

, Volume 109, Issue 5, pp 1064–1069 | Cite as

Development and validation of a PCR-based marker assay for negative selection of the HMW glutenin allele Glu-B1-1d (Bx-6) in wheat

Original Paper

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.

Notes

Acknowledgements

German wheat cultivars were kindly provided by Cebeco Saaten GmbH, Dr. J. Ackermann & Co., H.P. Peterson GmbH & Co. KG, IG Saatzucht GmbH & Co. KG, Lochow Petkus GmbH, Monsanto Deutschland GmbH, Nickerson GmbH, Nordsaat Saatzucht GmbH, Pflanzenzucht Dr. H.C. Carsten Inh. Erhardt Eger KG, Saatzucht Engelen-Büchling oHG, Saatzucht Firlbeck GmbH & Co. KG, Saatzucht Hadmersleben GmbH, Saatzucht Hans Schweiger & Co. oHG, Saatzucht Josef Breun GbR, Saatzuchtgesellschaft Streng’s Erben GmbH & Co. KG, Strube Saatzucht KG and Südwestdeutsche Saatzucht Dr. Hans-Rolf Späth. Durum wheat cultivars were kindly provided by Sai L.K. Hsam. We thank Berta Killermann (Section for Seed Certification of the Bavarian State Research Center for Agronomy) for providing the data for HMW glutenin subunit composition.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.EpiGene GmbHBiotechnology in Plant ProtectionFreisingGermany
  2. 2.Lehrstuhl für Pflanzenbau und Pflanzenzüchtung, Department Pflanzenwissenschaften, Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und UmweltTechnische Universität MünchenFreising-WeihenstephanGermany

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