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Multiplex-PCR typing of high molecular weight glutenin alleles in wheat

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Abstract

In Australian commercial cultivars, each high molecular weight glutenin (Glu-1) homoeologous locus consists of one of two predominant alleles: Glu-A1a (subunit Ax1) or Glu-A1b (subunit Ax2*) at the GluA1 locus, Glu-B1b (Bx7 and By8 subunits) or Glu-B1i (Bx17 and By18 subunits) at the Glu-B1 locus, and Glu-D1d (Dx5 and Dy10 subunits) or Glu-D1a (Dx2 and Dy12 subunits) at the Glu-D1 locus. PCR-based assays have been developed in this study to discriminate between these common alleles at each locus. Primers specific for the Glu-A1 Ax2* gene give a single fragment of 1319 bp only in the presence of this gene. Primers targeting the Glu-B1 locus resulted in a co-dominant marker for which the Bx7 genotype produced two fragments (630 bp and 766 bp) and the Bx17 genotype a single fragment (669 bp). The third pair of primers was specific for the Dx5 gene and resulted in a single band of 478 bp. A multiplexed PCR assay was established which permitted the discrimination of the major HMW glutenins in a single PCR reaction and agarose gel assay. As the HMW glutenin composition of a wheat line is extremely important in determining the functional properties of wheat gluten, these markers are useful for the purposes of marker-assisted breeding. These markers may also be useful for the purpose of DNA-based identification of wheat varieties.

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  1. Commonwealth Scientific and Industrial Research Organisation, Plant Industry, GPO Box 1600, Canberra, ACT, 2601, Australia

    W. Ma, W. Zhang & K.R. Gale

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  1. W. Ma
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  2. W. Zhang
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  3. K.R. Gale
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Ma, W., Zhang, W. & Gale, K. Multiplex-PCR typing of high molecular weight glutenin alleles in wheat. Euphytica 134, 51–60 (2003). https://doi.org/10.1023/A:1026191918704

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