Application of a high-throughput antibody-based assay for identification of the granule-bound starch synthase Wx-B1b allele in Australian wheat lines

Abstract

An enzyme-linked immunosorbent assay (ELISA) for the discrimination of Wx-B1a and Wx-B1b genotypes at the granule-bound starch synthase I (GBSSI) or waxy locus of hexaploid wheat (Triticum aestivum L.) was adapted to a high-throughput, 96-well microtitre plate format. This test is applicable to the direct analysis of starch, flour, or crushed grain and requires less than 1 grain to perform. Several hundred samples may be routinely analysed in one day. The assay was validated using quantitative trait locus (QTL) analysis of a doubled haploid mapping population of the cross Cranbrook (Wx-B1a)/Halberd (Wx-B1b). This demonstrated that the assay unambiguously identified 153 of 161 lines analysed,..