Allelic effects and variations for key bread-making quality genes in bread wheat using high-throughput molecular markers

We developed and validated high-throughput Kompetitive Allele-Specific PCR (KASP) assays for key genes underpinning bread-making quality, including the wbm gene on chromosome 7AL and over-expressed glutenin Bx7(OE) (Glu-B1 al) gene. Additionally, we used pre-existing KASP assay for Secl (1B.1R translocation) gene on chromosome 1B. The newly developed KASP assays were compared with gel-based markers for reliability and phenotypically validated in a diversity panel for Mixograph, Rapid Visco Analyzer (RVA) and Mixolab traits. Genotypes carrying the 1B.1R translocation had significantly lower Mixolab parameters than those without the translocation. Similarly, superior effects of the wbm+ and Bx7(OE) alleles on Mixograph and RVA properties and their extremely low frequencies in global wheat collections supported the idea of using these genes for bread making quality improvement. The allele frequencies of wbm+ and Bx7(OE) were extremely low in historical Chinese and CIMMYT wheat germplasm, but were relatively higher in synthetic hexaploid wheats and their breeding derivatives. In both the Vavilov and Watkins global landrace collections, the frequency of wbm+ was 6.4 and 3.5%, and frequency of Bx7(OE) was 3.2% and 7.0%, respectively. The high-throughput marker resources and large-scale global germplasm screening provided further opportunities to exploit these genes in wheat breeding to enhance bread-making quality.