Characterization of a near isogenic barley line with high grain β-amylase activity reveals a separation in the tight co-regulation of B-hordeins (Hor2) with endosperm-specific β-amylase (Bmy1).

GSHO 2096 is a near isogenic barley line with extremely high grain β-amylase activity, a desirable trait in the malting and brewing industry. High levels of grain β-amylase activity are caused by a surge in endosperm-specific β-amylase (Bmy1) gene expression during the early stages of grain development with high expression levels persisting throughout development. Origins of the high β-amylase activity trait are perplexing considering GSHO 2096 is not supposed to have grain β-amylase activity. GSHO 2096 is reported to be derived from a Bowman x Risø 1508 cross followed by recurrent backcrossing to Bowman (BC5). Risø 1508 carries a mutated form of the barley prolamin binding factor, which is responsible for Bmy1 expression during grain development. Thus, the pedigree of GSHO 2096 was explored to determine the potential origins of the high grain β-amylase trait. Genotyping using the barley 50k iSelect SNP array revealed Bowman and GSHO 2096 were very similar (95.4 %) and provided evidence that both Risø 56 and 1508 are in the pedigree. Risø mutants 56 and 1508 both have perturbed hordein gene expression leading to a discernable pattern using SDS-PAGE. GSHO 2096 and Risø 56 have the same hordein pattern whereas Bowman and Risø 1508 have unique patterns. RNAseq revealed that Hor2 (B-hordein) gene expression was completely downregulated making it unique as the only known line with Bmy1 expression without Hor2 co-expression. Regardless of pedigree, GSHO 2096 remains an extremely valuable high β-amylase activity line with potential utilization in breeding for malt quality.