Analysis of β-d-glucan biosynthetic genes in oat reveals glucan synthesis regulation by light

Abstract

Background and Aims: Oat (Avena sativa) has human health benefits when consumed as a whole-grain food, attributed to the high content of (1,3;1,4)-β-d-glucan (mixed-linkage glucan [MLG]), but little is known about the synthase genes and synthesis mechanism of MLG polysaccharides in this species. Methods: The concentration of oat MLGs under different light intensities was measured by a standard enzymatic approach and further verified by immunoelectron microscopy. The effect of light intensity on MLG synthase genes was examined by RT-qPCR and western blot analyses. The pattern of expression directed by the promoter of the oat MLG synthase gene was also investigated by histochemical β-glucuronidase (GUS) analysis. Key Results: The oat orthologues of genes implicated in the synthesis of MLG in other cereals, including cellulose synthase-like (Csl) F, H and J gene families, were defined. Transcript profiling of these genes across oat tissues indicated that AsCslF6 transcripts dominated. Under high light intensities, the expression of AsCslF6, a major isoform of the MLG synthase genes, increased to >30 % of the dark growth control. The amount of MLG in oat rose from 0.07 to 1.06 % with increased light intensity. Histochemical tests showed that the AsCslF6 gene promoter preferentially directs GUS expression under high light intensity conditions. Conclusions: Oat MLG synthesis is regulated by light. High light intensity upregulates the expression of the MLG synthase AsCslF6 gene, leading to an increase in the amount of MLG in oat leaves.