Identification and functional characterization of two acyl CoA:diacylglycerol acyltransferase 1 (DGAT1) genes from forage sorghum (Sorghum bicolor) embryo.

Elevating the lipid content in high-biomass forage crops has emerged as a new research platform for increasing energy density and improving livestock production efficiency associated with improved human health beneficial meat and milk quality. To gain insights of triacylglycerol (TAG) biosynthesis in forage sorghum, two type-1 diacylglycerol acyltransferase (designated as SbDGAT1-1 and SbDGAT1-2) were characterized for its in vivo function. SbDGAT1-2 is more abundantly expressed in embryo and bran during the early stage of the grain development in comparison to SbDGAT1-1. Heterologous expression of SbDGAT1 genes in TAG deficient H1246 strain restored the TAG accumulation capability with high substrate predilection towards 16:0, 16:1 and 18:1 fatty acids (FA). In parallel, we have identified N-terminal intrinsically disordered region (IDR) in SbDGAT1 proteins. To test the efficacy of the N-terminal region, truncated variants of SbDGAT1-1 (designated as SbDGAT1-1(39-515) and SbDGAT1-1(89-515)) were generated and expressed in yeast H1246 strain. Deletion in the N-terminal region resulted in decreased accumulation of TAG and FA (16:0 and 18:0) when compared to the SbDGAT1-1 variant expressed in yeast H1246 strain. The present study provides significant insight in forage sorghum DGAT1 gene function, useful for enhancing the green-forage TAG content through metabolic engineering.