Abstract
During the transformation of wild-type (WT) Arabidopsis thaliana, a T-DNA containing OLEOSIN-GFP (OLE1-GFP) was inserted by happenstance within the GBSS1 gene, resulting in significant reduction in amylose and increase in leaf oil content in the transgenic line (OG). The synergistic effect on oil accumulation of combining gbss1 with the expression of OLE1-GFP was confirmed by transforming an independent gbss1 mutant (GABI_914G01) with OLE1-GFP. The resulting OLE1-GFP/gbss1 transgenic lines showed higher leaf oil content than the individual OLE1-GFP/WT or single gbss1 mutant lines. Further stacking of the lipogenic factors WRINKLED1, Diacylglycerol O-Acyltransferase (DGAT1), and Cys-OLEOSIN1 (an engineered sesame OLEOSIN1) in OG significantly elevated its oil content in mature leaves to 2.3% of dry weight, which is 15 times higher than that in WT Arabidopsis. Inducible expression of the same lipogenic factors was shown to be an effective strategy for triacylglycerol (TAG) accumulation without incurring growth, development, and yield penalties.
Highlights
The oil triacylglycerol (TAG) is an energy-dense molecule that is essential for human nutrition and constitutes feedstocks for the production of biofuels and bioproducts [1].TAGs consist of three fatty acids esterified to a single molecule of glycerol
The transgenic line was generated by transforming Arabidopsis WT (WT)
At the borders of the insert site expression of GBSS1 in leaves of OLE1-GFP transgenic line (OG) showed that its expression was reduced by more than we identified a sequence encoding the 10th exon of GBSS1 (Figure 1D)
Summary
The oil triacylglycerol (TAG) is an energy-dense molecule that is essential for human nutrition and constitutes feedstocks for the production of biofuels and bioproducts [1]. TAGs consist of three fatty acids esterified to a single molecule of glycerol. Derived sugars provide carbon skeletons, energy, and reductant for fatty acid (FA) and TAG synthesis. Vegetative tissues, which constitute a major portion of plant biomass, have very low levels (typically less than 0.1% of dry weight) of TAG. Recent efforts in genetic and metabolic engineering have resulted in the successful accumulation of TAG in the leaves of both dicot and monocot crop plants [4,5]
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