Abstract
Increasing the yield of oilseed crops is an important objective for biotechnologists. A number of individual genes involved in triacylglycerol metabolism have previously been reported to enhance the oil content of seeds when their expression is altered. However, it has yet to be established whether specific combinations of these genes can be used to achieve an additive effect and whether this leads to enhanced yield. Using Arabidopsis (Arabidopsis thaliana) as an experimental system, we show that seed-specific overexpression of WRINKLED1 (a transcriptional regulator of glycolysis and fatty acid synthesis) and DIACYLGLYCEROL ACYLTRANSFERASE1 (a triacylglycerol biosynthetic enzyme) combined with suppression of the triacylglycerol lipase SUGAR-DEPENDENT1 results in a higher percentage seed oil content and greater seed mass than manipulation of each gene individually. Analysis of total seed yield per plant suggests that, despite a reduction in seed number, the total yield of oil is also increased.
Highlights
Increasing the yield of oilseed crops is an important objective for biotechnologists
Previous studies have shown that overexpression of the WRI1 transcription factor activates the expression of multiple genes associated with lower glycolysis and fatty acid synthesis (Baud et al, 2007; To et al, 2012) and can enhance oil content in seeds of Arabidopsis (Cernac and Benning, 2004)
To test whether combined expression of WRI1 and DIACYLGLYCEROLACYL TRANSFERASE1 (DGAT1) can have an additive effect on seed oil content, and whether this might be further augmented by blocking TAG turnover by RNA interference (RNAi) suppression of the lipase SUGAR-DEPENDENT1 (SDP1; Kelly et al, 2013a; Kim et al, 2014), transfer DNA (T-DNA) constructs were designed for the manipulation of each gene under the control of a seed-specific promoter (Fig. 1B)
Summary
Increasing the yield of oilseed crops is an important objective for biotechnologists. An alternative approach to targeting individual enzymes is to manipulate the expression of transcriptional master regulators that govern the expression of multiple enzymes in the metabolic pathway (Broun, 2004; Grotewold, 2008) This too has proven to be an effective means to enhance seed oil content (Cernac and Benning, 2004; Shen et al, 2010). There are currently no academic reports of gene stacking to enhance seed oil content, several recent studies have successfully applied this approach to manipulate oil metabolism in leaves (Sanjaya et al, 2011; Fan et al, 2013; Kelly et al, 2013b; Vanhercke et al, 2013, 2014; Winichayakul et al, 2013) In general these studies suggest that a push-pull-protect strategy may be effective (Fig. 1A).
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