Fig. 1. A schematic representation of the conventional VLC-PUFA biosynthetic pathway from linoleic and α-linolenic acid precursors. The alternative Δ8-pathway.

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Fig. 1. A schematic representation of the conventional VLC-PUFA biosynthetic pathway from linoleic and α-linolenic acid precursors. The alternative Δ8-pathway (not show in Fig. 1) utilizes a Δ9-elongase and Δ8-desaturase to produce DGLA and ETA, respectively. From: Metabolic engineering of the omega-3 long chain polyunsaturated fatty acid biosynthetic pathway into transgenic plants J Exp Bot. 2012;63(7):2397-2410. doi:10.1093/jxb/err454 J Exp Bot | © The Author [2012]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 2. A schematic representation of the main lipid classes and biochemical pathways involved in the production of triacylglycerols. The acyl-CoA pool is shown as the central route for TAG synthesis; however, it is clear that fatty acids are routed through PC/DAG and directly from PC; the influence of each route varying with species. The primary activities (see www. arabidopsisacyllipids.plantbiology.msu.edu/pathways for a full description of Arabidopsis acyl lipid metabolism) are: glycerol-3-phosphate acyltransferase (GPAT); 1-acylglycerol-3-phosphate acyltransferase (LPAAT); phosphatidate phosphatase (PP); phosphatidylcholine:diacylglycerol cholinephosphotransferase (PDCT); diacylglycerol cholinephosphotransferase (DAG-CPT); acyl-CoA:diacylglycerol acyltransferase (DGAT); phospholipid:diacylglycerol acyltransferase (PDAT); and 1-acylglycerol-3-phosphocholine acyltransferase (LPCAT). From: Metabolic engineering of the omega-3 long chain polyunsaturated fatty acid biosynthetic pathway into transgenic plants J Exp Bot. 2012;63(7):2397-2410. doi:10.1093/jxb/err454 J Exp Bot | © The Author [2012]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 3. The application of emerging technology to the analysis of lipids in Arabidopsis seed expressing the VLC-PUFA biosynthetic pathway. Firstly the separation and identification of acyl-CoA species by LC-MS/MS. (A) Total ion count for multireaction monitoring (MRM); (B) extracted MRM traces for EPA and DHA allowing the quantification of co-eluting acyl-CoA species (D). The analysis of the acyl-CoA pool allows unambiguous identification of individual acyl-CoA species and therefore the efficacy of both desaturase and elongase genes in planta (transgenic Arabidopsis plants are described in Sayanova et al., 2011b). Secondly the use of ESI-MS/MS systematically to characterize discrete lipid pools enumerates each iterative engineering attempt (C; PC head group positive precursor ion scan for engineered and wild-type mature Arabidopsis seed). From: Metabolic engineering of the omega-3 long chain polyunsaturated fatty acid biosynthetic pathway into transgenic plants J Exp Bot. 2012;63(7):2397-2410. doi:10.1093/jxb/err454 J Exp Bot | © The Author [2012]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com