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SUMMARY Bioengineering efforts to increase oil in non‐storage vegetative tissues, which constitute the majority of plant biomass, are promising sustainable sources of renewable fuels and feedstocks. While plants typically do not accumulate significant amounts of triacylglycerol (TAG) in vegetative tissues, we report here that the expression of a plastid‐localized phospholipase A1 protein, DEFECTIVE IN ANTHER DEHISCENCE1 (DAD1), led to a substantial increase in leaf TAG in Arabidopsis. Using an inducible system to control DAD1 expression circumvented growth penalties associated with overexpressing DAD1 and resulted in a rapid burst of TAG within several hours. The increase of TAG was accompanied by the formation of oil bodies in the leaves, petioles, and stems, but not in the roots. Lipid analysis indicated that the increase in TAG was negatively correlated with plastidial galactolipid concentration. The fatty acid (FA) composition of TAG predominantly consisted of 18:3. Expression of DAD1 in thefad3fad7fad8mutant, devoid of 18:3, resulted in comparable TAG accumulation with 18:2 as the major FA constituent, reflecting the flexiblein vivosubstrate use of DAD1. The transient expression of either Arabidopsis DAD1 orNicotiana benthamianaDAD1 (NbDAD1) inN. benthamianaleaves stimulated the accumulation of TAG. Similarly, transgenic soybeans expressing Arabidopsis DAD1 exhibited an accumulation of TAG in the leaves, showcasing the biotechnological potential of this technology. In summary, inducible expression of a plastidial lipase resulted in enhanced oil production in vegetative tissues, extending our understanding of lipid remodeling mediated by DAD1 and offering a valuable tool for metabolic engineering.
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Triacylglycerol stability limits futile cycles and inhibition of carbon capture in oil-accumulating leaves
Abstract Engineering plant vegetative tissue to accumulate triacylglycerols (TAG, e.g. oil) can increase the amount of oil harvested per acre to levels that exceed current oilseed crops. Engineered tobacco (Nicotiana tabacum) lines that accumulate 15% to 30% oil of leaf dry weight resulted in starkly different metabolic phenotypes. In-depth analysis of the leaf lipid accumulation and 14CO2 tracking describe metabolic adaptations to the leaf oil engineering. An oil-for-membrane lipid tradeoff in the 15% oil line (referred to as HO) was surprisingly not further exacerbated when lipid production was enhanced to 30% (LEAFY COTYLEDON 2 (LEC2) line). The HO line exhibited a futile cycle that limited TAG yield through exchange with starch, altered carbon flux into various metabolite pools and end products, and suggested interference of the glyoxylate cycle with photorespiration that limited CO2 assimilation by 50%. In contrast, inclusion of the LEC2 transcription factor in tobacco improved TAG stability, alleviated the TAG-to-starch futile cycle, and recovered CO2 assimilation and plant growth comparable to wild type but with much higher lipid levels in leaves. Thus, the unstable production of storage reserves and futile cycling limit vegetative oil engineering approaches. The capacity to overcome futile cycles and maintain enhanced stable TAG levels in LEC2 demonstrated the importance of considering unanticipated metabolic adaptations while engineering vegetative oil crops.
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- Award ID(s):
- 1829365
- PAR ID:
- 10495259
- Publisher / Repository:
- Oxford University Press
- Date Published:
- Journal Name:
- Plant Physiology
- ISSN:
- 0032-0889
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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