Running the numbers on plant synthetic biology solutions to global problems

Gelder, Kristen Van; Oliveira-Filho, Edmar R.; Messina, Carlos D.; Venado, Rafael E.; Wilker, Jennifer; Rajasekar, Shanmugam; Amthor, Jeffrey S.; Hanson, Andrew D.

doi:10.1016/j.plantsci.2023.111815

Cited by 7 publications

(2 citation statements)

References 63 publications

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“…Biodiesel derived from lipid accumulating energycane is an attractive approach to minimize the use of food and feed crops in the production of renewable transportation fuels. Additional feedstocks and engineering strategies will be needed to establish a robust pipeline for production of sustainable aviation fuel [ 96 ].…”

Section: Discussionmentioning

confidence: 99%

Intron-mediated enhancement of DIACYLGLYCEROL ACYLTRANSFERASE1 expression in energycane promotes a step change for lipid accumulation in vegetative tissues

Cao,

Luo,

Korynta

et al. 2023

Biotechnol Biofuels

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Background Metabolic engineering for hyperaccumulation of lipids in vegetative tissues is a novel strategy for enhancing energy density and biofuel production from biomass crops. Energycane is a prime feedstock for this approach due to its high biomass production and resilience under marginal conditions. DIACYLGLYCEROL ACYLTRANSFERASE (DGAT) catalyzes the last and only committed step in the biosynthesis of triacylglycerol (TAG) and can be a rate-limiting enzyme for the production of TAG. Results In this study, we explored the effect of intron-mediated enhancement (IME) on the expression of DGAT1 and resulting accumulation of TAG and total fatty acid (TFA) in leaf and stem tissues of energycane. To maximize lipid accumulation these evaluations were carried out by co-expressing the lipogenic transcription factor WRINKLED1 (WRI1) and the TAG protect factor oleosin (OLE1). Including an intron in the codon-optimized TmDGAT1 elevated the accumulation of its transcript in leaves by seven times on average based on 5 transgenic lines for each construct. Plants with WRI1 (W), DGAT1 with intron (Di), and OLE1 (O) expression (WDiO) accumulated TAG up to a 3.85% of leaf dry weight (DW), a 192-fold increase compared to non-modified energycane (WT) and a 3.8-fold increase compared to the highest accumulation under the intron-less gene combination (WDO). This corresponded to TFA accumulation of up to 8.4% of leaf dry weight, a 2.8-fold or 6.1-fold increase compared to WDO or WT, respectively. Co-expression of WDiO resulted in stem accumulations of TAG up to 1.14% of DW or TFA up to 2.08% of DW that exceeded WT by 57-fold or 12-fold and WDO more than twofold, respectively. Constitutive expression of these lipogenic “push pull and protect” factors correlated with biomass reduction. Conclusions Intron-mediated enhancement (IME) of the expression of DGAT resulted in a step change in lipid accumulation of energycane and confirmed that under our experimental conditions it is rate limiting for lipid accumulation. IME should be applied to other lipogenic factors and metabolic engineering strategies. The findings from this study may be valuable in developing a high biomass feedstock for commercial production of lipids and advanced biofuels. Graphical abstract

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Section: Discussionmentioning

confidence: 99%

Intron-mediated enhancement of DIACYLGLYCEROL ACYLTRANSFERASE1 expression in energycane promotes a step change for lipid accumulation in vegetative tissues

Cao,

Luo,

Korynta

et al. 2023

Biotechnol Biofuels

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“…This could make a significant contribution to meeting the increasing demand for diesel fuels with reduced CO 2 emissions. Additional feedstocks and synthetic biology strategies are required to provide a robust supply of sustainable aviation fuel (Van Gelder et al, 2023)…”

Section: Discussionmentioning

confidence: 99%

Triacylglycerol, total fatty acid, and biomass accumulation of metabolically engineered energycane grown under field conditions confirms its potential as feedstock for drop‐in fuel production

Cao,

Kannan,

Luo

et al. 2023

GCB Bioenergy

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Metabolic engineering for hyperaccumulation of lipids in vegetative tissues of high biomass crops promises a step change in oil yields for the production of advanced biofuels. Energycane is the ideal feedstock for this approach due to its exceptional biomass production and persistence under marginal conditions. Here, we evaluated metabolically engineered energycane with constitutive expression of the lipogenic factors WRINKLED1 (WRI1), DIACYLGLYCEROL ACYLTRANSFERASE1 (DGAT1), and OLEOSIN1 (OLE1) for the accumulation of triacylglycerol (TAG), total fatty acid (TFA), and biomass under field conditions at the University of Florida‐IFAS experiment station near Citra, Florida. TAG and TFA accumulation were highest in leaves (up to 9.9% and 12.9% of DW, respectively), followed by juice from crushed stems, stems, and roots. TAG and TFA accumulation increased up to harvest time and correlated highest with OLE1 and DGAT1 expression. Biomass dry weight, TAG, and TFA content differed greatly depending on DGAT1 and OLE1 expression in transgenic lines with similar WRI1 expression. Biomass did not significantly differ between WT and line L2 with DAGT1 and OLE1 expressed at low levels and TAG and TFA accumulating to 12‐ and 1.6‐fold that of WT leaves, respectively. In contrast, line L13, with intron‐mediated enhancement of DGAT1 expression, displayed a 245‐ to 330‐fold increase in TAG and a 4.75‐ to 6.45‐fold increase in TFA content compared with WT leaves and a biomass reduction of 52%. These results provide the basis for developing novel feedstocks for expanding plant lipid production and point to new prospects for advanced biofuels.

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Root mucilage nitrogen for rhizosphere microorganisms under drought

Nazari,

Bickel,

Kuzyakov

et al. 2024

Biol Fertil Soils

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Nitrogen (N) is a crucial nutrient for the growth and activity of rhizosphere microorganisms, particularly during drought conditions. Plant root-secreted mucilage contains N that could potentially nourish rhizosphere microbial communities. However, there remains a significant gap in understanding mucilage N content, its source, and its utilization by microorganisms under drought stress. In this study, we investigated the impact of four maize varieties (DH02 and DH04 from Kenya, and Kentos and Keops from Germany) on the secretion rates of mucilage from aerial roots and explored the origin of mucilage N supporting microbial life in the rhizosphere. We found that DH02 exhibited a 96% higher mucilage secretion rate compared to Kentos, while Keops showed 114% and 89% higher secretion rates compared to Kentos and DH04, respectively. On average, the four maize varieties released 4 μg N per root tip per day, representing 2% of total mucilage secretion. Notably, the natural abundance of 15N isotopes increased (higher δ15N signature) with mucilage N release. This indicates a potential dilution of the isotopic signal from biological fixation of atmospheric N by mucilage-inhabiting bacteria as mucilage secretion rates increase. We proposed a model linking mucilage secretion to a mixture of isotopic signatures and estimated that biological N fixation may contribute to 45 - 75% of mucilage N per root tip. The N content of mucilage from a single maize root tip can support a bacterial population ranging from 107 to 1010 cells per day. In conclusion, mucilage serves as a significant N-rich resource for microbial communities in the rhizosphere during drought conditions.

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Running the numbers on plant synthetic biology solutions to global problems

Cited by 7 publications

References 63 publications

Intron-mediated enhancement of DIACYLGLYCEROL ACYLTRANSFERASE1 expression in energycane promotes a step change for lipid accumulation in vegetative tissues

Intron-mediated enhancement of DIACYLGLYCEROL ACYLTRANSFERASE1 expression in energycane promotes a step change for lipid accumulation in vegetative tissues

Triacylglycerol, total fatty acid, and biomass accumulation of metabolically engineered energycane grown under field conditions confirms its potential as feedstock for drop‐in fuel production

Root mucilage nitrogen for rhizosphere microorganisms under drought

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