PPT1-mediated plastidic phosphoenolpyruvate import enhances fatty acid biosynthesis in sugar-rich tissues

Wang, Jiang; Lin, Yi‐Hsuan; Xue, Xueyi; Beuchat, Gabriel; Li, Yaxin; Li, Jiankun; Chen, Li‐Qing

doi:10.1111/nph.71160

Title: PPT1-mediated plastidic phosphoenolpyruvate import enhances fatty acid biosynthesis in sugar-rich tissues

Journal Article · Tue Apr 07 00:00:00 UTC 2026 · New Phytologist

DOI: https://doi.org/10.1111/nph.71160 · OSTI ID:3030837

^[1];

^[1]

Univ. of Illinois at Urbana-Champaign, IL (United States)

Metabolic engineering of vegetative tissues for lipid production holds transformative potential for sustainable biofuels, yet achieving sufficient yields remains challenging. Here, we present a strategy to enhance fatty acid synthesis by redirecting cytosolic phosphoenolpyruvate (PEP) into plastids through overexpression of the plastidial phosphoenolpyruvate/phosphate translocator (PPT1) in vegetative tissues of Arabidopsis thaliana. Integrated metabolomic and transcriptomic analyses revealed that AtPPT1 overexpression alleviated metabolite overaccumulation in high-sugar tissues, consistent with enhanced carbon flux coordination between the cytosol and chloroplast. Notably, phosphofructokinase activity, a key step in glycolysis, was elevated, linking plastidic PEP import to increased glycolytic throughput. In Arabidopsis, overexpression of AtPPT1 increased fatty acid content and lipid droplet accumulation in the sugar-accumulating sweet11;12;13 mutant, but not in wild-type Col-0. Together, these findings establish PEP redirection as an effective strategy to boost fatty acid and lipid production in sugar-rich vegetative tissues and provide a complementary metabolic module for future lipid-engineering efforts.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Univ. of Illinois at Urbana-Champaign, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018254; SC0018420

OSTI ID:: 3030837

Journal Information:: New Phytologist, Journal Name: New Phytologist; ISSN 0028-646X; ISSN 1469-8137

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

References (59)

A mass spectrometry‐based high‐throughput screening method for engineering fatty acid synthases with improved production of medium‐chain fatty acids Xue, Pu; Si, Tong; Mishra, Shekhar Biotechnology and Bioengineering, Vol. 117, Issue 7 https://doi.org/10.1002/bit.27343	journal	April 2020
The application of the anthrone reagent to the estimation of starch in cereals Clegg, K. M. Journal of the Science of Food and Agriculture, Vol. 7, Issue 1 https://doi.org/10.1002/jsfa.2740070108	journal	January 1956
Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method Livak, Kenneth J.; Schmittgen, Thomas D. Methods, Vol. 25, Issue 4 https://doi.org/10.1006/meth.2001.1262	journal	December 2001
ggplot2 Wickham, Hadley Use R! https://doi.org/10.1007/978-3-319-24277-4	book	January 2016
Ultrastructural effects in potato leaves due to antisense-inhibition of the sucrose transporter indicate an apoplasmic mode of phloem loading Schulz, Alexander; Kühn, Christina; Riesmeier, Jörg W. Planta, Vol. 206, Issue 4 https://doi.org/10.1007/s004250050430	journal	October 1998
Biosynthesis of triacylglycerols Lehner, Richard; Kuksis, A. Progress in Lipid Research, Vol. 35, Issue 2 https://doi.org/10.1016/0163-7827(96)00005-7	journal	June 1996
Metabolic engineering of vegetative organs for enhanced oil accumulation in biofuel feedstocks Khan, Maqsood; Zhang, Jingtao; Zhang, Yuhao The Crop Journal, Vol. 14, Issue 1 https://doi.org/10.1016/j.cj.2025.11.009	journal	February 2026
Molecular and functional characterization of the plastid-localized Phosphoenolpyruvate enolase (ENO1) from Arabidopsis thaliana Prabhakar, Veena; Löttgert, Tanja; Gigolashvili, Tamara FEBS Letters, Vol. 583, Issue 6 https://doi.org/10.1016/j.febslet.2009.02.017	journal	February 2009
BnaPPT1 is essential for chloroplast development and seed oil accumulation in Brassica napus Tang, Shan; Peng, Fei; Tang, Qingqing Journal of Advanced Research, Vol. 42 https://doi.org/10.1016/j.jare.2022.07.008	journal	December 2022
Emerging technologies for biofuel production: A critical review on recent progress, challenges and perspectives Ambaye, Teklit Gebregiorgis; Vaccari, Mentore; Bonilla-Petriciolet, Adrián Journal of Environmental Management, Vol. 290 https://doi.org/10.1016/j.jenvman.2021.112627	journal	July 2021
SnRK1 activation, signaling, and networking for energy homeostasis Crepin, Nathalie; Rolland, Filip Current Opinion in Plant Biology, Vol. 51 https://doi.org/10.1016/j.pbi.2019.03.006	journal	October 2019
Multifaceted functions of post-translational enzyme modifications in the control of plant glycolysis O’Leary, Brendan; Plaxton, William C. Current Opinion in Plant Biology, Vol. 55 https://doi.org/10.1016/j.pbi.2020.01.009	journal	June 2020
Turning Over a New Leaf in Lipid Droplet Biology Pyc, Michal; Cai, Yingqi; Greer, Michael S. Trends in Plant Science, Vol. 22, Issue 7 https://doi.org/10.1016/j.tplants.2017.03.012	journal	July 2017
A plastidial sodium-dependent pyruvate transporter Furumoto, Tsuyoshi; Yamaguchi, Teppei; Ohshima-Ichie, Yumiko Nature, Vol. 476, Issue 7361 https://doi.org/10.1038/nature10250	journal	August 2011
LipidBlast in silico tandem mass spectrometry database for lipid identification Kind, Tobias; Liu, Kwang-Hyeon; Lee, Do Yup Nature Methods, Vol. 10, Issue 8 https://doi.org/10.1038/nmeth.2551	journal	June 2013
MS-DIAL: data-independent MS/MS deconvolution for comprehensive metabolome analysis Tsugawa, Hiroshi; Cajka, Tomas; Kind, Tobias Nature Methods, Vol. 12, Issue 6 https://doi.org/10.1038/nmeth.3393	journal	May 2015
A moonlighting role for enzymes of glycolysis in the co-localization of mitochondria and chloroplasts Zhang, Youjun; Sampathkumar, Arun; Kerber, Sandra Mae-Lin Nature Communications, Vol. 11, Issue 1 https://doi.org/10.1038/s41467-020-18234-w	journal	September 2020
Mapping of the plant SnRK1 kinase signalling network reveals a key regulatory role for the class II T6P synthase-like proteins Van Leene, Jelle; Eeckhout, Dominique; Gadeyne, Astrid Nature Plants, Vol. 8, Issue 11 https://doi.org/10.1038/s41477-022-01269-w	journal	November 2022
Characterization of two functional phosphoenolpyruvate/phosphate translocator (PPT) genes in Arabidopsis-AtPPT1 may be involved in the provision of signals for correct mesophyll development Knappe, Silke; Lottgert, Tanja; Schneider, Anja The Plant Journal, Vol. 36, Issue 3 https://doi.org/10.1046/j.1365-313X.2003.01888.x	journal	November 2003
The phenotype of the Arabidopsis cue1 mutant is not simply caused by a general restriction of the shikimate pathway Voll, Lars; Häusler, Rainer E.; Hecker, Rolf The Plant Journal, Vol. 36, Issue 3 https://doi.org/10.1046/j.1365-313X.2003.01889.x	journal	October 2003
Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana Clough, Steven J.; Bent, Andrew F. The Plant Journal, Vol. 16, Issue 6, p. 735-743 https://doi.org/10.1046/j.1365-313x.1998.00343.x	journal	December 1998
Reevaluation of the accepted allosteric mechanism of phosphofructokinase from Bacillus stearothermophilus Kimmel, Jennifer L.; Reinhart, Gregory D. Proceedings of the National Academy of Sciences, Vol. 97, Issue 8 https://doi.org/10.1073/pnas.050588097	journal	April 2000
Robust paths to net greenhouse gas mitigation and negative emissions via advanced biofuels Field, John L.; Richard, Tom L.; Smithwick, Erica A. H. Proceedings of the National Academy of Sciences, Vol. 117, Issue 36 https://doi.org/10.1073/pnas.1920877117	journal	August 2020
A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis. Bell, E.; Creelman, R. A.; Mullet, J. E. Proceedings of the National Academy of Sciences, Vol. 92, Issue 19 https://doi.org/10.1073/pnas.92.19.8675	journal	September 1995
STAR: ultrafast universal RNA-seq aligner Dobin, Alexander; Davis, Carrie A.; Schlesinger, Felix Bioinformatics, Vol. 29, Issue 1 https://doi.org/10.1093/bioinformatics/bts635	journal	October 2012
featureCounts: an efficient general purpose program for assigning sequence reads to genomic features Liao, Y.; Smyth, G. K.; Shi, W. Bioinformatics, Vol. 30, Issue 7 https://doi.org/10.1093/bioinformatics/btt656	journal	November 2013
Trimmomatic: a flexible trimmer for Illumina sequence data Bolger, Anthony M.; Lohse, Marc; Usadel, Bjoern Bioinformatics, Vol. 30, Issue 15 https://doi.org/10.1093/bioinformatics/btu170	journal	April 2014
Co-silencing of PhENO1 and PhPPT alters anthocyanin production by reducing phosphoenolpyruvate supply in petunia flower Li, Xin; Cao, Jiahao; Jiang, Guiyun Horticulture Research, Vol. 12, Issue 5 https://doi.org/10.1093/hr/uhaf040	journal	February 2025
The role of transporters in supplying energy to plant plastids Flugge, U. -I.; Hausler, R. E.; Ludewig, F. Journal of Experimental Botany, Vol. 62, Issue 7 https://doi.org/10.1093/jxb/erq361	journal	April 2011
ALLENE OXIDE CYCLASE (AOC) gene family members of Arabidopsis thaliana: tissue- and organ-specific promoter activities and in vivo heteromerization* Stenzel, Irene; Otto, Markus; Delker, Carolin Journal of Experimental Botany, Vol. 63, Issue 17 https://doi.org/10.1093/jxb/ers261	journal	October 2012
Differential Sensitivity of Oleosins to Proteolysis During Oil Body Mobilization in Sunflower Seedlings Sadeghipour, Hamid Reza; Bhatla, Satish Chander Plant and Cell Physiology, Vol. 43, Issue 10 https://doi.org/10.1093/pcp/pcf142	journal	October 2002
Functional Diversification of Acyl-Coenzyme A Oxidases in Jasmonic Acid Biosynthesis and Action Schilmiller, Anthony L.; Koo, Abraham J. K.; Howe, Gregg A. Plant Physiology, Vol. 143, Issue 2 https://doi.org/10.1104/pp.106.092916	journal	December 2006
In Vivo Packaging of Triacylglycerols Enhances Arabidopsis Leaf Biomass and Energy Density Winichayakul, Somrutai; Scott, Richard William; Roldan, Marissa Plant Physiology, Vol. 162, Issue 2 https://doi.org/10.1104/pp.113.216820	journal	April 2013
Sugar Potentiation of Fatty Acid and Triacylglycerol Accumulation Zhai, Zhiyang; Liu, Hui; Xu, Changcheng Plant Physiology, Vol. 175, Issue 2 https://doi.org/10.1104/pp.17.00828	journal	August 2017
Pyruvate Orthophosphate Dikinase mRNA Organ Specificity in Wheat and Maize Aoyagi, Kazuko; Bassham, James A. Plant Physiology, Vol. 76, Issue 1 https://doi.org/10.1104/pp.76.1.278	journal	September 1984
Sugar Sensing and Signaling in Plants Rolland, Filip; Moore, Brandon; Sheen, Jen The Plant Cell, Vol. 14, Issue suppl 1 https://doi.org/10.1105/tpc.010455	journal	May 2002
The Phosphoenolpyruvate/Phosphate Translocator Is Required for Phenolic Metabolism, Palisade Cell Development, and Plastid-Dependent Nuclear Gene Expression Streatfield, Stephen J.; Weber, Andreas; Kinsman, Elizabeth A. The Plant Cell, Vol. 11, Issue 9 https://doi.org/10.1105/tpc.11.9.1609	journal	September 1999
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, Viet Dang; Kannan, Baskaran; Luo, Guangbin GCB Bioenergy, Vol. 15, Issue 12 https://doi.org/10.1111/gcbb.13107	journal	October 2023
Extensive expression regulation and lack of heterologous enzymatic activity of the Class II trehalose metabolism proteins fromArabidopsis thaliana Ramon, Matthew; De Smet, Ive; Vandesteene, Lies Plant, Cell & Environment, Vol. 32, Issue 8 https://doi.org/10.1111/j.1365-3040.2009.01985.x	journal	August 2009
WRINKLED1 encodes an AP2/EREB domain protein involved in the control of storage compound biosynthesis in Arabidopsis Cernac, Alex; Benning, Christoph The Plant Journal, Vol. 40, Issue 4, p. 575-585 https://doi.org/10.1111/j.1365-313X.2004.02235.x	journal	September 2004
Plant triacylglycerols as feedstocks for the production of biofuels Durrett, Timothy P.; Benning, Christoph; Ohlrogge, John The Plant Journal, Vol. 54, Issue 4, p. 593-607 https://doi.org/10.1111/j.1365-313X.2008.03442.x	journal	May 2008
Moss phosphofructokinase is less sensitive than other plant phosphofructokinases to inhibition by phosphoenolpyruvate Kelly, Grahame J.; Latzko, Erwin Physiologia Plantarum, Vol. 52, Issue 3 https://doi.org/10.1111/j.1399-3054.1981.tb06051.x	journal	July 1981
Sucrose rather than GA transported by AtSWEET13 and AtSWEET14 supports pollen fitness at late anther development stages Wang, Jiang; Xue, Xueyi; Zeng, Houqing New Phytologist, Vol. 236, Issue 2 https://doi.org/10.1111/nph.18368	journal	July 2022
Potato tuber expression of Arabidopsis WRINKLED1 increase triacylglycerol and membrane lipids while affecting central carbohydrate metabolism Hofvander, Per; Ischebeck, Till; Turesson, Helle Plant Biotechnology Journal, Vol. 14, Issue 9 https://doi.org/10.1111/pbi.12550	journal	March 2016
Up-regulation of lipid biosynthesis increases the oil content in leaves of Sorghum bicolor Vanhercke, Thomas; Belide, Srinivas; Taylor, Matthew C. Plant Biotechnology Journal, Vol. 17, Issue 1 https://doi.org/10.1111/pbi.12959	journal	July 2018
Engineering triacylglycerol accumulation in duckweed ( Lemna japonica ) Liang, Yuanxue; Yu, Xiao‐Hong; Anaokar, Sanket Plant Biotechnology Journal, Vol. 21, Issue 2 https://doi.org/10.1111/pbi.13943	journal	November 2022
Development of vegetative oil sorghum: From lab‐to‐field Park, Kiyoul; Quach, Truyen; Clark, Teresa J. Plant Biotechnology Journal, Vol. 23, Issue 2 https://doi.org/10.1111/pbi.14527	journal	November 2024
Araport11: a complete reannotation of the Arabidopsis thaliana reference genome Cheng, Chia-Yi; Krishnakumar, Vivek; Chan, Agnes P. The Plant Journal, Vol. 89, Issue 4 https://doi.org/10.1111/tpj.13415	journal	February 2017
Sucrose Efflux Mediated by SWEET Proteins as a Key Step for Phloem Transport Chen, L. -Q.; Qu, X. -Q.; Hou, B. -H. Science, Vol. 335, Issue 6065 https://doi.org/10.1126/science.1213351	journal	December 2011
Triacylglycerol Metabolism, Function, and Accumulation in Plant Vegetative Tissues Xu, Changcheng; Shanklin, John Annual Review of Plant Biology, Vol. 67, Issue 1 https://doi.org/10.1146/annurev-arplant-043015-111641	journal	April 2016
When SWEETs Turn Tweens: Updates and Perspectives Xue, Xueyi; Wang, Jiang; Shukla, Diwakar Annual Review of Plant Biology, Vol. 73, Issue 1 https://doi.org/10.1146/annurev-arplant-070621-093907	journal	May 2022
The Organization and Regulation of Plant Glycolysis Plaxton, William C. Annual Review of Plant Physiology and Plant Molecular Biology, Vol. 47, Issue 1 https://doi.org/10.1146/annurev.arplant.47.1.185	journal	June 1996
Bioconductor: open software development for computational biology and bioinformatics Gentleman, Robert C.; Carey, Vincent J.; Bates, Douglas M. Genome Biology, Vol. 5, Issue 10, p. R80 https://doi.org/10.1186/gb-2004-5-10-r80	journal	September 2004
Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 Love, Michael I.; Huber, Wolfgang; Anders, Simon Genome Biology, Vol. 15, Issue 12 https://doi.org/10.1186/s13059-014-0550-8	journal	December 2014
Intron-mediated enhancement of DIACYLGLYCEROL ACYLTRANSFERASE1 expression in energycane promotes a step change for lipid accumulation in vegetative tissues Cao, Viet Dang; Luo, Guangbin; Korynta, Shelby Biotechnology for Biofuels and Bioproducts, Vol. 16, Issue 1 https://doi.org/10.1186/s13068-023-02393-1	journal	October 2023
Biogenesis and functions of lipid droplets in plants: Thematic Review Series: Lipid Droplet Synthesis and Metabolism: from Yeast to Man Chapman, Kent D.; Dyer, John M.; Mullen, Robert T. Journal of Lipid Research, Vol. 53, Issue 2 https://doi.org/10.1194/jlr.R021436	journal	November 2011
A Modular Cloning System for Standardized Assembly of Multigene Constructs Weber, Ernst; Engler, Carola; Gruetzner, Ramona PLoS ONE, Vol. 6, Issue 2 https://doi.org/10.1371/journal.pone.0016765	journal	February 2011
CUE1: A Mesophyll Cell-Specific Positive Regulator of Light-Controlled Gene Expression in Arabidopsis Li, Hsou-min; Culligan, Kevin; Dixon, Richard A. The Plant Cell, Vol. 7, Issue 10 https://doi.org/10.2307/3870022	journal	October 1995
The Role of Triacylglycerol in Plant Stress Response Lu, Junhao; Xu, Yang; Wang, Juli Plants, Vol. 9, Issue 4 https://doi.org/10.3390/plants9040472	journal	April 2020

Similar Records

Data for Purple Acid Phosphatase2 Stimulates a Futile Cycle of Lipid Synthesis and Degradation, and Mitigates the Negative Growth Effects of Triacylglycerol Accumulation in Vegetative Tissues

Dataset · Mon Jul 18 00:00:00 UTC 2022 · OSTI ID:3014372

Cai, Yingqi; Zhai, Zhiyang; Blanford, Jantana; +5 more

Purple acid phosphatase2 stimulates a futile cycle of lipid synthesis and degradation, and mitigates the negative growth effects of triacylglycerol accumulation in vegetative tissues

Journal Article · Mon Jul 18 00:00:00 UTC 2022 · New Phytologist · OSTI ID:1878300

Cai, Yingqi; Zhai, Zhiyang; Blanford, Jantana; +5 more

Subcellular distribution of gluconeogenetic enzymes in germinating castor bean endosperm

Journal Article · Sun Jul 01 04:00:00 UTC 1979 · Plant Physiol.; (United States) · OSTI ID:5487919

Nishimura, M; Beevers, H

Related Subjects

Arabidopsis thaliana
PPT
SWEET
biofuel
metabolic engineering
phosphoenolpyruvate

Title: PPT1-mediated plastidic phosphoenolpyruvate import enhances fatty acid biosynthesis in sugar-rich tissues

Citation Formats

References (59)

Similar Records

Related Subjects