13C-labeling reveals how membrane lipid components contribute to triacylglycerol accumulation in Chlamydomonas

Young, Danielle Yvonne; Pang, Na; Shachar-Hill, Yair

doi:10.1093/plphys/kiac154

13C-labeling reveals how membrane lipid components contribute to triacylglycerol accumulation in Chlamydomonas

Journal Article · 04 April 2022 · Plant Physiology (Bethesda)

DOI:https://doi.org/10.1093/plphys/kiac154· OSTI ID:1979588

; ;

Abstract Lipid metabolism in microalgae has attracted much interest due to potential utilization of lipids as feedstocks for biofuels, nutraceuticals, and other high-value compounds. Chlamydomonas reinhardtii is a model organism for characterizing the synthesis of the neutral lipid triacylglycerol (TAG), from which biodiesel is made. While much of TAG accumulation under N-deprivation is the result of de novo fatty acid (FA) synthesis, recent work has revealed that approximately one-third of FAs, especially polyunsaturated FAs (PUFAs), come from preexisting membrane lipids. Here, we used 13C-isotopic labeling and mass spectrometry to analyze the turnover of glycerol backbones, headgroups, FAs, whole molecules, and molecular fragments of individual lipids. About one-third of the glyceryl backbones in TAG are derived from preexisting membrane lipids, as are approximately one-third of FAs. The different moieties of the major galactolipids turn over synchronously, while the FAs of diacylglyceryltrimethylhomoserine (DGTS), the most abundant extraplastidial lipid, turn over independently of the rest of the molecule. The major plastidic lipid monogalactosyldiacylglycerol (MGDG), whose predominant species is 18:3α/16:4, was previously shown to be a major source of PUFAs for TAG synthesis. This study reveals that MGDG turns over as whole molecules, the 18:3α/16:4 species is present in both DAG and TAG, and the positional distribution of these PUFAs is identical in MGDG, DAG, and TAG. We conclude that headgroup removal with subsequent acylation is the mechanism by which the major MGDG species is converted to TAG during N-deprivation. This has noteworthy implications for engineering the composition of microalgal TAG for food, fuel, and other applications.

Research Organization:: Michigan State Univ., East Lansing, MI (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

DOE Contract Number:: SC0018269

OSTI ID:: 1979588

Journal Information:: Plant Physiology (Bethesda), Vol. 189, Issue 3; ISSN 0032-0889

Publisher:: American Society of Plant Biologists

Country of Publication:: United States

Language:: English

References (48)

Assessing compartmentalized flux in lipid metabolism with isotopes Allen, Doug K. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1861, Issue 9 https://doi.org/10.1016/j.bbalip.2016.03.017	journal	September 2016
Tracking the metabolic pulse of plant lipid production with isotopic labeling and flux analyses: Past, present and future Allen, Doug K.; Bates, Philip D.; Tjellström, Henrik Progress in Lipid Research, Vol. 58 https://doi.org/10.1016/j.plipres.2015.02.002	journal	April 2015
Carbon and Acyl Chain Flux during Stress-induced Triglyceride Accumulation by Stable Isotopic Labeling of the Polar Microalga Coccomyxa subellipsoidea C169 Allen, James W.; DiRusso, Concetta C.; Black, Paul N. Journal of Biological Chemistry, Vol. 292, Issue 1 https://doi.org/10.1074/jbc.M116.760843	journal	January 2017
Tracing metabolic flux through time and space with isotope labeling experiments Allen, Doug K.; Young, Jamey D. Current Opinion in Biotechnology, Vol. 64 https://doi.org/10.1016/j.copbio.2019.11.003	journal	August 2020
Analysis of Acyl Fluxes through Multiple Pathways of Triacylglycerol Synthesis in Developing Soybean Embryos Bates, Philip D.; Durrett, Timothy P.; Ohlrogge, John B. Plant Physiology, Vol. 150, Issue 1 https://doi.org/10.1104/pp.109.137737	journal	March 2009
The Significance of Different Diacylgycerol Synthesis Pathways on Plant Oil Composition and Bioengineering Bates, Philip D.; Browse, John Frontiers in Plant Science, Vol. 3 https://doi.org/10.3389/fpls.2012.00147	journal	January 2012
Acyl Editing and Headgroup Exchange Are the Major Mechanisms That Direct Polyunsaturated Fatty Acid Flux into Triacylglycerols Bates, Philip D.; Fatihi, Abdelhak; Snapp, Anna R. Plant Physiology, Vol. 160, Issue 3 https://doi.org/10.1104/pp.112.204438	journal	August 2012
Three Acyltransferases and Nitrogen-responsive Regulator Are Implicated in Nitrogen Starvation-induced Triacylglycerol Accumulation in Chlamydomonas Boyle, Nanette R.; Page, Mark Dudley; Liu, Bensheng Journal of Biological Chemistry, Vol. 287, Issue 19 https://doi.org/10.1074/jbc.M111.334052	journal	March 2012
Differential effects of nitrogen and sulfur deprivation on growth and biodiesel feedstock production of Chlamydomonas reinhardtii Cakmak, Turgay; Angun, Pinar; Demiray, Yunus Emre Biotechnology and Bioengineering, Vol. 109, Issue 8 https://doi.org/10.1002/bit.24474	journal	March 2012
Properties of a fatty acyl hydrolase preferentially attacking monogalactosyldiacylglycerols in Dunaliella salina chloroplasts No authors listed Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, Vol. 878, Issue 3 https://doi.org/10.1016/0005-2760(86)90243-2	journal	October 1986
The role of triacylglycerol as a reservoir of polyunsaturated fatty acids for the rapid production of chloroplastic lipids in certain microalgae Cohen, Z.; Khozin-Goldberg, I.; Adlerstein, D. Biochemical Society Transactions, Vol. 28, Issue 6 https://doi.org/10.1042/bst0280740	journal	December 2000
Phospholipid:diacylglycerol acyltransferase: An enzyme that catalyzes the acyl-CoA-independent formation of triacylglycerol in yeast and plants Dahlqvist, A.; Stahl, U.; Lenman, M. Proceedings of the National Academy of Sciences, Vol. 97, Issue 12, p. 6487-6492 https://doi.org/10.1073/pnas.120067297	journal	May 2000
A chloroplast pathway for the de novo biosynthesis of triacylglycerol in Chlamydomonas reinhardtii Fan, Jilian; Andre, Carl; Xu, Changcheng FEBS Letters, Vol. 585, Issue 12, p. 1985-1991 https://doi.org/10.1016/j.febslet.2011.05.018	journal	May 2011
A Central Role for Triacylglycerol in Membrane Lipid Breakdown, Fatty Acid β -Oxidation, and Plant Survival under Extended Darkness Fan, Jilian; Yu, Linhui; Xu, Changcheng Plant Physiology, Vol. 174, Issue 3 https://doi.org/10.1104/pp.17.00653	journal	June 2017
A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues Folch, Jordi; Lees, M.; Stanley, G. H. Sloane Journal of Biological Chemistry, Vol. 226, Issue 1 https://doi.org/10.1016/S0021-9258(18)64849-5	journal	May 1957
The role of lipid metabolism in the acquisition of desiccation tolerance in Craterostigma plantagineum : a comparative approach Gasulla, Francisco; vom Dorp, Katharina; Dombrink, Isabel The Plant Journal, Vol. 75, Issue 5 https://doi.org/10.1111/tpj.12241	journal	June 2013
Lipids of Chlamydomonas reinhardtii. Incorporation of [14C]Acetate, [14C]Palmitate and [14C]Oleate into Different Lipids and Evidence for Lipid-Linked Desaturation of Fatty Acids Giroud, Christian; Eichenberger, Waldemar Plant and Cell Physiology, Vol. 30, Issue 1 https://doi.org/10.1093/oxfordjournals.pcp.a077705	journal	January 1989
Conversion of membrane lipid acyl groups to triacylglycerol and formation of lipid bodies upon nitrogen starvation in biofuel green algae Chlorella UTEX29 Goncalves, Elton C.; Johnson, Jodie V.; Rathinasabapathi, Bala Planta, Vol. 238, Issue 5 https://doi.org/10.1007/s00425-013-1946-5	journal	August 2013
Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Gorman, D. S.; Levine, R. P. Proceedings of the National Academy of Sciences, Vol. 54, Issue 6 https://doi.org/10.1073/pnas.54.6.1665	journal	December 1965
Analysis of genetically modified oils Hazebroek, J. Progress in Lipid Research, Vol. 39, Issue 6 https://doi.org/10.1016/S0163-7827(00)00012-6	journal	November 2000
Systems-Wide Analysis of Acclimation Responses to Long-Term Heat Stress and Recovery in the Photosynthetic Model OrganismChlamydomonas reinhardtii Hemme, Dorothea; Veyel, Daniel; Mühlhaus, Timo The Plant Cell, Vol. 26, Issue 11 https://doi.org/10.1105/tpc.114.130997	journal	November 2014
HEAT INDUCIBLE LIPASE1 Remodels Chloroplastic Monogalactosyldiacylglycerol by Liberating α-Linolenic Acid in Arabidopsis Leaves under Heat Stress Higashi, Yasuhiro; Okazaki, Yozo; Takano, Kouji The Plant Cell, Vol. 30, Issue 8 https://doi.org/10.1105/tpc.18.00347	journal	July 2018
Recycling of the major thylakoid lipid MGDG and its role in lipid homeostasis in Chlamydomonas reinhardtii Iwai, Masako; Yamada-Oshima, Yui; Asami, Kota Plant Physiology, Vol. 187, Issue 3 https://doi.org/10.1093/plphys/kiab340	journal	July 2021
Impacts of phosphorus availability on lipid production by Chlamydomonas reinhardtii Kamalanathan, Manoj; Gleadow, Roslyn; Beardall, John Algal Research, Vol. 12 https://doi.org/10.1016/j.algal.2015.08.021	journal	November 2015
Improving biodiesel fuel properties by modifying fatty ester composition Knothe, Gerhard Energy & Environmental Science, Vol. 2, Issue 7 https://doi.org/10.1039/b903941d	journal	January 2009
Lipidomic and transcriptomic analyses of Chlamydomonas reinhardtii under heat stress unveil a direct route for the conversion of membrane lipids into storage lipids : Lipid remodelling under heat stress in Légeret, B.; Schulz-Raffelt, M.; Nguyen, H. M. Plant, Cell & Environment, Vol. 39, Issue 4 https://doi.org/10.1111/pce.12656	journal	January 2016
A Galactoglycerolipid Lipase Is Required for Triacylglycerol Accumulation and Survival Following Nitrogen Deprivation in Chlamydomonas reinhardtii Li, Xiaobo; Moellering, Eric R.; Liu, Bensheng The Plant Cell, Vol. 24, Issue 11 https://doi.org/10.1105/tpc.112.105106	journal	November 2012
Choreography of Transcriptomes and Lipidomes of Nannochloropsis Reveals the Mechanisms of Oil Synthesis in Microalgae Li, Jing; Han, Danxiang; Wang, Dongmei The Plant Cell, Vol. 26, Issue 4 https://doi.org/10.1105/tpc.113.121418	journal	April 2014
Acyl-Lipid Metabolism Li-Beisson, Yonghua; Shorrosh, Basil; Beisson, Fred The Arabidopsis Book, Vol. 11 https://doi.org/10.1199/tab.0161	journal	January 2013
Metabolism of acyl‐lipids in Chlamydomonas reinhardtii Li‐Beisson, Yonghua; Beisson, Fred; Riekhof, Wayne The Plant Journal, Vol. 82, Issue 3 https://doi.org/10.1111/tpj.12787	journal	March 2015
Isolation and characterization of an Arabidopsis thaliana knockout line for phospholipid: diacylglycerol transacylase gene (At5g13640) Mhaske, Vandana; Beldjilali, Khedidja; Ohlrogge, John Plant Physiology and Biochemistry, Vol. 43, Issue 4 https://doi.org/10.1016/j.plaphy.2005.01.013	journal	April 2005
IsoCor: isotope correction for high-resolution MS labeling experiments Millard, Pierre; Delépine, Baudoin; Guionnet, Matthieu Bioinformatics, Vol. 35, Issue 21 https://doi.org/10.1093/bioinformatics/btz209	journal	March 2019
Changes in Transcript Abundance in Chlamydomonas reinhardtii following Nitrogen Deprivation Predict Diversion of Metabolism Miller, Rachel; Wu, Guangxi; Deshpande, Rahul R. Plant Physiology, Vol. 154, Issue 4 https://doi.org/10.1104/pp.110.165159	journal	October 2010
Freezing Tolerance in Plants Requires Lipid Remodeling at the Outer Chloroplast Membrane Moellering, Eric R.; Muthan, Bagyalakshmi; Benning, Christoph Science, Vol. 330, Issue 6001 https://doi.org/10.1126/science.1191803	journal	August 2010
Kinetic complexities of triacylglycerol accumulation in developing embryos from Camelina sativa provide evidence for multiple biosynthetic systems Pollard, Mike; Shachar-Hill, Yair Journal of Biological Chemistry, Vol. 298, Issue 1 https://doi.org/10.1016/j.jbc.2021.101396	journal	January 2022
Plastidic glucose-6-phosphate dehydrogenases are regulated to maintain activity in the light Preiser, Alyssa L.; Fisher, Nicholas; Banerjee, Aparajita Biochemical Journal, Vol. 476, Issue 10 https://doi.org/10.1042/BCJ20190234	journal	May 2019
Lipid droplet synthesis is limited by acetate availability in starchless mutant of Chlamydomonas reinhardtii Ramanan, Rishiram; Kim, Byung-Hyuk; Cho, Dae-Hyun FEBS Letters, Vol. 587, Issue 4 https://doi.org/10.1016/j.febslet.2012.12.020	journal	January 2013
Conversion of Monogalactosyldiacylglycerols to Triacylglycerols in Ozone-Fumigated Spinach Leaves Sakaki, Takeshi; Saito, Kazuki; Kawaguchi, Akihiko Plant Physiology, Vol. 94, Issue 2 https://doi.org/10.1104/pp.94.2.766	journal	October 1990
The Chlamydomonas heat stress response Schroda, Michael; Hemme, Dorothea; Mühlhaus, Timo The Plant Journal, Vol. 82, Issue 3 https://doi.org/10.1111/tpj.12816	journal	March 2015
Some evidence for the reversibility of the cholinephosphotransferasecatalysed reaction in developing linseed cotyledons in vivo Slack, C. Roger; Campbell, Lindsay C.; Browse, John A. Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, Vol. 754, Issue 1 https://doi.org/10.1016/0005-2760(83)90076-0	journal	November 1983
Cloning and Functional Characterization of a Phospholipid:Diacylglycerol Acyltransferase from Arabidopsis Ståhl, Ulf; Carlsson, Anders S.; Lenman, Marit Plant Physiology, Vol. 135, Issue 3 https://doi.org/10.1104/pp.104.044354	journal	July 2004
Biosynthesis of Triacylglycerol Molecules with a Tailored PUFA Profile in Industrial Microalgae Xin, Yi; Shen, Chen; She, Yiting Molecular Plant, Vol. 12, Issue 4 https://doi.org/10.1016/j.molp.2018.12.007	journal	April 2019
The metabolic origins of non-photorespiratory CO2 release during photosynthesis: a metabolic flux analysis Xu, Yuan; Fu, Xinyu; Sharkey, Thomas D. Plant Physiology, Vol. 186, Issue 1 https://doi.org/10.1093/plphys/kiab076	journal	February 2021
Galactolipid DGDG and Betaine Lipid DGTS Direct De Novo Synthesized Linolenate into Triacylglycerol in a Stress-Induced Starchless Mutant of Chlamydomonas reinhardtii Yang, Miao; Kong, Fantao; Xie, Xi Plant and Cell Physiology, Vol. 61, Issue 4 https://doi.org/10.1093/pcp/pcaa012	journal	February 2020
Phospholipid:Diacylglycerol Acyltransferase Is a Multifunctional Enzyme Involved in Membrane Lipid Turnover and Degradation While Synthesizing Triacylglycerol in the Unicellular Green Microalga Chlamydomonas reinhardtii Yoon, Kangsup; Han, Danxiang; Li, Yantao The Plant Cell, Vol. 24, Issue 9 https://doi.org/10.1105/tpc.112.100701	journal	September 2012
Large fluxes of fatty acids from membranes to triacylglycerol and back during N-deprivation and recovery in Chlamydomonas Young, Danielle Yvonne; Shachar-Hill, Yair Plant Physiology, Vol. 185, Issue 3 https://doi.org/10.1093/plphys/kiaa071	journal	December 2020
Modifications of the metabolic pathways of lipid and triacylglycerol production in microalgae Yu, Wei-Luen; Ansari, William; Schoepp, Nathan G. Microbial Cell Factories, Vol. 10, Issue 1 https://doi.org/10.1186/1475-2859-10-91	journal	January 2011
DGAT1 and PDAT1 Acyltransferases Have Overlapping Functions in Arabidopsis Triacylglycerol Biosynthesis and Are Essential for Normal Pollen and Seed Development Zhang, Meng; Fan, Jilian; Taylor, David C. The Plant Cell, Vol. 21, Issue 12 https://doi.org/10.1105/tpc.109.071795	journal	December 2009

Similar Records

Do betaine lipids replace phosphatidylcholine as fatty acid editing hubs in microalgae?

Journal Article · 2023 · Frontiers in Plant Science · OSTI ID:1909955

Hoffmann, Danielle Yvonne; Shachar-Hill, Yair

Kinetic complexities of triacylglycerol accumulation in developing embryos from Camelina sativa provide evidence for multiple biosynthetic systems

Journal Article · 2021 · Journal of Biological Chemistry · OSTI ID:1830024

Pollard, Mike; Shachar-Hill, Yair

Related Subjects

Plant Sciences

13C-labeling reveals how membrane lipid components contribute to triacylglycerol accumulation in Chlamydomonas

Citation Formats

References (48)

Similar Records

Related Subjects