Generation of camelina mid-oleic acid seed oil by identification and stacking of fatty acid biosynthetic mutants

Neumann, Nicholas G.; Nazarenus, Tara J.; Aznar-Moreno, Jose A.; Rodriguez-Aponte, Sergio A.; Mejias Veintidos, Valerie A.; Comai, Luca; Durrett, Timothy P.; Cahoon, Edgar B.

doi:10.1016/j.indcrop.2020.113074

Generation of camelina mid-oleic acid seed oil by identification and stacking of fatty acid biosynthetic mutants

Journal Article · Tue Nov 17 23:00:00 EST 2020 · Industrial Crops and Products

DOI:https://doi.org/10.1016/j.indcrop.2020.113074· OSTI ID:1851641

Neumann, Nicholas G. ^[1]; Nazarenus, Tara J. ^[2]; Aznar-Moreno, Jose A. ^[3]; Rodriguez-Aponte, Sergio A. ^[2]; Mejias Veintidos, Valerie A. ^[2]; Comai, Luca ^[4]; Durrett, Timothy P. ^[3]; Cahoon, Edgar B. ^[2]

Univ. of Kansas, Lawrence, KS (United States). Dept. of Biochemistry and Molecular Biophysics; Univ. of Kansas, Lawrence, KS (United States)
Univ. of Nebraska, Lincoln, NE (United States). Center for Plant Science Innovation. Dept. of Biochemistry
Univ. of Kansas, Lawrence, KS (United States). Dept. of Biochemistry and Molecular Biophysics
Univ. of California, Davis, CA (United States). Dept. of Plant Biology. Davis Genome Center

Modifying oilseeds to obtain a desired fatty acid composition is often necessary to enable use as feedstocks for specific applications such as food processing, biofuels, or biolubricants. A mutant population of camelina (Camelina sativa), an emerging specialty oilseed crop, was screened by high-throughput gas chromatography for lines with altered seed oil fatty acid composition. By leveraging knowledge of fatty acid synthesis in Arabidopsis thaliana, mutations in orthologs of FATTY ACID ELONGASE1 (FAE1), FATTY ACID DESATURASE2 (FAD2), FATTY ACID DESATURASE3 (FAD3), and β-KETO-ACYL-ACP SYNTHASE II (KASII; FAB1) were identified. The mutations altered conserved amino acid residues in the encoded proteins. The ability of the mutations in FAE1, FAD2 and FAD3 to affect enzyme function was demonstrated by comparing in vivo activities of wild-type and mutant alleles in yeast. In addition, expression of wild-type cDNA in camelina complemented fatty acid phenotypes of these mutants. As camelina has a hexaploid genome, the effect of a mutation in one of the three homeologs for each gene resulted in no or less severe growth phenotypes compared to similar mutations in Arabidopsis. Mid-oleic oils with nearly 40 % oleic acid and reduced very long-chain (≤C20) fatty acid content were obtained by crossing to obtain a fae1c/fad2a/fae1a/fad3a quadruple mutant. Little effect on total seed oil content was observed in the stacked mutant line. Finally, the resulting mid-oleic acid oil had improved oxidative stability due to reductions in polyunsaturated fatty acid content, increasing its utility for biofuels and other applications.

View Accepted Manuscript (DOE)

Research Organization:: Colorado State Univ., Fort Collins, CO (United States)

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

Grant/Contract Number:: SC0012459

OSTI ID:: 1851641

Alternate ID(s):: OSTI ID: 1809660

Journal Information:: Industrial Crops and Products, Journal Name: Industrial Crops and Products Journal Issue: C Vol. 159; ISSN 0926-6690

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

References (45)

Camelina-derived jet fuel and diesel: Sustainable advanced biofuels Shonnard, David R.; Williams, Larry; Kalnes, Tom N. Environmental Progress & Sustainable Energy, Vol. 29, Issue 3 https://doi.org/10.1002/ep.10461	journal	June 2010
Mutants of Arabidopsis with alterations in seed lipid fatty acid composition Lemieux, B.; Miquel, M.; Somerville, C. Theoretical and Applied Genetics, Vol. 80, Issue 2 https://doi.org/10.1007/BF00224392	journal	August 1990
Isolation of EMS-induced mutants in Arabidopsis altered in seed fatty acid composition James, D. W.; Dooner, H. K. Theoretical and Applied Genetics, Vol. 80, Issue 2 https://doi.org/10.1007/BF00224393	journal	August 1990
Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation Lu, Chaofu; Kang, Jinling Plant Cell Reports, Vol. 27, Issue 2 https://doi.org/10.1007/s00299-007-0454-0	journal	September 2007
Genetic engineering of medium-chain-length fatty acid synthesis in Dunaliella tertiolecta for improved biodiesel production Lin, Huixin; Lee, Yuan Kun Journal of Applied Phycology, Vol. 29, Issue 6 https://doi.org/10.1007/s10811-017-1210-7	journal	July 2017
De novo transcriptome analysis of an imminent biofuel crop, Camelina sativa L. using Illumina GAIIX sequencing platform and identification of SSR markers Mudalkar, Shalini; Golla, Ramesh; Ghatty, Sreenivas Plant Molecular Biology, Vol. 84, Issue 1-2 https://doi.org/10.1007/s11103-013-0125-1	journal	September 2013
Structure indices in FA chemistry. How relevant is the iodine value? Knothe, Gerhard Journal of the American Oil Chemists' Society, Vol. 79, Issue 9 https://doi.org/10.1007/s11746-002-0569-4	journal	September 2002
Dependence of oil stability index of fatty compounds on their structure and concentration and presence of metals Knothe, Gerhard; Dunn, Robert O. Journal of the American Oil Chemists' Society, Vol. 80, Issue 10 https://doi.org/10.1007/s11746-003-0814-x	journal	October 2003
Evaluation of Biodiesel Derived from Camelina sativa Oil Soriano, Nestor U.; Narani, Akash Journal of the American Oil Chemists' Society, Vol. 89, Issue 5 https://doi.org/10.1007/s11746-011-1970-1	journal	November 2011
Oxidative stabilities of soybean oils with elevated palmitate and reduced linolenate contents Shen, Nuo; Fehr, Walter; Johnson, Lawrence Journal of the American Oil Chemists' Society, Vol. 74, Issue 3 https://doi.org/10.1007/s11746-997-0140-y	journal	March 1997
Trans -free margarine from highly saturated soybean oil Kok, Li Lee; Fehr, Walter R.; Hammond, Earl G. Journal of the American Oil Chemists' Society, Vol. 76, Issue 10 https://doi.org/10.1007/s11746-999-0092-5	journal	October 1999
Arabidopsis mutants deficient in polyunsaturated fatty acid synthesis. Biochemical and genetic characterization of a plant oleoyl-phosphatidylcholine desaturase. Miquel, M.; Browse, J. Journal of Biological Chemistry, Vol. 267, Issue 3 https://doi.org/10.1016/S0021-9258(18)45974-1	journal	January 1992
Rapid method for the determination of fatty acid profiles from fats and oils using trimethylsulphonium hydroxide for transesterification Butte, W. Journal of Chromatography A, Vol. 261 https://doi.org/10.1016/S0021-9673(01)87931-0	journal	January 1983
Web-based primer design for single nucleotide polymorphism analysis Neff, Michael M.; Turk, Edward; Kalishman, Michael Trends in Genetics, Vol. 18, Issue 12 https://doi.org/10.1016/S0168-9525(02)02820-2	journal	December 2002
Camelina sativa: An ideal platform for the metabolic engineering and field production of industrial lipids Bansal, Sunil; Durrett, Timothy P. Biochimie, Vol. 120 https://doi.org/10.1016/j.biochi.2015.06.009	journal	January 2016
Biotechnology tools and applications for development of oilseed crops with healthy vegetable oils Msanne, Joseph; Kim, Hyojin; Cahoon, Edgar B. Biochimie, Vol. 178 https://doi.org/10.1016/j.biochi.2020.09.020	journal	November 2020
Brassicaceae germplasm diversity for agronomic and seed quality traits under drought stress Enjalbert, Jean-Nicolas; Zheng, Shusong; Johnson, Jerry J. Industrial Crops and Products, Vol. 47 https://doi.org/10.1016/j.indcrop.2013.02.037	journal	May 2013
Molecular cloning and characterization of the genes encoding a microsomal oleate Δ12 desaturase (CsFAD2) and linoleate Δ15 desaturase (CsFAD3) from Camelina sativa Rodríguez-Rodríguez, Manuel Fernando; Salas, Joaquín J.; Venegas-Calerón, Mónica Industrial Crops and Products, Vol. 89 https://doi.org/10.1016/j.indcrop.2016.05.038	journal	October 2016
Extending the story of very-long-chain fatty acid elongation Haslam, Tegan M.; Kunst, Ljerka Plant Science, Vol. 210 https://doi.org/10.1016/j.plantsci.2013.05.008	journal	September 2013
New approaches to facilitate rapid domestication of a wild plant to an oilseed crop: Example pennycress (Thlaspi arvense L.) Sedbrook, John C.; Phippen, Winthrop B.; Marks, M. David Plant Science, Vol. 227 https://doi.org/10.1016/j.plantsci.2014.07.008	journal	October 2014
Identification of three genes encoding microsomal oleate desaturases (FAD2) from the oilseed crop Camelina sativa Kang, Jinling; Snapp, Anna R.; Lu, Chaofu Plant Physiology and Biochemistry, Vol. 49, Issue 2 https://doi.org/10.1016/j.plaphy.2010.12.004	journal	February 2011
Mutagenesis of the FAE1 genes significantly changes fatty acid composition in seeds of Camelina sativa Ozseyhan, Mehmet E.; Kang, Jinling; Mu, Xiaopeng Plant Physiology and Biochemistry, Vol. 123 https://doi.org/10.1016/j.plaphy.2017.11.021	journal	February 2018
Life cycle assessment of camelina oil derived biodiesel and jet fuel in the Canadian Prairies Li, Xue; Mupondwa, Edmund Science of The Total Environment, Vol. 481 https://doi.org/10.1016/j.scitotenv.2014.02.003	journal	May 2014
Strict EU ruling on gene-edited crops squeezes science Wight, Andrew J. Nature, Vol. 563, Issue 7729 https://doi.org/10.1038/d41586-018-07166-7	journal	October 2018
The emerging biofuel crop Camelina sativa retains a highly undifferentiated hexaploid genome structure Kagale, Sateesh; Koh, Chushin; Nixon, John Nature Communications, Vol. 5, Issue 1 https://doi.org/10.1038/ncomms4706	journal	April 2014
A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers Konieczny, Andrzej; Ausubel, Frederick M. The Plant Journal, Vol. 4, Issue 2 https://doi.org/10.1046/j.1365-313X.1993.04020403.x	journal	August 1993
A KAS2 cDNA complements the phenotypes of the Arabidopsis fab1 mutant that differs in a single residue bordering the substrate binding pocket Carlsson, Anders S.; LaBrie, Samuel T.; Kinney, Anthony J. The Plant Journal, Vol. 29, Issue 6 https://doi.org/10.1046/j.1365-313X.2002.01253.x	journal	March 2002
Toward production of jet fuel functionality in oilseeds: identification of FatB acyl-acyl carrier protein thioesterases and evaluation of combinatorial expression strategies in Camelina seeds Kim, Hae Jin; Silva, Jillian E.; Vu, Hieu Sy Journal of Experimental Botany, Vol. 66, Issue 14 https://doi.org/10.1093/jxb/erv225	journal	May 2015
Large-Scale Discovery of Induced Point Mutations With High-Throughput TILLING Till, B. J. Genome Research, Vol. 13, Issue 3 https://doi.org/10.1101/gr.977903	journal	March 2003
A Condensing Enzyme from the Seeds of Lesquerella fendleri That Specifically Elongates Hydroxy Fatty Acids Moon, Hangsik; Smith, Mark A.; Kunst, Ljerka Plant Physiology, Vol. 127, Issue 4 https://doi.org/10.1104/pp.010544	journal	December 2001
Cloning of Higher Plant [omega]-3 Fatty Acid Desaturases Yadav, N. S.; Wierzbicki, A.; Aegerter, M. Plant Physiology, Vol. 103, Issue 2 https://doi.org/10.1104/pp.103.2.467	journal	October 1993
A Mutant of Arabidopsis Deficient in the Elongation of Palmitic Acid Wu, J.; James Jr, D. W.; Dooner, H. K. Plant Physiology, Vol. 106, Issue 1 https://doi.org/10.1104/pp.106.1.143	journal	September 1994
Discovery of Rare Mutations in Populations: TILLING by Sequencing Tsai, Helen; Howell, Tyson; Nitcher, Rebecca Plant Physiology, Vol. 156, Issue 3 https://doi.org/10.1104/pp.110.169748	journal	April 2011
TILLING: practical single-nucleotide mutation discovery Comai, Luca; Henikoff, Steven The Plant Journal, Vol. 45, Issue 4 https://doi.org/10.1111/j.1365-313X.2006.02670.x	journal	February 2006
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
Camelina seed transcriptome: a tool for meal and oil improvement and translational research Nguyen, Huu T.; Silva, Jillian E.; Podicheti, Ram Plant Biotechnology Journal, Vol. 11, Issue 6 https://doi.org/10.1111/pbi.12068	journal	April 2013
The Regulatory Status of Genome‐edited Crops Wolt, Jeffrey D.; Wang, Kan; Yang, Bing Plant Biotechnology Journal, Vol. 14, Issue 2 https://doi.org/10.1111/pbi.12444	journal	August 2015
Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa , using CRISPR/Cas9 gene editing Jiang, Wen Zhi; Henry, Isabelle M.; Lynagh, Peter G. Plant Biotechnology Journal, Vol. 15, Issue 5 https://doi.org/10.1111/pbi.12663	journal	January 2017
Selective gene dosage by CRISPR-Cas9 genome editing in hexaploid Camelina sativa Morineau, Céline; Bellec, Yannick; Tellier, Frédérique Plant Biotechnology Journal, Vol. 15, Issue 6 https://doi.org/10.1111/pbi.12671	journal	April 2017
The developmental transcriptome atlas of the biofuel crop Camelina sativa Kagale, Sateesh; Nixon, John; Khedikar, Yogendra The Plant Journal, Vol. 88, Issue 5 https://doi.org/10.1111/tpj.13302	journal	December 2016
Identification, characterization and field testing of Brassica napus mutants producing high‐oleic oils Bai, Shuangyi; Engelen, Steven; Denolf, Peter The Plant Journal, Vol. 98, Issue 1 https://doi.org/10.1111/tpj.14195	journal	January 2019
Map-based cloning of a gene controlling omega-3 fatty acid desaturation in Arabidopsis Arondel, V.; Lemieux, B.; Hwang, I. Science, Vol. 258, Issue 5086 https://doi.org/10.1126/science.1455229	journal	November 1992
Polyploid genome of Camelina sativa revealed by isolation of fatty acid synthesis genes Hutcheon, Carolyn; Ditt, Renata F.; Beilstein, Mark BMC Plant Biology, Vol. 10, Issue 1 https://doi.org/10.1186/1471-2229-10-233	journal	January 2010
Transcriptome profiling of Camelina sativa to identify genes involved in triacylglycerol biosynthesis and accumulation in the developing seeds Abdullah, Hesham M.; Akbari, Parisa; Paulose, Bibin Biotechnology for Biofuels, Vol. 9, Issue 1 https://doi.org/10.1186/s13068-016-0555-5	journal	July 2016
Predicting the Functional Effect of Amino Acid Substitutions and Indels Choi, Yongwook; Sims, Gregory E.; Murphy, Sean PLoS ONE, Vol. 7, Issue 10 https://doi.org/10.1371/journal.pone.0046688	journal	October 2012

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Generation of camelina mid-oleic acid seed oil by identification and stacking of fatty acid biosynthetic mutants

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