NC1203: Lipids In Plants: Improving and Developing Sustainability of Crops ("LIPIDS of Crops")

(Multistate Research Project)

Status: Active

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SAES-422 Reports

Annual/Termination Reports:

[09/30/2022] [10/16/2023] [08/30/2024] [10/18/2025]

Date of Annual Report: 09/30/2022

Report Information

Annual Meeting Dates: 09/17/2022 - 09/18/2022
Period the Report Covers: 10/01/2021 - 09/30/2022

Participants

Cahoon, Edgar - University of Nebraska (UNL); Clemente, Thomas - UNL; Hoffmann-Benning, Susanne - Michigan State University; Kosma, Dylan - University of Nevada Reno (UNR); Lee, Young-Jin - Iowa State University; Louis, Joe - University of Nebraska-Lincoln; Minton, Ernie - Kansas State University (KSU); Schrick, Kathrin - KSU; Thelen, Jay - University of Missouri - Columbia; Welti, Ruth - KSU; Dhankher, Om Parkash - University of Massachusetts Amherst; Koo, Abraham - MU; Narayanan, Sruthi - Clemson University; Wang, Xuemin (Sam) - Donald Danforth Center; Yandeau-Nelson, Marna - Iowa State University; Van Doren, Steven - University of Missouri; Bates, Phil - Washington State University; Tamborindeguy, Cecilia - Texas A&M

Brief Summary of Minutes

Brief summary minutes of annual meeting: The 2022 NC-1203 meeting was held as a hybrid Zoom-in person meeting on September 17 at Michigan State University. The meeting was opened by the host Susanne Hoffmann-Benning. A time of introductions was followed by presentations and discussions for each of the individual project aims. Multistate Project Administrative Advisor Ernie Minton joined later and gave comments about the current status of the group and the need for the group to find another Multistate Project Administrative Advisor. Accomplishments for each of the aims are summarized below. This was followed by a discussion of future plans for each of the aims and the timing of the 2023 annual meeting hosted by Tom Clemente at the University of Nebraska Lincoln and establishment of future meeting sites; in 2024 the meeting will be held at the University of Nevada Reno (hosted by Dylan Kosma) and in 2025 in Kansas City (alternatively, at Kansas State University) (hosted by Kathrin Schrick).

Accomplishments

A pdf file containing the entire report and publications will be attached under "publications"

Publications

Impact Statements

Date of Annual Report: 10/16/2023

Report Information

Annual Meeting Dates: 08/14/2023 - 08/14/2023
Period the Report Covers: 10/01/2022 - 08/14/2023

Participants

Allen, Doug – Donald Danforth Center; Cahoon, Edgar - University of Nebraska, Lincoln (UNL); Clemente, Thomas - UNL; Durrett, Timothy - Kansas State University (KSU); Hoffmann-Benning, Susanne - Michigan State University; Huang, Chien-Yu – Louisiana State University; Lawrence-Dill, Carolyn, Iowa State University (ISU); Louis, Joe - UNL; Rosten, Rebecca – UNL; Schrick, Kathrin – KSU; Thelen, Jay - University of Missouri (MU), Columbia; Welti, Ruth - KSU; Dhankher, Om Parkash - University of Massachusetts, Amherst; Koo, Abraham - MU; Narayanan, Sruthi - Clemson University; Wang, Xuemin (Sam) - Donald Danforth Center; Yandeau-Nelson, Marna - ISU; Van Doren, Steven - MU; Bates, Phil - Washington State University; Tamborindeguy, Cecilia - Texas A&M

Brief Summary of Minutes

The 2023 NC-1203 meeting was held as a hybrid in person and Zoom meeting on August 14, 2023, at the University of Nebraska, Lincoln. The meeting was opened by the host Tom Clemente. A time of introductions was followed by presentations by the participants. Multistate Project Administrative Advisor Carolyn Lawrence-Dill joined later to provide a summary of information corresponding to multistate projects and opportunities for additional funding support. Accomplishments for each of the objectives are summarized below. The presentations were followed by a discussion of future plans for each of the aims and the timing of the 2024 annual meeting co-hosted by Joe Louis (President) and Rebecca Roston (Secretary) at the University of Nebraska Lincoln one day ahead of the ISPL meeting in July 2024. Future meeting sites: In 2025 we plan to hold the annual meeting in Kansas City (alternatively, at Kansas State University) (hosted by Kathrin Schrick, who will serve as Vice President in 2024), and in 2026 the meeting will be held at the University of Nevada, Reno (hosted by Dylan Kosma, who will serve as Vice President in 2025). As part of the itinerary of the next year’s 2024 annual meeting, a new Secretary will be elected for 2025.

Accomplishments

A pdf file containing the entire report and publications will be attached under "publications"

Publications

Impact Statements

Date of Annual Report: 08/30/2024

Report Information

Annual Meeting Dates: 07/14/2024 - 07/19/2024
Period the Report Covers: 08/15/2023 - 07/19/2024

Participants

Allen, Doug - USDA/Donald Danforth Plant Science Center; Cahoon, Edgar - University of Nebraska, Lincoln (UNL); Clemente, Thomas - UNL; Durrett, Timothy - Kansas State University (KSU); Hoffmann-Benning, Susanne - Michigan State University; Huang, Chien-Yu - Louisiana State University; Louis, Joe - UNL; Roston, Rebecca - UNL; Schrick, Kathrin - KSU; Thelen, Jay - University of Missouri (MU), Columbia; Welti, Ruth - KSU; Dhankher, Om Parkash Dhankher - University of Massachusetts, Amherst; Koo, Abraham - MU; Narayanan, Sruthi - Clemson University; Wang, Xuemin (Sam) - Donald Danforth Center; Yandeau-Nelson, Marna - Iowa State University (ISU); Van Doren, Steven - MU; Bates, Phil - Washington State University; Tamborindeguy, Cecilia - Texas A&M;, Scott Peck - MU; Young-Jin Lee - ISU; Adam Yokom - MU;

Brief Summary of Minutes

The 2024 NC-1203 meeting was held as a hybrid in-person and Zoom meeting on July 16, 2024, at the University of Nebraska, Lincoln, to take advantage of the International Symposium on Plant Lipids occurring at the same location. Presentations by the participants occurred throughout the society meeting. We have plans to request a new academic advisor from Kansas State, and proposed addition of several future members. Future meeting sites: In 2025 we plan to hold the annual meeting in Kansas City (alternatively, at Kansas State University) hosted by Kathrin Schrick, in 2026 the meeting will be held at the University of Nevada, Reno (hosted by Dylan Kosma), and in 2027 the meeting will be held in conjunction with the 2027 Gordon Research Conference on Plant Lipids, likely in California, with more than half of attendees planning to attend. Philip Bates will host the 2027 business meeting. Hosting beyond 2026 will depend on renewal of the project. Timothy Durrett volunteered to organize writing of the renewal with Ruth Welti, Susanne Hoffmann-Benning, and Rebecca Roston volunteering to assist.

Accomplishments

Activities and accomplishments related to each of the project’s three objectives are described below. The report also summarizes the collaborative research efforts of LIPIDS of Crops members including the establishment of new collaborations as well as the outcomes of existing collaborations in the form of publications and grant awards. <h1>Objective 1: Improve and extend methods for lipid characterization and measurement</h1> The Welti and Durrett groups (Kansas) collaborated to create a novel semi-targeted mass spectral approach for lipid analysis, capable of yielding over 100,000 mass spectral intensities representing both known and unknown lipids. This method was applied to camelina plants, with the resulting data correlated with transcriptomic analyses by Trupti Joshi's group (Missouri). This interdisciplinary effort not only produced valuable insights into lipid metabolism during seed development but also set the stage for future collaborative projects aimed at refining lipid data annotation techniques. In addition to advancing lipid analysis methods, Welti’s group (Kansas) has been instrumental in establishing a global Plant Lipid Interest Group under the International Lipidomics Society. This collaboration involves plant lipidomics labs from Germany, the UK, France, the US, and other countries. The group's first major initiative, a ring trial comparing lipid analyses across different labs, exemplifies the collaborative spirit fostered by this grant. The ongoing work aims to standardize lipidomics practices and make common reference materials available to the global plant lipidomics community, ensuring consistency and reliability in lipid research. The Lee group (Iowa) has also made significant contributions, developing mass spectrometry imaging with isotope labeling to study lipid biosynthesis in various plant species, including duckweed, Arabidopsis, and maize. Their spatiotemporal analysis of lipid labeling patterns has provided new insights into membrane lipid restructuring and biosynthesis, with findings published in multiple papers. These advancements are a direct result of collaborative efforts supported within NC-1203, highlighting its role in pushing the boundaries of lipid characterization and measurement. <h1>Objective 2: Identify lipid-related mechanisms to increase agricultural resilience</h1> Several labs have focused on the role of lipids in plant signaling and metabolism. The Hoffmann-Benning lab (Michigan) developed an innovative optogenetics method to study phloem lipids and lipid-binding proteins, enabling the monitoring of these proteins' movement within plants. This work is being complemented by the Schrick lab (Kansas), which has been investigating lipid-binding transcription factors and their role in gene expression regulation. Through collaboration with the Roeder group (New York) at Cornell, the Schrick lab is exploring how lipids interact with transcription factors to influence plant development. These studies collectively enhance our understanding of lipid-mediated signaling and its impact on plant growth. Another area of focus has been the relationship between lipid composition and plant resilience to environmental stress. The Allen (USDA) and Bates labs (Washington), in collaboration with the Gehan lab (Missouri) at the Danforth Center, studied how lipid-engineered tobacco plants respond to temperature changes, revealing that increased lipid levels in guard cells can affect stomatal function and leaf temperature regulation. Similarly, the Roston lab (Nebraska), in collaboration with the Welti (Kansas) and Durrett labs (Kansas), has been investigating the role of lipids in cold tolerance, profiling plant responses to low temperatures. These studies highlight the importance of lipid stability and composition in enhancing crop resilience to various environmental challenges. The metabolic pathways and biosynthesis of lipids in crops have also been a key focus, with the Bates lab (Washington) working with USDA scientists to discover a novel metabolic pathway, "triacylglycerol remodeling," in the Brassicaceae species. This pathway influences oil composition in oilseed crops, and their findings are poised to impact agricultural practices. Similarly, the Yandeau-Nelson and Nikolau teams (Iowa) have employed synthetic biology approaches to study cuticle synthesis in maize, focusing on how gene mutations affect cuticular wax composition and its protective functions. Their work, in collaboration with Joe Louis (Nebraska), is expected to extend to biotic stress resistance, further enhancing crop resilience. The role of lipids in plant-pathogen interactions has been explored by the Tamborindeguy lab (Texas), which has identified bacterial lipoproteins that manipulate plant defenses, setting the stage for better disease management strategies. This theme is echoed in the work of the Huang lab (Louisiana), which has been studying the lipid droplets in soybean and fungal pathogens, identifying inhibitors that suppress fungal growth. Both labs are contributing to the development of lipid-based approaches for managing plant diseases. Finally, the Welti lab (Kansas), in collaboration with Durrett (Kansas), Schrick (Kansas), and Trupti Joshi (Missouri), is advancing our understanding of lipid metabolism by characterizing genes involved in cuticle biosynthesis and sequencing the transcriptome of Chinese elm. Their research is uncovering new insights into lipid functions in plants, with potential applications in crop improvement. <h1>Objective 3: Develop crops with improved yield and/or functionality</h1> Multiple collaborations originating in the NC1203 have significantly advanced research on improving crop yield and functionality through lipid production. For instance, the Durrett lab's work (Kansas) on cloning and characterizing novel seed-specific promoters from camelina, alongside their collaboration with the Allen group (USDA), resulted in the generation of soybean lines with improved amino acid compositions, contributing to a better understanding of gene expression's impact on metabolic pathways. Similarly, the Bates lab leveraged triacylglycerol remodeling mechanisms to engineer camelina for enhanced fatty acid compositions, and their findings are now being tested for broader applications. The Thelen lab (Missouri), in collaboration with the Koo (Missouri), Bates (Washington), and Allen (USDA) labs, used global profiling methods to study the metabolic effects of enhancing acetyl-CoA carboxylase activity in Brassicaceae. This collaborative effort uncovered unexpected insights into fatty acid turnover and catabolic pathways, which could inform future strategies for improving oil content in crops. In another notable partnership, the Cahoon lab (Nebraska) teamed up with the Hongfei Lin lab at Washington State University to develop biodesigned camelina oils for sustainable aviation fuel (SAF) production. Their work optimized oilseed feedstock for SAF production, achieving high alkane yields that closely mimic Jet A composition, contributing to the SAF Grand Challenge. Further, the Dhankher lab's (Massachusetts) work on engineering camelina for increased oil yield and fatty acid composition, through transcriptomic, metabolomic, and lipidomic approaches, identified key genes responsible for these traits. Their findings have led to significant increases in seed yield and oil content in engineered camelina lines, providing a strong foundation for future crop improvement efforts. The Yokom (Missouri), Van Doren (Missouri), and Thelen (Missouri) labs collaborated to study the structural aspects of acetyl-CoA carboxylase activity in pennycress and other species, focusing on the protein-protein interactions that drive lipid production. Their work has advanced our understanding of the regulation of de novo fatty acid synthesis, offering new targets for crop engineering. The Koo lab's (Missouri) collaboration with the Welti (Kansas) and Allen (USDA) labs resulted in significant progress in metabolic engineering to increase biomass oil content, with findings under review for publication. Similarly, the Clemente (Nebraska) lab's research on the genetic basis of soybean seed protein accumulation led to the identification of gene edits that impact seed protein content and maturity, with ongoing studies aimed at further elucidating these genetic pathways. In summary, NC-1203 facilitated critical partnerships that advanced the scientific understanding of lipid metabolism and crop yield improvement and laid the groundwork for future research and development efforts, underscoring the impact on the plant lipid field. <h1>2024 Milestones</h1> <ul> <li>Development of in vivo isotope labeling for mass spectrometry imaging of plant metabolites.</li> <li>Analysis of post-translational regulation of  lipid-binding transcription factors during growth and development</li> <li>Quantification of protein, lipid, and carbohydrate contents of soybean seed with altered protein amino acid quality.</li> <li>Analysis of oil, protein and amino acid content revealed similar oil content between transgenic lines engineered to suppress enzymes associated with cys and met turnover and wild-type control plants, elevated total protein levels (including increased protein-bound cys and met) in transgenic lines, as well as increased free cys and met.</li> <li>Wound-healing and lipidomic measurements of engineered potato and camelina lines completed.</li> <li>Field trials of DHA-producing soybean lines were conducted. </li> <li>Metabolomics of first-iteration of improved camelina astaxanthin lines completed.</li> <li>Field trials conducted on second generation of astaxanthin-producing camelina.</li> <li>Phenotypic and genotypic measurements of second-generation of sorghum vegetative oil lines completed.</li> <li>Single-nuclei sequencing on developing glabra2 (gl2) mutant versus wild-type seed to determine cell- and tissue-specific gene expression differences correlated with higher seed oil production.</li> <li>Analysis of the chromatin-remodeling mechanism that lipid-binding HD-Zip IV transcription factors utilize in controlling gene expression.</li> <li>Proximity labeling of a lipid sensing HD-Zip IV transcription factor followed by MS-based proteomics to determine the protein complex involved in regulation of gene expression.</li> <li>Characterization of the mechanism underlying subcellular localization of transcription factors in response to lipid changes. A predicted nuclear localization signal (NLS) was found to be necessary and sufficient for nuclear localization of two HD-Zip IV transcription factors. While the NLS overlaps with the DNA binding domain, mutant analysis shows that the two functions can be separated. Protein-protein interaction studies and mutant analysis indicate that an alpha importin is required for nuclear import. This work was completed and the paper has been published (Ahmad et al. 2024). <h1>Grants awarded</h1> PI: Doug Allen. Co-PI(s): Veena Veena, Timothy. Durrett. Agency: United Soybean Board. Title: Engineering Increased Protein and Oil in Soybeans for Improved Seed Value. Dates 10/1/2023 – 9/30/2024. Total cost: $ 207,183. PI: Edgar Cahoon; Co-PI: Tom E. Clemente, Nebraska Soybean Board. Title: Biotechnological Development of Optimized Soybean Germplasm for Aquaculture Feedstock. Dates 10/1/2023-9/30/2024. Total cost: $75,000 PI: Edgar Cahoon; Co-PIs: Ruth Welti, Kent Chapman, Marisol Berti, Sanju Sanjaya. USDA-NIFA. Title: 26th International Symposium on Plant Lipids and 1st International Camelina Conference. 3/1/2024-3/28/2025. Total cost: $31,000. PI: Edgar Cahoon; Co-PIs: Ruth Welti, Kent Chapman, Sanju Sanjaya. NSF. Title: 26th International Symposium on Plant Lipids. 3/1/2024-3/28/2025. Total cost: $21,900. PI: Edgar Cahoon; Co-PIs: Ruth Welti, Kent Chapman, Sanju Sanjaya. DOE. Title: 26th International Symposium on Plant Lipids. 3/1/2024-3/28/2025. Total cost: $10,000. PI: Edgar Cahoon; Co-PI: Erich Grotewold. DOE. Title:1st International Camelina Conference. 8/15/2024-8/14/2025. Total cost: $8,000 PI Rebecca Roston. Co-PIs Toshi Obata, James Schnable, Frank Harmon. NSF-PGRP “RESEARCH-PGR: Cycling to low-temperature tolerance” 05/2024 - 04/2027 Total cost: $1,800,000 PI: Steven Van Doren, co-Is: Jay Thelen, Philip Bates. Environmental Molecular Sciences Laboratory and Joint Genome Institute FICUS program. “Structural mechanisms of enzyme regulation to open the tap of plant oil synthesis” 10/2024 – 9/30/2026. In-kind value in 2024: $80,000 PI: Steven Van Doren. University of Missouri Research Council. “Enzyme Controlling Synthesis of Oils and Biofuel in Crops: Validation of Structural Models” 11/2023 - 12/2024 Total cost: $14,993 PI: Steven Van Doren. co-I Adam Yokom. MU CAFNR Joy of Discovery program. “A Braking Mechanism at Initiation of Oil Synthesis by a New Winter Cover Crop” 4/2024 - 3/2026 Total cost: $19,998 <h1>Patents</h1>  Kim H, Park K, Cahoon EB (2023) Methods and compositions for making ketocarotenoids. Application Date 10/09/2023   </li> </ul>

Publications

<h1>Publications</h1> Abdullah, H.M., Pang, N.,  Chilcoat, B., Shachar-Hill, Y., Schnell, D.J., and Dhankher, O.P.. Overexpression of the Phosphatidylcholine: Diacylglycerol Cholinephosphotransferase (PDCT) Gene Increases Carbon Flux Towards Triacylglycerol (TAG) Synthesis in Camelina sativa Seeds. Plant Physiology & Biochemistry, 208: 108470 (2024). https://doi.org/10.1016/j.plaphy.2024.108470 Alexander, L.E., Winkelman, D., Stenback, K.E., Lane, M., Campbell, K.R., Trost, E., Flyckt, K., Schelling, M.A., Rizhsky, L., Yandeau-Nelson, M.D., Nikolau, B.J. 2024. The impact of GLOSSY2 and GLOSSY2-LIKE BAHD-proteins in affecting the product profile of the maize fatty acid elongase. Front Plant Sci. 15: 1403779. doi: 10.3389/fpls.2024.1403779 Ahmad, B., Lerma-Reyes, R., Mukherjee, T., Nguyen, H.V., Weber, A.L., Cummings, E.E., Schulze, W.X., Comer, J.R., Schrick, K. 2024. Nuclear localization of HD-Zip transcription factor GLABRA2 is driven by Importin alpha. J. Exp. Bot. erae326. doi:10.1093/jxb/erae326. Azeez A, Bates PD (2024) Self-incompatibility based functional genomics for rapid phenotypic characterization of seed metabolism genes. Plant Biotechnology Journal. doi:<a href="https://doi.org/10.1111/pbi.14383">https://doi.org/10.1111/pbi.14383</a>  Chen, K., Bhunia, R.K., Wendt, M.M., Campidilli, G., McNinch, C., Hassan, A., Li, L., Nikolau, B.J., Yandeau-Nelson, M.D. 2024a. Cuticle development and the underlying transcriptome-metabolome associations during early seedling establishment. J Exp Bot.  erae311. doi: 10.1093/jxb/erae311. Chen, K., Alexander, L.E., Mahgoub, U., Okazaki, Y., Higashi, Y., Perera, A.M., Showman, L.J., Loneman, D., Dennison, T.S., Lopez, M., Claussen, R., Peddicord, L., Saito, K., Lauter, N., Dorman, K.S., Nikolau, B.J., Yandeau-Nelson, M.D. 2024b. Dynamic relationships among pathways producing hydrocarbons and fatty acids of maize silk cuticular waxes. Plant Physiol. 195(3): 2234-2255. doi: 10.1093/plphys/kiae150 Chen M, Wang S, Zhang Y, Fang D, Thelen JJ. (2023) Plastid Phosphatidylglycerol Homeostasis Influences Polar Lipid Synthesis in Arabidopsis. Metabolites. 13:318. Esterhuizen, L., Ampimah, N., Yandeau-Nelson, M.D., Nikolau, B.J., Sparks, E.E., Saha, R. AraRoot-A comprehensive genome-scale metabolic model for the Arabidopsis root system. Preprint in bioRxiv; doi: 10.1101/2024.07.28.605515 Hoffmann-Benning, S. and Simon-Plas, F. (2024). Editorial: Lipid signaling in plant physiology. Plant Science 334.<a href="https://doi.org/10.1016/j.plantsci.2024.112088"> https://doi.org/10.1016/j.plantsci.2024.112088</a> Holtsclaw RE, Mahmud S, Koo AJ. (2024) Identification and characterization of GLYCEROLIPASE A1 for wound-triggered JA biosynthesis in Nicotiana benthamiana leaves. Plant Mol Biol. 114:4 doi: 10.1007/s11103-023-01408-7. Johnson BS, Allen DK, Bates PD (2024) Triacylglycerol stability limits futile cycles and inhibition of carbon capture in oil-accumulating leaves. Plant Physiology. doi: 10.1093/plphys/kiae121  Kenchanmane Raju SK, Zhang Y, Mahboub S, Ngu DW, Qiu Y, Harmon FG, Schnable JC, Roston RL. Rhythmic lipid and gene expression responses to chilling in panicoid grasses. Journal of Experimental Botany. 2024 May 29:erae247. Kim, P., S. Mahboob, H.T. Nguyen, S. Eastman*, O. Meyer*, M. Sousek, R.E. Gaussoin, J.L. Brungardt, T.A. Jackson-Ziems, R. Roston, J.A. Alfano, T.E. Clemente, and M.Guo 2024. Characterization of soybean events with enhanced expression of the microtubule-associated protein 65-1 (MAP65-1). Mol. Plant-Microbe Int. 37:62-71. DOI: https://doi.org/10.1094/MPMI-09-23-0134-R Kimberlin AN, Mahmud S, Holtsclaw RE, Walker A, Conrad K, Morley SA, Welti R, Allen DK, and Koo AJ. Increasing oil production in leaves by engineering plastidial phospholipase A1. Under review. Kulke, M., Kurtz, E., Boren, D., Olson, D. M., Koenig, A. M., Hoffmann-Benning, S., & Vermaas, J. V. (2024). PLAT Domain Protein 1 (PLAT1/PLAFP) Binds to the Arabidopsis thaliana Plasma Membrane and Inserts a Lipid. Plant science 338. <a href="https://doi.org/10.1016/j.plantsci.2023.111900">https://doi.org/10.1016/j.plantsci.2023.111900</a> Lee, Y.J., Hapuarachchige, P., Larson, E., Le, N.goc; Forsman, Trevor, 2024, Visualizing 13C-labeled Metabolites in Maize Root Tips with Mass Spectrometry Imaging, J. Am. Soc. Mass Spectrom. 35, 7. <a href="https://doi.org/10.1021/jasms.4c00042">https://doi.org/10.1021/jasms.4c00042</a> Li-Beisson Y, Roston R. Plant and Algal Lipids: In All Their States and On All Scales. Plant and Cell Physiology. 2024 May, pcae061. Muthan B, Wang J, Welti R, Kosma DK, Yu L, Deo B, Khatiwada S, Vulavala VKR, Childs KL, Xu C, Durrett TP, Sanjaya SA. Mechanisms of Spirodela polyrhiza tolerance to FGD wastewater-induced heavy-metal stress: Lipidomics, transcriptomics, and functional validation. J Hazard Mater. 2024 May 5;469:133951. doi: 10.1016/j.jhazmat.2024.133951.  Na, S., Lee, Y.J., 2024, Mass Spectrometry Imaging of Arabidopsis thaliana with in vivo D2O Labeling, Front. Plant Sci. 15:1379299, <a href="https://doi.org/10.3389/fpls.2024.1379299">https://doi.org/10.3389/fpls.2024.1379299</a>. Neumann N, Harman M, Kuhlman A, Durrett TP. 2024. Arabidopsis diacylglycerol acyltransferase1 mutants require fatty acid desaturation for normal seed development. Plant J. 119: 916-926.  doi: 10.1111/tpj.16805 Neumann N, Fei T, Wang T, Durrett TP. 2023. Defining the physical properties of blends of acetyl-triacylglycerols derived from transgenic oil seeds. J Am Oil Chem Soc. 101(2): 197–204. doi: 10.1002/aocs.12746 Nguyen D, Groth N, Mondloch K, Cahoon EB, Jones K, Busta L (2024) Project ChemicalBlooms: Collaborating with citizen scientists to survey the chemical diversity and phylogenetic distribution of plant epicuticular wax blooms. 8 (5), e588 https://doi.org/10.1002/pld3.588 Osinuga A, Solís AG, Cahoon RE, Al-Siyabi A, Cahoon EB, Saha R (2024) Deciphering Sphingolipid Biosynthesis Dynamics in Arabidopsis thaliana cell cultures: Quantitative Analysis Amidst Data Variability. iScience DOI: <a href="https://doi.org/10.1016/j.isci.2024.110675">https://doi.org/10.1016/j.isci.2024.110675</a> Parchuri P, Bhandari S, Azeez A, Chen G, Johnson K, Shockey J, Smertenko A, Bates PD (2024) Identification of triacylglycerol remodeling mechanism to synthesize unusual fatty acid containing oils. Nature Communications 15 (1):3547. doi:10.1038/s41467-024-47995-x  Qin, P.,Chen, P.,Zhou, Y.,Zhang, W.,Zhang, Y.,Xu, J.,Gan, L.,Liu, Y.,Romer, J.,Dormann, P.,Cahoon, E. B. & Zhang, C. (2024) Vitamin E biofortification: enhancement of seed tocopherol concentrations by altered chlorophyll metabolism, Front Plant Sci. 15, 1344095 [10.3389/fpls.2024.1344095]. Quach, T., Nguyen, H., Meyer, O., S.J. Sato, Clemente, T.E., and Guo, M. 2023. Introduction of genome editing reagents and genotyping of derived edited alleles in soybean (Glycine max (L.) Merr.) Plant Genome Engineering: Methods & Protocols <a href="https://doi.org/10.1007/978-1-0716-3131-7_17">https://doi.org/10.1007/978-1-0716-3131-7_17</a> Quach, T.N., Sato, S.J., Behrens, M.R., Black, P.N., DiRusso, C.C., Cerutti, H.D. and Clemente, T.E.. 2023. A facile Agrobacterium-mediated transformation method for the model unicellular green algae Chlamydomonas reinhardtii. In Vitro Cellular & Mol Biol.-Plant 59:671-683. Schrick, K., Ahmad, B., Nguyen, H.V. 2023. HD-Zip IV transcription factors: Drivers of epidermal cell fate integrate metabolic signals. Curr Opin Plant Biol. 75:e102407. doi:10.1016/j.pbi.2023.102417 Shomo ZD, Mahboub S, Vanviratikul H, McCormick M, Tulyananda T, Roston RL, Warakanont J. All members of the Arabidopsis DGAT and PDAT acyltransferase families operate during high and low temperatures. Plant Physiology. 2024 May;195(1):685-97. Shomo ZD, Li F, Smith CN, Edmonds SR, Roston RL. From Sensing to Acclimation: The Role of Membrane Lipid Remodeling in Plant Responses to Low Temperatures. Plant Physiology. 2024 Jul 19:kiae382. Singh, G., Le, H., Ablordeppey, K., Long, S., Minocha, R., and Dhankher, O.P.. Overexpression of gamma-Glutamyl Cyclotransferases 2;1 (CsGGCT2;1) Reduces Arsenic Toxicity and Accumulation in Camelina sativa (L.). Plant Cell Reports, 43:14 (2024). https://doi.org/10.1007/s00299-023-03091-w Spivey WW, Rustgi S, Welti R, Roth MR, Burow MD, Bridges WC Jr, Narayanan S. Lipid modulation contributes to heat stress adaptation in peanut. Front Plant Sci. 2023 Dec 18;14:1299371. doi: 10.3389/fpls.2023.1299371. Surber SM, Thien Thao NP, Smith CN, Shomo ZD, Barnes AC, Roston RL. Exploring cotton SFR2’s conundrum in response to cold stress. Plant Signaling & Behavior. 2024 Dec 31;19(1):2362518. Tat, V.T., Lee, Y.J., 2024, Spatiotemporal Study of Galactolipid Biosynthesis in Duckweed with Mass Spectrometry Imaging and in vivo Isotope Labeling, Plant and Cell Physiology, 65(6), 986–998. <a href="https://doi.org/10.1093/pcp/pcae032">https://doi.org/10.1093/pcp/pcae032</a> Villalobos, J. A.,Cahoon, R. E.,Cahoon, E. B. & Wallace, I. S. (2024) Glucosylceramides impact cellulose deposition and cellulose synthase complex motility in Arabidopsis, Glycobiology. 34 [10.1093/glycob/cwae035]. Wang M, Garneau MG, Poudel AN, Lamm D, Koo AJ, Bates PD, Thelen JJ. (2022) Overexpression of pea α-carboxyltransferase in Arabidopsis and Camelina increases fatty acid synthesis leading to improved seed oil content. Plant J. 110:1035-1046. Wang S, Blume RY, Zhou ZW, Nazarenus TJ, Blume YB, Cahoon EB*, Chen L*, Liang G* (2024) Chromosome-level assembly and analysis of Camelina neglecta – a novel diploid model for camelina biotechnology research. Biotechnology for Biofuels and Bioproducts 17 (1), 17 (*Co-corresponding authors) https://doi.org/10.1186/s13068-024-02466-9 Vadde, B.V.L., Russell, N.J., Bagde, S.R., Askey, B., Saint-Antoine, M.M., Brownfield, B.A., Mughal, S., Apprill, L.E., Khosla, A., Clark, F.K., Schwarz, E.M., Alseekh, S., Fernie, A.R., Singh, A., Schrick, K., Fromme, J.C., Skirycz, A., Formosa-Jordan, P., Roeder, A.H.K. 2024. The transcription factor ATML1 maintains giant cell identity by inducing synthesis of its own long-chain fatty acid-containing ligands. Preprint in bioRxiv; doi:10.1101/2024.03.14.584694. Wojciechowska, I., Mukherjee, T., Knox-Brown, P., Hu, X., Khosla, A., Subedi, B., Ahmad, B., Mathews, G.L., Panagakis, A.A., Thompson, K.A., Peery, S.T.,  Szlachetko, J., Thalhammer, A., Hincha, D.K., Skirycz, A., Schrick, K. 2024. Arabidopsis PROTODERMAL FACTOR2 binds lysophosphatidylcholines and transcriptionally regulates phospholipid metabolism. New Phytol. (published online 7-1-2024). doi:10:1111/nph.19917 Xu, C.,Shaw, T.,Choppararu, S. A.,Lu, Y.,Farooq, S. N.,Qin, Y.,Hudson, M.,Weekley, B.,Fisher, M.,He, F.,Da Silva Nascimento, J. R.,Wergeles, N.,Joshi, T.,Bates, P. D.,Koo, A. J.,Allen, D. K.,Cahoon, E. B.,Thelen, J. J. & Xu, D. (2024) FatPlants: a comprehensive information system for lipid-related genes and metabolic pathways in plants, Database (Oxford). 2024 [10.1093/database/baae074].

Impact Statements

The LIPIDS of Crops multi-state research project has an overarching goal to increase the value of crop oilseeds by increasing seed oil content, making unusual and economically important fatty acids, finding new markets for existing or future vegetable oils and oilseed crops (e.g., camelina), and also adding value to the defatted meal particularly for niche crops like camelina. Each of these goals has the potential to impact the economy and move towards renewable energy independence. Additionally, LIPIDS of Crops is working to improve crop resilience to environmental stresses, including those associated with climate change. The NC-1203 group has interacted collaboratively to achieve project milestones during the year as indicated by milestones and 40 publications, 10 grant proposals funded, and 1 patents listed below, as well as standards and protocols that have been shared among participants. Future work will focus on completing the remaining and future milestones.

Date of Annual Report: 10/18/2025

Report Information

Annual Meeting Dates: 09/13/2025 - 09/14/2025
Period the Report Covers: 10/01/2024 - 09/30/2025

Participants

Allen, Doug - Donald Danforth Plant Science Center; Cahoon, Edgar - University of Nebraska, Lincoln (UNL); Durrett, Timothy - Kansas State University (KSU); Hoffmann-Benning, Susanne - Michigan State University; Huang, Chien-Yu - Louisiana State University; Kosma, Dylan – UNR; Louis, Joe - UNL; Parchuri, Prasad – KSU; Roston, Rebecca - UNL; Schrick, Kathrin - KSU; Thelen, Jay - University of Missouri (MU), Columbia; Welti, Ruth - KSU; Dhankher, Om Parkash - University of Massachusetts, Amherst; Koo, Abraham - MU; Mukherjee, Thiya – Texas A&M University San Antonio; Narayanan, Sruthi - Clemson University; Wang, Xuemin (Sam) - Donald Danforth Center; Yandeau-Nelson, Marna - Iowa State University (ISU); Van Doren, Steven - MU; Bates, Phil - Washington State University; Tamborindeguy, Cecilia - Texas A&M;, Scott Peck - MU; Young-Jin Lee - ISU; Adam Yokom - MU.

Brief Summary of Minutes

The 2025 NC-1203 meeting was held as a hybrid in-person and Zoom meeting on September 13-14, 2025, at Kansas State University in Manhattan, Kansas. Presentations of research progress and future directions were presented by the participants throughout the society meeting. Overall future research directions and writing of the renewal during Fall 2025 was discussed. Future meeting sites: In 2026 the meeting will be held at the University of Nevada, Reno (hosted by Dylan Kosma), and in 2027 the meeting will be held in conjunction with the 2027 Gordon Research Conference on Plant Lipids, in Pomona California, with more than half of the NC-1203 members planning to attend. Philip Bates will host the 2027 business meeting. The 2028 meeting will be held at the University of Missouri and be hosted by Jay Thelen.

Accomplishments

Research Activities and outputs:   Objective 1: Improve and extend methods for lipid characterization and measurement Significant progress was made this year toward enhancing lipid and protein characterization techniques across diverse plant systems, with innovations that support both fundamental research and applied breeding. Researchers developed and refined analytical platforms that improve speed, increase the specificity of compound identification, enhance quantification and standardization, and extend approaches to measure additional plant tissue types. Bates lab developed GC-FID based derivatization methods for rapid characterization of whole seed fatty acid composition and quantity for a variety of current and emerging oilseed crops including Camelina sativa, Thlaspi avernse (pennycress), Cuphea viscosissima, Brassica napus (var. Canola), Limnanthes alba (Meadowfoam) and Cannabis sativa L. (hemp) (Garneau, et al. 2025). This method will allow quicker screening of seed lipid content during research and breeding applications. The Lee group developed a new mass spectrometry imaging (MSI) technique that can determine carbon-carbon double bond positions of PC lipids, called OzMALDI, by introducing ozone gas into MALDI source (Rensner, et al., 2025). Using this method, we could successfully visualize PC double-bond isomers in camelina and soybean seeds, engineered by Cahoon’s lab. Also demonstrated by the group is the use of unsupervised machine learning for the MSI data analysis with in vivo isotope labeling (Johnson, et al., 2025). The Kosma Lab developed a GC-MS-based method for a complete characterization of external (pollenkitt) and internal fatty acids and specialized metabolites of pollen that requires a relatively small amount of pollen (~4-5 mg). This method has been used to investigate commercial and wild (Great Basin native) bee pollinated plant species. This will enable studies aimed at understanding pollen nutrition for pollinators in both ecological and agricultural contexts. The Thelen lab has developed a multiplexed AQUA-MRM, LC-MS/MS assay for absolute quantitation of Arabidopsis acetyl-CoA carboxylase catalytic and effector proteins and demonstrated the utility of this approach to study the spatiotemporal regulation of this multienzyme complex that catalyzes the committed step of de novo fatty acid synthesis.  The Welti lab is working with the Plant and Algal Lipid Interest Group of the International Lipidomics Society toward standardizing lipidomics results and creating reference mixtures. The group members have begun a multi-stage ring trial, in which they are analyzing the same samples. Together, these advances position the project to deliver robust, scalable, and integrative analytical tools that support breeding, metabolic engineering, ecological research, and global data harmonization.   Objective 2: Identify lipid-related mechanisms to increase agricultural resilience This year’s research advanced our understanding of how lipid metabolism contributes to plant resilience under environmental stress, nutrient limitation, and biotic pressure. Through multi-institutional collaborations, investigators explored genetic, enzymatic, structural, and signaling mechanisms that shape lipid function in plant defense, development, and stress adaptation. A major focus was uncovering metabolic bottlenecks and adaptations triggered by genetic modification. Genetic changes due to breeding or bioengineering can lead to unexpected metabolic adaptations that may create bottlenecks to the desired metabolic accumulation. Understanding these metabolic adaptations is the first step in developing metabolism based rational engineering or breeding approaches. In 2025 three publications from multi-state collaborations investigated metabolic adaptations due to mutation or engineering of lipid metabolism. The Allen and Bates labs collaborated to investigate bottlenecks to lipid accumulation in vegetative tissues in tobacco. Key results indicate that oil-starch futile cycling can inhibit photosynthesis limiting total plant growth and oil accumulation, but stabilization of oil content limits the futile cycle (Johnson et. al., 2025). Additionally, heat stress can limit total oil accumulation (Murphy et. al., 2025). The Thelen, Allen, Koo, and Bates labs took a multi-omic approach to understand the effect of mutations of negative regulators of acetyl-CoA carboxylase the key regulatory point of fatty acid biosynthesis. Key results indicate that large changes to central carbon metabolism were induced that ultimately led to increases in both seed oil and protein content in the model plant Arabidopsis (Kataya et. al., 2025). Since activity of the biotin carboxylase subunit (BC) of acetyl-CoA carboxylase by effectors regulates fatty acid synthesis, Yokom determined the structure of the dimer of BC by cryo-EM. The Van Doren group quantified the accessibility of the BC surface using mass spectra collected by the Thelen group. Van Doren used crosslinks of BC identified by mass spec at PNNL to dock two of Yokom’s dimer of BC into a tetramer. Three of four potential binding sites appear to be available for effectors to bind, in principle, in the tetramer. The Bates and Parchuri labs collaborated to characterize novel functionalities of key enzymes of oil biosynthesis DGAT1 and DGAT2 in a wide range of oil accumulating plants including Camelina sativa, pennycress, cotton, peanut, tung tree, castor. The results indicate never before recognized diversity in substrate selectivity for the sn-1,2 and sn-2,3 enantiomers of diacylglycerol the precursor to triacylglycerol. These results imply that different species may use a variety of mechanism in producing the final oil composition including triacylglycerol remodeling, and thus future bioengineering strategies to control oil composition will likely need to be crop specific. Additionally, the Bates lab discovered the key regulatory elements for DGAT1 expression in pollen, allowing for the first time complete replacement of endogenous oil biosynthetic enzymes with exogenous enzymes (McGuire et. al., 2025). The Koo lab is investigating how GLUTAMATE RECEPTOR LIKE (GLR) proteins mediate the systemic wound response for jasmonate (JA) biosynthesis. Through a structure-function analysis, they identified a region that, when deleted, eliminates systemic JA biosynthesis. In collaboration with the Peck lab, they confirmed the correct targeting of the mutated GLR to the plasma membrane, validating the results of the GLR variants. In addition, the Koo and Peck labs collaborated to discover that a higher order mutant of a group of calcium signaling proteins may be involved in systemic wound response. Separately, the Koo lab has identified a novel chemical inhibitor that targets JA pathway and suppresses the wound response.   The Wang Lab investigated the role of membrane phospholipid remodeling in rapeseed and camelina response to phosphate deficiency, The results show that the phosphate starvation induced phospholipase C, NPC4, promotes not only membrane lipid remodeling , but also P remobilization from old, senescent, source tissues to young, growing, sink tissues under P deficiency (Li et al., 2025). Manipulations of the pathway have potential to enhance crop plant growth and seed production, particularly under P-deficient conditions (Yang et al., 2025). Welti lab and Turkish collaborators investigated the effect of salicylic acid priming of wheat seeds on wheat tolerance for cadmium and showed that the positive effects of priming do not seem to be mediated through lipid alterations (Colak et al., 2025). The Roston lab made progress in understanding freezing-activated galactolipid remodeler SFR2 as a reporter to uncover how plants gauge cold severity, employing yeast two hybrid analysis to identify yet another kinase potentially involved that was not previously associated with cold tolerance. Current data support a kinase/phosphatase pathway that modulates membrane repair in proportion to stress intensity. A future collaboration is planned with the Thelen lab to investigate the role of the kinases more broadly. In sorghum, time-bounded multi-omics found that while some cold responses to acclimation are conserved, many metabolite shifts are line-specific; photosynthetic readouts were reproducible only when measured within ~3 h of treatment, revealing strong time-of-day constraints, and lipid results were different between two tolerant lines. These findings motivated an updated model—daily temperature cycles synchronize lipid and soluble-metabolite programs essential for tolerance—which the Roston lab is now testing in collaboration with others at Nebraska and Frank Harmon of USDA. The Louis Lab, in collaboration with the Cahoon and Welti Labs, recently identified a key role for triacylglycerols (TAGs), whose major constituents are fatty acids, in sorghum defense against the sugarcane aphid (SCA; Melanaphis sacchari). To investigate this, we employed electrospray ionization mass spectrometry (ESI-MS) to compare lipid profile alterations in SCA-resistant (SC265) and SCA-susceptible (SC1345) sorghum genotypes before and after aphid feeding. Our preliminary analyses revealed that sorghum lines differing in resistance also exhibit distinct lipid signatures. Notably, SCA feeding caused significant changes in TAG accumulation. While basal TAG levels were higher in the susceptible SC1345 line, seven days of SCA feeding led to a sharp reduction in several molecular species of TAGs in this genotype. In contrast, the resistant SC265 line maintained relatively stable TAG profiles under infestation. These findings suggest that TAG metabolism, and possibly TAG-derived metabolites, play a critical role in sorghum defense against SCA. Ongoing work, in collaboration with the Cahoon Lab at UNL, is focused on elucidating the specific mechanisms underlying this TAG-mediated defense.  The Hoffmann-Benning lab has made progress in their Molecular Dynamics (MD) Modeling approach. They were able to show that PLAFP not only inserts into the membrane to detect and remove phosphatidic acid, it also dissociates from the membrane once the lipid is bound. This supports the hypothesis that protein and lipid jointly serve as a long-distance signal. To further investigate, they designed light-regulated gene expression for PLAFP and two control genes (FT, GRIP1) and were able to show movement of PLAFP throughout the plant. Gene-expression studies are currently in progress to support those findings. The Schrick lab initiated MD simulations of lysophospholipid binding to the steroidogenic acute regulatory protein (StAR)-related lipid transfer (START) domain from homeodomain leucine-zipper (HD-Zip) IV transcription factor PDF2. These studies follow experimental data showing that PDF2 recruits lysophosphatidylcholine 18:1 in vivo and in vitro (Wojciechowska et al., 2024). The MD simulations address both lipid and membrane binding activities of the START domain and suggest that membrane contact occurs through key arginine residues in predicted loops on the periphery of the binding pocket. A separate study found that nuclear localization of HD-Zip IV proteins occurs through Importin alpha-mediated recognition of a classic nuclear localization sequence that overlaps with the DNA binding domain (Ahmad et al., 2024). The Huang Lab is studying the role of the lipid droplets during plant-microbe interactions of important fungal diseases of soybean, tomato and strawberry. The group screened Plant-derived lipid biosynthesis inhibitors and identified a batch of inhibitors, including flavonoids, phthalides, fatty acids, and terpenes, that exhibited antimicrobial or toxin-suppressing effects. The tested fungal pathogens include Cercospora kikuchii, C. cf. flagellaris, and C. sojina, which cause Cercospora Leaf Blight and Frogeye Leaf Spot on soybeans. Additionally, Botrytis cinerea and Agroathelia rolfsii, which cause gray mold and southern blight diseases, respectively, have a broad host range; aggressive Neopestalotiopsis sp., which was reported as an emerging strawberry disease in recent years. A future collaboration is planned with Sanjaya and Welti to explore the functions of lipids in defense. The Yandeau-Nelson lab, in partial collaboration with previous NC-1203 member Basil Nikolau, have assessed the role of the genetic redundancy in cuticular wax biosynthesis in maize, focusing on the 28 genes encoding ketoacyl-CoA synthetases. Taking a synthetic biology approach, they have tested different iterations of the fatty acid elongase pathway from maize (i.e. ZmFAE) in yeast and have demonstrated differences in very long chain fatty acid profiles among the different ZmFAE combinations. The Yandeau-Nelson team has also functionally characterized maize glossy2 and glossy2-like within the ZmFAE systems, and also in single and double mutants in maize.  Importantly, cuticular wax structure changes in gl2; gl2-like double mutants and is accompanied by a change in the cuticle’s function as a water barrier. A future collaboration is planned with the Louis lab to assess whether these mutants experience changed aphid-plant interactions. Together, these discoveries highlight the central role of lipid pathways in plant resilience—from membrane integrity and nutrient remobilization to stress signaling and defense. The integration of structural biology, multi-omics, and synthetic biology continues to drive innovation in crop improvement, positioning lipid metabolism as a powerful lever for agricultural sustainability.   Objective 3: Develop crops with improved yield and/or functionality This year’s collaborative research advanced the frontier of crop improvement by harnessing lipid metabolism to enhance yield, oil content, stress resilience, and value-added traits across a range of species. Through strategic metabolic engineering, gene discovery, and translational field studies, the project demonstrated how lipid pathways can be leveraged to meet the growing demands for food, fuel, and climate-resilient agriculture. A central theme was the enhancement of oil accumulation and composition in both seeds and vegetative tissues. The Bates lab demonstrated that recently discovered triacylglycerol remodeling enzymes from Physaria fendleri could be utilized to enhance both total oil accumulation, and accumulation of valuable hydroxylated fatty acids in Arabidopsis thaliana and Camelina sativa. The Mukherjee, Durrett, and Allen labs described the changes in soybean seed size as a consequence of SDP1 suppression, which have now been verified in multiple generations. Importantly, observed increases in seed size do not negatively impact seed number; investigations as to the underlying mechanism are ongoing. Ongoing collaborative work will generate and characterize stable mutant lines that investigate carbon partitioning and lipid turnover in developing soybean seeds. The Allen lab in collaboration with Koo and Thelen reported the beta oxidation of fatty acids including some that were lipid-derived in a recent report (Koley et al., 2025). The study indicated that in multiple seed and a high-oil leaf system, that fat fatty acid oxidation occurs across development and appears to be, in collaboration with Koo and Thelen, reported the beta oxidation of fatty acids, including some that were lipid-derived, a regular part of metabolism. The Koo lab collaborated with the Welti and Allen labs to boost plant biomass oil production using pathway engineering. The inducible expression of a plastid-localized phospholipase A1 (PLA1) called DEFECTIVE AND ANTHER DEHISCENCE 1 (DAD1) has led to the accumulation of TAGs in leaves, forming lipid droplets (LDs) (Kimberlin et al., 2025). Lipidomics and FAME analyses revealed that the accumulated TAGs were primarily composed of 18:3 fatty acids coming from MGDG and DGDG. Similar results were observed when DAD1 or its Nicotiana benthamiana homolog, NbDAD1, were transiently expressed in N. benthamiana. A stable transgenic soybean line harboring the inducible PLA1 also successfully increased leaf TAGs. Building on this work, the Koo lab is developing genetic tools and new lines with greater oil content.  In collaboration with the Allen and Lee labs, the Durrett lab metabolically engineered camelina and pennycress lines to accumulate almost pure levels of acetyl-TAG, an unusually structured lipid with useful properties (Alkotami et al., 2024). The trace amounts of endogenous TAG remaining were localized to the embryonic axis of the seed. Future work will investigate the source and role of this residual TAG. The Kosma lab recently developed a variety of potato that has reduced sprouting without compromising overall tuber yield (Vulavala et al., 2024). This unexpected discovery came about through CRISPR-Cas9 gene editing targeting transcription factors that regulate specialized lipid metabolism (wound suberin) during tuber wound healing. The Cahoon lab successfully engineered oilseed camelina to produce the high-value ketocarotenoid astaxanthin, a red pigment with exceptional antioxidant properties. Utilizing genes from the plant Adonis aestivalis, their strategy led to the nearly complete conversion of β-carotene to ketocarotenoids, primarily astaxanthin. Field trials demonstrated a maximal accumulation of approximately 135 μg/g seed weight, with astaxanthin making up about 47 μg/g. The research also showed that the extracted oil from these seeds had enhanced oxidative stability, making it useful for food applications like oleogels. This breakthrough provides a new, sustainable plant-based source for a commercially valuable pigment used in aquaculture and the food industry, reducing reliance on less efficient methods (Kim et al., 2025). The Cahoon and Clemente labs successfully engineered high-yielding sorghum to accumulate energy-dense vegetable oil in its leaves and stems, a novel approach for producing renewable diesel and sustainable aviation fuel. Using a "push-pull-protect" genetic strategy, the engineered sorghum accumulated oil in its leaves and stems to amounts of up to 5.5% DW and 3.5% DW respectively under field conditions. These results represented a 78-fold increase in leaves and a 58-fold increase in stems compared to wild-type plants. Their work highlights the effectiveness of a lab-to-field pipeline for developing high-vegetative-oil biomass crops. This achievement demonstrates a powerful new approach to creating feedstocks for sustainable fuels, offering a promising solution to strengthen American energy independence (Park et al., 2025). The Dhankher Lab cloned and characterized MGAT1 genes encoding monoacylglycerol acyltransferase from Camelina sativa. Overexpression of CsMGAT1 in Camelina under the control of seed-specific BcNA1 (napin) promoter from Brassica napus, resulted in a significant increase in seed yield and oil contents. Camelina transgenics exhibited a 33-57% increase in seed yield, 8–10% seed oil content, >20% oil yields per plant, and altered polyunsaturated fatty acid (PUFA) content, compared to their parental wild-type (WT) plants. Results from [14C]acetate labeling of Camelina developing embryos expressing CsMGAT in culture indicated increased rates of radiolabeled fatty acid incorporation into glycerolipids compared to WT embryos. These findings suggest that overexpression of MGAT positively impacts fatty acid flux into lipids, contributing to increased oil accumulation and potentially enhancing seed yield in Camelina. Narayanan Lab investigated the expression of the genes regulating lipid metabolic changes contributing to heat tolerance in peanut genotypes. They conducted a comprehensive lipidome analysis of 52 peanut recombinant inbred lines (RILs) of F6 population derived from a cross between the heat-tolerant genotype ICGS76 and the heat-susceptible TamrunOL02, under optimum (29/20°C) and heat stress conditions (38/28°C). They found that the sequestration of unsaturated acyl chains from membrane lipids to triacylglycerols (TG) and sterol esters (SE) helps to reduce the unsaturation levels in membrane lipids which in turn helps to maintain optimal membrane fluidity and integrity under heat stress conditions. They further investigated the expression patterns of key genes involved in the identified lipid remodeling. The tested genes were diacylglycerol acyltransferases (DGAT1-2, DGAT3-3), fatty acid desaturase (FAD3-2), phospholipid:diacylglycerol acyltransferase (PDAT), acyl-coA:sterol acyltransferase (ASAT), phospholipid:sterol acyl transferase (PSAT) and heat inducible lipase (HIL1). Gene expression analysis revealed an upregulation of ASAT, PSAT, DGAT3-3 and PDAT, that uniquely regulate the acylation of sterols and TGs. This result confirmed the role of TGs and SEs in heat stress tolerance through acyl sequestration. FAD3-2 which converts 18:2 fatty acids to 18:3, showed decreased expression, potentially contributing to reduced fatty acid unsaturation levels in membrane lipids by lowering the 18:3 fatty-acid amount under heat stress. The identified genes (ASAT, PSAT, DGAT3-3, PDAT, and FAD3-2) and associated lipid-related mechanisms of heat-stress tolerance will help develop heat-tolerant peanut varieties. Further, the genes will help develop molecular markers associated with heat tolerance that will speed up the peanut breeding programs for heat tolerance. These integrated efforts underscore the potential of lipid-based strategies to improve crop performance, resilience, and market value across diverse agricultural systems.   Milestones: <ul> <li>Developed method for rapid characterization of whole seed fatty acid composition and quantity for a variety of current and emerging oilseed crops (Garneau et al. 2025).</li> <li>Developed a new MSI technique to visualize carbon-carbon double bonds of lipids (Rensner et al., 2025).</li> <li>Demonstrated the use of unsupervised machine learning for the data analysis of MSI with in vivo isotope labeling (Johnson et al., 2025).</li> <li>Identified DIACYLGLYCEROL ACYLTRANSFERASE 1 regulatory elements essential for expression in pollen, and key to ultimate replacement of endogenous acyltransferase with those leading to higher yields or altered fatty acid compositions (McGuire et al. 2025).</li> <li>Engineered a new plant-based platform using the oilseed camelina to produce the high-value pigment astaxanthin for use in food and aquaculture (Kim et al., 2025).</li> <li>Identified metabolic adaptations to mutations and genetic engineering that lipid total oil accumulation in seeds and leaves (Kataya et al., 2025; Murphy et. al., 2025; Johnson et. al., 2025; Kimberlin et al., 2025). This knowledge will be the basis for new hypothesis driven approaches to enhance lipid accumulation in different plant tissues.</li> <li>Demonstrated fatty acid catabolism is active throughout development in filling seed and transgenic high-oil leaves (Koley et al., 2025).</li> <li>Developed a new pathway to create high-yielding sorghum capable of producing large amounts of vegetable oil in its leaves and stems for renewable diesel and sustainable aviation fuel (Park et al., 2025).</li> <li>Identified a lysophospholipid ligand for a transcriptional regulator that plays a role in epidermal development and response to phosphate limitation (Wojciechowska et al., 2024).</li> <li>Characterized the nuclear localization mechanism of a lipid-binding transcription factor implicated in regulation of seed oil levels (Ahmad et al., 2024)</li> <li>Developed a new variety of potato with reduced sprouting during cold storage that stands to significantly reduce postharvest potato tuber yield losses (Vulavala et al., 2024).</li> <li>Identified that membrane fluidity changes in response to temperature gated rice colonization by Magnaporthe oryzae (Richter et al. 2024).</li> <li>Identified lipid traits key to improving maize cold tolerance (Ojeda-Rivera et al. 2025).</li> <li>Narayanan spent three months of her sabbatical (March-May 2025) in the Allen Lab and established a new collaborative program that focuses on carbon metabolism for enhancing the quality of oilseed crops under optimal and heat-stress conditions.</li> </ul>

Publications

<ol> <li>Ahmad, B., Lerma-Reyes, R., Mukherjee, T., Nguyen, H.V., Weber, A.L., Cummings, E.E., Schulze, W.X., Comer, J.R., Schrick, K. 2024. Nuclear localization of HD-Zip transcription factor GLABRA2 is driven by Importin α. J Exp Bot. 75(20):6441-6461. doi:10.1093/jxb/erae326.</li> <li>Garneau MG, Parchuri P, Zander N, Bates PD (2025) Rapid quantification of whole seed fatty acid amount, composition, and shape phenotypes from diverse oilseed species with large differences in seed size. Plant Methods 21: 67</li> <li>McGuire ST, Shockey J, Bates PD (2025) The first intron and promoter of Arabidopsis DIACYLGLYCEROL ACYLTRANSFERASE 1 exert synergistic effects on pollen and embryo lipid accumulation. New Phytologist 245: 263-281</li> <li>Kataya A, Nascimento JRS, Xu C, Garneau MG, Koley S, Kimberlin A, Mukherjee T, Mooney BP, Xu D, Bates PD, Allen DK, Koo AJ, Thelen JJ (2025) Comparative Omics Reveals Unanticipated Metabolic Rearrangements in a High-Oil Mutant of Plastid Acetyl-CoA Carboxylase. Journal of Proteome Research 24: 2675-2688</li> <li>Kim, H.,Liu, L.,Han, L.,Park, K.,Kim, H. J.,Nguyen, T.,Nazarenus, T. J.,Cahoon, R. E.,Haslam, R. P.,Ciftci, O.,Napier, J. A. & Cahoon, E. B. (2025) Oilseed-based metabolic engineering of astaxanthin and related ketocarotenoids using a plant-derived pathway: Lab-to-field-to-application, Plant Biotechnology Journal. 23, 3451-3464 [<a href="https://doi.org/10.1111/pbi.70148">https://doi.org/10.1111/pbi.70148</a>].</li> <li>Park, K.,Quach, T.,Clark, T. J.,Kim, H.,Zhang, T.,Wang, M.,Guo, M.,Sato, S.,Nazarenus, T. J.,Blume, R.,Blume, Y.,Zhang, C.,Moose, S. P.,Swaminathan, K.,Schwender, J.,Clemente, T. E. & Cahoon, E. B. (2025) Development of vegetative oil sorghum: From lab-to-field, Plant Biotechnology Journal. 23, 660-673 [https://doi.org/10.1111/pbi.14527].</li> <li>Koley S, Jyoti P, Lingwan M, Wei M, Xu C, Chu KL,  Williams RB, Koo AJ, Thelen JJ, Xu D, Allen DK (2025) Persistent fatty acid catabolism during plant oil synthesis. Cell Reports 44(4):115492 (2025)</li> <li>Rhee, S. Y.,Anstett, D. N.,Cahoon, E. B.,Covarrubias-Robles, A. A.,Danquah, E.,Dudareva, N.,Ezura, H.,Gilbert, K. J.,Gutiérrez, R. A.,Heck, M.,Lowry, D. B.,Mittler, R.,Muday, G.,Mukankusi, C.,Nelson, A. D. L.,Restrepo, S.,Rouached, H.,Seki, M.,Walker, B.,Way, D. & Weber, A. P. M. (2025) Resilient plants, sustainable future, Trends in Plant Science. 30, 382-388 [10.1016/j.tplants.2024.11.001].</li> <li>Blume, R. Y.,Hotsuliak, V. Y.,Nazarenus, T. J.,Cahoon, E. B. & Blume, Y. B. (2024) Genome-wide identification and diversity of FAD2, FAD3 and FAE1 genes in terms of biotechnological importance in Camelina species, BMC Biotechnology. 24, 107 [10.1186/s12896-024-00936-4].</li> <li>Colak N, Kurt-Celebi A, Roth MR, Welti R, Torun H, Ayaz FA. (2025) Salicylic acid priming before cadmium exposure increases wheat growth but does not uniformly reverse cadmium effects on membrane glycerolipids. Plant Biol (Stuttg). 27(1):79-91. doi: 10.1111/plb.13736.</li> </ol> <ol start="11"> <li>Dong, J., Croslow, S. W., Lane, S. T., Castro, D. C.,Blanford, J., Zhou, S.,Park, K., Burgess, S., Root, M., Cahoon, E. B., Shanklin, J., Sweedler, J. V., Zhao, H. & Hudson, M. E. 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Impact Statements

The LIPIDS of Crops multi-state research project has an overarching goal to increase the value of crop oilseeds by increasing seed oil content, making unusual and economically important fatty acids, finding new markets for existing or future vegetable oils and oilseed crops (e.g., camelina), and also adding value to the defatted meal particularly for niche crops like camelina. Each of these goals has the potential to impact the economy and move towards renewable energy independence. Additionally, LIPIDS of Crops is working to improve crop resilience to environmental stresses, including those associated with climate change. The NC-1203 group has interacted collaboratively to achieve project milestones during the year as indicated by milestones and 31 publications, 9 grant proposals funded, and 1 patent filed listed below, as well as standards and protocols that have been shared among participants.
Grants awarded: 1. PI: Philip Bates. Co-PI: Prasad Parchuri. Agency: USDA-NIFA. Title: Elucidation of triacylglycerol remodeling mechanisms in Brassicaceae crops for designer seed oils. Dates: 9/1/2025 – 8/31/2028. Total cost: $649,993. 2. PI. Timothy Durrett. Co-PIs: Ruth Welti, Kathrin Schrick, Vara Prasad, Michael Chao. Agency: NSF. Title: MRI: Track 1 Acquisition of a quadrupole time-of-flight mass spectrometer and accessories for the Kansas Lipidomics Research Center. Dates: 9/15/2025 - 9/14/2028. Total cost: $1,096,696 3. PI: Doug Allen. Co-PIs: Veena Veena, Timothy. Durrett. Agency: United Soybean Board. Title: Engineering Increased Protein and Oil in Soybeans for Improved Seed Value. Dates 10/1/2024 – 9/30/2025. Total cost: $ 211,652. 4. PI: Ozan Ciftci. Co-PIs: Edgar Cahoon, Julia McQuillan, Tracy Niday. Agency: National Science Foundation. Title: NSF Global Centers: Food Innovation and Diversification to Advance the Bioeconomy (FoodID). Dates: 1/1/2025-12/31/2027. Total cost: $2,000,000. 5. PI: Edgar Cahoon. Agency: Nebraska Soybean Board. Title: Genetic Enhancement of Soybean Oil Content and Quality. Dates: 10/1/2024-9/30/2025. Total cost: $87,365. 6. PI: Rebecca Roston. Agency: NSF-PGRP “RESEARCH-PGR: Cycling to low-temperature tolerance” with co-PIs Toshi Obata, James Schnable, Frank Harmon Dates: 5/1/2024 - 4/30/2027. Total cost: $1,800,000 7. PI: Chien-Yu. Co-PI: Sara Thomas-Sharma. Agency: Louisiana Soybean & Grain Research & Promotion Board. Title: Implement Plant-Derived Natural Molecules to Manage Important Fungal Diseases in Soybeans. Dates: 5/1/2024 - 4/30/2025. Total cost: $27,662. 8. PI: Raghuwinder Singh. Co-PIs: Chien-Yu Huang, Congliang Zhou, Mary Helen Ferguson, Clark Robertson. Agency: Louisiana Department of Agriculture and Forestry. Title: Improving disease detection, forecasting and developing best management practices to mitigate Neopestalotiopsis leaf, fruit and crown disease of strawberry. Dates: 10/10/2025 - 5/31/2027. Total cost: $74,277. 9. PI: Om Parkash Dhankher, Co-PIs: Baoshan Xing. Agency: Advanced Research Projects Agency- Energy (ARPAe under DOE). Title: Camelina sativa for Hyperaccumulation of Nickel (CaSH-Ni). Dates: 12/01/2024- 11/30/2027. Total cost: $1,297,055.
Patent filed: Vulavala VK, Kosma DK, Santos P, inventors; University of Nevada, Reno, assignee. Potato variety named'unr-01'. United States patent application US 18/739,142. 2024 Dec 26.