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Title: Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil

Abstract

Modified fatty acids (mFA) have diverse uses, e.g., cyclopropane fatty acids (CPA) are feedstocks for producing coatings, lubricants, plastics, and cosmetics. The expression of mFA-producing enzymes in crop and model plants generally results in lower levels of mFA accumulation than in their natural-occurring source plants. In order to further our understanding of metabolic bottlenecks that limit mFA accumulation, we generated transgenic Camelina sativa lines co-expressing Escherichia coli cyclopropane synthase (EcCPS) and Sterculia foetida lysophosphatidic acid acyltransferase (SfLPAT). In contrast to transgenic CPA-accumulating Arabidopsis, CPA accumulation in camelina caused only minor changes in seed weight, germination rate, oil accumulation, and seedling development. CPA accumulated to much higher levels in membrane than storage lipids, comprising more than 60% of total fatty acid in both phosphatidylcholine (PC) and phosphatidylethanolamine (PE) versus 26% in diacylglycerol (DAG) and 12% in triacylglycerol (TAG) indicating bottlenecks in the transfer of CPA from PC to DAG and from DAG to TAG. Upon coexpression of SfLPAT with EcCPS, di-CPA-PC increased by ~50% relative to lines expressing EcCPS alone with the di-CPA-PC primarily observed in the embryonic axis and mono-CPA-PC primarily in cotyledon tissue. EcCPS-SfLPAT lines revealed a redistribution of CPA from the sn-1 to sn-2 positions within PC andmore » PE that was associated with a doubling of CPA accumulation in both DAG and TAG. Finally, the identification of metabolic bottlenecks in acyl transfer between site of synthesis (phospholipids) and deposition in storage oils (TAGs) lays the foundation for the optimizing CPA accumulation through directed engineering of oil synthesis in target crops.« less

Authors:
 [1];  [2];  [3];  [4];  [1];  [5];  [4];  [3];  [2];  [6];  [6]
  1. Stony Brook Univ., NY (United States). Dept. of Chemistry. Biochemistry and Cell Biology Dept.
  2. Univ. of Nebrask. Center for Plant Science Innovation and Dept. of Biochemistry
  3. Univ. of North Texas, Denton, TX (United States). Center for Advanced Scientific Computing and Modeling (CASCaM). Dept. of Biological Sciences
  4. Brookhaven National Lab. (BNL), Upton, NY (United States). Biology Dept.
  5. Stony Brook Univ., NY (United States). Biochemistry and Cell Biology Dept.
  6. Stony Brook Univ., NY (United States). Dept. of Chemistry. Biochemistry and Cell Biology Dept.; Brookhaven National Lab. (BNL), Upton, NY (United States). Biology Dept.
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Science Foundation (NSF)
OSTI Identifier:
1412647
Report Number(s):
BNL-114333-2017-JA
Journal ID: ISSN 1467-7644
Grant/Contract Number:  
SC0012704; FG02-09ER64812; KC0304000; DBI 1117680
Resource Type:
Accepted Manuscript
Journal Name:
Plant Biotechnology Journal
Additional Journal Information:
Journal Volume: 16; Journal Issue: 4; Journal ID: ISSN 1467-7644
Publisher:
Society for Experimental Biology; Association of Applied Biology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; cyclopropane fatty acid; Camelina sativa; unusual fatty acid; lipid metabolism; triacylglycerol; lipid synthesis

Citation Formats

Yu, Xiao-Hong, Cahoon, Rebecca E., Horn, Patrick J., Shi, Hai, Prakash, Richa Rawat, Cai, Yuanheng, Hearney, Maegan, Chapman, Kent D., Cahoon, Edgar B., Schwender, Jorg, and Shanklin, John. Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil. United States: N. p., 2017. Web. doi:10.1111/pbi.12839.
Yu, Xiao-Hong, Cahoon, Rebecca E., Horn, Patrick J., Shi, Hai, Prakash, Richa Rawat, Cai, Yuanheng, Hearney, Maegan, Chapman, Kent D., Cahoon, Edgar B., Schwender, Jorg, & Shanklin, John. Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil. United States. doi:10.1111/pbi.12839.
Yu, Xiao-Hong, Cahoon, Rebecca E., Horn, Patrick J., Shi, Hai, Prakash, Richa Rawat, Cai, Yuanheng, Hearney, Maegan, Chapman, Kent D., Cahoon, Edgar B., Schwender, Jorg, and Shanklin, John. Wed . "Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil". United States. doi:10.1111/pbi.12839. https://www.osti.gov/servlets/purl/1412647.
@article{osti_1412647,
title = {Identification of bottlenecks in the accumulation of cyclic fatty acids in camelina seed oil},
author = {Yu, Xiao-Hong and Cahoon, Rebecca E. and Horn, Patrick J. and Shi, Hai and Prakash, Richa Rawat and Cai, Yuanheng and Hearney, Maegan and Chapman, Kent D. and Cahoon, Edgar B. and Schwender, Jorg and Shanklin, John},
abstractNote = {Modified fatty acids (mFA) have diverse uses, e.g., cyclopropane fatty acids (CPA) are feedstocks for producing coatings, lubricants, plastics, and cosmetics. The expression of mFA-producing enzymes in crop and model plants generally results in lower levels of mFA accumulation than in their natural-occurring source plants. In order to further our understanding of metabolic bottlenecks that limit mFA accumulation, we generated transgenic Camelina sativa lines co-expressing Escherichia coli cyclopropane synthase (EcCPS) and Sterculia foetida lysophosphatidic acid acyltransferase (SfLPAT). In contrast to transgenic CPA-accumulating Arabidopsis, CPA accumulation in camelina caused only minor changes in seed weight, germination rate, oil accumulation, and seedling development. CPA accumulated to much higher levels in membrane than storage lipids, comprising more than 60% of total fatty acid in both phosphatidylcholine (PC) and phosphatidylethanolamine (PE) versus 26% in diacylglycerol (DAG) and 12% in triacylglycerol (TAG) indicating bottlenecks in the transfer of CPA from PC to DAG and from DAG to TAG. Upon coexpression of SfLPAT with EcCPS, di-CPA-PC increased by ~50% relative to lines expressing EcCPS alone with the di-CPA-PC primarily observed in the embryonic axis and mono-CPA-PC primarily in cotyledon tissue. EcCPS-SfLPAT lines revealed a redistribution of CPA from the sn-1 to sn-2 positions within PC and PE that was associated with a doubling of CPA accumulation in both DAG and TAG. Finally, the identification of metabolic bottlenecks in acyl transfer between site of synthesis (phospholipids) and deposition in storage oils (TAGs) lays the foundation for the optimizing CPA accumulation through directed engineering of oil synthesis in target crops.},
doi = {10.1111/pbi.12839},
journal = {Plant Biotechnology Journal},
number = 4,
volume = 16,
place = {United States},
year = {2017},
month = {9}
}

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