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Specialized lysophosphatidic acid acyltransferases contribute to unusual fatty acid accumulation in exotic Euphorbiaceae seed oils

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Abstract

Main conclusion

In vivo and in vitro analyses of Euphorbiaceae species’ triacylglycerol assembly enzymes substrate selectivity are consistent with the co-evolution of seed-specific unusual fatty acid production and suggest that many of these genes will be useful for biotechnological production of designer oils.

Many exotic Euphorbiaceae species, including tung tree (Vernicia fordii), castor bean (Ricinus communis), Bernardia pulchella, and Euphorbia lagascae, accumulate unusual fatty acids in their seed oils, many of which have valuable properties for the chemical industry. However, various adverse plant characteristics including low seed yields, production of toxic compounds, limited growth range, and poor resistance to abiotic stresses have limited full agronomic exploitation of these plants. Biotechnological production of these unusual fatty acids (UFA) in high yielding non-food oil crops would provide new robust sources for these valuable bio-chemicals. Previous research has shown that expression of the primary UFA biosynthetic gene alone is not enough for high-level accumulation in transgenic seed oils; other genes must be included to drive selective UFA incorporation into oils. Here, we use a series of in planta molecular genetic studies and in vitro biochemical measurements to demonstrate that lysophosphatidic acid acyltransferases from two Euphorbiaceae species have high selectivity for incorporation of their respective unusual fatty acids into the phosphatidic acid intermediate of oil biosynthesis. These results are consistent with the hypothesis that unusual fatty acid accumulation arose in part via co-evolution of multiple oil biosynthesis and assembly enzymes that cooperate to enhance selective fatty acid incorporation into seed oils over that of the common fatty acids found in membrane lipids.

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Abbreviations

DGAT:

Diacylglycerol acyltransferase

ESA:

Eleostearic acid

FADX:

Tung tree fatty acid conjugase X

FAH:

Castor fatty acid hydroxylase

FAME:

Fatty acid methyl ester

FID:

Flame ionization detection

GPAT:

Glycerol-3-phosphate acyltransferase

HFA:

Hydroxy fatty acids

HPLC:

High-performance liquid chromatography

GC:

Gas chromatography

LPA:

Lysophosphatidic acid

LPAT:

Lysophosphatidic acid acyltransferase

PCR:

Polymerase chain reaction

PDAT:

Phospholipid: diacylglycerol acyltransferase

PDCT:

Phosphatidylcholine:diacylglycerol cholinephosphotransferase

TLC:

Thin layer chromatography

UFA:

Unusual fatty acid

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Acknowledgements

The authors would like to thank Ms. Tien Thuy Vuong for technical assistance. This work was supported by the U.S. Department of Agriculture, Agricultural Research Service Current Research Information System project number 6054-41000-102-00D (to JS and CM) and the National Science Foundation (Directorate for Biological Sciences, Division of Molecular and Cellular Bioscience, award #1613923, to PDB and JS).

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  1. Hari Kiran Kotapati & Philip D. Bates

    Present address: Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA

Authors and Affiliations

  1. United States Department of Agriculture, Agricultural Research Service, Southern Regional Research Center, New Orleans, LA, 70124, USA

    Jay Shockey & Catherine Mason

  2. Department of Plant Breeding, Swedish University of Agricultural Sciences, 230 53, Alnarp, Sweden

    Ida Lager & Sten Stymne

  3. Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS, 39406, USA

    Hari Kiran Kotapati, Jennifer Sheffield & Philip D. Bates

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  1. Jay Shockey
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Shockey, J., Lager, I., Stymne, S. et al. Specialized lysophosphatidic acid acyltransferases contribute to unusual fatty acid accumulation in exotic Euphorbiaceae seed oils. Planta 249, 1285–1299 (2019). https://doi.org/10.1007/s00425-018-03086-y

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