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A fatty acid condensing enzyme from Physaria fendleri increases hydroxy fatty acid accumulation in transgenic oilseeds of Camelina sativa

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Abstract

Main conclusion

Co-expression of a lesquerella fatty acid elongase and the castor fatty acid hydroxylase in camelina results in higher hydroxy fatty acid containing seeds with normal oil content and viability.

Producing hydroxy fatty acids (HFA) in oilseed crops has been a long-standing goal to replace castor oil as a renewable source for numerous industrial applications. A fatty acid hydroxylase, RcFAH, from Ricinus communis, was introduced into Camelina sativa, but yielded only 15 % of HFA in its seed oil, much lower than the 90 % found in castor bean. Furthermore, the transgenic seeds contained decreased oil content and the germination ability was severely affected. Interestingly, HFA accumulation was significantly increased in camelina seed when co-expressing RcFAH with a fatty acid condensing enzyme, LfKCS3, from Physaria fendleri, a native HFA accumulator relative to camelina. The oil content and seed germination of the transgenic seeds also appeared normal compared to non-transgenics. LfKCS3 has been previously characterized to specifically elongate the hydroxylated ricinoleic acid to lesquerolic acid, the 20-carbon HFA found in lesquerella oil. The elongation reaction may facilitate the HFA flux from phosphatidylcholine (PC), the site of HFA formation, into the acyl-CoA pool for more efficient utilization in triacylglycerol (TAG) biosynthesis. This was demonstrated by increased HFA accumulation in TAG concurrent with reduced HFA content in PC during camelina seed development, and increased C20-HFA in HFA-TAG molecules. These effects of LfKCS3 thus may effectively relieve the bottleneck for HFA utilization in TAG biosynthesis and the feedback inhibition to fatty acid synthesis, result in higher HFA accumulation and restore oil content and seed viability.

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Abbreviations

DAG:

Diacylglycerol

ER:

Endoplasmic reticulum

FAE1:

Fatty acid elongation 1

FAMEs:

Fatty acid methyl esters

GC–MS:

Gas chromatography–mass spectrometry

HFA:

Hydroxy fatty acid

LfKCS3:

Lesquerella (Physaria) fendleri fatty acid condensing enzyme

MAG:

Monoacylglycerol

ODP:

Oleate derivative proportion

PC:

Phosphatidylcholine

RcFAH:

Ricinus communis fatty acid hydroxylase

TAG:

Triacylglycerol

VLCFA:

Very-long-chain fatty acid (C20 and longer)

X:Y:

A fatty acid containing X carbons with Y double bonds

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Acknowledgments

The authors thank Dr. Ljerka Kunst (University of British Columbia) for kindly providing the lesquerella LfKCS3 gene construct MHS15. This work was supported by grants from U.S. Department of Agriculture and the Plant Genome Research Program of the U.S. National Science Foundation.

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  1. Department of Plant Sciences and Plant Pathology, Montana State University, Bozeman, MT, 59717-3150, USA

    Anna R. Snapp, Jinling Kang, Xiaoli Qi & Chaofu Lu

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  1. Anna R. Snapp
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  2. Jinling Kang
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  4. Chaofu Lu
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Correspondence to Chaofu Lu.

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Snapp, A.R., Kang, J., Qi, X. et al. A fatty acid condensing enzyme from Physaria fendleri increases hydroxy fatty acid accumulation in transgenic oilseeds of Camelina sativa . Planta 240, 599–610 (2014). https://doi.org/10.1007/s00425-014-2122-2

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