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Castor patatin-like phospholipase A IIIβ facilitates removal of hydroxy fatty acids from phosphatidylcholine in transgenic Arabidopsis seeds

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Castor patatin-like phospholipase A IIIβ facilitates the exclusion of hydroxy fatty acids from phosphatidylcholine in developing transgenic Arabidopsis seeds.

Abstract

Hydroxy fatty acids (HFAs) are industrial useful, but their major natural source castor contains toxic components. Although expressing a castor OLEATE 12-HYDROXYLASE in Arabidopsis thaliana leads to the synthesis of HFAs in seeds, a high proportion of the HFAs are retained in phosphatidylcholine (PC). Thus, the liberation of HFA from PC seems to be critical for obtaining HFA-enriched seed oils. Plant phospholipase A (PLA) catalyzes the hydrolysis of PC to release fatty acyl chains that can be subsequently channeled into triacylglycerol (TAG) synthesis or other metabolic pathways. To further our knowledge regarding the function of PLAs from HFA-producing plant species, two class III patatin-like PLA cDNAs (pPLAIIIβ or pPLAIIIδ) from castor or Physaria fendleri were overexpressed in a transgenic line of A. thaliana producing C18-HFA, respectively. Only the overexpression of RcpPLAIIIβ resulted in a significant reduction in seed HFA content with concomitant changes in fatty acid composition. Reductions in HFA content occurred in both PC and TAG indicating that HFAs released from PC were not incorporated into TAG. These results suggest that RcpPLAIIIβ may catalyze the removal of HFAs from PC in the developing seeds synthesizing these unusual fatty acids.

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Acknowledgements

The authors thank Dr. John Browse of Washington State University for kindly providing Arabidopsis CL7 line. This work was supported by the Canada Research Chairs (R.J.W. and G.C.), Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grants (RGPIN-2016-05926 to G.C. and RGPIN-2014-04585 to R.J.W.), and Alberta Innovates Bio Solutions (R.J.W.). The infrastructure used in this work was funded by the Canadian Foundation for Innovation and Research Capacity Program of Alberta Enterprise and Advanced Education.

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R.J.W. oversaw the project; R.J.W. and G.C. conceived the project; Y.L., G.C., E.M. and R.J.W. designed the experiments; R.J.W., G.C. and S.D.S. supervised the experiments; Y.L. performed most of the experiments and data analysis; G.C. conducted some of the experiments and data analysis; Z.S. performed qRT-PCR, K.C. and Y.L. made constructs for gene transformation; J.D., M.S., and T.M. generated important plant materials, genes, and gene libraries. Y.L. and G.C. wrote the initial draft of the article. All authors participated in interpretation of the data and were instrumental in the preparation of the final article.

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Correspondence to Guanqun Chen or Randall J. Weselake.

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Lin, Y., Chen, G., Mietkiewska, E. et al. Castor patatin-like phospholipase A IIIβ facilitates removal of hydroxy fatty acids from phosphatidylcholine in transgenic Arabidopsis seeds. Plant Mol Biol 101, 521–536 (2019). https://doi.org/10.1007/s11103-019-00915-w

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  • DOI: https://doi.org/10.1007/s11103-019-00915-w

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