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Engineering plants with increased levels of the antioxidant chlorogenic acid

Abstract

The trend to view many foods not only as sustenance but also as medicine, so-called functional foods, is increasing. Phenolics are the most widespread dietary antioxidants, and among these, chlorogenic acid (CGA) accumulates to high levels in some crop plants. CGA acts as an antioxidant in plants and protects against degenerative, age-related diseases in animals when supplied in their diet. cDNA clones encoding the enzyme that synthesizes CGA, hydroxycinnamoyl-CoA quinate: hydroxycinnamoyl transferase (HQT), were characterized from tomato and tobacco. Gene silencing proved HQT to be the principal route for accumulation of CGA in solanaceous species. Overexpression of HQT in tomato caused plants to accumulate higher levels of CGA, with no side-effects on the levels of other soluble phenolics, and to show improved antioxidant capacity and resistance to infection by a bacterial pathogen. Tomatoes with elevated CGA levels could be used in foods with specific benefits for human health.

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Figure 1: Proposed pathways for the synthesis of chlorogenic acid in plants.
Figure 2: Analysis of the predicted HQT amino acid sequences and HQT gene expression.
Figure 3: Activity of recombinant HQT expressed in E. coli.
Figure 4: Modulation of HQT activity by transient transformation of N. benthamiana.
Figure 5: Correlation of increased HQT expression and gene silencing of HQT with the changes in HQT enzyme activity and content of CGA in transgenic tomato lines.
Figure 6: Increased levels of CGA result in better protection against oxidative stress and bacterial infection.

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Acknowledgements

We thank Alison Smith, Nick Walton and Adrian Parr for advice on enzyme purification and assay, Lionel Hill, Melinda Meyer and Fred Mellon for advice on HPLC and LC-MS, Mike Naldrett and Andrew Bottrill for provision of the Q-ToF-MS data, Phil Mullineaux and Baldeep Kumar for advice on measuring oxidative stress in plants, Max Dow for advice on pathogen infection of tomato, Mary Parker for lignin analysis, David Hopwood for comments on the manuscript, and Keith Waldron, Charlotte Parker, Stephen Bornemann and Andrew Smith for stimulating discussions on strategies to manipulate phenylpropanoid metabolism. This work was supported by award 218/D11645 from the Biological and Biotechnological Science Research Council (BBSRC) Agri-Food Committee, by the EU FP5 PROFOOD project (QLK1-CT-2001-01080) and both C.M. and A.J.M. are supported by core strategic grants from BBSRC.

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Correspondence to Cathie Martin.

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The authors have filed a patent application in the United Kingdom based on the technology described in this paper.

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Niggeweg, R., Michael, A. & Martin, C. Engineering plants with increased levels of the antioxidant chlorogenic acid. Nat Biotechnol 22, 746–754 (2004). https://doi.org/10.1038/nbt966

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