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Combining an in vitro reporter gene assay with metabolomics to identify tomato phytochemicals responsible for inducing electrophile-responsive element (EpRE)-mediated gene transcription

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Abstract

The electrophile-responsive element (EpRE) is a transcriptional enhancer involved in cancer-chemoprotective gene expression effects of certain dietary compounds. In this study we measured the ability of extracts of glycosidase treated tomato fruits from 97 different accessions to induce EpRE-mediated luciferase expression using EpRE-LUX reporter cells and analyzed the same extracts using LC–MS-based untargeted metabolomics profiling. We were able to pinpoint those tomato compounds that were most correlated with EpRE-mediated luciferase induction, by combining reporter gene assay data with the metabolic profiles of the same extracts. Flavonoids were the compounds showing the strongest positive correlation with EpRE-LUX activity. These results were validated using a transgenic tomato line accumulating high levels of flavonoids. Results obtained corroborated that flavonoids are an important determinant of the ability of tomato fruit extracts to induce EpRE-mediated beneficial health effects. Overall, these results indicate that combining untargeted metabolomics with reporter gene assays provides a powerful tool for nutritionists, plant breeders and food chemists towards identification of potential health-beneficial constituents of tomato fruits, as well as of other crops and products derived thereof.

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Acknowledgments

This work was financially supported by the Food and Nutrition Delta Grant no. FND07007. We thank Syngenta for providing the tomato accessions. We thank Bert Schipper for excellent handling of the LC–MS instrument. AGB, YMT and RCHdV acknowledge the Centre for Biosystems Genomics, while RCHdV also acknowledges the Netherlands Metabolomics Centre for additional funding. Both Centers are part of the Netherlands Genomics Initiative/Netherlands Organization for Scientific Research.

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Authors and Affiliations

  1. Division of Toxicology, Wageningen University, Tuinlaan 5, 6703 HE, Wageningen, The Netherlands

    Henriëtte D. L. M. van Eekelen, Linda Gijsbers, Laura H. J. de Haan, Jac M. M. J. G. Aarts & Ivonne M. C. M. Rietjens

  2. Plant Research International, P.O. Box 16, 6700 AA, Wageningen, The Netherlands

    Henriëtte D. L. M. van Eekelen, Chris A. Maliepaard, Robert A. M. Vreeburg, Richard Finkers, Yury M. Tikunov, Victoria M. Gomez Roldan, Ric C. H. de Vos & Arnaud G. Bovy

  3. Centre for Biosystems Genomics, P.O. Box 98, 6708 PB, Wageningen, The Netherlands

    Yury M. Tikunov, Ric C. H. de Vos & Arnaud G. Bovy

  4. Netherlands Metabolomics Centre, Einsteinweg 55, 2333 CC, Leiden, The Netherlands

    Ric C. H. de Vos

Authors
  1. Henriëtte D. L. M. van Eekelen
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  2. Linda Gijsbers
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  3. Chris A. Maliepaard
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  4. Robert A. M. Vreeburg
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  7. Victoria M. Gomez Roldan
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  8. Laura H. J. de Haan
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  9. Ric C. H. de Vos
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  11. Ivonne M. C. M. Rietjens
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  12. Arnaud G. Bovy
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Corresponding author

Correspondence to Henriëtte D. L. M. van Eekelen.

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Supplemental Fig. 1

Principal components analysis of untargeted LC–MS data from the all tomato accessions. Tomato accessions that showed either highest or lowest EpRE-mediated luciferase induction are marked red and green, respectively. Clustering of these tomato accessions along the first three principal components (PC): PC1, PC2, and PC3 explains 34.8, 9.8, and 6.5 % of the total metabolic variation (JPEG 40 kb)

Supplemental Fig. 2

Scatter plots of log-transformed and mean-centered peak intensities over all 93 tomato accessions of quercetin-glycosides versus quercetin aglycone (JPEG 62 kb)

Supplemental Fig. 3

Scatter plot of log-transformed and mean-centred peak intensities over all 93 tomato accessions of tri caffeoylquinic acid versus quercetin aglycone (JPEG 34 kb)

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van Eekelen, H.D.L.M., Gijsbers, L., Maliepaard, C.A. et al. Combining an in vitro reporter gene assay with metabolomics to identify tomato phytochemicals responsible for inducing electrophile-responsive element (EpRE)-mediated gene transcription. Metabolomics 11, 302–311 (2015). https://doi.org/10.1007/s11306-014-0694-2

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  • DOI: https://doi.org/10.1007/s11306-014-0694-2

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