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Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols

Abstract

Tomatoes are an excellent source of the carotenoid lycopene, a compound that is thought to be protective against prostate cancer. They also contain small amounts of flavonoids in their peel (5–10 mg/kg fresh weight), mainly naringenin chalcone and the flavonol rutin, a quercetin glycoside. Flavonols are very potent antioxidants, and an increasing body of epidemiological data suggests that high flavonoid intake is correlated with a decreased risk for cardiovascular disease. We have upregulated flavonol biosynthesis in the tomato in order to generate fruit with increased antioxidant capacity and a wider range of potential health benefit properties. This involved transformation of tomato with the Petunia chi-a gene encoding chalcone isomerase. Resulting transgenic tomato lines produced an increase of up to 78 fold in fruit peel flavonols, mainly due to an accumulation of rutin. No gross phenotypical differences were observed between high-flavonol transgenic and control lines. The phenotype segregated with the transgene and demonstrated a stable inheritance pattern over four subsequent generations tested thus far. Whole-fruit flavonol levels in the best of these lines are similar to those found in onions, a crop with naturally high levels of flavonol compounds. Processing of high-flavonol tomatoes demonstrated that 65% of flavonols present in the fresh fruit were retained in the processed paste, supporting their potential as raw materials for tomato-based functional food products.

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Figure 1: Flavonoid biosynthetic pathway.
Figure 2: Accumulation of rutin (dashed line) and naringenin chalcone (solid line) in developing tomato peel.
Figure 3: Northern analysis of developing tomato fruit.
Figure 4: Flavonoid levels in the peel of transgenic tomato fruit.
Figure 5: Chromatogram recorded at 370 nm, of nonhydrolyzed peel of control and pBBC50-transformed fruit.
Figure 6: Flavonol levels of field trial transgenic fruit.

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References

  1. Shintani, D. & DellaPenna, D. Elevating the vitamin E content of plants through metabolic engineering. Science 282, 2098–2100 (1998).

    Article  CAS  Google Scholar 

  2. Ye, X. et al. Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287, 303–305 (2000).

    Article  CAS  Google Scholar 

  3. Romer, S. et al. Elevation of the provitamin A content of transgenic tomato plants. Nat. Biotechnol. 18, 666–669 (2000).

    Article  CAS  Google Scholar 

  4. Middleton, E. & Kandaswami, C. The impact of plant flavonoids on mammalian biology: implications for immunity, inflammation and cancer. In The flavonoids: advances in research since 1986. (ed. Harbourne, J.B.) 619–652 (Chapman and Hall, London; 1994).

    Google Scholar 

  5. Hertog, M.G.L. et al. Flavonoid intake and long term risk of coronary heart disease and cancer in the 7 Countries study. Arch. Intern. Med. 155, 381–386 (1995).

    Article  CAS  Google Scholar 

  6. Hertog, M.G.L., Feskens E.J.M., Hollman P.C.H., Katan M.B. & Kromhout D. Dietary antioxidant flavonols and risk of coronary heart disease risk—the Zutphen elderly study. Lancet 342, 1007–1011 (1993).

    Article  CAS  Google Scholar 

  7. Knekt, P., Jarvinen, R., Reunanen, A. & Maatela, J. Flavonoid intake and coronary mortality in Finland: a cohort study. Br. Med. J. 312, 478–481 (1996).

    Article  CAS  Google Scholar 

  8. Weisshaar, B. & Jenkins, G.I. Phenylpropanoid biosynthesis and its regulation. Curr. Opin. Plant Biol. 1, 251–257 (1998).

    Article  CAS  Google Scholar 

  9. Mol, J., Grotewold, E. & Koes, R. How genes paint flowers and seeds. Trends Plant Sci. 3, 212–217 (1998).

    Article  Google Scholar 

  10. Dixon, R.A. & Steele, C.L. Flavonoids and isoflavonoids—a goldmine for metabolic engineering. Trends Plant Sci. 4, 394–400 (1999).

    Article  CAS  Google Scholar 

  11. Krause, M. & Galensa, R. Determination of naringenin and naringenin chalcone in tomato skins by HPLC after solid phase extraction. Z. Lebensmittel 194, 29–32 (1992).

    Article  CAS  Google Scholar 

  12. Burbulis, I.E. & Winkel-Shirley, B. Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Proc. Natl. Acad. Sci. USA 96, 12929–12934 (1999).

    Article  CAS  Google Scholar 

  13. Ewald, C., Fjelkner-Modig, S., Johansson, K., Sjoholm, I. & Akesson, B. Effect of processing on major flavonoids in processed onions, green beans, and peas. Food Chem. 64, 231–235 (1999).

    Article  CAS  Google Scholar 

  14. van Tunen, A.J. et al. Cloning of the 2 chalcone flavanone isomerase genes from Petunia—co-ordinate light-regulated and differential expression of flavonoid genes. EMBO J. 7, 1257–1263 (1988).

    Article  CAS  Google Scholar 

  15. Becker, D., Kemper, E., Schell, J. & Masterson, R. New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol. Biol. 20, 1195–1197 (1992).

    Article  CAS  Google Scholar 

  16. Fillatti, J.J., Kiser, J., Rose, R. & Comai, L. Efficient transfer of a glyphosate tolerance gene into tomato using a binary Agrobacterium tumefaciens vector. Bio Technology 5, 726–730 (1987).

    CAS  Google Scholar 

  17. Hertog, M.G.L., Hollman, P.C.H. & Venema, D.P. Optimization of a quantitative HPLC determination of potentially anticarcinogenic flavonoids in vegetables and fruits. J. Agric. Food Chem. 40, 1591–1598 (1992).

    Article  CAS  Google Scholar 

  18. Sambrook, J. et al. Molecular cloning. A laboratory manual. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 1989).

    Google Scholar 

Download references

Acknowledgements

The authors wish to thank Dr. Adel Elsheikh and Dr. Jinguo Hu (Lipton Innovation Centre, Stockton, CA) for their assistance with transgenic field trials. We also thank Bob Cowper, Carl Jarman, Tracey Macdonald, and Katherine Redwood (URC, Sharnbrook, UK) for their technical contributions.

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Correspondence to Martine E. Verhoeyen.

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Muir, S., Collins, G., Robinson, S. et al. Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols. Nat Biotechnol 19, 470–474 (2001). https://doi.org/10.1038/88150

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