Location via proxy:   [ UP ]  
[Report a bug]   [Manage cookies]                
Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Improving the nutritional value of Golden Rice through increased pro-vitamin A content

Nature Biotechnology volume 23, pages 482–487 (2005)Cite this article

Abstract

'Golden Rice' is a variety of rice engineered to produce β-carotene (pro-vitamin A) to help combat vitamin A deficiency1, and it has been predicted that its contribution to alleviating vitamin A deficiency would be substantially improved through even higher β-carotene content2. We hypothesized that the daffodil gene encoding phytoene synthase (psy), one of the two genes used to develop Golden Rice, was the limiting step in β-carotene accumulation. Through systematic testing of other plant psys, we identified a psy from maize that substantially increased carotenoid accumulation in a model plant system. We went on to develop 'Golden Rice 2' introducing this psy in combination with the Erwinia uredovora carotene desaturase (crtI) used to generate the original Golden Rice1. We observed an increase in total carotenoids of up to 23-fold (maximum 37 μg/g) compared to the original Golden Rice and a preferential accumulation of β-carotene.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Expression of a psy transgene increases the carotenoid content of maize callus.
Figure 2: Carotenoid enhancement of the rice endosperm by transformation with psy orthologues and crtI.

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

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

    Article  CAS  Google Scholar 

  2. Zimmerman, R. & Qaim, M. Potential health benefits of Golden Rice: a Philippines case study. Food Policy 29, 147–168 (2004).

    Article  Google Scholar 

  3. Yeum, K.J. & Russell, R.M. Carotenoid bioavailability and bioconversion. Ann. Rev. Nutr. 22, 483–504 (2002).

    Article  CAS  Google Scholar 

  4. West, K.P. Jr. & Darnton-Hill, I. Vitamin A deficiency. in Nutrition and health in developing countries (eds. Semba, R.D. & Bloem, M.W.) 267–306 (Humana Press, Totowa, NJ, 2001).

    Chapter  Google Scholar 

  5. Dawe, D., Robertson, R. & Unnevehr, L. Golden Rice: what role could it play in alleviation of VAD? Food Policy 27, 541–560 (2002).

    Article  Google Scholar 

  6. Datta, K. et al. Bioengineered 'golden' indica rice cultivars with β-carotene metabolism in the endosperm with hygromycin and mannose selection systems. Plant Biotechnol. J. 1, 81–90 (2003).

    Article  CAS  Google Scholar 

  7. Hoa, T.T.C., Al-Babili, S., Schaub, P., Potrykus, I. & Beyer, P. Golden Indica and Japonica rice lines amenable to deregulation. Plant Physiol. 113, 161–169 (2003).

    Article  Google Scholar 

  8. Fraser, P.D., Truesdale, M., Bird, C.R., Schuch, W. & Bramley, P.M. Carotenoid biosynthesis during tomato fruit development. Plant Physiol. 105, 405–413 (1994).

    Article  CAS  Google Scholar 

  9. Ronen, G., Cohen, M., Zamir, D. & Hirschberg, J. Regulation of carotenoid biosynthesis during tomato fruit development: expression of the gene for lycopene epsilon-cyclase is down-regulated during ripening and is elevated in the mutant. Delta. Plant J. 17, 341–351 (1999).

    Article  CAS  Google Scholar 

  10. Fraser, P.D. et al. Evaluation of transgenic tomato plants expressing an additional phytoene synthase in a fruit-specific manner. Proc. Natl. Acad. Sci. USA 99, 1092–1097 (2002).

    Article  CAS  Google Scholar 

  11. Shewmaker, C.K., Sheehy, J.A., Daley, M., Colburn, S. & Yang Ke, D. Seed-specific overexpression of phytoene synthase: increase in carotenoids and other metabolic effects. Plant J. 20, 401–412 (1999).

    Article  CAS  Google Scholar 

  12. Burkhardt, P.K. et al. Transgenic rice (Oryza sativa) endosperm expressing daffodil (Narcissus pseudonarcissus) phytoene synthase accumulates phytoene, a key intermediate of provitamin A biosynthesis. Plant J. 11, 1071–1078 (1997).

    Article  CAS  Google Scholar 

  13. Camara, B., Hugueney, P., Bouvier, F., Kuntz, M. & Moneger, R. Biochemistry and molecular biology of chromoplast development. Int. Rev. Cytol. 163, 175–247 (1995).

    Article  CAS  Google Scholar 

  14. Rabbani, S., Beyer, P., Lintig, J., Hugueney, P. & Kleinig, H. Induced beta–carotene synthesis driven by triacylglycerol deposition in the unicellular alga Dunaliella bardawil. Plant Physiol. 116, 1239–1248 (1998).

    Article  CAS  Google Scholar 

  15. Keappler, H.F., Somers, D.A., Rines, H.W. & Cockburn, A.F. Silicon carbide fiber-mediated stable transformation of plant cells. Theor. Appl. Genet. 84, 560–566 (1992).

    Article  Google Scholar 

  16. Buckner, B., San Miguel, P., Janick-Buckner, D. & Bennetzen, J.L. The y1 gene of maize codes for phytoene synthase. Genetics 143, 479–488 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Bartley, G.E., Viitanen, P.V., Bacot, K.O. & Scolnik, P.A. A tomato gene expressed during fruit ripening encodes an enzyme of the carotenoid biosynthesis pathway. J. Biol. Chem. 267, 5036–5039 (1992).

    CAS  PubMed  Google Scholar 

  18. Romer, S., Hugueney, P., Bouvier, F., Camara, B. & Kuntz, M. Expression of the genes encoding the early carotenoid biosynthetic enzymes in Capsicum annuum. Biochem. Biophys. Res. Commun. 196, 1414–1421 (1993).

    Article  CAS  Google Scholar 

  19. Scolnik, P.A. & Bartley, G.E. Nucleotide sequence of an Arabidopsis cDNA for phytoene synthase. Plant Physiol. 104, 1471–1472 (1994).

    Article  CAS  Google Scholar 

  20. Schledz, M. et al. Phytoene synthase from Narcissus pseudonarcissus: functional expression, galactolipid requirement, topological distribution in chromoplasts and induction during flowering. Plant J. 10, 781–792 (1996).

    Article  CAS  Google Scholar 

  21. Pandit, J. et al. Crystal structure of human squalene synthase. A key enzyme in cholesterol biosynthesis. J. Biol. Chem. 275, 30610–30617 (2000).

    Article  CAS  Google Scholar 

  22. Palaisa, K.A., Morgante, M., Williams, M. & Rafalski, A. Contrasting effects of selection on sequence diversity and linkage disequilibrium at two phytoene synthase loci. Plant Cell 15, 1795–1806 (2003).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  24. Institute of Medicine Dietary Reference Intakes for Vitamin A, Vitamin K, Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum, Nickel, Silicon, Vanadium, and Zinc (National Academy Press, Washington, DC, 2001).

  25. Goff, S.A. et al. A draft sequence of the rice genome (Oryza sativa L. ssp japonica). Science 296, 92–100 (2002).

    Article  CAS  Google Scholar 

  26. Misawa, N. et al. Functional expression of the Erwinia uredovora carotenoid biosynthesis gene crtI in transgenic plants showing an increase of β-carotene biosynthesis activity and resistance to the bleaching herbicide norflurazon. Plant J. 4, 833–40 (1993).

    Article  CAS  Google Scholar 

  27. Tanaka, A. et al. Enhancement of foreign gene expression by a dicot intron in rice but not in tobacco is correlated with an increased level of mRNA and an efficient splicing of the intron. Nuc. Acids Res. 18, 6767–6770 (1990).

    Article  CAS  Google Scholar 

  28. Negrotto, D., Jolley, M., Beer, S., Wenck, A.R. & Hansen, G. The use of phosphomannose isomerase as a selectable marker to recover transgenic maize plants (Zea mays L.) via Agrobacterium transformation. Plant Cell Reports 19, 798–803 (2000).

    Article  CAS  Google Scholar 

  29. Hiei, Y., Ohta, S., Komari, T. & Kumashiro, T. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6, 271–282 (1994).

    Article  CAS  Google Scholar 

  30. Zhang, J., Xu, R.-J., Elliott, M.C. & Chen,, D.-F. Agrobacterium-mediated transformation of elite japonica and indica rice varieties. Mol. Biotechnol. 8, 223–231 (1997).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Will Parish, Erik Dunder, Dong Fang Chen and Annalisa Tiozzo for tissue culture, Karen Bacon and Fasica Woldeyes for plant growth, Melanie Watkins for plant assessment, Elek Bolygo for analytical advice, Ebun Eno-Amooquaye for western blot analysis, Keith Ward for advice on statistics and others who gave technical support to the research. We would also like to thank Peter Beyer, Lu Liu and John Ray for plasmids. We thank Peter Beyer for discussion on the manuscript.

Author information

Authors and Affiliations

  1. Syngenta, Jealott's Hill International Research Centre, Bracknell, RG42 6EY, Berkshire, UK

    Jacqueline A Paine, Catherine A Shipton, Sunandha Chaggar, Rhian M Howells, Mike J Kennedy, Gareth Vernon, Susan Y Wright & Rachel Drake

  2. Syngenta CTL, Alderley Park, Alderley Edge, Macclesfield, SK10 4TJ, Cheshire, UK

    Edward Hinchliffe

  3. Syngenta Biotechnology Inc., PO Box 12257, Research Triangle Park, 27709, North Carolina, USA

    Jessica L Adams & Aron L Silverstone

Authors
  1. Jacqueline A Paine
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Catherine A Shipton
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sunandha Chaggar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rhian M Howells
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mike J Kennedy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gareth Vernon
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Susan Y Wright
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Edward Hinchliffe
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jessica L Adams
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Aron L Silverstone
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Rachel Drake
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rachel Drake.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Carotenoid biosynthesis in plants (PDF 64 kb)

Supplementary Fig. 2

Expression of CRTI and maize PSY proteins in rice endosperm (PDF 192 kb)

Supplementary Table 1

Carotenoid composition of maize callus expressing a Psy transgene (PDF 61 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Paine, J., Shipton, C., Chaggar, S. et al. Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nat Biotechnol 23, 482–487 (2005). https://doi.org/10.1038/nbt1082

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1082

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing