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The beet R locus encodes a new cytochrome P450 required for red betalain production

Nature Genetics volume 44, pages 816–820 (2012)Cite this article

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Abstract

Anthocyanins are red and violet pigments that color flowers, fruits and epidermal tissues in virtually all flowering plants. A single order, Caryophyllales, contains families in which an unrelated family of pigments, the betalains, color tissues normally pigmented by anthocyanins1. Here we show that CYP76AD1 encoding a novel cytochrome P450 is required to produce the red betacyanin pigments in beets. Gene silencing of CYP76AD1 results in loss of red pigment and production of only yellow betaxanthin pigment. Yellow betalain mutants are complemented by transgenic expression of CYP76AD1, and an insertion in CYP76AD1 maps to the R locus2,3 that is responsible for yellow versus red pigmentation. Finally, expression of CYP76AD1 in yeast verifies its position in the betalain biosynthetic pathway. Thus, this cytochrome P450 performs the biosynthetic step that provides the cyclo-DOPA moiety of all red betacyanins. This discovery will contribute to our ability to engineer this simple, nutritionally valuable pathway4 into heterologous species.

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Figure 1: Betalain biosynthetic pathway.
Figure 2: CYP76AD1 is required for red betalains.
Figure 3: Complementation of yellow to red with CYP76AD1.
Figure 4: Expression of beet BvDODA1 and CYP76AD1 in yeast.

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Acknowledgements

The authors dedicate this manuscript to Tom Mabry and thank him for inspiration. The authors thank K. Keller for help with pigment analysis. This work was supported by USDA grant 2008-01111 and US National Science Foundation (NSF) grant MCB-1122179.

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Author notes

  1. Gregory J Hatlestad and Rasika M Sunnadeniya: These authors contributed equally to this work.

Authors and Affiliations

  1. Section of Molecular Cell and Developmental Biology, Institute for Cell and Molecular Biology, The University of Texas at Austin, Austin, Texas, USA

    Gregory J Hatlestad, Rasika M Sunnadeniya, Neda A Akhavan, Antonio Gonzalez & Alan M Lloyd

  2. Department of Horticulture, University of Wisconsin–Madison, Madison, Wisconsin, USA

    Irwin L Goldman

  3. US Department of Agriculture–Agricultural Research Service (USDA-ARS), Sugarbeet and Bean Research Unit, East Lansing, Michigan, USA

    J Mitchell McGrath

  4. Department of Crop and Soil Sciences, Michigan State University, East Lansing, Michigan, USA

    J Mitchell McGrath

Authors
  1. Gregory J Hatlestad
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  4. Antonio Gonzalez
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  7. Alan M Lloyd
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Contributions

G.J.H., R.M.S., N.A.A., A.G. and A.M.L. performed experiments. G.J.H., R.M.S. and A.M.L. wrote the paper. I.L.G., J.M.M., G.J.H., R.M.S. and A.M.L. helped conceive of the project, analyze data and edit the manuscript. I.L.G. and J.M.M. contributed critical resources and reagents.

Corresponding author

Correspondence to Alan M Lloyd.

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Competing interests

I.L.G. has a financial interest in PhytoColorants, LLC, which is involved in developing natural commercial food colors from sources such as beet.

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Supplementary Table 1 and Supplementary Figures 1–4 (PDF 412 kb)

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Hatlestad, G., Sunnadeniya, R., Akhavan, N. et al. The beet R locus encodes a new cytochrome P450 required for red betalain production. Nat Genet 44, 816–820 (2012). https://doi.org/10.1038/ng.2297

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