Transgenic Research

, Volume 26, Issue 1, pp 13–23 | Cite as

Reconstruction of the astaxanthin biosynthesis pathway in rice endosperm reveals a metabolic bottleneck at the level of endogenous β-carotene hydroxylase activity

  • Chao Bai
  • Judit Berman
  • Gemma Farre
  • Teresa Capell
  • Gerhard Sandmann
  • Paul Christou
  • Changfu Zhu
Original Paper


Astaxanthin is a high-value ketocarotenoid rarely found in plants. It is derived from β-carotene by the 3-hydroxylation and 4-ketolation of both ionone end groups, in reactions catalyzed by β-carotene hydroxylase and β-carotene ketolase, respectively. We investigated the feasibility of introducing an extended carotenoid biosynthesis pathway into rice endosperm to achieve the production of astaxanthin. This allowed us to identify potential metabolic bottlenecks that have thus far prevented the accumulation of this valuable compound in storage tissues such as cereal grains. Rice endosperm does not usually accumulate carotenoids because phytoene synthase, the enzyme responsible for the first committed step in the pathway, is not present in this tissue. We therefore expressed maize phytoene synthase 1 (ZmPSY1), Pantoea ananatis phytoene desaturase (PaCRTI) and a synthetic Chlamydomonas reinhardtii β-carotene ketolase (sCrBKT) in transgenic rice plants under the control of endosperm-specific promoters. The resulting grains predominantly accumulated the diketocarotenoids canthaxanthin, adonirubin and astaxanthin as well as low levels of monoketocarotenoids. The predominance of canthaxanthin and adonirubin indicated the presence of a hydroxylation bottleneck in the ketocarotenoid pathway. This final rate-limiting step must therefore be overcome to maximize the accumulation of astaxanthin, the end product of the pathway.


Rice (Oryza sativa L.) Astaxanthin Ketocarotenoids Carotenoids β-carotene ketolase Multigene transformation 



This work was supported by the Ministerio de Economía y Competitividad, Spain (BIO2014-54441-P; BIO2014-54426-P; BIO2011-23324; BIO2011-22525; PIM2010PKB-00746); the Plan Nacional de I + D + I (2008-2011); the IBERCAROT network (112RT0445); PROGRAMA ESTATAL DE INVESTIGACIÓN CIENTÍFICA Y TÉCNICA DE EXCELENCIA, Spain (BIO2015-71703-REDT); COST action (EUROCAROTEN, OC-2015-1-19780); European Union Framework 7 European Research Council IDEAS Advanced Grant (to P.C.) Program-BIOFORCE and ERC Proof of Concept Grant (to P.C.), Catalan Government 2014 SGR 1296 Agricultural Biotechnology Research Group. C.B. is the recipient of a Ph.D. fellowship from the Universitat de Lleida, Spain.

Author contributions

T.C., G.S., P.C. and C.Z. conceived and designed research. C.B., J.B., G.F., T.C., G.S., P.C. and C.Z. conducted experiments. C.B., J.B., G.F. and C.Z. contributed new reagents or analytical tools. C.B., T.C., G.S., P.C. and C.Z. analyzed data. C.B., T.C., G.S., P.C. and C.Z. wrote the manuscript. All authors read and approved the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11248_2016_9977_MOESM1_ESM.doc (95 kb)
Supplementary material 1 (DOC 95 kb)


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Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  • Chao Bai
    • 1
  • Judit Berman
    • 1
  • Gemma Farre
    • 1
  • Teresa Capell
    • 1
  • Gerhard Sandmann
    • 2
  • Paul Christou
    • 1
    • 3
  • Changfu Zhu
    • 1
  1. 1.Department of Plant Production and Forestry Science, ETSEAUniversity of Lleida-Agrotecnio CenterLleidaSpain
  2. 2.Institute of Molecular BioscienceJ. W. Goethe UniversityFrankfurt am MainGermany
  3. 3.ICREA, Catalan Institute for Research and Advanced StudiesBarcelonaSpain

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