Plant Cell Reports

, Volume 36, Issue 6, pp 933–945 | Cite as

The Arabidopsis ORANGE (AtOR) gene promotes carotenoid accumulation in transgenic corn hybrids derived from parental lines with limited carotenoid pools

  • Judit Berman
  • Uxue Zorrilla-López
  • Vicente Medina
  • Gemma Farré
  • Gerhard Sandmann
  • Teresa Capell
  • Paul Christou
  • Changfu Zhu
Original Article


Key message

The AtOR gene enhances carotenoid levels in corn by promoting the formation of plastoglobuli when the carotenoid pool is limited, but has no further effect when carotenoids are already abundant.


The cauliflower orange (or) gene mutation influences carotenoid accumulation in plants by promoting the transition of proplastids into chromoplasts, thus creating intracellular storage compartments that act as metabolic sink. We overexpressed the Arabidopsis OR gene under the control of the endosperm-specific wheat LMW glutenin promoter in a white corn variety that normally accumulates only trace amounts of carotenoids. The total endosperm carotenoid content in the best-performing AtOR transgenic corn line was 32-fold higher than wild-type controls (~25 µg/g DW at 30 days after pollination) but the principal carotenoids remained the same, suggesting that AtOR increases the abundance of existing carotenoids without changing the metabolic composition. We analyzed the expression of endogenous genes representing the carotenoid biosynthesis and MEP pathways, as well as the plastid fusion/translocation factor required for chromoplast formation, but only the DXS1 gene was upregulated in the transgenic corn plants. The line expressing AtOR at the highest level was crossed with four transgenic corn lines expressing different carotenogenic genes and accumulating different carotenoids. The introgression of AtOR increased the carotenoid content of the hybrids when there was a limited carotenoid pool in the parental line, but had no effect when carotenoids were already abundant in the parent. The AtOR gene therefore appears to enhance carotenoid levels by promoting the formation of carotenoid-sequestering plastoglobuli when the carotenoid pool is limited, but has no further effect when carotenoids are already abundant because high levels of carotenoids can induce the formation of carotenoid-sequestering plastoglobuli even in the absence of AtOR.


Orange gene Carotenoids Plastoglobuli Corn Transgenic Hybrids 



Research at the Universitat de Lleida is supported by MINECO, Spain (BIO2014-54426-P; BIO2014-54441-P), by the Catalan Government (2014 SGR 1296 Agricultural Biotechnology Research Group), and by European Union Framework 7, European Research Council IDEAS Advanced Grant BIOFORCE and POC Grant (to PC). G. Farré is supported by a J de la C fellowship.

Author’s contribution

CZ, GS, TC, and PC designed research; JB, UZ, and GF performed research, analyzed data, and wrote the article; VM did TEM and analyzed these data.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

299_2017_2126_MOESM1_ESM.docx (115 kb)
Supplementary material 1 Supplementary Figure 1—RNA blot analysis of AtOR mRNA in wild type (M37W) and two different transgenic lines (OR1 and OR2) transformed with the AtOR gene driven by the wheat low-molecular-weight glutelin promoter. Each lane was loaded with 25 μg total RNA isolated from endosperm tissue Ribosomal RNA stained with ethidium bromide is shown as a loading control. Supplementary Figure 2—Transcript accumulation normalized against ACTIN mRNA in wild-type and transgenic lines presented as means of three technical replicates. Standard error bars were not included due to the use of technical replicates rather than biological replicates (the aim was to confirm transgene expression rather than to compare different transcript profiles). Supplementary Figure 3—Schematic representation of the transgenes expressed in our transgenic plants lines and hybrids. Supplementary Table 1—Carotenoid content and composition in wild-type M37W, transgenic lines OR2, CARO1, CARO2, KETO1 and KETO2, and hybrids ORxCARO1, ORxCARO2, ORxKETO1 and ORxKETO2 (T1 plants) at 30 (*) and 60 DAP (**) (µg/g DW±SE) (n = 3–5 seeds). Abbreviations: Phyt, phytoene; Lyco, lycopene; βcryp, β-cryptoxanthin; βcaro, β-carotene; Lut, lutein; Zea, zeaxanthin; Anthe, antheraxanthin; Viola, violaxanthin; CAROT, carotenoids. Supplementary Table 2—Ketocarotenoid content and composition of wild-type M37W, transgenic lines OR2, CARO1, CARO2, KETO1 and KETO2, and hybrids ORxCARO1, ORxCARO2, ORxKETO1 and ORxKETO2 T1 at 30 (*) and 60 DAP (**) (µg/g DW±SE) (n = 3–5 seeds). Abbreviations: Asta, astaxanthin; Cantha, canthaxanthin; Adonir, adonirubin; Adonix, adonixanthin; 3OHechi, 3-OH-echinenone; KETO, ketocarotenoids. Supplementary Table 3—Oligonucleotide sequences of corn ACTIN, endogenous carotenogenic genes and transgenes for real-time quantitative RT-PCR analysis. (DOCX 114 KB)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Judit Berman
    • 1
  • Uxue Zorrilla-López
    • 1
  • Vicente Medina
    • 1
  • Gemma Farré
    • 1
  • Gerhard Sandmann
    • 2
  • Teresa Capell
    • 1
  • Paul Christou
    • 1
    • 3
  • Changfu Zhu
    • 1
  1. 1.Department of Plant Production and Forestry ScienceUniversity of Lleida-Agrotecnio CenterLleidaSpain
  2. 2.Biosynthesis Group, Molecular BiosciencesJohann Wolfgang Goethe UniversitätFrankfurtGermany
  3. 3.ICREA, Catalan Institute for Research and Advanced StudiesBarcelonaSpain

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