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Overexpression of the rice carotenoid cleavage dioxygenase 1 gene in Golden Rice endosperm suggests apocarotenoids as substrates in planta

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Abstract

Carotenoids are converted by carotenoid cleavage dioxygenases that catalyze oxidative cleavage reactions leading to apocarotenoids. However, apocarotenoids can also be further truncated by some members of this enzyme family. The plant carotenoid cleavage dioxygenase 1 (CCD1) subfamily is known to degrade both carotenoids and apocarotenoids in vitro, leading to different volatile compounds. In this study, we investigated the impact of the rice CCD1 (OsCCD1) on the pigmentation of Golden Rice 2 (GR2), a genetically modified rice variety accumulating carotenoids in the endosperm. For this purpose, the corresponding cDNA was introduced into the rice genome under the control of an endosperm-specific promoter in sense and anti-sense orientations. Despite high expression levels of OsCCD1 in sense plants, pigment analysis revealed carotenoid levels and patterns comparable to those of GR2, pleading against carotenoids as substrates in rice endosperm. In support, similar carotenoid contents were determined in anti-sense plants. To check whether OsCCD1 overexpressed in GR2 endosperm is active, in vitro assays were performed with apocarotenoid substrates. HPLC analysis confirmed the cleavage activity of introduced OsCCD1. Our data indicate that apocarotenoids rather than carotenoids are the substrates of OsCCD1 in planta.

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Abbreviations

CCD1:

Carotenoid cleavage dioxygenase 1

GR:

Golden Rice

Gt1:

Glutelin 1 promoter

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Acknowledgments

This work was funded by The HarvestPlus research consortium (http://www.harvestplus.org) and by the Deutsche Forschungsgemeinschaft (DFG) Grant AL 892/1-4. We are indebted to Dr. Rhodora Aldemita for valuable discussions.

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  1. Faculty of Biology, Institute of Biology II, Albert-Ludwigs University of Freiburg, Schaenzlestr. 1, 79104, Freiburg, Germany

    Andrea Ilg, Qiuju Yu, Patrick Schaub, Peter Beyer & Salim Al-Babili

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  1. Andrea Ilg
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  2. Qiuju Yu
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Correspondence to Salim Al-Babili.

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Ilg, A., Yu, Q., Schaub, P. et al. Overexpression of the rice carotenoid cleavage dioxygenase 1 gene in Golden Rice endosperm suggests apocarotenoids as substrates in planta . Planta 232, 691–699 (2010). https://doi.org/10.1007/s00425-010-1205-y

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