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Cloning and functional characterization of the maize carotenoid isomerase and β-carotene hydroxylase genes and their regulation during endosperm maturation

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Abstract

In order to gain further insight into the partly-characterized carotenoid biosynthetic pathway in corn (Zea mays L.), we cloned cDNAs encoding the enzymes carotenoid isomerase (CRTISO) and β-carotene hydroxylase (BCH) using endosperm mRNA isolated from inbred line B73. For both enzymes, two distinct cDNAs were identified mapping to different chromosomes. The two crtiso cDNAs (Zmcrtiso1 and Zmcrtiso2) mapped to unlinked genes each containing 12 introns, a feature conserved among all crtiso genes studied thus far. ZmCRTISO1 was able to convert tetra-cis prolycopene to all-trans lycopene but could not isomerize the 15-cis double bond of 9,15,9′-tri-cis-ζ-carotene. ZmCRTISO2 is inactivated by a premature termination codon in B73 corn, but importantly the mutation is absent in other corn cultivars and the active enzyme showed the same activity as ZmCRTISO1. The two bch cDNAs (Zmbch1 and Zmbch2) mapped to unlinked genes each coding sequences containing five introns. ZmBCH1 was able to convert β-carotene into β-cryptoxanthin and zeaxanthin, but ZmBCH2 was able to form β-cryptoxanthin alone and had a lower overall activity than ZmBCH1. All four genes were expressed during endosperm development, with mRNA levels rising in line with carotenoid accumulation (especially zeaxanthin and lutein) until 25 DAP. Thereafter, expression declined for three of the genes, with only Zmcrtiso2 mRNA levels maintained by 30 DAP. We discuss the impact of paralogs with different expression profiles and functions on the regulation of carotenoid synthesis in corn.

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Acknowledgments

This work was supported by grants from the Spanish Ministry of Science and Innovation (MICINN) (BFU2007-61413), European Research Council Advanced Grant (BIOFORCE) to PC, Aciones complementarias, BIO2007-30738-E MICINN, Spain, a grant from the National Natural Science Foundation of China (grant no. 30870222) and the Foundation from Science and Technology Agency of Jilin Province (grant no. 20050543). G.F. and S.N. were supported by MICINN PhD fellowships. We would like to thank Dr. Ana Ana Butrón and Amando Ordás in Misión Biológica de Galicia, Consejo Superior de Investigaciones Cientificas, Apartado 28, 36080 Pontevedra, Spain, for supplying corn seeds.

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Authors and Affiliations

  1. School of Life Sciences, Northeast Normal University, 130024, Changchun, China

    Qunrui Li & Changfu Zhu

  2. Departament de Producció Vegetal i Ciència Forestal, Universitat de Lleida, Av. Alcalde Rovira Roure, 191, 25198, Lleida, Spain

    Gemma Farre, Shaista Naqvi, Georgina Sanahuja, Chao Bai, Teresa Capell, Paul Christou & Changfu Zhu

  3. Biosynthesis Group, Molecular Biosciences, J.W. Goethe Universität, Biocampus 213, P.O. Box 111932, 60054, Frankfurt, Germany

    Jürgen Breitenbach & Gerhard Sandmann

  4. Institucio Catalana de Recerca i Estudis Avancats, Passeig Llúis Companys, 23, 08010, Barcelona, Spain

    Paul Christou

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  1. Qunrui Li
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  2. Gemma Farre
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  3. Shaista Naqvi
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  4. Jürgen Breitenbach
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  6. Chao Bai
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  7. Gerhard Sandmann
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  8. Teresa Capell
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  9. Paul Christou
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  10. Changfu Zhu
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Correspondence to Changfu Zhu.

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Qunrui Li and Gemma Farre contributed equally to this work.

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Li, Q., Farre, G., Naqvi, S. et al. Cloning and functional characterization of the maize carotenoid isomerase and β-carotene hydroxylase genes and their regulation during endosperm maturation. Transgenic Res 19, 1053–1068 (2010). https://doi.org/10.1007/s11248-010-9381-x

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  • DOI: https://doi.org/10.1007/s11248-010-9381-x

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