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Isolation and characterization of carotenoid cleavage dioxygenase 4 genes from different citrus species

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Abstract

In plants, the carotenoid cleavage dioxygenase 4 (CCD4) could target on plastoglobules and cleave specific carotenoids, producing apocarotenoids and volatile compounds. These compounds are important for color and aroma formation in fruits and flowers. In this study, five CCD4 gene members (CCD4a, b, c, d, and e) were investigated in different citrus species including mandarin, pummelo, and sweet orange. Sequence analysis showed that the CCD4 genes from all the species examined exhibited extensive allelic variability (including SNPs and frame-shift mutations). Furthermore, the distribution of the CCD4 allelic mutation sites supported our previous hypothesis that the sweet orange originated from the hybridization of mandarin and pummelo. A derived cleaved amplified polymorphic sequence (dCAPs) marker was then successfully developed based on the allelic polymorphism of CCD4c, providing an ideal molecular marker for studying the genetic relationship between citrus species. Quantitative RT-PCR analysis identified differential expression patterns for the CCD4 genes in tissues/organs, and CCD4b was shown to have a high-level expression in citrus fruit flavedos (especially those with a deep orange-reddish color). HPLC-based detection of a key component (i.e., β-citraurin) for orange-reddish flavedo formation in different citrus revealed a positive correlation between CCD4b expression levels and the presence of β-citraurin, suggesting that CCD4b may be responsible for β-citraurin biosynthesis in flavedo. In summary, this study not only reinforced the anticipated roles of CCD4 genes in flavedo color formation in citrus, but also provided new information about gene expression patterns, allelic polymorphism characteristics, and sequence variability for this gene subfamily.

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Acknowledgments

This work was supported by the National Basic Research Program of China (973 Program, 2011CB100601), and National Natural Science Foundation of China (Nos. 31201612, 31328018, 31221062).

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    Authors and Affiliations

    1. Key Laboratory of Horticultural Plant Biology of Ministry of Education, MOA, Wuhan, China

      Xiongjie Zheng, Zongzhou Xie, Kaijie Zhu, Qiang Xu, Xiuxin Deng & Zhiyong Pan

    2. Key Laboratory of Horticultural Crop Biology and Genetic Improvement (Central Region), MOA, Wuhan, China

      Xiongjie Zheng, Zongzhou Xie, Kaijie Zhu, Qiang Xu, Xiuxin Deng & Zhiyong Pan

    3. College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

      Xiongjie Zheng, Zongzhou Xie, Kaijie Zhu, Qiang Xu, Xiuxin Deng & Zhiyong Pan

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    1. Xiongjie Zheng
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    2. Zongzhou Xie
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    Correspondence to Zhiyong Pan.

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    Communicated by S. Hohmann.

    X. Zheng and Z. Xie contributed equally to this work.

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    Zheng, X., Xie, Z., Zhu, K. et al. Isolation and characterization of carotenoid cleavage dioxygenase 4 genes from different citrus species. Mol Genet Genomics 290, 1589–1603 (2015). https://doi.org/10.1007/s00438-015-1016-8

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    • DOI: https://doi.org/10.1007/s00438-015-1016-8

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