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Functional characterization of Citrus PSY gene in Hongkong kumquat (Fortunella hindsii Swingle)

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Abstract

Citrus, rich in carotenoids, is the most important fruit crop based on the total annual production. In the carotenoid biosynthesis pathway, phytoene synthase (PSY, EC 2.5.1.32) catalyzes the dimerization of two molecules of geranylgeranyl pyrophosphate (GGPP) to phytoene and has been shown to be a rate-limiting enzyme for the synthesis of carotenoids. In this study, we investigated catalytic activity of CsPSY from Cara Cara navel orange (Citrus sinensis Osbeck) by heterologous expression in Escherichia coli containing a GGPP-producing plasmid. Moreover, the effects of CsPSY overexpression on carotenoid accumulation were also functionally analyzed in transgenic Hongkong kumquat (Fortunella hindsii Swingle). The resulting transgenic plants produced orange fruits, and extracts from the fruits of four overexpressing plants had a 2.5-fold average increase of phytoene with the content approximately 71.38 μg/g fresh weight. Lycopene, β-carotene, and β-cryptoxanthin in transgenic fruits were also markedly increased, whereas the levels of lutein and violaxanthin kept nearly unchanged with 1.1–1.3 folds variation. Transcript levels of carotenoid biosynthetic genes in the CsPSY overexpressed plants remained unaltered except that PDS and ZDS showed a minor increase. This study suggests that CsPSY plays a crucial role in citrus carotenoid biosynthesis and could be used as a means of engineering fruit crop for the production of carotenoids.

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Abbreviations

PSY:

Phytoene synthase

cDNA:

Complementary DNA

GGPP:

Geranylgeranyl pyrophosphate

HPLC:

High-performance liquid chromatography

BA:

6-Benzyladenine

IBA:

Indole butyric acid

KT:

Kinetin

NAA:

α-Napthaleneacetic acid

NPT II:

Neomycin phosphotransferase II

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Acknowledgments

We are grateful to Dr. Norihiko Misawa (Marine Biotechnology Institute, Iwate, Japan) for the kindly providing us the plasmids. This research was supported by NSFC (30830078).

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, 430070, Wuhan, People’s Republic of China

    Jiancheng Zhang, Nengguo Tao, Qiang Xu, Wenjing Zhou, Hongbo Cao, Juan Xu & Xiuxin Deng

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  1. Jiancheng Zhang
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  2. Nengguo Tao
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  3. Qiang Xu
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  4. Wenjing Zhou
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  5. Hongbo Cao
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  6. Juan Xu
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  7. Xiuxin Deng
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Correspondence to Xiuxin Deng.

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Communicated by Y. Lu.

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Zhang, J., Tao, N., Xu, Q. et al. Functional characterization of Citrus PSY gene in Hongkong kumquat (Fortunella hindsii Swingle). Plant Cell Rep 28, 1737–1746 (2009). https://doi.org/10.1007/s00299-009-0774-3

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  • DOI: https://doi.org/10.1007/s00299-009-0774-3

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