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Cloning of phytoene desaturase and expression analysis of carotenogenic genes in persimmon (Diospyros kaki L.) fruits

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Abstract

Persimmon is a commercially important fruit crop, and the fruit is rich in different kinds of bioactive compounds, among which carotenoids contribute significantly to its color and nutritional value. In this study, the cDNA of phytoene desaturase gene (PDS) was isolated by rapid amplification of cDNA ends (RACE) technique. Sequence analysis indicated that the full-length cDNA of PDS was 2064 bp, encoding 586 amino acids and containing one open reading frame (ORF) of 1761 bp. Homology analysis showed that DkPDS, which had been submitted in GenBank with accession number GU112527, shared high similarities of 80–86% with PDS cloned from other plants. Prediction of deduced proteins showed that there was no signal peptide and transmembrane topological structure in DkPDS. It was a hydrophilic and stable protein, and located in chloroplast. To examine the specific expression patterns of carotenogenic genes we had cloned from persimmon, including phytoene synthase (DkPSY), DkPDS, ζ-carotene desaturase (DkZDS), lycopene β-cyclase (DkLCYB) and β-carotene hydroxylase (DkBCH), real-time quantitative PCR (Q-PCR) was performed in flesh at five different developmental stages. The results revealed that the expression levels of DkPSY, DkPDS and DkZDS gradually increased. Nevertheless, the expression level of DkLCYB was very low and maintained relatively stable. The expression level of DkBCH was also at a low level from stage 1 to 4, and then reached the maximum at stage 5. In addition, the expression level of DkZDS was higher than that of other genes. Carotenoid detection demonstrated that both β-cryptoxanthin and total carotenoids increased with fruit development, and zeaxanthin had little change, but with a sudden increase in final stage.

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Abbreviations

ABA:

Abscisic acid

BCH:

β-Carotene hydroxylase

Ct:

Threshold cycle

Da:

Dalton

FAO:

Food and Agriculture Organization

HPLC:

High performance liquid chromatography

LCYB:

Lycopene β-cyclase

LCYE:

Lycopene ε-cyclase

ORF:

Open reading frame

PCR:

Polymerase chain reaction

PDS:

Phytoene desaturase

pI:

Isoelectric point

PSY:

Phytoene synthase

Q-PCR:

Real-time quantitative PCR

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Semi-quantitative reverse transcription polymerase chain reaction

UTR:

Untranslated region

ZDS:

ζ-Carotene desaturase

ZEP:

Zeaxanthin epoxidase

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Acknowledgments

This work was financially supported by the Natural Science Foundation of China (30771491), Postdoctoral Science Foundation of Jiangsu Province (0901055C) and China Postdoctoral Science Foundation (20100471401). We also thank Institute of Agrobiology and Environmental Science, Zhejiang University providing the carotenoid analysis.

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  1. College of Horticulture and Plant Protection, Yangzhou University, Yangzhou, 225009, People’s Republic of China

    Daqiu Zhao, Chunhua Zhou, Fen Kong & Jun Tao

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  1. Daqiu Zhao
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  2. Chunhua Zhou
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Correspondence to Jun Tao.

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Zhao, D., Zhou, C., Kong, F. et al. Cloning of phytoene desaturase and expression analysis of carotenogenic genes in persimmon (Diospyros kaki L.) fruits. Mol Biol Rep 38, 3935–3943 (2011). https://doi.org/10.1007/s11033-010-0510-7

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