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Molecular Cloning and Expression of Phytoene Synthase, Lycopene Beta-cyclase, and Beta-carotene Hydroxylase Genes in Persimmon (Diospyros kaki L.) Fruits

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Abstract

Persimmon is not only an important fruit, which is eaten as a fresh fruit, but also traditionally used for many medicinal purposes, among which carotenoids contribute significantly to the color and nutritional value of persimmon fruit. Phytoene synthase (PSY), lycopene beta-cyclase (LCYB), and beta-carotene hydroxylase (BCH) are three important enzymes of carotenoid biosynthesis. In this study, cDNA and genomic sequences of DkPSY, DkLCYB, and DkBCH were isolated by rapid amplification of cDNA ends and polymerase chain reaction (PCR) using RNA and DNA isolated from the flesh tissues of persimmon used as template. Sequence analysis demonstrated that the individual cDNA and deduced proteins showed high identities to those of three carotenogenic cDNAs and proteins from other plants. Real-time quantitative PCR analysis revealed the tissue-specific expression patterns of DkPSY, DkLCYB, and DkBCH genes in fruit of persimmon, in which expression level of DkBCH was strongest among three genes analyzed. And, in different tissues, the expression level of peel tissue was higher than that of flesh and pedicel tissues.

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Abbreviations

aa:

Amino acid

BCH:

Beta-carotene hydroxylase

Ct:

Threshold cycle

ECH:

ε-Carotene hydroxylase

FAO:

Food and Agriculture Organization

LCYB:

Lycopene beta-cyclase

ORF:

Open reading frame

PCR:

Polymerase chain reaction

PDS:

Phytoene desaturase

pI:

Theoretical isoelectric point

PSY:

Phytoene synthase

Q-PCR:

Real-time quantitative PCR

RACE:

Rapid amplification of cDNA ends

UTR:

Untranslated region

ZDS:

ζ-Carotene desaturase

References

  • Achiwa Y, Hibasami H, Katsuzaki H, Imai K, Komiya T (1997) Inhibitory effects of persimmon (Diospyros kaki) extract and related polyphenol compounds on growth of human lymphoid leukemia cells. Biosci Biotechnol Biochem 61(7):1099–1101

    Article  PubMed  CAS  Google Scholar 

  • Ahn HS, Jeon TI, Lee JY, Hwang SG, Lim Y, Park DK (2002) Antioxidative activity of persimmon and grape seed extract: in vitro and in vivo. Nutr Res 22:1265–1273

    Article  CAS  Google Scholar 

  • Buckner B, Miguel PS, Janick-Buckner D, Bennetzen JL (1996) The y1 gene of maize codes for phytoene synthase. Genetics 143:479–488

    PubMed  CAS  Google Scholar 

  • Chen SH, Gao QC (1995) Comprehensive utilization of persimmon skin. Agriculture Products Development 5:35–36 (in Chinese)

    CAS  Google Scholar 

  • Chen DM, Zhang SL, Jin YF (1997) A method for genomic DNA preparation of woody fruit crops. Journal of Zhejiang Agricultural University 23:621–624 (in Chinese)

    CAS  Google Scholar 

  • Cunningham FX Jr, Gantt E (1998) Genes and enzymes of carotenoid biosynthesis in plants. Ann Rev Plant Physiol Plant Mol Biol 49:557–583

    Article  CAS  Google Scholar 

  • Feng TK, Li SG, Wang YL, Luo YP (2007) Cloning and sequence analysis of β-carotene hydroxylase gene from nanfeng Orange (Citrus reticulata Blanco var. kinokuni (Tanaka) H. H. Hu). Acta Botanica Boreali-Occidentalla Sinica 27:0238–0243 (in Chinese)

    CAS  Google Scholar 

  • Fraser PD, Bramley PM (2004) The biosynthesis and nutritional uses of carotenoids. Prog Lipid Res 43:228–265

    Article  PubMed  CAS  Google Scholar 

  • Fukuda T, Shibata H (1994) Persimmon calyx extracts as anticonvulsants and to alleviate the side effects of barbituric acid compounds. Patent-Japan Kokai Tokkyo Koho 06:649

    Google Scholar 

  • Giuliano G, Tavazza R, Diretto G, Beyer P, Taylor MA (2008) Metabolic engineering of carotenoid biosynthesis in plants. Trends Biotechnol 26:139–145

    Article  PubMed  CAS  Google Scholar 

  • Gorinstein S, Zachwieja Z, Folta M, Barton H, Piotrowicz J, Zemser M (2001) Comparative contents of dietary fiber, total phenolics, and minerals in persimmons and apples. J Agric Food Chem 49(2):952–957

    Article  PubMed  CAS  Google Scholar 

  • Gu HF, Li CM, Xu YJ, Hu WF, Chen MH, Wan QH (2008) Structural features and antioxidant activity of tannin from persimmon flesh. Food Res Int 41:208–217

    Article  CAS  Google Scholar 

  • Harker M, Hirschberg J (1998) Molecular biology of carotenoid biosynthetic organisms. Methods Enzymol 297:244–263

    Article  CAS  Google Scholar 

  • Joseph H (2001) Carotenoid biosynthesis in flowering plants. Plant Biol 4:210–218

    Google Scholar 

  • Kawase M, Motohashi N, Satoh K, Sakagami H, Nakashima H (2003) Biological activity of persimmon (Diospyros kaki) peel extracts. Phytother Res 17(5):495–500

    Article  PubMed  Google Scholar 

  • Kim IJ, Ko KC, Kim CS, Chung WI (2001a) Isolation and characterization of cDNAs encoding β-carotene hydroxylase in Citrus. Plant Sci 161:1005–1010

    Article  CAS  Google Scholar 

  • Kim IJ, Ko KC, Kim CS, Chung WI (2001b) Isolation and expression patterns of a cDNA encoding phytoene synthase in Citrus. J Plant Physiol 158:795–800

    Article  CAS  Google Scholar 

  • Kudritskaya SE, Fishman GM, Chikovani DM (1984) Carotenoids of the fruit of the subtropical persimmon, variety khachia. Chem Nat Compd 20:369

    Article  Google Scholar 

  • Mallavadhani UV, Panda AK, Rao YR (1998) Pharmacology and chemotaxonomy of Diospyros. Phytochemistry 49:901–951

    Article  PubMed  CAS  Google Scholar 

  • Nakagawa T, Nakatsuka A, Yano K, Yasugahira S, Nakamura R, Sun N, Itai A, Suzuki T, Itamura H (2008) Expressed sequence tags from persimmon at different developmental stages. Plant Cell Rep 27:931–938

    Article  PubMed  CAS  Google Scholar 

  • Niikawa T, Suzuki T, Ozeki T, Kato M, Ikoma Y (2007) Characteristics of carotenoid accumulation during maturation of the Japanese Persimmon ‘Fuyu’. Hortic Res (Japan) 6(2):251–256 (In Japanese)

    Article  CAS  Google Scholar 

  • Pecker I, Gabbay R, Cunningham FX, Hirschberg J (1996) Cloning and characterization of the cDNA for lycopene β-cyclase from tomato reveals decrease in its expression during fruit ripening. Plant Mol Biol 30:807–819

    Article  PubMed  CAS  Google Scholar 

  • Ramos A, Coesel S, Marques A, Rodrigues M, Baumgartner A, Noronha J, Rauter A, Brenig B, Varela J (2008) Isolation and characterization of a stress-inducible Dunaliella salina LCYB gene encoding a functional lycopene β-cyclase. Appl Microbiol Biotechnol 79:819–828

    Article  PubMed  CAS  Google Scholar 

  • Robert V, Jana J, Maja MP, Franci S, Valentina S (2010) Comparative study of primary and secondary metabolites in 11 cultivars of persimmon fruit (Diospyros kaki L.). Food Chem 119:477–483

    Article  Google Scholar 

  • Ronen G, Cohen M, Zamir D, Hirschberg J (1999) Regulation of carotenoid biosynthesis during tomato fruit development: expression of the gene for lycopene epsilon-cyclase is down-regulated during ripening and is elevated in the mutant Delta. Plant J 17:341–351

    Article  PubMed  CAS  Google Scholar 

  • Sakanaka S, Tachibana Y, Okada Y (2005) Preparation and antioxidant properties of extracts of Japanese persimmon leaf tea (Kakinohacha). Food Chem 89:569–575

    Article  CAS  Google Scholar 

  • Salvini M, Bernini A, Fambrini M, Pugliesi C (2005) cDNA cloning and expression of the phytoene synthase gene in sunflower. J Plant Physiol 162:479–484

    Article  PubMed  CAS  Google Scholar 

  • Shela G, Elzbieta B, Gustaw K, Marina Z, Simon T (1998) Dietary persimmon improves lipid metabolism in rats fed diets containing cholesterol. J Nutr 128(11):2023–2027

    Google Scholar 

  • Tao J, Zhang SL (2003) Metabolism of carotenoid and its regulation in garden crop. Journal of Zhejiang University (Agriculture and Life Science Editon) 29:585–590 (in Chinese)

    CAS  Google Scholar 

  • Thomas P, Chen TS (1998) Quantitative analyses of major carotenoid fatty acid esters in fruits by liquid chromatography: persimmon and papaya. Food Sci 53(6):1720–1722

    Google Scholar 

  • Xu CJ, Chen KS, Zhang SL, Qing QP (2003) Cloning of β-carotene hydroxylase cDNA from orange (Citrus sinensis) and its transient antisense expression. Journal of Agricultural Biotechnology 11:593–597 (in Chinese)

    Google Scholar 

  • Xu CJ, Chen KS, Zhang B, Wang QJ, Ye WJ (2004) A study on methods for RNA extraction from Citrus tissues. Journal of Fruit Science 21:136–140 (in Chinese)

    Google Scholar 

  • Yan Y, Jiang JG, Song DL (2004) Cloning and sequencing of genomic DNA encoding phytoene synthase from Dunaliell Salina. Sea-Lake Salt and Chemical Industry 34:22–24 (in Chinese)

    Google Scholar 

  • Ye ZW, Jiang JG, Wu GH (2008) Biosynthesis and regulation of carotenoids in Dunaliella: progresses and prospects. Biotechnol Adv 26:352–360

    Article  PubMed  CAS  Google Scholar 

  • Yuan B, Xu HL, Leng P (2006) Content and chemical composition of carotenoids in persimmon fruit. Chinese Agricultural Science Bulletin 22:277–280 (in Chinese)

    Google Scholar 

  • Zhu CF, Yamamura S, Koiwa H, Nishihara M, Sandmann G (2002) cDNA cloning and expression of carotenogenic genes during flower development in Gentiana lutea. Plant Mol Biol 48:277–2851

    Article  PubMed  CAS  Google Scholar 

  • Zhu CF, Chen X, Wang YD (2004) Carotenoid biosynthesis in plants and application of its relative genes in gene engineering. Journal of Plant Physiology and Molecular Biology 30:609–618 (in Chinese)

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

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).

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Authors and Affiliations

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

    Daqiu Zhao, Chunhua Zhou, Yanle Sheng & Jun Tao

  2. Key Laboratory of Plant Functional Genomics of Ministry of Education, College of Agronomy, Yangzhou University, Yangzhou, 225009, People’s Republic of China

    Guohua Liang

Authors
  1. Daqiu Zhao
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  2. Chunhua Zhou
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  3. Yanle Sheng
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  4. Guohua Liang
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  5. Jun Tao
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Corresponding author

Correspondence to Jun Tao.

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Table S1

Primers used for isolation of the full-length PSY, LCYB, and BCH cDNAs from persimmon fruits (DOC 53 kb)

Table S2

Primers used for isolation of the genomic sequence and gene expression analysis (DOC 57 kb)

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Zhao, D., Zhou, C., Sheng, Y. et al. Molecular Cloning and Expression of Phytoene Synthase, Lycopene Beta-cyclase, and Beta-carotene Hydroxylase Genes in Persimmon (Diospyros kaki L.) Fruits. Plant Mol Biol Rep 29, 345–351 (2011). https://doi.org/10.1007/s11105-010-0238-5

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