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Synthesis and accumulation of anthocyanins in sour cherries during ripening in accordance with antioxidant capacity development and chalcone synthase expression


Phytonutrient metabolism in sour cherries takes place during fruit ripening. This study demonstrated that total phenolic–flavonoid contents decline during ripening while total anthocyanin content significantly increases at the same period. There were no detectable anthocyanins in green cherry fruits. Anthocyanin biosynthesis started concurrently with color formation. Cyanidin-3-glucosylrutinoside and cyanidin-3-rutinoside were the early anthocyanins accumulated in fruits and were accompanied with cyanidin-3-sophoroside and cyanidin-3-glucoside in fully ripe fruits. Phenylpropanoid pathway involves anthocyanin biosynthesis, and chalcone synthase (CHS) is one of the key enzymes regulating the pathway. Three CHS genes (PcChs1, PcChs2, PcChs3) were isolated from sour cherry genome, and their transcription profiles were determined using the RT-PCR approach. There was no CHS gene expression before breaker stage, and PcChs1, PcChs2, PcChs3 transcription were upregulated in parallel with pigmentation in sour cherry cells and PcChs1 had the highest transcripts in fully ripe fruits.

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  1. 1.

    Kang SY, Seeram NP, Nair MG, Bourquin LD (2003) Tart cherry anthocyanins inhibit tumor development in ApcMin mice and reduce proliferation of human colon cancer cells. Cancer Lett 194:13–19

    Article  CAS  Google Scholar 

  2. 2.

    Wang H, Nair MG, Strasburg GM, Chang YC, Booren AM, Gray JI, Dewitt DL (1999) Antioxidant and antiinflammatory activities of anthocyanins and their aglycon, cyanidin, from tart cherries. J Nat Prod 62:294–296

    Article  CAS  Google Scholar 

  3. 3.

    Connolly DAJ, McHugh MP, Padilla-Zakour OL (2006) Efficacy of a tart cherry juice blend in preventing the symptoms of muscle damage. Br J Sports Med 40:679–683

    Article  CAS  Google Scholar 

  4. 4.

    Kuehl KS, Perrier ET, Elliot DL, Chesnutt JC (2010) Efficacy of tart cherry juice in reducing muscle pain during running: a randomized controlled trial. J Int Soc Sports Nutr 7:7–17

    Article  Google Scholar 

  5. 5.

    Kelley DS, Rasooly R, Jacob RA, Kader AA, Mackey BE (2006) Consumption of bing sweet cherries lowers circulating concentrations of inflammation markers in healthy men and women. J Nutr 136:981–986

    CAS  Google Scholar 

  6. 6.

    Howatson G, Bell PG, Tallent J, Middleton B, McHugh MP, Ellis J (2012) Effect of tart cherry juice (Prunus cerasus) on melatonin levels and enhanced sleep quality. Eur J Nutr 51:909–916

    Article  CAS  Google Scholar 

  7. 7.

    Heller W, Hahlbrock K (1980) Highly purified flavanone synthase from parsley catalyzes the formation of naringenin chalcone. Arch Biochem Biophys 200:617–619

    Article  CAS  Google Scholar 

  8. 8.

    Jiang C, Kim SY, Suh D-Y (2008) Divergent evolution of the thiolase superfamily and chalcone synthase family. Mol Phylogenet Evol 49:691–701

    Article  CAS  Google Scholar 

  9. 9.

    Poll L, Petersen M, Nielsen GS (2003) Influence of harvest year and harvest time on soluble solids, titrateable acid, anthocyanin content and aroma components in sour cherry (Prunus cerasus L. cv. “Stevnsbær”). Eur Food Res Technol 216(3):212–216

    CAS  Google Scholar 

  10. 10.

    Lodhi MA, Guang-Ning Y, Weeden NF, Reisch BI (1994) A simple and efficient method for DNA extraction from grapevine cultivars, vitis species and ampelopsis. Plant Mol Biol 12:6–13

    Article  CAS  Google Scholar 

  11. 11.

    Lopez-Gomez R, Gomez-Lim MA (1992) A method for extracting intact RNA from fruits rich in polysaccharides using ripe mango mesocarp. HortScience 27:440–442

    CAS  Google Scholar 

  12. 12.

    Liu M, Li XQ, Weber C, Lee CY, Brown J, Liu RH (2002) Antioxidant and antiproliferative activities of raspberries. J Agric Food Chem 50:2926–2930

    Article  CAS  Google Scholar 

  13. 13.

    Lee J, Durst RW, Wrolstad RE (2005) Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: collaborative study. J AOAC Int 88:1269–1278

    CAS  Google Scholar 

  14. 14.

    Karaaslan M, Yilmaz FM, Cesur Ö, Vardin H, Ikinci A, Dalgiç AC (2014) Drying kinetics and thermal degradation of phenolic compounds and anthocyanins in pomegranate arils dried under vacuum conditions. Int J Food Sci Technol 49(2):595–605

    Article  CAS  Google Scholar 

  15. 15.

    Re R, Pellegrini N, Proteggente A, Pannala AS, Yang M, Rice-Evans CA (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26:1231–1237

    Article  CAS  Google Scholar 

  16. 16.

    Blando F, Gerardi C, Nicoletti I (2004) Sour cherry anthocyanins as ingredients for functional foods. J Biomed Biotechnol 5:253–258

    Article  Google Scholar 

  17. 17.

    Benzie IF, Strain J (1996) The ferric reducing ability of plasma (FRAP) as a measure of “Antioxidant Power”: the FRAP assay. Anal Biochem 239:70–76

    Article  CAS  Google Scholar 

  18. 18.

    Garofulić IE, Dragović-Uzelac V, Jambrak AR, Jukić M (2013) The effect of microwave assisted extraction on the isolation of anthocyanins and phenolic acids from sour cherry Marasca (Prunus cerasus var. Marasca). J Food Eng 117:437–442

    Article  CAS  Google Scholar 

  19. 19.

    Kirakosyan A, Seymour EM, Urcuyo Llanes DE, Kaufman PB, Bolling SF (2009) Chemical profile and antioxidant capacities of tart cherry products. Food Chem 115:20–25

    Article  CAS  Google Scholar 

  20. 20.

    Simunic V, Kovac S, Gaso-Sokac D, Pffannhauser W, Murkovic M (2005) Determination of anthocyanins in four Croatian cultivars of sour cherries. Eur Food Res Technol 220:575–578

    Article  CAS  Google Scholar 

  21. 21.

    Tomàs-Barberàn FA, Gil MI, Cremin P, Waterhouse AL, Hess-Pierce B, Kader AA (2001) HPLC–DAD–ESIMS analysis of phenolic compounds in nectarines, peaches, and plums. J Agric Food Chem 49:4748–4760

    Article  CAS  Google Scholar 

  22. 22.

    el Amira A, Behija SE, Beligh M, Lamia L, Manel I, Mohamed H, Lotfi A (2012) Effects of ripening stage on phenolic profile, phytochemical composition and antioxidant activity at date palm fruit. J Agric Food Chem 60:10896–10902

    Article  CAS  Google Scholar 

  23. 23.

    Myhara RM, Karkalas J, Taylor MS (1999) The composition of maturing Omani dates. J Sci Food Agric 79:1345–1350

    Article  CAS  Google Scholar 

  24. 24.

    Amiot JM, Tacchini M, Aubert SY, Oleszek W (1995) Influence of cultivar, maturity stage and storage conditions on phenolic composition and enzymatic browning of pear fruit. J Agric Food Chem 43:1132–1137

    Article  CAS  Google Scholar 

  25. 25.

    Allaith AAA (2008) Antioxidant activity of Bahraini date palm (Phoenix dactylifera L.) fruit of various cultivars. Int J Food Sci Technol 43:1033–1040

    Article  CAS  Google Scholar 

  26. 26.

    Awad MA, Al-Qurashia AD, Mohamed SA (2011) Antioxidant capacity, antioxidant compounds and antioxidant enzyme activities in five date cultivars during development and ripening. Sci Hort 129:688–693

    Article  CAS  Google Scholar 

  27. 27.

    Prior RL, Wu X, Schaich K (2005) Standardized methods for the determination of antioxidant capacity and phenolics in foods, and dietary supplements. J Agric Food Chem 53:4290–4302

    Article  CAS  Google Scholar 

  28. 28.

    Wiczkowski W, Szawara-Nowak D, Topolska J (2015) Changes in the content and composition of anthocyanins in red cabbage and its antioxidant capacity during fermentation, storage and stewing. Food Chem 167:115–123

    Article  CAS  Google Scholar 

  29. 29.

    Huang D, Ou B, Hampsch-Woodill M, Flanagan JA, Prior RL (2002) High-throughput assay of oxygen radical absorbance capacity (ORAC) using a multichannel liquid handling system coupled with a microplate fluorescence reader in 96-well format. J Agric Food Chem 50:4437–4444

    Article  CAS  Google Scholar 

  30. 30.

    Ou B, Hampsch-Woodill M, Prior RL (2001) Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe. J Agric Food Chem 49:4619–4626

    Article  CAS  Google Scholar 

  31. 31.

    Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Byrne DH (2006) Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. J Food Compos Anal 19:669–675

    Article  CAS  Google Scholar 

  32. 32.

    Proteggente AR, Pannala AS, Paganga G, Van Buren L, Wagner E, Wiseman S, Van De Put F, Dacombe C, Rice-Evans CA (2002) The antioxidant activity of regularly consumed fruit and vegetables reflects their phenolic and vitamin C composition. Free Radic Res 36:217–233

    Article  CAS  Google Scholar 

  33. 33.

    Lister CE, Wilson PE, Sutton KH, Morrison SC (2002) Understanding the health benefits of blackcurrants. Acta Hort 585:443–449

    Article  CAS  Google Scholar 

  34. 34.

    Schlesier K, Harwat M, Böhm V, Bitsch R (2002) Assessment of antioxidant activity by using different in vitro methods. Free Radic Res 36:177–187

    Article  CAS  Google Scholar 

  35. 35.

    Chaovanalikit A, Wrolstad RE (2004) Anthocyanin and polyphenolic composition of fresh and processed cherries. J Food Sci 69:73–83

    Google Scholar 

  36. 36.

    Eid NM, Al-Awadi B, Vauzour D, Oruna-Concha MJ, Spencer JP (2013) Effect of cultivar type and ripening on the polyphenol content of date palm fruit. J Agric Food Chem 61:2453–2460

    Article  CAS  Google Scholar 

  37. 37.

    Delgado R, Martín P, del Álamo M, González M-R (2004) Changes in the phenolic composition of grape berries during ripening in relation to vineyard nitrogen and potassium fertilisation rates. J Sci Food Agric 84:623–630

    Article  CAS  Google Scholar 

  38. 38.

    Liang Z, Sang M, Fan P, Wu B, Wang L, Duan W, Li S (2011) Changes of polyphenols, sugars, and organic acid in 5 Vitis genotypes during berry ripening. J Food Sci 76:1231–1238

    Article  CAS  Google Scholar 

  39. 39.

    Holoton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071–1083

    Article  Google Scholar 

  40. 40.

    Tsuda T, Yamaguchi M, Honda C, Moriguchi T (2004) Expression of anthocyanin biosynthesis genes in the skin of peach and nectarine fruit. J Am Soc Hortic Sci 129:857–862

    CAS  Google Scholar 

  41. 41.

    Boss PK, Davies C, Robinson SP (1996) Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifera L. cv Shiraz grape berries and the implications for pathway regulation. Plant Physiol 111:1059–1066

    CAS  Google Scholar 

  42. 42.

    Boss PK, Davies C, Robinson SP (1996) Expression of anthocyanin biosynthesis pathway genes in red and white grapes. Plant Mol Biol 32:565–569

    Article  CAS  Google Scholar 

  43. 43.

    Honda CN, Kotoda M, Wada S, Kondo S, Kobayashi J, Soejima Z, Zhang T, Tsuda T, Moriguchi T (2002) Anthocyanin biosynthetic genes are coordinately expressed during red coloration in apple skin. Plant Physiol Biochem 40:955–962

    Article  CAS  Google Scholar 

  44. 44.

    Kobayashi S, Ishimaru M, Ding CK, Yakushiji H, Goto N (2001) Comparison of UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT) gene sequences between white grapes (Vitis vinifera) and their sports with red skin. Plant Sci 160:543–550

    Article  CAS  Google Scholar 

  45. 45.

    Kondo S, Hiraoka K, Kobayashi S, Honda C, Terahara N (2002) Changes in the expression of anthocyanin biosynthetic genes during apple development. J Am Soc Hortic Sci 127:971–976

    CAS  Google Scholar 

  46. 46.

    Li Y, Sakiyama R, Maruyama H, Kawabata S (2001) Regulation of anthocyanin biosynthesis during fruit development in ‘Nyoho’ strawberry. J Jpn Soc Hortic Sci 70:28–32

    Article  CAS  Google Scholar 

  47. 47.

    Moyano E, Portero-Robles I, Medina-Escobar N, Valpuesta V, Munoz-Blanco J, Caballero JL (1998) A fruit-specific putative dihydroflavonol 4-reductase gene is differentially expressed in strawberry during the ripening process. Plant Physiol 117:711–716

    Article  CAS  Google Scholar 

  48. 48.

    Parker JL (2010) What makes a cherry red: an investigation into flavonoid pathway regulation in sweet cherry (Prunus avium L.) fruit. Ph.D. Thesis, The University of Adelaide

  49. 49.

    Mori T, Sakurai M, Sakuta M (2001) Effects of conditioned medium on activities of PAL, CHS, DAHP synthase (DS-Co and DS-Mn) and anthocyanin production in suspension cultures of Fragaria ananassa. Plant Sci 160:355–360

    Article  CAS  Google Scholar 

  50. 50.

    Davis G, Ananga A, Krastanova S, Sutton S, Ochieng JW, Leong S, Tsolova V (2012) Elevated gene expression in chalcone synthase enzyme suggests an increased production of flavonoids in skin and synchronized red cell cultures of North American native grape berries. DNA Cell Biol 31:939–945

    Article  CAS  Google Scholar 

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The research support from TÜBİTAK (Project# 112O001) is gratefully acknowledged by the authors.

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Correspondence to Mehmet Karaaslan.

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Karaaslan, M., Yılmaz, F.M., Karaaslan, A. et al. Synthesis and accumulation of anthocyanins in sour cherries during ripening in accordance with antioxidant capacity development and chalcone synthase expression. Eur Food Res Technol 242, 189–198 (2016).

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  • Sour cherry
  • Anthocyanins
  • Phenolics
  • Chalcone synthase
  • HPLC