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Isolation and functional characterization of a novel gene coding for flavonoid 3′-hydroxylase from globe artichoke

  • Original Papers
  • Published:
Biologia Plantarum

Abstract

Globe artichoke (Cynara cardunculus L. var. scolymus) is rich in flavonoids which contribute to its health-promoting properties. With the aim of understanding the genetic control of flavonoid accumulation in artichoke, we isolated an artichoke full-length cDNA sequence encoding flavonoid 3′-hydroxylase (F3′H), a major enzyme of the flavonoid hydroxylation pattern. In silico studies confirmed that the deduced amino acid sequence of CcF3′H is highly similar to F3′Hs isolated from other Asteraceae. The Northern blot analysis demonstrated that CcF3′H was highly expressed in leaves and in specific parts of the heads. Its expression differed slightly among artichoke cultivars. The overexpression of CcF3′H in tobacco plants led to the accumulation of flavonoids and to an increase of flower colour intensity, thus identifying CcF3′H as promising candidate for genetic engineering. CcF3′H represents the first structural gene of the flavonoid biosynthesis isolated from C. cardunculus, and its characterization sheds light on the accumulation of flavonoids.

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Abbreviations

ANS:

anthocyanidin synthase

CGA:

chlorogenic acid

DFR:

dihydroflavonol 4-reductase

FHT:

flavanone 3-hydroxylase

FLS:

flavonol synthase

FNSII:

flavone synthase II

F3′H:

flavonoid 3′-hydroxylase

F3′5′H:

flavonoid 3′,5′-hydroxylase

nptII:

neomycin phosphotransferase gene

ORF:

open reading frame

RACE:

rapid amplification of cDNA ends

RT-PCR:

reverse transcription polymerase chain reaction

UTR:

untranslated regions

WT:

wild type

References

  • Bogs, J., Ebadi, A., McDavid, D., Robinson, S.P.: Identification of the flavonoid hydroxylases from grapevine and their regulation during fruit development. — Plant Physiol. 140: 279–291, 2006.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Brown, J.E., Rice-Evans, C.A.: Luteolin-rich artichoke extract protects low density lipoprotein from oxidation in vitro. — Free Radical Res. 29: 247–255, 1998.

    Article  CAS  Google Scholar 

  • Brugliera, F., Barri-Rewell, G., Holton, T.A., Mason, J.G.: Isolation and characterization of a flavonoid 3′-hydroxylase cDNA clone corresponding to the Ht1 locus of Petunia hybrida. — Plant J. 19: 441–451, 1999.

    Article  CAS  PubMed  Google Scholar 

  • Chen, S.M., Li, C.H., Zhu, X.R., Deng, Y.M., Sun, W., Wang, L.S., Chen, F.D., Zhang, Z.: The identification of flavonoids and the expression of genes of anthocyanin biosynthesis in the chrysanthemum flowers. — Biol. Plant. 56: 458–464, 2012.

    Article  CAS  Google Scholar 

  • Chomeczynski, P., Sacchi, N.: Single-step method of RNA isolation by acid guanidinium thiocyanate phenol chloroform extraction. — Anal. Biochem. 162: 156–159, 1987.

    Google Scholar 

  • Comino, C., Hehn, A., Moglia, A., Menin, B., Bourgaud, F., Lanteri, S., Portis, E.: The isolation and mapping of a novel hydroxycinnamoyltransferase in the globe artichoke chlorogenic acid pathway. — BMC Plant Biol. 9: 30, 2009.

    Article  PubMed Central  PubMed  Google Scholar 

  • Comino, C., Lanteri, S., Portis, E., Acquadro, A., Romani, A., Hehn, A., Larbat, R., Bourgaud, F.: Isolation and functional characterization of a cDNA coding a hydroxycinnamoyltransferase involved in phenylpropanoid biosynthesis in Cynara cardunculus L. — BMC Plant Biol. 7: 14, 2007.

    Article  PubMed Central  PubMed  Google Scholar 

  • Croft, K.D.: The chemistry and biological effects of flavonoids and phenolic acids. — Ann. N.Y. Acad. Sci. 854: 435–442, 1998.

    Article  CAS  PubMed  Google Scholar 

  • Crosby, K.C., Pietraszewska-Bogiel, A., Gadella, T.W. Jr., Winkel, B.S.: Förster resonance energy transfer demonstrates a flavonoid metabolon in living plant cells that displays competitive interactions between enzymes. — FEBS Lett. 585: 2193–2198, 2011.

    Article  CAS  PubMed  Google Scholar 

  • De Paolis, A., Pignone, D., Morgese, A., Sonnante, G.: Characterization and differential expression analysis of artichoke phenylalanine ammonia-lyase coding sequences. — Physiol. Plant. 132: 33–43, 2008.

    PubMed  Google Scholar 

  • Dereeper, A., Guignon, V., Blanc, G., Audic, S., Buffet, S., Chevenet, F., Dufayard, J.F., Guindon, S., Lefort, V., Lescot, M., Claverie, J.M., Gascuel, O.: Phylogeny.fr: robust phylogenetic analysis for the non-specialist. — Nucl. Acids Res. 1(Suppl.): W465–W469, 2008.

    Article  Google Scholar 

  • Forkmann, G.: Flavonoids as flower pigments: the formation of the natural spectrum and its extension by genetic engineering. — Plant Breed. 106: 1–26. 1991.

    Article  CAS  Google Scholar 

  • Fratianni, F., Tucci, M., De Palma, M., Pepe, R., Nazzaro, F.: Polyphenolic composition in different parts of some cultivars of globe artichoke Cynara cardunculus L. var. scolymus (L.) Fiori. — Food Chem. 104: 1282–1286, 2007.

    Article  CAS  Google Scholar 

  • Gebhardt, R.: Inhibition of cholesterol biosynthesis in primary cultured rat hepatocytes by artichoke (Cynara scolymus L.) extracts. — J. Pharmacol. exp. Therapy 286: 1122–1128, 1998.

    CAS  Google Scholar 

  • Guindon, S., Dufayard, J.F., Lefort, V., Anisimova, M., Hordijk, W., Gascuel, O.: New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. — System. Biol. 59: 307–321, 2010.

    Article  CAS  Google Scholar 

  • Halbwirth, H.: The creation and physiological relevance of divergent hydroxylation patterns in the flavonoid pathway. — Int. J. mol. Sci. 11: 595–621, 2010.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Han, Y., Vimolmangkang, S., Soria-Guerra, R.E., Rosales-Mendoza, S., Zheng, D., Lygin, A.V., Korban, S.S.: Ectopic expression of apple F3′H genes contributes to anthocyanin accumulation in the Arabidopsis tt7 mutant grown under nitrogen stress. — Plant Physiol. 153: 806–820, 2010.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Harborne, J.B., Williams, C.A.: Advances in flavonoid research since 1992. — Phytochemistry 55: 481–504, 2000.

    Article  CAS  PubMed  Google Scholar 

  • Hofgen, R., Willmitzer, L.: Storage of competent cells for Agrobacterium transformation. — Nucl. Acids Res. 16: 9877, 1988.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Holton, T.A., Brugliera, F., Tanaka, Y.: Cloning and expression of flavonol synthase from Petunia hybrida. — Plant J. 4: 1003–1010, 1993.

    Article  CAS  PubMed  Google Scholar 

  • Holton, T.A., Cornish, E.C.: Genetics and biochemistry of anthocyanin biosynthesis. — Plant Cell 7: 1071–1083, 1995.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Horsh, R.B., Fry, J.E., Hoffmann, N.L., Eicholtz, D., Rogers, S.H., Fraley, R.T.: A simple and general method for transferring genes in plants. — Science 227: 1229–1231, 1987.

    Google Scholar 

  • Ishiguro, K., Taniguchi, M., Tanaka, Y.: Functional analysis of Antirrhinum kelloggii flavonoid 3′-hydroxylase and flavonoid 3′,5′-hydroxylase genes; critical role in flower color and evolution in the genus Antirrhinum. — J. Plant Res. 125: 451–456, 2012.

    Article  CAS  PubMed  Google Scholar 

  • Kitada, C., Gong, Z., Tanaka, Y., Yamazaki, M., Saito, K.: Differential expression of two cytochrome P450s involved in the biosynthesis of flavones and anthocyanins in chemovarietal forms of Perilla frutescens. — Plant Cell Physiol. 42: 1338–1344, 2001.

    CAS  PubMed  Google Scholar 

  • Lattanzio, V., Kroon, P.A., Linsalata, V., Carinali, A.: Globe artichoke: a functional food and source of nutraceutical ingredients. — J. Funct. Foods 1: 131–144, 2009.

    Article  CAS  Google Scholar 

  • Lombardo, S., Pandino, G., Mauromicale, G., Knodler, M., Carle, R., Schieber, A.: Influence of genotype, harvest time and plant part on polyphenolic composition of globe artichoke (Cynara cardunculus L.var. scolymus L. Fiori). — Food Chem. 119: 1175–1181, 2010.

    Article  CAS  Google Scholar 

  • Martens, S., Mithofer, A.: Flavones and flavone synthases. — Phytochemistry 66: 2399–2407, 2005.

    Article  CAS  PubMed  Google Scholar 

  • Mateus, N., Silva, A.M.S., Santos-Buelga, C., Rivas-Gonzalo, J.C., De Freitas, V.: Identification of anthocyanin-flavanol pigments in red wines by NMR and mass spectrometry. — J. Agr. Food. Chem. 50: 2110–2116, 2002.

    Article  CAS  Google Scholar 

  • Moglia, A., Comino, C., Portis, E., Acquadro, A., De Vos, R.C., Beekwilder, J., Lanteri, S.: Isolation and mapping of a C3’H gene (CYP98A49) from globe artichoke, and its expression upon UV-C stress. — Plant Cell Rep. 28: 963–974, 2009.

    Article  CAS  PubMed  Google Scholar 

  • Murakami, K., Mihara, K., Omura, T.: The transmembrane region of microsomal cytochrome P450 identified as the endoplasmic reticulum retention signal. — J. Biochem. 116: 164–175, 1994.

    CAS  PubMed  Google Scholar 

  • Nakatsuka, T., Nishihara, M., Mishiba, K., Yamamura, S.: Temporal expression of flavonoid biosynthesis-related genes regulates flower pigmentation in gentian plants. — Plant Sci. 168: 1309–1318, 2005.

    Article  CAS  Google Scholar 

  • Nakatsuka, T., Nishihara, M., Mishiba, K., Yamamura, S.: Heterologous expression of two gentian cytochrome P450 genes can modulate the intensity of flower pigmentation in transgenic tobacco plants. — Mol. Breed. 17: 91–99, 2006.

    Article  CAS  Google Scholar 

  • Nishihara, M., Nakatsuka, T.: Genetic engineering of flavonoid pigments to modify flower color in floricultural plants. — Biotechnol. Lett. 33: 433–441, 2011.

    Article  CAS  PubMed  Google Scholar 

  • Noda, N., Kanno, Y., Kato, N., Kazuma, K., Suzuki, M.: Regulation of gene expression involved in flavonol and anthocyanin biosynthesis during petal development in lisianthus (Eustoma grandiflorum). — Physiol. Plant. 122: 305–313, 2004.

    Article  CAS  Google Scholar 

  • Sato, T., Sakaguchi, M., Mihara, K., Omura, T.: The aminoterminal structures that determine topological orientation of cytochrome P-450 in microsomal membrane. — Embo J. 9: 2391–2397, 1990.

    CAS  PubMed Central  PubMed  Google Scholar 

  • Schardl, C., Byrd, A.D., Benzion, G.B., Altschuler, M.A., Hildebrand, D.F., Hunt, A.G.: Design and construction of a versatile system for the expression of foreign genes in plants. — Gene 61: 1–11, 1987.

    Article  CAS  PubMed  Google Scholar 

  • Schlangen, K., Miosic, S., Thill, J., Halbwirth, H.: Cloning, functional expression, and characterization of a chalcone 3-hydroxylase from Cosmos sulphureus. — J. exp. Bot. 61: 3451–3459, 2010.

    Article  CAS  PubMed  Google Scholar 

  • Schoenbohm, C., Martens, S., Eder, C., Forkmann, G., Weisshaar, B.: Identification of the Arabidopsis thaliana flavonoid 3′-hydroxylase gene and functional expression of the encoded P450 enzyme. — Biol. Chem. 381: 749–753, 2000.

    Article  CAS  PubMed  Google Scholar 

  • Schutz, K., Kammerer, D., Carle, R., Schieber, A.: Identification and quantification of caffeoylquinic acids and flavonoids from artichoke (Cynara scolymus L.) heads, juice, and pomace by HPLC-DAD-ESI/MS. — J. Agr. Food Chem. 52: 4090–4096, 2004.

    Article  Google Scholar 

  • Schutz, K., Persike, M., Carle, R., Schieber, A.: Characterization and quantification of anthocyanins in selected artichoke (Cynara scolymus L.) cultivars by HPLC-DADESI-MS. — Anal. Bioanal. Chem. 384: 1511–1517, 2006.

    Article  PubMed  Google Scholar 

  • Seitz, C., Eder, C., Deiml, B., Kellner, S., Martens, S., Forkmann, G.: Cloning, functional identification and sequence analysis of flavonoid 3′-hydroxylase and flavonoid 3′,5′-hydroxylase cDNA reveals independent evolution of flavonoid 3′,5′-hydroxylase in the Asteraceae family. — Plant mol. Biol. 61: 365–381, 2006.

    Article  CAS  PubMed  Google Scholar 

  • Sonnante, G., D’Amore, R., Blanco, E., Pierri, C.L., De Palma, M., Luo, J., Tucci, M., Martin, C.: Novel hydroxycinnamoyl-coenzyme A quinate transferase genes from artichoke are involved in the synthesis of chlorogenic acid. — Plant Physiol. 153: 1224–1238, 2010.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Soriano, I.R., Asenstorfer, R.E., Schmidt, O., Riley, I.T.: Inducible flavone in oats (Avena sativa) is a novel defense against plant-parasitic nematodes. — Phytopathology 94: 1207–1214, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Treutter, D.: Significance of flavonoids in plant resistance and enhancement of their biosynthesis. — Plant Biol. 7: 581–591, 2005.

    Article  CAS  PubMed  Google Scholar 

  • Tsuda, T.: Dietary anthocyanin-rich plants: biochemical basis and recent progress in health benefits studies. — Mol. Nutr. Food Res. 56: 159–170, 2012.

    Article  CAS  PubMed  Google Scholar 

  • Ueyama, U., Suzuki, K., Fukuchi-Mizutani, M., Fukui, Y., Miyazaki, K., Ohkawa, H., Kusumi, T., Tanaka, Y.: Molecular and biochemical characterization of torenia flavonoid 3′-hydroxylase and flavone synthase II and modification of flower color by modulating the expression of these genes. — Plant Sci. 163: 253–263, 2002.

    Article  CAS  Google Scholar 

  • Wang, M., Simon, J.E., Aviles, I.F., He, K., Zheng, Q.Y., Tadmor, Y.: Analysis of antioxidative phenolic compounds in artichoke (Cynara scolymus L.). — J. Agr. Food Chem. 51: 601–608, 2003.

    Article  CAS  Google Scholar 

  • Wassenegger, M., Pélissier, T.: A model for RNA-mediated gene silencing in higher plants. — Plant mol. Biol. 37: 349–362, 1998.

    Article  CAS  PubMed  Google Scholar 

  • Winkel, B.S.: Metabolic channeling in plants. — Annu. Rev. Plant. Biol. 55: 85–107, 2004.

    Article  CAS  PubMed  Google Scholar 

  • Winkel-Shirley, B.: Flavonoid biosynthesis: a colorful model for genetics, biochemistry, cell biology, and biotechnology. — Plant Physiol. 126: 485–493, 2001.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Winkel-Shirley, B.: Biosynthesis of flavonoids and effects of stress. — Curr. Opin. Plant Biol. 5: 218–223, 2002.

    Article  CAS  PubMed  Google Scholar 

  • Yamazaki, S., Sato, K., Suhara, K., Sakaguchi, M., Mihara, K., Omura, T.: Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s. — J. Biochem. 114: 652–657, 1993.

    CAS  PubMed  Google Scholar 

  • Zhang, J., Subramanian, S., Zhang, Y., Yu, O.: Flavone synthases from Medicago truncatula are flavanone-2-hydroxylases and are important for nodulation. — Plant Physiol. 144: 741–751, 2007.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhang, Q., Su, L.J., Chen, J.W., Zeng, X.Q., Sun, B.Y., Peng, C.L.: The antioxidative role of anthocyanins in Arabidopsis under high-irradiance. — Biol. Plant. 56: 97–104, 2012.

    Article  Google Scholar 

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Authors and Affiliations

  1. IBBR Institute of Biosciences and Bioresources, CNR, I-80055, Portici, Italy

    M. De Palma & M. Tucci

  2. Institute of Food Science and Technology, CNR, I-83100, Avellino, Italy

    F. Fratianni & F. Nazzaro

Authors
  1. M. De Palma
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  2. F. Fratianni
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  3. F. Nazzaro
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  4. M. Tucci
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Correspondence to M. Tucci.

Additional information

Acknowledgements: We thank Dr. R. Pepe from the Agricultural Research Council, the Research Centre for Vegetable Crops (CRAORT) for providing globe artichoke cultivars. We are also grateful to Mr. R. Nocerino, A. Scafarto, and G. Guarino from the CNR-IBBR for an excellent technical assistance, and to Dr. G. Caruso from the Agronomy Department, University of Naples Federico II, for help with the statistical analysis. This research was partly supported by grants from the Italian Ministry for University and Scientific and Technological Research (DD 1291 15/12/2003, project 161 and DD n. 62/Ric 08/10/2012, project PON02_00395_3215002).

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Palma, M.D., Fratianni, F., Nazzaro, F. et al. Isolation and functional characterization of a novel gene coding for flavonoid 3′-hydroxylase from globe artichoke. Biol Plant 58, 445–455 (2014). https://doi.org/10.1007/s10535-014-0424-7

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  • DOI: https://doi.org/10.1007/s10535-014-0424-7

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