, 230:1141 | Cite as

Transcriptional regulation of anthocyanin biosynthesis in red cabbage

  • Youxi Yuan
  • Li-Wei Chiu
  • Li LiEmail author
Original Article


The color of red cabbage (Brassica oleracea var. capitata) is due to anthocyanin accumulation. To investigate the regulatory control of anthocyanin production in red cabbage, the expression of anthocyanin biosynthetic and regulatory genes from eight commercial cultivars was examined. While the four green varieties had negligible amount of anthocyanins under normal growth condition, the four red cultivars contained up to 1.60 mg g−1 fresh weight. HPLC analysis of the four red cultivars revealed that they produced similar composition of various forms of cyanidin glucosides but at different concentrations. Molecular analysis indicated that all the red cabbage shared common mechanism of regulatory control for anthocyanin biosynthesis. Except CHI which showed similar expression levels between green and red cultivars, the other structural genes, CHS, F3H, F3′H, DFR, LDOX, and GST, were constitutively up-regulated during all stages of vegetative growth in red varieties. The expression of these structural genes was also dramatically increased in green and red cabbage under nutrient stresses. The increased expression of the structural genes coincided with a coordinated increase in transcript levels of a bHLH gene, BoTT8, and a MYB transcription factor, BoMYB2. These results indicate that activation of these two regulatory factors by unknown mechanisms constitutively up-regulates nearly the entire pathway genes for the onset of anthocyanin biosynthesis in red cabbage. Moreover, the amount of total anthocyanins in red cabbage was found to be positively correlated with total antioxidant power, implicating the potential health benefit of red cabbage to human health.


Anthocyanin biosynthesis Red cabbage Transcriptional regulation Nutrient deficiency 



Phenylalanine ammonia lyase


Chalcone synthase


Chalcone isomerases


Flavonone 3-hydroxylase


Flavonoid 3′-hydroxylase


Dihydroflavonol reductase


Anthocyanidin synthase




Transparent Testa Glabra1


Transparent Testa8


Production of Anthocyanin Pigment1




Enhancer of Glabra3


  1. Allan AC, Hellens RP, Laing WA (2008) MYB transcription factors that colour our fruit. Trends Plant Sci 13:99–102CrossRefPubMedGoogle Scholar
  2. Arapitsas P, Sjoberg PJR, Turner C (2008) Characterisation of anthocyanins in red cabbage using high resolution liquid chromatography coupled with photodiode array detection and electrospray ionization-linear ion trap mass spectrometry. Food Chem 109:219–226CrossRefGoogle Scholar
  3. Benzie IFF, Strain JJ (1999) Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. In: Lester P (ed) Methods in enzymology: oxidants and antioxidants part A. Academic Press, pp 15–27Google Scholar
  4. Borevitz JO, Xia Y, Blount J, Dixon RA, Lamb C (2000) Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis. Plant Cell 12:2383–2393CrossRefPubMedGoogle Scholar
  5. Broun P (2005) Transcriptional control of flavonoid biosynthesis: a complex network of conserved regulators involved in multiple aspects of differentiation in Arabidopsis. Curr Opin Plant Biol 8:272CrossRefPubMedGoogle Scholar
  6. Burr FA, Burr B, Scheffler BE, Blewitt M, Wienand U, Matz EC (1996) The maize repressor-like gene intensifier1 shares homology with the r1/b1 multigene family of transcription factors and exhibits missplicing. Plant Cell 8:1249–1259CrossRefPubMedGoogle Scholar
  7. Castellarin SD, Di Gaspero G (2007) Transcriptional control of anthocyanin biosynthetic genes in extreme phenotypes for berry pigmentation of naturally occurring grapevines. BMC Plant Biol 7:46CrossRefPubMedGoogle Scholar
  8. Chandler VL, Radicella JP, Robbins TP, Chen J, Turks D (1989) Two regulatory genes of the maize anthocyanin pathway are homologous: isolation of B utilizing R genomic sequences. Plant Cell 1:1175–1183CrossRefPubMedGoogle Scholar
  9. Charron CS, Clevidence BA, Britz SJ, Novotny JA (2007) Effect of dose size on bioavailability of acylated and nonacylated anthocyanins from red cabbage (Brassica oleracea L. var. capitata). J Agric Food Chem 55:5354–5362CrossRefPubMedGoogle Scholar
  10. Cone KC, Cocciolone SM, Burr FA, Burr B (1993) Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant. Plant Cell 5:1795–1805CrossRefPubMedGoogle Scholar
  11. Dixon RA, Xie DY, Sharma SB (2005) Proanthocyanidins—a final frontier in flavonoid research? New Phytologist 165:9–28CrossRefPubMedGoogle Scholar
  12. Espley RV, Hellens RP, Putterill J, Stevenson DE, Kutty-Amma S, Allan AC (2007) Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant J 49:414–427CrossRefPubMedGoogle Scholar
  13. Feyissa D, Løvdal T, Olsen K, Slimestad R, Lillo C (2009) The endogenous GL3, but not EGL3, gene is necessary for anthocyanin accumulation as induced by nitrogen depletion in Arabidopsis rosette stage leaves. Planta 230:747–754CrossRefPubMedGoogle Scholar
  14. Gao L, Mazza G (1994) Quantitation and distribution of simple and acylated anthocyanins and other phenolics in blueberries. J Food Sci 59:1057–1059CrossRefGoogle Scholar
  15. Gonzalez A, Zhao M, Leavitt JM, Lloyd AM (2008) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827CrossRefPubMedGoogle Scholar
  16. Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780CrossRefPubMedGoogle Scholar
  17. Grotewold E, Drummond BJ, Bowen B, Peterson T (1994) The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell 76:543–553CrossRefPubMedGoogle Scholar
  18. Hagiwara A, Yoshino H, Ichihara T, Kawabe M, Tamano S, Aoki H, Koda T, Nakamura M, Imaida K, Ito N, Shirai T (2002) Prevention by natural food anthocyanins, purple sweet potato color and red cabbage color, of 2-amino-1-methyl-6-phenylimidazo [4, 5-b]pyridine (PhIP)-associated colorectal carcinogenesis in rats initiated with 1, 2-dimethylhydrazine. J Toxicol Sci 27:57–68CrossRefPubMedGoogle Scholar
  19. Harborne JB, Williams CA (2000) Advances in flavonoid research since 1992. Phytochemistry 55:481–504CrossRefPubMedGoogle Scholar
  20. Heim MA, Jakoby M, Werber M, Martin C, Weisshaar B, Bailey PC (2003) The basic helix-loop-helix transcription factor family in plants: a genome-wide study of protein structure and functional diversity. Mol Biol Evol 20:735–747CrossRefPubMedGoogle Scholar
  21. Hoagland DR, Arnon D (1950) The water culture method for growing plants without soil. Calif Agr Expt Sta Circ 347:1–32Google Scholar
  22. Hou DX (2003) Potential mechanisms of cancer chemoprevention by anthocyanins. Curr Mol Med 3:149–159CrossRefPubMedGoogle Scholar
  23. Hrazdina G, Marx GA, Hoch HC (1982) Distribution of secondary plant metabolites and their biosynthetic enzymes in pea (Pisum sativum L.) leaves: anthocyanins and flavonol glycosides. Plant Physiol 70:745–748CrossRefPubMedGoogle Scholar
  24. Jiang C, Gao X, Liao L, Harberd NP, Fu X (2007) Phosphate starvation root architecture and anthocyanin accumulation responses are modulated by the gibberellin-DELLA signaling pathway in Arabidopsis. Plant Physiol 145:1460–1470CrossRefPubMedGoogle Scholar
  25. Keck AS, Finley JW (2004) Cruciferous vegetables: cancer protective mechanisms of glucosinolate hydrolysis products and selenium. Integr Cancer Ther 3:5–12CrossRefPubMedGoogle Scholar
  26. Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10:236–242CrossRefPubMedGoogle Scholar
  27. Kubo H, Peeters AJM, Aarts MGM, Pereira A, Koornneef M (1999) ANTHOCYANINLESS2, a homeobox gene affecting anthocyanin distribution and root development in Arabidopsis. Plant Cell 11:1217–1226CrossRefPubMedGoogle Scholar
  28. Lea US, Slimestad R, Smedvig P, Lillo C (2007) Nitrogen deficiency enhances expression of specific MYB and bHLH transcription factors and accumulation of end products in the flavonoid pathway. Planta 225:1245–1253CrossRefPubMedGoogle Scholar
  29. Lila MA (2004) Anthocyanins and human health: an in vitro investigative approach. J Biomed Biotech 2004:306–313CrossRefGoogle Scholar
  30. Lu S, Van EJ, Zhou X, Lopez AB, O’Halloran DM, Cosman KM, Conlin BJ, Paolillo DJ, Garvin DF, Vrebalov J, Kochian LV, Kupper H, Earle ED, Cao J, Li L (2006) The cauliflower Or gene encodes a DnaJ cysteine-rich domain-containing protein that mediates high levels of beta-carotene accumulation. Plant Cell 18:3594–3605CrossRefPubMedGoogle Scholar
  31. Ludwig SR, Wessler SR (1990) Maize R gene family: tissue-specific helix-loop-helix proteins. Cell 62:849–851CrossRefPubMedGoogle Scholar
  32. Mano H, Ogasawara F, Sato K, Higo H, Minobe Y (2007) Isolation of a regulatory gene of anthocyanin biosynthesis in tuberous roots of purple-fleshed sweet potato. Plant Physiol 143:1252–1268CrossRefPubMedGoogle Scholar
  33. Martin C, Prescott A, Mackay S, Bartlett J, Vrijlandt E (1991) Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. Plant J 1:37–49CrossRefPubMedGoogle Scholar
  34. McDougall GJ, Fyffe S, Dobson P, Stewart D (2007) Anthocyanins from red cabbage—stability to simulated gastrointestinal digestion. Phytochemistry 68:1285–1294CrossRefPubMedGoogle Scholar
  35. Mertens-Talcott SU, Rios J, Jilma-Stohlawetz P, Pacheco-Palencia LA, Meibohm B, Talcott ST, Derendorf H (2008) Pharmacokinetics of anthocyanins and antioxidant effects after the consumption of anthocyanin-rich acai juice and pulp (Euterpe oleracea Mart.) in human healthy volunteers. J Agric Food Chem 56:7796–7802CrossRefPubMedGoogle Scholar
  36. Murillo G, Mehta RG (2001) Cruciferous vegetables and cancer prevention. Nutri Cancer 41:17–28CrossRefGoogle Scholar
  37. Nesi N, Debeaujon I, Jond C, Pelletier G, Caboche M, Lepiniec L (2000) The TT8 gene encodes a basic helix-loop-helix domain protein required for expression of DFR and BAN genes in Arabidopsis siliques. Plant Cell 12:1863–1878CrossRefPubMedGoogle Scholar
  38. Ogawa K, Sakakibara H, Iwata R, Ishii T, Sato T, Goda T, Shimoi K, Kumazawa S (2008) Anthocyanin composition and antioxidant activity of the Crowberry (Empetrum nigrum) and other berries. J Agric Food Chem 56:4457–4462CrossRefPubMedGoogle Scholar
  39. Palapol Y, Ketsa S, Lin-Wang K, Ferguson IB, Allan AC (2009) A MYB transcription factor regulates anthocyanin biosynthesis in mangosteen (Garcinia mangostana L.) fruit during ripening. Planta 229:1323–1334CrossRefPubMedGoogle Scholar
  40. Payne CT, Zhang F, Lloyd AM (2000) GL3 encodes a bHLH protein that regulates trichome development in Arabidopsis through interaction with GL1 and TTG1. Genetics 156:1349–1362PubMedGoogle Scholar
  41. Paz-Ares J, Ghosal D, Wienand U, Peterson P, Saedler H (1987) The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J 6:3553PubMedGoogle Scholar
  42. Quattrocchio F, Wing JF, Leppen HTC, Mol JNM, Koes RE (1993) Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes. Plant Cell 5:1497–1512CrossRefPubMedGoogle Scholar
  43. Quattrocchio F, Wing J, van der Woude K, Souer E, de Vetten N, Mol J, Koes R (1999) Molecular analysis of the anthocyanin2 gene of petunia and its role in the evolution of flower color. Plant Cell 11:1433–1444CrossRefPubMedGoogle Scholar
  44. Rapisarda P, Fanella F, Maccarone E (2000) Reliability of analytical methods for determining anthocyanins in blood orange juices. J Agric Food Chem 48:2249–2252CrossRefPubMedGoogle Scholar
  45. Spelt C, Quattrocchio F, Mol JNM, Koes R (2000) Anthocyanin1 of petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes. Plant Cell 12:1619–1632CrossRefPubMedGoogle Scholar
  46. Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456CrossRefPubMedGoogle Scholar
  47. Sweeney MT, Thomson MJ, Pfeil BE, McCouch S (2006) Caught red-handed: Rc encodes a basic helix-loop-helix protein conditioning red pericarp in rice. Plant Cell 18:283–294CrossRefPubMedGoogle Scholar
  48. Walker AR, Lee E, Bogs J, McDavid DAJ, Thomas MR, Robinson SP (2007) White grapes arose through the mutation of two similar and adjacent regulatory genes. Plant J 49:772–785CrossRefPubMedGoogle Scholar
  49. Wu X, Prior RL (2005) Identification and characterization of anthocyanins by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry in common foods in the United States: vegetables, nuts, and grains. J Agric Food Chem 53:3101–3113CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Plant Breeding and Genetics, Robert W. Holley Center for Agriculture and Health, USDA-ARSCornell UniversityIthacaUSA

Personalised recommendations