Molecular Breeding

, Volume 18, Issue 3, pp 219–228 | Cite as

High glucosinolate broccoli: a delivery system for sulforaphane

  • Golge Sarikamis
  • Julietta Marquez
  • Ruth Maccormack
  • Richard N. Bennett
  • Jeremy Roberts
  • Richard Mithen
Open Access


The development of hybrid broccoli genotypes with enhanced levels of 4-methylsulphinylbutyl glucosinolate, the precursor of anticarcinogenic isothiocyanate sulforaphane (SF), by introgressing genomic segments from the wild ancestor Brassica villosa is described. We demonstrate that to obtain enhanced levels of either 3-methylsulphinylpropyl or 4-methylsulphinylbutyl glucosinolate it is necessary to have B. villosa alleles in either a homozygous or heterozygous state at a single quantitative trait locus (QTL) on O2. The ratio of these two glucosinolates, and thus whether iberin or SF is generated upon hydrolysis, is determined by the presence or absence of B. villosa alleles at this QTL, but also at an additional QTL2 on O5. We further demonstrate that following mild cooking high glucosinolate broccoli lines generate about three fold higher levels of SF than conventional varieties. Commercial freezing processes and storage of high glucosinolate broccoli maintains the high level of glucosinolates compared to standard cultivars, although the blanching process denatures the endogenous myrosinase activity.


Broccoli Glucosinolates Isothiocyanates Sulforaphane Iberin Breeding 



We thank the staff at the University of Nottingham for help with field work. We also thank Sarah Pettitt, Food and Consumer Division, Christian Salvesen, for processing the frozen broccoli, Martyn Watling and his staff for assistance with the broccoli cultivation at ADAS Terrington, and Martin Brittain and his staff for assistance with broccoli cultivation and harvesting near Boston, UK. Funding was provided by the University of Nottingham, Seminis Inc and the Biotechnology and Biological Sciences Research Council.


  1. Abercrombie JM, Farnham MW, Rushing JW (2005) Genetic combining ability of glucoraphanin level and other horticultural traits of broccoli. Euphytica 143:145–151CrossRefGoogle Scholar
  2. Bennett R, Mellon FA, Rosa E, Perkins L, Kroon PA (2004) Profiling glucosinolates, flavonoids, alkaloids and other secondary metabolites in tissues of Azima tetracantha L. (Salvadoraceae). J Agri Food Chem (in press)Google Scholar
  3. Bonnesen C, Eggleston IM, Hayes JD (2001) Dietary indoles and isothiocyanates that are generated from cruciferous vegetables can both stimulate apoptosis and confer protection against DNA damage in human colon cell lines. Cancer Res 61:6120–6130PubMedGoogle Scholar
  4. Chen YR, Wang W, Kong AN, Tan TH (1998) Molecular mechanisms of c-Jun N-terminal kinase-mediated apoptosis induced by anticarcinogenic isothiocyanates. J Biol Chem 273:1769–1775PubMedCrossRefGoogle Scholar
  5. Chiao JW, Chung FL, Kancherla R, Ahmed T, Mittelman A, Conaway CC (2002) Sulforaphane and its metabolite mediate growth arrest and apoptosis in human prostate cancer cells. Int J Oncol 20:631–636PubMedGoogle Scholar
  6. Conaway CC, Getahun SM, Liebes LL, Pusateri DJ, Topham DK, Botero-Omary M, Chung FL (2000) Disposition of glucosinolates and sulforaphane in humans after ingestion of steamed and fresh broccoli. Nutr Cancer 38:168–178PubMedCrossRefGoogle Scholar
  7. de Quiros HC, Magrath R, McCallum D, Kroymann J, Scnabelrauch D, Mitchell-Olds T, Mithen R (2000) alpha-keto acid elongation and glucosinolate biosynthesis in Arabidopsis thaliana. Theor Appl Genet 101:429–437CrossRefGoogle Scholar
  8. Dinkova-Kostova AT, Holtzclaw WD, Cole RN, Itoh K, Wakabayashi N, Katoh Y, Yamamoto M, Talalay P (2002) Direct evidence that sulfhydryl groups of keap1 are the sensors regulating induction of phase 2␣enzymes that protect against carcinogens and oxidants. Proc Natl Acad Sci USA 99:11908–11913PubMedCrossRefGoogle Scholar
  9. Farnham MW, Stephenson KK, Fahey JW (2005) Glucoraphanin level in broccoli seed is largely determined by genotype. Hortscience 40:50–53Google Scholar
  10. Farnham MW, Wilson PE, Stephenson KK, Fahey JW (2004) Genetic and environmental effects on glucosinolate content and chemoprotective potency of broccoli. Plant Breed 123:60–65CrossRefGoogle Scholar
  11. Faulkner K, Mithen R, Williamson G (1998) Selective increase of the potential anticarcinogen 4- methylsulphinylbutyl glucosinolate in broccoli. Carcinogenesis 19:605–609PubMedCrossRefGoogle Scholar
  12. Fimognari C, Nusse M, Berti F, Iori R, Cantelli-Forti G, Hrelia P (2002a) Cyclin D3 and p53 mediate sulforaphane-induced cell cycle delay and apoptosis in non-transformed human T lymphocytes. Cell Mol Life Sci 59:2004–2012CrossRefGoogle Scholar
  13. Fimognari C, Nusse M, Cesari R, Iori R, Cantelli-Forti G, Hrelia P (2002b) Growth inhibition, cell-cycle arrest and apoptosis in human T-cell leukemia by the isothiocyanate sulforaphane. Carcinogenesis 23:581–586CrossRefGoogle Scholar
  14. Finley JW (2005) Proposed criteria for assessing the efficacy of cancer reduction by plant foods enriched in carotenoids, glucosinolates, polyphenols and selenocompounds. Ann Bot (Lond) 95:1075–1096CrossRefGoogle Scholar
  15. Foo HL, Gronning LM, Goodenough L, Bones AM, Danielsen BE, Whiting DA, Rossiter JT (2000) Purification and characterisation of epithiospecifier protein from Brassica napus: enzymic intramolecular sulphur addition within alkenyl thiohydroximates derived from alkenyl glucosinolate hydrolysis. FEBS Lett 468:243–246PubMedCrossRefGoogle Scholar
  16. Gamet-Payrastre L, Li P, Lumeau S, Cassar G, Dupont MA, Chevolleau S, Gasc N, Tulliez J, Terce F (2000) Sulforaphane, a naturally occurring isothiocyanate, induces cell cycle arrest and apoptosis in HT29 human colon cancer cells. Cancer Res 60:1426–1433PubMedGoogle Scholar
  17. Getahun SM, Chung FL (1999) Conversion of glucosinolates to isothiocyanates in humans after ingestion of cooked watercress. Cancer Epidemiol Biomarkers Prev 8:447–451
  18. Hu R, Kim BR, Chen C, Hebbar V, Kong AN (2003) The roles of JNK and apoptotic signaling pathways in PEITC-mediated responses in human HT-29 colon adenocarcinoma cells. Carcinogenesis 24:1361–1367PubMedCrossRefGoogle Scholar
  19. Jackson SJ, Singletary KW (2003) Sulforaphane: a naturally occurring mammary carcinoma mitotic inhibitor which disrupts tubulin polymerization. CarcinogenesisGoogle Scholar
  20. Jeffery EH, Brown AF, Kurilich AC, Keck AS, Matusheski N, Klein BP, Juvik JA (2003) Variation in content of bioactive components in broccoli. J Food Compost Anal 16:323–330CrossRefGoogle Scholar
  21. Joseph MA, Moysich KB, Freudenheim JL, Shields PG, Bowman ED, Zhang Y, Marshall JR, Ambrosone CB (2004) Cruciferous vegetables, genetic polymorphisms in glutathione s-transferases m1 and t1, and prostate cancer risk. Nutr Cancer 50:206–213PubMedCrossRefGoogle Scholar
  22. Kim BR, Hu R, Keum YS, Hebbar V, Shen G, Nair SS, Kong AN (2003) Effects of glutathione on antioxidant response element-mediated gene expression and apoptosis elicited by sulforaphane. Cancer Res 63:7520–7525PubMedGoogle Scholar
  23. Kore AM, Jeffery EH, Wallig MA (1993) Effects of 1-isothiocyanato-3-(methylsulfinyl)-propane on xenobiotic metabolizing enzymes in rats. Food Chem Toxicol 31:723–9PubMedCrossRefGoogle Scholar
  24. Lambrix V, Reichelt M, Mitchell-Olds T, Kliebenstein DJ, Gershenzon J (2001) The Arabidopsis epithiospecifier protein promotes the hydrolysis of glucosinolates to nitriles and influences Trichoplusia ni herbivory. Plant Cell 13:2793–2807PubMedCrossRefGoogle Scholar
  25. Lin HJ, Probst-Hensch NM, Louie AD, Kau IH, Witte JS, Ingles SA, Frankl HD, Lee ER, Haile RW (1998) Glutathione transferase null genotype, broccoli, and lower prevalence of colorectal adenomas. Cancer Epidemiol Biomarkers Prev 7:647–652PubMedGoogle Scholar
  26. Lowe AJ, Moule C, Trick M, Edwards KJ (2004) Efficient large-scale development of microsatellites for marker and mapping applications in Brassica crop species. Theor Appl Genet 108:1103–1112PubMedCrossRefGoogle Scholar
  27. Magrath R, Bano F, Morgner M, Parkin I, Sharpe A, Lister C, Dean C, Turner J, Lydiate D, Mithen R (1994) Genetics of aliphatic glucosinolates.1. side-chain elongation in Brassica napus and Arabidopsis thaliana. Heredity 72:290–299Google Scholar
  28. Matusheski NV, Juvik JA, Jeffery EH (2004) Heating decreases epithiospecifier protein activity and increases sulforaphane formation in broccoli. Phytochemistry 65:1273–1281PubMedCrossRefGoogle Scholar
  29. Mithen R, Faulkner K, Magrath R, Rose P, Williamson G, Marquez J (2003) Development of isothiocyanate-enriched broccoli, and its enhanced ability to induce phase 2 detoxification enzymes in mammalian cells. Theor Appl Genet 106:727–734PubMedGoogle Scholar
  30. Munday R, Munday CM (2004) Induction of phase II detoxification enzymes in rats by plant-derived isothiocyanates: comparison of allyl isothiocyanate with sulforaphane and related compounds. J Agric Food Chem 52:1867–1871PubMedCrossRefGoogle Scholar
  31. Nachshon-Kedmi M, Fares FA, Yannai S (2004) Therapeutic activity of 3,3′-diindolylmethane on prostate cancer in an in vivo model. Prostate 61:153–160PubMedCrossRefGoogle Scholar
  32. Spitz MR, Duphorne CM, Detry MA, Pillow PC, Amos CI, Lei L, de Andrade M, Gu XJ, Hong WK, Wu XF (2000) Dietary intake of isothiocyanates: evidence of a joint effect with glutathione S-transferase polymorphisms in lung cancer risk. Cancer Epidemiol Biomarkers Prev 9:1017–1020PubMedGoogle Scholar
  33. Wang L, Liu D, Ahmed T, Chung FL, Conaway C, Chiao JW (2004) Targeting cell cycle machinery as a molecular mechanism of sulforaphane in prostate cancer prevention. Int J Oncol 24:187–192PubMedGoogle Scholar
  34. Wang W, Wang S, Howie AF, Beckett GJ, Mithen R, Bao Y (2005) Sulforaphane, erucin, and iberin up-regulate thioredoxin reductase 1 expression in human MCF-7 cells. J Agric Food Chem 53:1417–1421PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Golge Sarikamis
    • 1
  • Julietta Marquez
    • 1
  • Ruth Maccormack
    • 3
  • Richard N. Bennett
    • 2
  • Jeremy Roberts
    • 4
  • Richard Mithen
    • 2
  1. 1.Division of Agricultural and Environmental SciencesUniversity of NottinghamSutton BonningtonUK
  2. 2.Phytochemicals and Health ProgrammeInstitute of Food ResearchNorwichUK
  3. 3.John Innes CentreNorwichUK
  4. 4.Division of Plant ScienceUniversity of NottinghamSutton BonningtonUK

Personalised recommendations