Brassica pp 317-338 | Cite as

Biochemical Genetics of Glucosinolate Biosynthesis in Brassica

  • R. Mithen
  • R. Parker
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 54)


Glucosinolates are the major class of secondary metabolites found in Brassica crops. The molecule comprises a β-thioglucose moiety, a sulphonated oxime moiety and a variable side chain, derived from one of several amino acids (Fig. 1). The economic importance of glucosinolates in Brassica crops is due to the biological activity of their degradation products, which include isothiocyanate, nitriles and a range of indole compounds (Figs. 2 and 3). These compounds are generated by the action of the endogenous plant enzyme myrosinase, or by thioglucosidases from gut microbes following consumption of intact glucosinolates. The nature and biological activity of these products is dependent on the glucosinolate chain structure, reviewed in detail below. The importance of these compounds is threefold. Firstly, certain degradation products reduce the nutritional quality of rape meal. This is largely due to 5-vinyloxazolidine2-thione (Fig. 2), derived from 2-hydroxy-3-butenyl glucosinolate (“progoitrin”), which, as the name suggests, has goitrogenic activity. Isothiocyanates and other glucosinolate-derived compounds can also reduce palatability of the meal. Secondly, isothiocyanates are important flavour compounds in cruciferous vegetable crops. These compounds, along with indolyl glucosinolate degradation products, have also been shown to have anticarcinogenic activity. Thirdly, isothiocyanate and other degradation products mediate plantherbivore and tritrophic interactions. This chapter concentrates on the genetic control of glucosinolate biosynthesis. Factors determining the nature of the degradation products and their biological activity have been reviewed elsewhere (Mithen 2001).


Glucosinolate Content Biochemical Genetic Glucosinolate Biosynthesis Alkenyl Glucosinolates Indolyl Glucosinolates 
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  1. Bak S, Feyereisen R (2001) The involvement of two P450 enzymes, CYP83B1 and CYP83A1, in auxin homeostasis and glucosinolate biosynthesis. Plant Physiol 127: 108–118PubMedCrossRefGoogle Scholar
  2. Bak S, Nielsen HL, Halkier BA (1998) The presence of CYP79 homologues in glucosinolateproducing plants shows evolutionary conservation of the enzymes in the conversion of amino acid to aldoxime in the biosynthesis of cyanogenic glycosides and glucosinolates. Plant Mol Biol 38: 725–734PubMedCrossRefGoogle Scholar
  3. Bak S, Olsen CE, Petersen BL, Moller BL, Halkier BA (1999) Metabolic engineering of phydroxybenzylglucosinolate in Arabidopsis by expression of the cyanogenic CYP79A1 from Sorghum bicolor. Plant J 20: 663–671PubMedCrossRefGoogle Scholar
  4. Bak S, Tax FE, Feldmann KA, Galbraith DW, Feyereisen R (2001) CYP83B1, a cytochrome P450 at the metabolic branch point in auxin and indole glucosinolate biosynthesis in Arabidopsis. Plant Cell 13: 101–111PubMedGoogle Scholar
  5. Bernardi R, Negri A, Ronchi S, Palmieri S (2000) Isolation of the epithiospecifier protein from oil-rape (Brassica napus ssp. oleifera) seed and its characterization. FEBS Lett 467: 296–298PubMedCrossRefGoogle Scholar
  6. Bones AM, Rossiter JT (1996) The myrosinase-glucosinolate system, its organisation and biochemistry. Physiol Plant 97: 194–208CrossRefGoogle Scholar
  7. Campos de Quiros H, Magrath R, McCallum D, Kroymann J, Scnabelrauch D, Mitchell-Olds T, Mithen R (2000) a-Keto acid elongation and glucosinolate biosynthesis in Arabidopsis thaliana. Theor Appl Genet 101: 429–437Google Scholar
  8. Chapple CCS, Glover JR, Ellis BE (1990) Purification and characterisation of methionineglyoxalate aminotransferase from Brassica carinata and Brassica napus. Plant Physiol 94: 1887–1896PubMedCrossRefGoogle Scholar
  9. Chisholm MD, Wetter LR (1964) Biosynthesis of mustard oil glucosides. IV. The administration of methionine-14C and related compounds to horseradish. Can J Biochem 42: 1033–1040PubMedCrossRefGoogle Scholar
  10. Downey RK, Craig BM, Young CG (1969). Breeding rapeseed for oil and meal quality. J Am Oil Chem Soc 46: 121–123CrossRefGoogle Scholar
  11. Du LC, Halkier BA (1998) Biosynthesis of glucosinolates in the developing walls and seeds of Sinapis alba. Phytochemistry 48: 1145–1150CrossRefGoogle Scholar
  12. Du LC, Lykkesfeldt J, Olsen CE, Halkier BA (1995) Involvement of cytochrome P450 in oxime production in glucosinolate biosynthesis as demonstrated by an in vitro microsomal enzyme system isolated from jasmonic acid-induced seedlings of Sinapis alba. Proc Natl Acad Sci USA 92: 12505–2509PubMedCrossRefGoogle Scholar
  13. Ettinger MG, Lundeen AJ (1956) The structure of sinigrin and sinalbin: an enzymic rearrangement. J Am Chem Soc 78: 4172–4173CrossRefGoogle Scholar
  14. Ettlinger MG, Lundeen AJ (1957) First synthesis of a mustard oil glucoside: the enzymatic Lossen arrangement. J Am Chem Soc 79: 1764–1765CrossRefGoogle Scholar
  15. Fahey JW, Zalcmann AT, Talalay P (2001) The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. Phytochemistry 56: 5–51PubMedCrossRefGoogle Scholar
  16. Faulkner K, Mithen RF, Williamson G (1998) Selective increase of the potential anticarcinogen 4-methylsulphinylbutyl glucosinolate in broccoli. Carcinogenesis 19: 605–609PubMedCrossRefGoogle Scholar
  17. Ferriera ME, Williams PH, Osborn TC (1994) RFLP mapping of Brassica napus using double-haploid lines. Theor Appl Genet 89: 615–621Google Scholar
  18. Fischer H, Chen L, Wallisch S (1996) The evolution of angiosperm seed proteins: a methioninerich legumin subfamily present in lower angiosperm clades. J Mol Evol 43: 399–404PubMedCrossRefGoogle Scholar
  19. Foo HL, Grønning LM, Goodenough L, Bones AM, Danielsen B-E, Whiting DA, Rossiter JT (2000) Purification and characterisation of epithiospecifer protein from Brassica napus, enzymic intramolecular sulphur addition within alkenyl thiohydroximates derived from alkenyl glucosinolate hydrolysis. FEBS Lett 468: 243–246PubMedCrossRefGoogle Scholar
  20. Giamoustaris A, Mithen RF (1996) Genetics of aliphatic glucosinolates. IV. Side chain modifications in Brassica oleracea. Theor Appl Genet 93: 1006–1010CrossRefGoogle Scholar
  21. Graser G, Schneider B, ldham NJ, Gershenzon J (2000) The methionine chain elongation pathway in the biosynthesis of glucosinolates in Eruca sativa ( Brassicaceae ). Arch Biochem Biophys 378: 411–419Google Scholar
  22. GrootWassink JW, Reed W, Kolenovsky AD (1994) Immunopurification and immunocharacterisation of the glucosinolate biosynthesis enzyme thiohydroximate S-glucosyltransferase. Plant Physiol 105: 425–433PubMedGoogle Scholar
  23. Hall C, McCallum D, Prescott A, Mithen R (2000) Biochemical genetics of glucosinolate chain modification in Arabidopsis and Brassica. Theor Appl Genet 102: 369–374CrossRefGoogle Scholar
  24. Hansen CH, Wittstock U, Olsen CE, Hick AJ, Pickett JA, Halkier BA (2001a) Cytochrome P450 CYP79F1 from Arabidopsis catalyzes the conversion of dihomomethionine and trihomomethionine to the corresponding aldoximes in the biosynthesis of aliphatic glucosinolates. J Biol Chem 14: 11078–11085CrossRefGoogle Scholar
  25. Hansen CH, Du L, Naur P, Olsen CE,Axelsen KB, Hick AJ, Pickett JA, Halkier BA (2001b) CYP83B1 is the oxime-metabolising enzyme in the glucosinolate pathway in Arabidopsis. J Biol Chem 27: 24790–24796Google Scholar
  26. Haughn GW, Davin L, Giblin M, Underhill EW (1991) Biochemical genetics of plant secondaryGoogle Scholar
  27. metabolism in Arabidopsis thaliana. The glucosinolates. Plant Physiol 97:217–226Google Scholar
  28. Heaney RK, Fenwick GR (1993) Methods for glucosinolate analysis. In: Waterman PG (ed)Google Scholar
  29. Methods for plant biochemistry. Academic Press, London, pp 531–550Google Scholar
  30. Hull AK, Vij R, Celenza JL (2000) Arabidopsis cytochrome P450s that catalyse the first step of tryptophan-dependent indole-3-acetic acid biosynthesis. Proc Natl Acad Sci USA 97: 2379–2384Google Scholar
  31. Josefsson E (1971) Studies of the biochemical background to differences in glucosinolate content in Brassica napus L. II. Administration of some sulphur-35 and carbon-14 compounds and localization of metabolic blocks. Physiol Plant 24: 161–175CrossRefGoogle Scholar
  32. Josefsson E (1973) Studies of the biochemical background to differences in glucosinolate content in Brassica napus L. III. Further studies to localize metabolic blocks. Physiol Plant 29: 28–32CrossRefGoogle Scholar
  33. Josefsson E, Appelqvist L-Ã (1968) Glucosinolates in seed of rape and turnip rape as affected by variety and environment. J Sci Food Agric 19: 564–570CrossRefGoogle Scholar
  34. Kahn RA, Fahrendorf T, Halkier BA, Moller BL (1999) Substrate specificity of the cytochrome P450 enzymes CYP79A1 and CYP71E1 involved in the biosynthesis of the cyanogenic glucoside dhurrin Sorghum bicolor ( L.) Moench. Arch Biochem Biophys 363: 9–18Google Scholar
  35. Kiddle GA, Bennett RN, Hick AJ, Wallsgrove RM (1999) C-S lyase activities in leaves of crucifers and non-crucifers, and the characterisation of three classes of C-S lyase activities from oilseed rape (Brassica napus L.). Plant Cell Environ 22: 433–445CrossRefGoogle Scholar
  36. Kliebenstein DJ, Kroymann J, Brown P, Figuth A, Pedersen D, Gershenzon J, Mitchell-Olds T (2001a) Genetic control of natural variation in Arabidopsis glucosinolate accumulation. Plant Physiol 126: 811–825PubMedCrossRefGoogle Scholar
  37. Kliebenstein DJ, Lambrix VM, Reichelt M, Gershenzon J, Mitchell-Olds T (2001b) Gene duplication in the diversification of secondary metabolism: tandem 2-oxoglutarate-dependent dioxygenases control glucosinolate biosynthesis in Arabidopsis. Plant Cell 13: 681–693PubMedGoogle Scholar
  38. Kondra ZP, Stefansson BR (1970) Inheritance of the major glucosinolates of rapeseed (Brassica napus) meal. Can J Plant Sci 50: 643–647CrossRefGoogle Scholar
  39. Kroymann J, Textor S, Tokuhisa JG, Falk KL, Bartam S, Gershenzon J, Mitchell-Olds T (2001) A gene controlling variation in Arabidopsis glucosinolate composition is part of the methionine chain elongation pathway. Plant Physiol 127: 1077–1088PubMedCrossRefGoogle Scholar
  40. Kushad MM, Brown AF, Kurilich AC, Juvik JA, Klein BP, Wallig MA, Jeffery EH (1999) Variation of glucosinolates in vegetable crops of Brassica oleracea. J Agric Food Chem 47:1541– 1548Google Scholar
  41. Lambrix V, Reichelt M, Mitchell-Olds T, Kliebenstein D, Gershenzon J (2001) The Arabidopsis epithiospecifier protein promotes the hydrolysis of the glucosinolates to nitriles and influences Trichoplusia ni herbivory. Plant Cell 13: 2793–2807PubMedGoogle Scholar
  42. Magrath R, Bano F, Morgner M, Parkin I, Sharpe A, Lister C, Dean C, Lydiate D, Mithen RF (1994) Genetics of aliphatic glucosinolates. I. Side chain elongation in Brassica napus and Arabidopsis thaliana. Heredity 72: 290–299CrossRefGoogle Scholar
  43. Matsuo M, Kirkland DF, Underhill EW (1972) 1-Nitro-2-phenylethane, a possible intermediate in the biosynthesis of benzylglucosinolate. Phytochemistry 11: 697–701Google Scholar
  44. Mikkelsen MD, Hansen CH, Wittstock U, Halkier BA (2000) Cytochrome P450 CYP79B2 from Arabidopsis catalyses the conversion of tryptophane to indole-3-acetaldoxime, a precursor of indole glucosinolates and indole-3-acetic acid. J Biol Chem 275: 33712–33717PubMedCrossRefGoogle Scholar
  45. Mithen RF (2001) Glucosinolates and their degradation products. Adv Bot Res 35:213–262 Mithen RF, Campos H (1996) Genetic variation of aliphatic glucosinolates in Arabidopsis thaliana and prospects for map based gene cloning. Entomol Exp Appl 80: 202–205CrossRefGoogle Scholar
  46. Mithen RF, Clarke J, Lister C, Dean C (1995) Genetics of aliphatic glucosinolates. III. Side chain modifications in Arabidopsis thaliana. Heredity 74: 210–215CrossRefGoogle Scholar
  47. Mithen RF, 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
  48. Parkin I, Magrath R, Keith D, Sharpe A, Mithen RF, Lydiate D (1994) Genetics of aliphatic glucosinolates. II. Hydroxylation of alkenyl glucosinolates in Brassica napus. Heredity 72: 594–598CrossRefGoogle Scholar
  49. Rask L, Andréasson E, Ekbom B, Erksson S, Pontoppidan B, Meijer J (2000) Myrosinase, gene family evolution and herbivore defence in Brassicaceae. Plant Mol Biol 42: 92–113CrossRefGoogle Scholar
  50. Reintanz B, Lehnen M, Reichelt M, Gershenzon J, Kowalczyk M, Sandberg G, Godde M, Uhl R, Palme K (2001) bus, a bushy Arabidopsis CYP79F1 knockout mutant with abolished synthesis of short-chain aliphatic glucosinolates. Plant Cell 13: 351–367Google Scholar
  51. Rodman JE, Soltis PS, Soltis DE, Sytsma KJ, Karol KG (1998) Parallel evolution of glucosinolate biosynthesis inferred from congruent nuclear and plastid gene phylogenies. Am J Bot 85: 997–1006PubMedCrossRefGoogle Scholar
  52. Sebastian RL, Howell EC, King GJ, Marshall DF, Kearsey MJ (2000) An integrated AFLP and RFLP Brassica oleracea linkage map from two morphologically distinct doubled-haploid mapping populations. Theor Appl Genet 100: 75–81CrossRefGoogle Scholar
  53. Strassman M, Ceci LN (1963) Enzymic formation of a-isopropylmalic acid, an intermediate in leucine biosynthesis. J Biol Chem 238: 2445–2452PubMedGoogle Scholar
  54. Tookey HL (1973) Separation of a protein required for epithiobutane formation. Can J Biochem 51: 1654–1660PubMedCrossRefGoogle Scholar
  55. Toroser D, Thormann CE, Osborn TC, Mithen RF (1995) RFLP mapping of quantitative trait loci controlling seed aliphatic glucosinolate content in oilseed rape (Brassica napus). Theor Appl Genet 91: 802–808CrossRefGoogle Scholar
  56. Uda Y, Kurata T, Arakawa N (1986) Effects of pH and ferrous ions on the degradation of glucosinolates by myrosinase. Agric Biol Chem 50: 2735–2740CrossRefGoogle Scholar
  57. Umbarger HE (1997) Biosynthesis of branched-chain amino acids. In: Neidhardt FC (ed) Escherichia coli and Salmonella cellular and molecular biology, vol 1, 2nd edn. ASM Press, Washington, DC, pp 422–456Google Scholar
  58. Underhill EW, Chisholm MD, Wetter LR (1962) Biosynthesis of mustard oil glucosides. Administration of14C-labelled compounds to horseradish, nasturtium and watercress. Can J Biochem Physiol 40: 1505–1514PubMedCrossRefGoogle Scholar
  59. Uzunova M, Ecke W, Weissleder K, Röbbelen G (1995) Mapping the genome of rapeseed (Bras-sica napus L). I. Construction of an RFLP linkage map and localisation of QTLs for seed glucosinolate content. Theor Appl Genet 90: 194–204Google Scholar
  60. Wasser LR, Watson WH (1963) Crystal structure of sinigrin. Nature 198: 127–1298Google Scholar
  61. Wittstock U, Halkier BA (2000). Cytochrome P450 CYP79A2 from Arabidopsis thaliana L. catalyzes the conversion of L-phenylalanine to phenylacetaldoxime in the biosynthesis of benzyl glucosinolate. J Biol Chem 275: 14659–14666PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2004

Authors and Affiliations

  • R. Mithen
    • 1
  • R. Parker
    • 2
  1. 1.Institute of Food ResearchNorwichUK
  2. 2.School of BiosciencesUniversity of NottinghamSutton Bonington, LoughboroughUK

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