, Volume 196, Issue 2, pp 239–244 | Cite as

Developmental regulation of aldoxime formation in seedlings and mature plants of Chinese cabbage (Brassica campestris ssp. pekinensis) and oilseed rape (Brassica napus): Glucosinolate and IAA biosynthetic enzymes

  • Richard Bennett
  • Jutta Ludwig-Muller
  • Guy Kiddle
  • Willy Hilgenberg
  • Roger Wallsgrove


The first steps in the biosynthesis of glucosinolates and indole-3-acetic acid (IAA) in oilseed rape (Brassica napus L.) and Chinese cabbage (Brassica campestris ssp. pekinensis) involve the formation of aldoximes. In rape the formation of aldoximes from chain-extended amino acids, for aromatic and aliphatic glucosinolate biosynthesis, is catalysed by microsomal flavin-containing monooxygenases. The formation of indole-3-aldoxime from l-tryptophan, the potential precursor of both indole-3-acetic acid and indolyl-glucosinolates, is catalysed by several microsomal peroxidases. The biosynthesis of glucosinolates and indole-3-acetic acid was shown to be under developmental control in oilseed rape and Chinese cabbage. No monooxygenase activities were detected in cotyledons or old leaves of either species. The highest monooxygenase activities were found in young expanding leaves; as the leaves reached full expansion and matured the activities decreased rapidly. The indole-aldoxime-forming activity was found in all of the tissues analysed, but there was also a clear decrease in foliar activity with maturity in leaves of rape and Chinese cabbage. Partial characterisation of the Chinese cabbage monooxygenases showed that they have essentially identical properties to the previously characterised rape enzymes; they are not cytochrome P450-type enzymes, but resemble flavin-containing monooxygenases. No monooxygenase inhibitors were detected in microsomes prepared from either cotyledons or old leaves.

Key words

Aldoxime biosynthesis Auxin biosynthesis Brassica Glucosinolate 





flavin-containing monooxygenase




indole-3-acetic acid














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  1. Baldwin, I.T. (1994) Chemical changes rapidly induced by folivory. In: Insect-plant interactions, vol. V, pp. 1–23, Bernays, E.A., ed. CRC Press, LondonGoogle Scholar
  2. Bennett, R., Donald, A., Dawson, G., Hick, A., Wallsgrove, R. (1993) Aldoxime-forming microsomal enzyme systems involved in the biosynthesis of glucosinolates in oilseed rape (Brassica napus) leaves. Plant Physiology 102, 1307–1312Google Scholar
  3. Bodnaryk, R.P. (1992) Effects of wounding on glucosinolates in the cotyledons of oilseed rape and mustard. Phytochemistry 31, 2671–2677Google Scholar
  4. Bodnaryk, R.P. (1994) Potent effect of jasmonates on indole glucosinolates in oilseed rape and mustard. Phytochemistry 35, 301–305Google Scholar
  5. Bowles, D.J. (1990) Defence-related proteins in higher plants. Annu. Rev. Biochem. 59, 873–907Google Scholar
  6. Chew, F.S. (1988) Biological effects of glucosinolates. In: Biologically active natural products for potential use in agriculture, pp. 151–181, Cutler, H., ed. American Chemical Society, WashingtonGoogle Scholar
  7. Clossais-Besnard, N., Lahrer, F. (1991) Physiological role of glucosinolates in Brassica napus. Concentrations and distribution pattern of glucosinolates among organs during a complete life cycle. J. Sci. Food Agric. 56, 25–38Google Scholar
  8. Coecke, S., Mertens, K., Segaert, A., Callaerts, A., Vercruysse, A., Rogiers, V. (1992) Spectrophotometric measurement of flavincontaining monooxygenase activity in freshly isolated rat hepatocytes and their cultures. Anal. Biochem. 205, 285–288Google Scholar
  9. Conn, E.E. (1981) The biochemistry of plants: a comprehensive treatise vol. 7, Secondary plant products. Academic Press, LondonGoogle Scholar
  10. Dawson, G.W., Hick, A.J., Bennett, R.N., Donald, A., Pickett, J.A., Wallsgrove, R.M. (1993) Synthesis of glucosinolate precursors and investigations into the biosynthesis of phenylalkyl and methylthioalkylglucosinolates. J. Biol. Chem. 268, 27154–27159Google Scholar
  11. Doughty, K.J., Porter, A.J.R., Morton, A.M., Kiddle, G., Bock, C.H., Wallsgrove, R.M. (1991) Variation in glucosinolate content of oilseed rape (Brassica napus L.) leaves. II Response to infection by Alternaria brassicae (Berk.) Sacc. Ann. Appl. Biol. 118, 469–477Google Scholar
  12. Felton, G.W., Donato, K., Del Vecchio, R.J., Duffey, S.S. (1989) Activation of plant oxidases by insect feeding reduces the nutritive quality of foliage to noctuid herbivores. J. Chem. Ecol. 15, 2667–2671Google Scholar
  13. Fieldsend, J., Milford, G.F.J. (1994) Changes in glucosinolates during crop development in single and double-low genotypes of winter oilseed rape. I. Profiles and tissue water concentration in vegetative tissues and developing pods. Ann. Appl. Biol. 124, in pressGoogle Scholar
  14. Glen, D.M., Jones, H., Fieldsend, J.K. (1990) Damage to oilseed rape by the field slug Deroceras reticulatum in relation to glucosinolate concentration. Ann. Appl. Biol. 117, 197–207Google Scholar
  15. Halkier, B.A., Møller, B.L. (1991) Involvement of cytochromes P450 in the biosynthesis of dhurrin in Sorghum bicolor (L.) Moench. Plant Physiol. 96, 10–17Google Scholar
  16. Helmlinger, J., Rausch, T., Hilgenberg, W. (1987) A soluble protein factor from Chinese cabbage converts indole-3-acetaldoxime to IAA. Phytochemistry 26, 615–618Google Scholar
  17. Jain, J.C., Michalyuk, M.R., GrootWassink, J.W.D., Underhill, E.W. (1989) Distribution of enzymes catalysing the glucosylation and sulfation steps of glucosinolate biosynthesis in Brassica juncea seedlings and cultured cells. Plant Sci. 64, 25–29Google Scholar
  18. Kjaer, A. (1974) The natural distribution of glucosinolates: a uniform class of sulfur-containing glucosides. In: Chemistry in botanical classification, pp. 229–234, Bendz, G., Santesson, J., eds. Academic Press, LondonGoogle Scholar
  19. Koshiba, T., Matsuyama, H. (1993) An in vitro system of indole-3 acetic acid formation from tryptophan in maize (Zea mays) coleoptile extracts. Plant Physiol. 102, 1319–1324Google Scholar
  20. Lea, P.J. (1993) Methods in plant biochemistry, vol. 9: Enzymes of secondary metabolism. Academic Press, LondonGoogle Scholar
  21. Lykkesfeldt, J., Møller, B.L. (1993) Synthesis of benzylglucosinolate in Tropaeolum majus L. Plant Physiol. 102, 609–613Google Scholar
  22. Ludwig-Muller, J., Hilgenberg, W. (1988) A plasma membranebound enzyme oxidises l-Trp to indole-3-aldoxime. Physiol. Plant. 74, 240–250Google Scholar
  23. Ludwig-Muller, J., Rausch, T., Lang, S., Hilgenberg, W. (1990) Plasma membrane bound high pH peroxidase isoenzymes convert tryptophan to indole-3-aldoxime. Phytochemistry 29, 1397–1400Google Scholar
  24. Masters, B.S.S., Williams Jr., C.H., Kamin, H. (1967) The preparation and properties of microsomal TPNH-cytochrome C reductase from pig liver. In: Methods in enzymology, vol. X, pp. 565–573, Estabrook, R.W., Pullman, M.E., eds. Academic Press, LondonGoogle Scholar
  25. Milford, G.F.J., Fieldsend, J.K., Porter, A.J.R., Rawlinson, C.J., Evans, E.J., Bilsborrow, P.E. (1989) Changes in glucosinolate concentration during vegetative growth of single and doublelow cultivars of winter oilseed rape. Aspects Appl. Biol. 23, 83–90Google Scholar
  26. Normanly, J., Cohen, J.D., Fink, G.R. (1993) Arabidopsis thaliana auxotrophs reveal a tryptophan-independent biosynthetic pathway for indole-3-acetic acid. Proc. Natl. Acad. Sci. USA 90, 10355–10359Google Scholar
  27. Porter, A.J.R., Morton, A.M., Kiddle, G., Doughty, K.J., Wallsgrove, R.M. (1991) Variation in the glucosinolate content of oilseed rape (Brassica napus L.) leaves. I Effect of leaf age and position. Ann. Appl. Biol. 118, 461–467Google Scholar
  28. Poulton, J.E., M011er, B.L. (1993) Glucosinolates. In: Methods in plant biochemistry, vol. 19, pp. 209–238, Lea, P.J., ed. Academic Press, LondonGoogle Scholar
  29. Ross, C.W. (1992) Hormones and growth regulators: auxins and gibberellins. In: Plant physiology, Salisbury, F.B., Ross, C.W., eds. Wadsworth Publishing Company, CaliforniaGoogle Scholar
  30. Sorenson, H. (1991) Glucosinolates: structure — properties — function. In: Canola and rapeseed, pp. 149–172, Shahidi, F., ed. Van Nostrand Rheinhold, New YorkGoogle Scholar
  31. Vance, C.P., Kirk, T.K., Sherwood, R.T. (1980) Lignification as a mechanism of disease resistance. Annu. Rev. Phytopath. 18, 259–288CrossRefPubMedGoogle Scholar
  32. Wallsgrove, R.M., Bennett, R., Donald, A., Kiddle, G., Porter, A., Doughty, K. (1993) The biochemical basis for the differential response of oilseed rape varieties to infection and stress. Asp. Appl. Biol. 34, 155–161Google Scholar
  33. Wiermann, R. (1981) Secondary plant products and cell and tissue differentiation. In: The biochemistry of plants: a comprehensive treatise, vol. 7, pp. 86–117, Conn, E.E., ed. Academic Press, LondonGoogle Scholar
  34. Ziegler, D.M. (1988) Flavin-containing monooxygenases: catalytic mechanism and substrate specificities. Drug Metab. Rev. 19, 1–33Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Richard Bennett
    • 1
  • Jutta Ludwig-Muller
    • 2
  • Guy Kiddle
    • 1
  • Willy Hilgenberg
    • 2
  • Roger Wallsgrove
    • 1
  1. 1.Biochemistry & Physiology DepartmentIACR Rothamsted Experimental StationHarpendenUK
  2. 2.Botanisches Institut, J.W. Goethe UniversitätFrankfurt am Main 11Germany

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