Original Article

Heredity (1991) 66, 105–115; doi:10.1038/hdy.1991.13

The cyanogenic polymorphism in Trifolium repens L. (white clover)

M A Hughes1

1Department of Biochemistry and Genetics, The Medical School, The University, Newcastle upon Tyne NE2 4HH

Received 16 May 1990.

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Abstract

The cyanogenic polymorphism in white clover is controlled by alleles of two independently segregating loci. Biochemical studies have shown that non-functional alleles of the Ac locus, which controls the level of cyanoglucoside produced in leaf tissue, result in the loss of several steps in the biosynthetic pathway. Alleles of the Li locus control the synthesis of the hydrolytic enzyme, linamarase, which is responsible for HCN release following tissue damage. Studies on the selective forces and the distribution of the cyanogenic morphs of white clover are discussed in relation to the quantitative variation in cyanogenesis revealed by biochemical studies. Molecular studies reveal considerable restriction fragment length polymorphism for linamarase homologous genes.

Keywords:

cyanogenesis, polymorphism, Trifolium repens, white clover

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References

  1. Angseeing, J A P. 1974. Selective eating of the acyanogenic form of Trifolium repens. Heredity, 32, 73–83.
  2. Armstrong, H E, Armstrong, E F, and Horton, E. 1913. Herbage studies II - Variation in Lotus corniculatus and Trifolium repens (cyanophoric plants). Roy Soc (Lond) Proc B, 86, 262–269.
  3. Askew, H O. 1933. Determination of hydrocyanic acid in white clover. NZJ Sci Techno I B, 14, 359–365.
  4. Bishop, J A, and Korn, M E. 1969. Natural selection and cyanogenesis in white clover, Trifolium repens. Heredity, 24, 423–430.
  5. Boersma, P, Kakes, P, and Schram, A W. 1983. Linamarase and beta-glucosidase activity in natural populations of Trifolium repens. Acta Bot Neerl, 32, 39–47.
  6. Burgess, R S L, and Ennos, R A. 1987. Selective grazing of acyanogenic white clover: Variation in behaviour among populations of the slug Deroceras reticulatum. Oecologia, 73, 432–435.
  7. Caradus, J R, MacKay, A C, Woodfield, D R, Van Den Bosch, J, and Wewala, S. 1989. Classification of a world collection of white clover cultivars. Euphytica, 42, 183–196.
  8. Carvalho, F J P De C. 1981. Ph.D. Dissertation, University of California, Davis, CA, USA.
  9. Chen, C-C, and Gibson, P B. 1972. Chromosome relationships of Trifolium uniflorum to T. repens and T. occidentale. Can J Genet Cytol, 14, 591–595.
  10. Collinge, D B, and Hughes, M A. 1982a. In vitro characterisation of the Ac locus in white clover (Trifolium repens L.). Arch Biochem Biophys, 218, 38–45.
  11. Collinge, D B, and Hughes, M A. 1982b. Developmental and physiological studies on the cyanogenic glucosides of white clover (Trifolium repens L.). J Exp Bot, 33, 154–161.
  12. Collinge, D B, and Hughes, M A. 1984. Evidence that linamarin and lotaustralin, the two cyanogenic glucosides of Trifolium repens L., are synthesised by a single set of microsomal enzymes controlled by the Ac/ac locus. Plant Sci Lett, 34, 119–125.
  13. Coop, I E. 1940. Cyanogenesis in white clover III Study of linamarase. NZJ Sci Technol B, 22–23, 71–83.
  14. Corkill, L. 1940. Cyanogenesis in white clover I Cyanogenesis in single plants. NZ J Sci Technol B, 22–23, 65–67.
  15. Corkill, L. 1942. Cyanogenesis in white clover (Trifolium repens L.) V. The inheritance of cyanogenesis. NZ J Sci Technol B, 23, 178–193.
  16. Crawford-Sidebothom, T J. 1972. The role of slugs and snails in the maintenance of the cyanogenesis polymorphisms of Lotus corniculatus and Trifolium repens. Heredity, 28, 405–411.
  17. Daday, H. 1954a. Gene frequencies in wild populations of Trifolium repens I. Distribution by altitude. Heredity, 8, 61–78. | ISI |
  18. Daday, H. 1954b. Gene frequencies in wild populations of Trifolium repens II. Distribution by altitude. Heredity, 8, 377–384.
  19. Daday, H. 1958. Gene frequencies in wild populations of Trifolium repens III. World distribution. Heredity, 12, 169–184.
  20. Daday, H. 1965. Gene frequencies in wild populations of Trifolium repens L. IV. Mechanism of natural selection. Heredity, 20, 355–365.
  21. De Araujo, A M. 1976. The relationship between altitude and cyanogenesis in white clover (Trifolium repens L.) Heredity, 37, 291–293.
  22. Dirzo, R. 1984. Herbivory: A phytocentric view in perspectives. In: Dirzo, R. and Sarukhan, J. (eds) Plant Population Ecology, Sinauer, USA, pp. 141–156.
  23. Dirzo, R, and Harper, J L. 1982a. Experimental studies on slug-plant interactions III. Differences in the acceptability of individual plants of Trifolium repens to slugs and snails. JEcol, 70, 101–117.
  24. Dirzo, R, and Harper, J L. 1982b. Experimental studies on slug-plant interactions IV. The performance of cyanogenic and acyanogenic morphs of Trifolium repens in the field. J Ecol, 70, 119–138.
  25. Dommee, B, Breakfield, P M, and MacNair, M R. 1980. Differential root growth of the cyanogenic phenotypes of Trifolium repens L. Oecol Plant, 1, 367–370.
  26. Dritschilo, W, Krummel, J, Nafus, D, and Pimentel, D. 1979. Herbivorous insects colonising cyanogenic and acyanogenic Trifolium repens. Heredity, 42, 49–56.
  27. Dunn, M A, Hughes, M A, and Sharif, A L. 1988. Synthesis of the cyanogenic beta-glucosidase, linamarase, in white clover. Arch Biochem Biophys, 243, 361–373.
  28. Ennos, R A. 1981a. Manifold effects of the cyanogenic loci in white clover. Heredity, 46, 127–132.
  29. Ennos, R A. 1981b. Detection of selection in populations of white clover (Trifolium repens L.). Biol J Linn Soc, 15, 75–82.
  30. Ennos, R A. 1982. Association of the cyanogenic loci in white clover. Genet Res, 40, 65–72.
  31. Ennos, R A. 1985. Measuring the effects of genetic variation of plant fitness. In: Haeck, J. and Woldendorp, J. P. (eds) Structure and Functioning of Plant Populations, North-Holland Publishing Co., Amsterdam, pp. 153–160.
  32. Foulds, W, and Grime, J P. 1972. The influence of soil moisture on the frequency of cyanogenic plants in populations of Trifolium repens and Lotus corniculatus. Heredity, 28, 143–146.
  33. Foulds, W, and Young, L. 1977. Effect of frosting, moisture stress and potassium cyanide on the metabolism of cyanogenic and acyanogenic phenotypes of Lotus corniculatus L. and Trifolium repens L. Heredity, 38, 19–24.
  34. Frazer, J, and Nowak, J. 1988. Studies on variability in white clover: Growth habits and cyanogenic glucosides. Ann Bot, 61, 311–318.
  35. Gibson, P B, Barnett, O W, and Gillingham, J T. 1972. Cyano-glucoside and hydrolysing enzyme in species related to Trifolium repens. Crop Sci, 12, 708–709.
  36. Gliddon, C, and Trathan, P. 1985. Interactions between white clover and perennial ryegrass in an old permanent pasture. In: Haeck, J. and Woldendorp, J. W. (eds) Structure and Functioning of Plant Populations, North-Holland Publishing Co., Amsterdam, pp. 161–169.
  37. Hahlbrock, K, and Conn, E E. 1971. Evidence for the formation of linamarin and lotaustralin in flax seedlings by the same glucosyltransferase. Phytochemistry, 10, 1019–1023.
  38. Halkier, B A, Olsen, C-E, and Moller, B L. 1989. The bio-synthesis of cyanogenic glucosides in higher plants J Biol Chem, 264, 19487–19494.
  39. Horrill, J C, and Richards, J A. 1986. Differential grazing by the mollusc Arion hortensis Fer. on cyanogenic and acyanogenic seedlings of the white clover, Trifolium repens L. Heredity, 56, 277–281.
  40. Hughes, M A. 1968. Studies on the beta-glucosidase system of Trifolium repens L. J Exp Bot, 19, 427–434.
  41. Hughes, M A, and Conn, E E. 1976. Cyanoglucoside biosynthesis in white clover (Trifolium repens L.). Phytochemistry, 15, 687–701.
  42. Hughes, M A, and Dunn, M A. 1982. Biochemical characterisation of the Li locus, which controls the activity of the cyanogenic beta-glucosidase in Trifolium repens L. Plant Mol Biol, 1, 169–181.
  43. Hughes, M A, Dunn, M A, and Pearson, J R. 1985. A regulatory element controlling the synthesis of the cyanogenic beta-glucosidase (linamarase) of white clover. Heredity, 55, 387–391.
  44. Hughes, M A, Sharif, A L, Dunn, M A, and Oxtoby, E. 1988. The molecular biology of cyanogenesis. In: Cyanide Compounds in Biology. CIBA Foundation Symposium 140, J. Wiley & Sons, Chichester, pp. 111–130.
  45. Hughes, M A, Sharif, A L, Dunn, M A, Oxtoby, E, and Pancoro, A. 1990. Restriction fragment length polymorphism segregation analysis of the Li locus in Trifolium repens L. Plant Mol Biol, 14, 407–414.
  46. Hughes, M A, and Stirling, J D. 1982. A study of dominance at the locus controlling cyanoglucoside production in Trifolium repens L. Euphytica, 31, 477–483.
  47. Hughes, M A, Stirling, J D, and Collinge, D B. 1984. The inheritance of cyanoglucoside content in Trifolium repens L. Biochem Genet, 22, 139–151.
  48. Jarvis, S S, and Hatch, D J. 1987. Differential effects of low concentration of aluminium on the growth of four genotypes of white clover. Plant Soil, 99, 241–253.
  49. Jones, D A. 1981. Cyanide and coevolution. In: Vennesland, B., Conn, E. E., Knowles, C. J., Westley, J. and Wissing, F. (eds) Cyanide and Biology, Academic Press, New York, pp. 509–516.
  50. Kakes, P. 1985. Linamarase and other beta3-glucosidases are present in the cell walls of Trifolium repens L. leaves. Planta, 166, 156–160.
  51. Kakes, P. 1987. On the polymorphisms for cyanogenesis in natural populations of Trifolium repens L. in the Netherlands I. Distribution of the genes Ac and Li. Acta Bot Neerl, 36, 59–69.
  52. Kakes, P. 1989. An analysis of the costs and benefits of the cyanogenic system in Trifolium repens. Theoret Appl Genet, 77, 111–118.
  53. Kakes, P, and Eeltink, H. 1985. The presence of a specialised beta-glucosidase, linamarase, in the leaves of Trifolium repens L. is controlled by the gene Li. Z Naturforsch Sect C Biosci, 40, 509–513.
  54. Lieberei, R, Biehl, B, Giesemann, A, and Junqueira, N T V. 1989. Cyanogenesis inhibits active defence reactions in plants. Plant Physiol, 90, 33–36.
  55. Maher, E P, and Hughes, M A. 1973. Studies on the nature of the Li locus in white clover II. The effects of genotype on enzyme activity and properties. Biochem Genet, 8, 13–26.
  56. Melville, J, and Doak, B W. 1940. Cyanogenesis in white clover II. Isolation of the glucoside constituents. NZJ Sci Technol B, 22, 67–70.
  57. Miller, J D, Gibson, P B, Cope, W A, and Knight, E E. 1975. Herbivore feeding on cyanogenic and acyanogenic white clover seedlings. Crop Sci, 15, 90–91.
  58. Mirande, M. 1912. Sur la presence de l'acide cyanohydrique dans le trefle rampant (Trifolium repens L.). Compt Rend Acad Sci (Paris), 155, 651–653.
  59. Mowat, D J, and Shakeel, M A. 1989. The effect of different cultivars of clover on numbers of, and leaf damage by, some invertebrate species. Grass Forage Sci, 44, 11–18.
  60. Nahrstedt, A, and Davis, R H. 1986. Uptake of linamarin and lotaustralin from their food plant by larvae of Zygaena trifolii. Phytochemistry, 25, 2299–2302.
  61. Paim, N R, and Dean, C E. 1975. Characteristics of cyanogenic and acyanogenic white clover plants. Soil Crop Sci Soc Forida Proc, 35, 18–21.
  62. Poesi, I, Kiss, L, Hughes, M A, and Nanasi, P. 1989. Kinetic investigation of the substrate specificity of the cyanogenic beta-D-glucosidase of white clover. Arch Biochem Biophys, 272, 496–506.
  63. Poulton, J E. 1988. Localisation and catabolism of cyanogenic glycosides. In: Cyanide Compounds in Biology, CLBA Foundation Symposium 140, J. Wiley & Sons, Chichester, pp. 67–91.
  64. Raubenheimer, D. 1989. Cyanoglucoside gynocardin from Acraea horta L. J Chem Ecol, 15, 2177–2189. | Article | ISI | ChemPort |
  65. Rogers, C F, and Frykolm, O C. 1937. Observations on the variations in cyanogenic power of white clover plants. J Agric Res, 55, 533–537.
  66. Selmar, D, Leiberei, R, Biehl, B, and Conn, E E. 1989. alpha;-Hydroxynitrile lyase in Hevea brasiliensis and its significance for rapid cyanogenesis. Physiol Plant, 75, 97–101.
  67. Till, I. 1987. Variability of expression of cyanogenesis in white clover (Trifolium repens L.). Heredity, 59, 265–271.
  68. Till-Bottraud, I, Kakes, P, and Dommee, B. 1988. Variable phenotypes and stable distribution of the cyanotypes of Trifolium repens L. in Southern France. Acya Oecologica, 9, 393–404.
  69. Turkington, R, Chan, M A, Vardy, A, and Harper, J L. 1979. The growth, distribution and neighbourhood relationships of T. repens in a permanent pasture III. The establishment and growth of T. repens in natural and perturbed sites. J Ecol, 67, 231–243.
  70. Van Wyk, B-E. 1989. The taxonomic significance of cyanogenesis in Lotononis and related genera. Biochem Syst Ecol, 17, 297–303.
  71. Vickery, P J, Wheeler, J L, and Mulcahy, C. 1987. Factors affecting the hydrogen cyanide potential of white clover (Trifolium repens L.). Aust J Agric Res, 38, 1053–1059.
  72. Ware, W M. 1925. Experiments and observations on forms and strains of Trifolium repens L. J Agri Sci, 15, 47–50.
  73. Whitman, R J. 1973. Herbivore feeding and cyanogenesis in Trifolium repens L. Heredity, 30, 241–245.
  74. Wilkinson, H T, and Millar, R L. 1978. Cyanogenic potential of Trifolium repens L. in relation to pepper spot caused by Stemphylium sarciniforme. Can J Bot, 56, 2491–2496.
  75. Zohary, M, and Heller, D. 1984. The Genus Trifolium. Israel Academy of Science, Jerusalem.