, Volume 178, Issue 3, pp 342–352 | Cite as

Comparative biochemical and immunological studies of the glycine betaine synthesis pathway in diverse families of dicotyledons

  • Elizabeth A. Weretilnyk
  • Sebastian Bednarek
  • Kent F. McCue
  • David Rhodes
  • Andrew D. Hanson


Members of the Chenopodiaceae can accumulate high levels (>100 μmol·(g DW)-1) of glycine betaine (betaine) in leaves when salinized. Chenopodiaceae synthesize betaine by a two-step oxidation of choline (choline→betaine aldehyde→ betaine), with the second step catalyzed by betaine aldehyde dehydrogenase (BADH, EC High betaine levels have also been reported in leaves of species from several distantly-related families of dicotyledons, raising the question of whether the same betaine-synthesis pathway is used in all cases.

Fast atom bombardment mass spectrometry showed that betaine levels of >100 μmol·(g DW)-1 are present in Lycium ferocissimum Miers (Solanaceae), Helianthus annuus L. (Asteraceae), Convolvulus arvensis L. (Convolvulaceae), and Amaranthus caudatus L. (Amaranthaceae), that salinization promotes betaine accumulation in these plants, and that they can convert supplied choline to betaine aldehyde and betaine. Nicotiana tabacum L. and Lycopersicon lycopersicum (L.) Karst. ex Farw. (Solanaceae), Lactuca sativa L. (Asteraceae) and Ipomoea purpurea L. (Convolvulaceae) also contained betaine, but at a low level (0.1–0.5 μmol·(g DW)-1. Betaine aldehyde dehydrogenase activity assays, immunotitration and immunoblotting demonstrated that the betaine-accumulating species have a BADH enzyme recognized by antibodies raised against BADH from Spinacia oleracea L. (Chenopodiaceae), and that the Mr of the BADH monomer is in all cases close to 63 000. These data indicate that the choline→betaine aldehyde→betaine pathway may have evolved by vertical descent from an early angiosperm ancestor, and might be widespread (albeit not always strongly expressed) among flowering plants. Consistent with these suggestions, Magnolia x soulangiana was found to have a low level of betaine, and to express a protein of Mr 63 000 which cross-reacted with antibodies to BADH from Spinacia oleracea.

Key words

Glycine betaine pathway Betaine aldehyde dehydrogenase Dicotyledons (glycine betaine) Fast atom bombardment mass spectrometry Salt stress 



Betaine aldehyde dehydrogenase


desorption chemical ionization mass spectrometry


fast atom bombardment mass spectrometry


relative molecular mass


polyacrylamide gel electrophoresis


sodium dodecyl sulfate


thin-layer chromatography


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Andresen, P.A., Kaasen, I., Styrvold, O.B., Boulnois, G., Strom, A.R. (1988) Molecular cloning, physical mapping and expression of the bet genes governing the osmoregulatory choline-glycine betaine pathway of Escherichia coli. J. Gen. Microbiol. 134, 1737–1746Google Scholar
  2. Bagnasco, S., Balaban, R., Fales, H.M., Yang, Y.-M., Burg, M. (1986) Predominant osmotically active organic solutes in rat and rabbit renal medullas. J. Biol. Chem. 261, 5872–5877Google Scholar
  3. Bowman, M.S., Rohringer, R. (1970) Formate metabolism and betaine formation in rust-infected wheat. Can. J. Bot. 48, 803–811Google Scholar
  4. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254Google Scholar
  5. Brouquisse, R., Weigel, P., Rhodes, D., Yocum, C.F., Hanson, A.D. (1989) Evidence for a ferredoxin-dependent choline monooxygenase from spinach chloroplast stroma. Plant Physiol. (in press)Google Scholar
  6. Burbidge, N.T., Gray, M. (1970) Flora of the A.C.T. Australian National University Press, Canberra, A.C.T.Google Scholar
  7. Byerrum, R.U., Sato, C.S., Ball, C.D. (1956) Utilization of betaine as a methyl group donor in tobacco. Plant Physiol. 31, 374–377Google Scholar
  8. Cronquist, A. (1968) The evolution and classification of flowering plants. Houghton Mifflin Co., Boston, Mass, USAGoogle Scholar
  9. Fowden, L. (1972) Amino acid complement of plants. Phytochemistry 11, 2271–2276Google Scholar
  10. Hanson, A.D., Hitz, W.D. (1982) Metabolic responses of mesophytes to plant water deficits. Annu. Rev. Plant Physiol. 33, 163–203Google Scholar
  11. Hoagland, D.R., Arnon, D.I. (1950) The water-culture method for growing plants without soil. Calif. Agric. Expt. Stn. Circ. No. 347Google Scholar
  12. Hitz, W.D., Rhodes, D., Hanson, A.D. (1981) Radiotracer evidence implicating phosphoryl and phosphatidyl bases in betaine synthesis by water stressed barley leaves. Plant Physiol. 68, 814–822Google Scholar
  13. Kelley, W.A., Adams, R.P. (1977) Preparation of extracts from juniper leaves for electrophoresis. Phytochemistry 16, 513–516Google Scholar
  14. Kessler, S.W. (1975) Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J. Immunol. 115, 1617–1624Google Scholar
  15. Ladyman, J.A.R., Hitz, W.D., Hanson, A.D. (1980) Translocation and metabolism of glycine betaine by barley plants in relation to water stress. Planta 150, 191–196Google Scholar
  16. Lerma, C., Hanson, A.D., Rhodes, D. (1988) Oxygen-18 and deuterium labeling studies of choline oxidation by spinach and sugar beet. Plant Physiol. 88, 695–702Google Scholar
  17. Le Rudulier, D., Strom, A.R., Dandekar, A.M., Smith, L.T., Valentine, R.C. (1984) Molecular biology of osmoregulation. Science 224, 1064–1068Google Scholar
  18. Mackay, M.A., Norton, R.S., Borowitzka, L.J. (1984) Organic osmoregulatory solutes in cyanobacteria. J. Gen. Microbiol. 130, 2177–2191Google Scholar
  19. Poljakoff-Mayber, A., Symon, D.E., Jones, G.P., Naidu, B.P., Paleg, L.G. (1987) Nitrogenous compatible solutes in native South Australian plants. Aust. J. Plant Physiol. 14, 341–350Google Scholar
  20. Rhodes, D., Rich, P.J., Myers, A.C., Reuter, C.C., Jamieson, G.C. (1987) Determination of betaines by fast atom bombardment mass spectrometry. Identification of glycine betaine deficient genotypes of Zea mays. Plant Physicl. 84, 781–788Google Scholar
  21. Skiba, W.E., Taylor, M.P., Wells, M.S., Mangum, J.H., Awad, W.M. (1982) Human hepatic methionine biosynthesis. Purification and characterization of betaine: homocysteine S-methyltransferase. J. Biol. Chem. 257, 14944–14948Google Scholar
  22. Stewart, G.R., Larher, F. (1980) Accumulation of amino acids and related compounds in relation to environmental stress. In: The biochemistry of plants, vol. 5: Amino acids and derivatives, pp. 609–635, Stumpf, P.K., Conn, E.E., eds. Academic Press, New York etc.Google Scholar
  23. Takhtajan, A. (1969) Flowering plants — origin and dispersal. Smithsonian Institution Press, Washington, D.C., USAGoogle Scholar
  24. Weigel, P., Weretilnyk, E.A., Hanson, A.D. (1986) Betaine aldehyde oxidation by spinach chloroplasts. Plant Physiol. 82, 753–759Google Scholar
  25. Weretilnyk, E.A., Hanson, A.D. (1988) Betaine aldehyde dehydrogenase polymorphism in spinach: genetic and biochemical characterization. Biochem. Genet. 26, 143–151Google Scholar
  26. Weretilnyk, E.A., Hanson, A.D. (1989) Betaine aldehyde dehydrogenase from spinach leaves: purification, in vitro translation of the mRNA, and regulation by salinity. Arch. Biochem. Biophys. (in press)Google Scholar
  27. Wyn Jones, R.G. (1984) Phytochemical aspects of osmotic adaptation. Recent Adv. Phytochem. 18, 55–78Google Scholar
  28. Wyn Jones, R.G., Storey, R. (1981) Betaines. In: The physiology and biochemistry of drought resistance in plants, pp. 171–204, Paleg, L.G., Aspinall, D., eds. Academic Press, Sydney New York London Toronto San FranciscoGoogle Scholar
  29. Yancey, P.H., Clark, M.E., Hand, S.C., Bowlus, R.D., Somero, G.N. (1982) Living with water stress: evolution of osmolyte systems. Science 217, 1214–1222Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Elizabeth A. Weretilnyk
    • 1
  • Sebastian Bednarek
    • 1
  • Kent F. McCue
    • 1
  • David Rhodes
    • 2
  • Andrew D. Hanson
    • 1
  1. 1.MSU-DOE Plant Research LaboratoryMichigan State UniversityEast LansingUSA
  2. 2.Department of HorticulturePurdue UniversityWest LafayetteUSA

Personalised recommendations