Abstract
The activity, uptake and metabolism of cytokinin metabolites was determined in soybean (Glycine max (L.) Merr.) callus tissue. The following activity sequence was established: zeatin riboside (ZR)>zeatin (Z)>O-glucosides of Z, ZR and their dihydro derivatives>lupinic acid (an alanine conjugate of Z)>7- and 9-glucosides of Z which were almost inactive. The 7- and 9-glucosides and lupinic acid were taken up very slowly by the callus tissue and showed great metabolic stability, but some degradation to 7-glucosyladenine, 9-glucosyladenine and the 9-alanine conjugate of adenine occurred. Compared with its aglycone, O-glucosyl-ZR exhibited slow uptake and greatly enhanced stability but gas chromatographic-mass spectrometric analysis showed that appreciable amounts were hydrolyzed to ZR in the tissue. Both ZR and O-glucosyl-ZR were metabolised extensively, with adenine, adenosine, and adenine nucleotide(s) as the major metabolites. A diversity of minor metabolites of ZR were identified, including O-glucosides, lupinic acid and dihydrolupinic acid. The metabolism of ZR was suppressed by 3-isobutyl-1-methylxanthine. When compared with the soybean callus line normally used for cytokinin bioassays (cv. Acme, cotyledonary callus), related callus lines exhibited greatly differing growth responses to cytokinin: however, these were not reflected in marked differences in metabolism.
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Abbreviations
- GC-MS:
-
gas chromatography-mass spectrometry
- HPLC:
-
high-performance liquid chromatography
- LA:
-
lupinic acid
- OGZR:
-
O-β-D-glucopyranosylzeatin riboside
- TLC:
-
thin-layer chromatography
- IMX:
-
3-isobutyl-1-methylxanthine
- Z:
-
zeatin
- ZR:
-
zeatin riboside
References
Conrad, V.K. (1974) Ein sensibilisierter Amaranthus-Cytokinintest (AT74). Biochem. Physiol. Pflanz. 165, 531–535
Cowley, D.E., Duke, C.C., Liepa, A.J., MacLeod, J.K., Letham, D.S. (1978) The structure and synthesis of cytokinin metabolites. I. The 7- and 9-β-D-glucofuranosides and pyranosides of zeatin and 6-benzylaminopurine. Aust. J. Chem. 31, 1095–1111
Davoll, J., Lythgoe, B., Todd, A.R. (1946) Experiments on the synthesis of purine nucleosides. XII. The configuration at the glycosidic centre in natural and synthetic pyrimidine and purine nucleosides. J. Chem. Soc 2, 833–838
Duke, C.C., MacLeod, J.K., Summons, R.E., Letham, D.S., Paker, C.W. (1978) The structure and synthesis of cytokinin metabolites. II. Lupinic acid and O-β-D-glucopyranosylzeatin from Lupinus angustifolius. Aust. J. Chem. 31, 1291–1301
Duke, C.C., Letham, D.S., Parker, C.W., MacLeod, J.K., Summons, R.E. (1979) The structure and synthesis of cytokinin metabolites. IV. The complex of O-glucosylzeatin derivatives formed in Populus species. Phytochemistry 18, 819–824
Elliott, D.C., Thompson, M.J. (1982) The identity of the major metabolite of benzylaminopurine in soybean cultures, and the inhibition of its formation by aminophylline. Plant Sci. Lett. 28, 29–38
Entsch, B., Parker, C.W., Letham, D.S., Summons, R.E. (1979) Preparation and characterisation, using high-performance liquid chromatography, of an enzyme forming glucosides of cytokinins. Biochim. Biophys. Acta. 570, 124–139
Gordon, M.E., Letham, D.S., Parker, C.W. (1974) The metabolism and translocation of zeatin in intact radish seedlings. Ann. Bot. (London) 38, 809–825
Horgan, R. (1975) A new cytokinin metabolite. Biochem. Biophys. Res. Commun. 65, 358–363
Kaminek, M., Hadačová, V., Luštinec, J. (1981) Plastids and the origin of cytokinin autonomy in tobacco tissue cultures. In: Metabolism and molecular activities of cytokinins, pp. 241–251, Guern, J., Péaud-Lenoël, C., eds. Springer, Berlin Heidelberg New York
Letham, D.S., Young, H. (1971) The synthesis of radioisotopically labelled zeatin. Phytochemistry 10, 2077–2081
Letham, D.S. (1978) Cytokinins. In: Phytohormones and related compounds — a comprehensive treatise, vol. 1, pp. 205–263, Letham, D.S., Goodwin, P.B., Higgins, T.J.V., eds. Elsevier/North-Holland, Amsterdam
Letham, D.S., Palni, L.M.S. (1983) The biosynthesis and metabolism of cytokinins. Annu. Rev. Plant Physiol. 34, 163–197
Letham, D.S., Palni, L.M.S., Tao, G.-Q., Gollnow, B.I., Bates, C. (1983) Regulators of cell division in plant tissues. XXIX. The activities of cytokinin glucosides and alanine conjugates in cytokinin bioassays. J. Plant Growth Regul 2, 103–115
Letham, D.S., Tao, G.-Q., Parker, C.W. (1982) An overview of cytokinin metabolism. In: Plant growth substances 1982, pp. 143–153, Wareing, P.F., ed. Academic Press, London New York
Miller, C.O. (1968) Naturally occurring cytokinins. In: Biochemistry and physiology of plant growth substances, pp. 33–45, Wightman, F., Setterfield, G., eds. Runge Press, Ottawa
Newton, C., Morgan, C.B., Morgan, D.G. (1980) Evaluation of a bioassay for cytokinins using soybean hypocotyl sections. J. Exp. Bot. 31, 721–729
Palmer, M.V., Scott, I.M., Horgan, R. (1981) Cytokinin metabolism in Phaseolus vulgaris L. II. Comparative metabolism of exogenous cytokinins by detached leaves. Plant Sci. Lett. 22, 187–195
Palni, L.M.S., Horgan, R. (1983) Cytokinin biosynthesis in crown gall tissue of vinca rosea: metabolism of isopentenyladenin. Phytochemistry 22, 1597–1601
Palni, L.M.S., Summons, R.E., Letham, D.S. (1983) Mass spectrometric analysis of cytokinins in plant tissues. V. Identification of the cytokinin complex of Datura innoxia crown gall tissue. Plant Physiol. 72, 858–863
Parker, C.W., Letham, D.S., Gollnow, B.I., Summons, R.E., Duke, C.C., MacLeod, J.K. (1978) Metabolism of zeatin by lupin seedlings. Planta 142, 239–251
Schmitz, R.Y., Skoog, F., Hecht, S.M., Bock, R.M., Leonard, N.J. (1972) Comparison of cytokinin activities of naturally occurring ribonucleosides and corresponding bases. Phytochemistry 11, 1603–1610
Scott, I.M., McGaw, B.A., Horgan, R., Williams, P.E. (1982) Biochemical studies on cytokinins in Vinca rosea crown gall tissue. In: Plant growth substances 1982, pp. 165–174, Wareing, P.F., ed. Academic Press, London
Shelton, K.R., Clark, J.M. (1967) A proton exchange between purines and water and its application to biochemistry. Biochemistry 6, 2735–2739
Summons, R.E., Duke, C.C., Eichholzer, J.V., Entsch, B., Letham, D.S., MacLeod, J.K., Parker, C.W. (1979) Mass spectrometric analysis of cytokinins in plant tissues. II. Quantitation of cytokinins in Zea mays kernels using deuterium labelled standards. Biomed. Mass Spectrom. 6, 407–413
Summons, R.E., Entsch, B., Letham, D.S., Gollnow, B.I., MacLeod, J.K. (1980) Metabolites of zeatin in sweetcorn kernels: purification and identifications using high-performance liquid chromatography and chemical-ionisation mass spectrometry. Planta 147, 422–434
Summons, R.E., Letham, D.S., Gollnow, B.I., Parker, C.W., Entsch, B., Johnson, L.P., MacLeod, J.K., Rolfe, B.G. (1981) Cytokinin translocation and metabolism in species of the leguminosae. In: Metabolism and molecular activities of cytokinins, pp. 56–65, Guern, J., Péaud-Lenoël, C., eds. Springer, Berlin Heidelberg New York
Summons, R.E., Palni, L.M.S., Letham, D.S. (1983) Determination of intact zeatin nucleotide by direct chemical ionisation mass spectrometry. FEBS Lett. 151, 122–126
van Staden, J., Papaphilippou, A.P. (1977) Biological activity of O-β-D-glucopyranosylzeatin. Plant Physiol. 60, 649–650
van Staden, J., Hutton, M.J. (1982) Metabolism of [8-14C]-zeatin in soybean callus. Z. Pflanzenphysiol. 106, 355–365
Wang, T.L. (1979) The sensitivity of soybean tissue cultures to the thymidine analogue, 5-bromodeoxyuridine. Plant Sci Lett. 17, 123–128
Yeoman, M.M., MacLeod, A.J. (1977) Tissue (callus) cultures — techniques. In: Plant tissue and cell culture, pp. 31–59, Street, H.E., ed. Blackwell, Oxford
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Palni, L.M.S., Palmer, M.V. & Letham, D.S. The stability and biological activity of cytokinin metabolites in soybean callus tissue. Planta 160, 242–249 (1984). https://doi.org/10.1007/BF00402861
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DOI: https://doi.org/10.1007/BF00402861