The stability and biological activity of cytokinin metabolites in soybean callus tissue

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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.