Separation and identification of the cytokinins in mulberry, Morus alba L.

YANAGISAWA YUKIO, SHIOIRI HIDENARI, ITO KOICHI, SUZUKI AKIRA, TANAKA KEIJI

doi:10.11416/kontyushigen1930.54.439

Endogenous cytokinins in mulberry leaves were identified by several assay methods. In extracts of mulberry leaves three substances with Rf 0.16-0.28, 0.60-0.68, and 0.88-0.92 were separated by HPTLC (high performance thin layer chromatography) and further assayed by the 265nm reflection scan of Shimazu CS-930. One of these substances was considered to be ZR (zeatin riboside), as its chromatographic pattern was similar to that of authentic ZR. The ZR was further assayed by HPLC (high performance liquid chromatography) and in this case the retention time of the separated ZR corresponded to that of authentic ZR. Mulberry leaf extract was fractioned by column chromatography with PVP (insoluble polyvinylpyrrolidone). When each fraction was further assayed by HPTLC, ZR, Z (zeatin), and 2iP (isopentenyl adenine) were detected in the fraction number 7, 8-16, 17 (ZR), 7, 8-21-24 (Z), and 5, 6-7, 8 (2iP), respectively. Bioassay for cytokinins showed the occurrence of three distinct regions of cytokinin activity. The activity was detected at Rf 0.20-0.30, 0.60-0.70, and 0.90-1.00. Cytokinin activity at Rf 0.90-1.00 at least partly corresponded to that of ZR. In addition, the cytokinin level in the axillary bud on the branch which had elongated after the summer pruning in the previous year was higher than that in the axillary bud on the stump of the branch base which had been formed by spring pruning and both cytokinin levels increased with the sprouting.

Separation and identification of the cytokinins in mulberry, Morus alba L.

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