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Articles

Cytokinins in tRNA Obtained from Spinacia oleracea L. Leaves and Isolated Chloroplasts ¹

Santhanam Swaminathan

Norimoto Murai

Western Regional Research Center, United States Department of Agriculture, Agricultural Research Service, Berkeley, California 94710, Department of Human Oncology, University of Wisconsin Medical School, 2-C University Hospitals, Madison, Wisconsin 53706, Institute of Plant Development, Birge Hall, University of Wisconsin, Madison, Wisconsin 53706

Cytokinin-active ribonucleosides have been isolated from tRNA of whole spinach (Spinacia oleracea L.) leaves and isolated spinach chloroplasts. The tRNA from spinach leaf blades contained: 6-(4-hydroxy-3-methyl-2-butenylamino)-9--D-ribofuranosylpurine (cis and trans isomers), 6-(3-methyl-2-butenylamino)-9--D-ribofuranosylpurine, and 6-(4-hydroxy-3-methyl-2-butenylamino)-2-methylthio-9--D -ribofuranosylpurine (cis and trans isomers). A method for isolation of large amounts of intact chloroplasts was developed and subsequently used for the isolation of chloroplast tRNA. The chloroplast tRNA contained 6-(3-methyl-2-butenylamino)-9--D-ribofuranosylpurine and 6-(4-hydroxy-3-methyl-2-butenylamino)-2-methylthio-9--D -ribofuranosylpurine (the cis isomer only). The structures of these compounds were assigned on the basis of their chromatographic properties and mass spectra of trimethylsilyl derivatives which were identical with those of the corresponding synthetic compounds. The results of this study indicate that ribosylzeatin was present in spinach leaf tRNA, but absent from the purified chloroplast tRNA preparation.

¹ This research was performed when H. J. V. was a National Research Council Postdoctoral Research Associate.

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