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Subcellular Localization of Cytokinins in Transfer Ribonucleic Acid ¹

Folke Skoog^b

^a Department of Biochemistry, Laboratory of Molecular Biology, University of Wisconsin, Madison, Wisconsin 53706, Institute of Plant Development, Birge Hall, University of Wisconsin, Madison, Wisconsin 53706

Evidence on the localization of cytokinins in chloroplast tRNA was obtained by comparison of Euglena gracilis var. bacillaris light-grown and dark-grown wild type cultures and chloroplast-bleached mutant strains. The several cytokinins characteristic of tRNA were separated by Sephadex LH-20 column chromatography of the hydrolysates and were quantitatively determined by tobacco bioassays of the eluates. The results indicate that 6-(3-methyl-2-butenylamino)-9--D-ribofuranosylpurine (i⁶ A) is formed in both the cytoplasmic and chloroplast tRNA, whereas 6-(4-hydroxy-3-methyl-cis-2-butenylamino)-9--D- ribofurano-sylpurine (c-io⁶A) is produced mainly in the cytoplasmic tRNA and 6-(4-hydroxy-3-methyl-2-butenylamino)-2-methylthio-9--D- ribofurano-sylpurine (ms²io⁶A) is localized exclusively in chloroplast tRNA. The restriction of the methiolation reaction to the chloroplast is supported by results of radioisotope experiments showing that ³⁵S-labeled MgSO₄ is incorporated into ms²io⁶A in the wild type cultures, but not in the chloroplast-bleached mutant strains.

¹ This work was supported by funds from the Wisconsin Alumni Research Foundation, and by National Science Foundation Research Grant BMS72-02226 to F. S.

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