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Plant and Cell Physiology, 2001, Vol. 42, No. 7 677-685
© 2001 Oxford University Press
Identification of Plant Cytokinin Biosynthetic Enzymes as Dimethylallyl Diphosphate:ATP/ADP Isopentenyltransferases
Department of Biology, Graduate School of Science, Osaka University, and Recognition and Formation, PRESTO, JST, Osaka, 560-0043 Japan
It has been believed that the key step in cytokinin biosynthesis is the addition of a 5-carbon chain to the N6 of AMP. To identify cytokinin biosynthesis enzymes that catalyze the formation of the isopentenyl side chain of cytokinins, the Arabidopsis genomic sequence was searched for genes that could code for isopentenyltransferases. This resulted in the identification of nine putative genes for isopentenyltransferases. One of these, AtIPT4, was subjected to detailed analysis. Overexpression of AtIPT4 caused cytokinin-independent shoot formation on calli. As shoot formation on calli normally occurs only when cytokinins are applied, it suggested that this gene product catalyzed cytokinin biosynthesis in plants. Recombinant AtIPT4 catalyzed the transfer of an isopentenyl group from dimethylallyl diphosphate to the N6 of ATP and ADP, but not to that of AMP. AtIPT4 did not exhibit the DMAPP:tRNA isopentenyltransferase activity. These results indicate that cytokinins are, at least in part, synthesized from ATP and ADP in plants.
1 Corresponding author: E-mail, kakimoto@bio.sci.osaka-u.ac.jp; Fax, +81-6-6850-5420; Phone, +81-6-6850-5421.
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