In addition to their role as plant hormones, cytokinins are also found as structural components in tRNA. Six different tRNA cytokinins have been found in plants, but most other organisms, including humans, have only one–isopentenyladenosine. In an attempt to probe if the different forms have different functionality, we attempted to alter tRNA cytokinin composition by expressing the human tRNA isopentenyltransferase gene (EC 5.1.2.8) in tobacco [Nicotiana tabacum (L.) cv. Wisconsin 38]. The resulting transgenics had ~40% more isopentenyladenosine in tRNA, and an altered phenotype characterised by reduced internode length, increased stem diameter and rigidity, greener leaves, increased axillary bud outgrowth, abnormal flower morphology, and reduced seed viability. The levels of the two other major isoprene adenines of tRNA, cis-zeatin and 2-methyltiolated cis-zeatin, were also increased, but to a lower degree. Nearly all of the increase in isopentenyladenosine was in a single tRNA species. Two quantitatively minor isopentenyladenosine-containing tRNAs had also increased strongly. IPPT: Dimethylallylpyrophosphate: