MATERIALS AND METHODSCytokinin activity based on two bioassays was at least 100-fold higher in Psychotria punctata leaf discs with bacterial nodules than in discs without them. Nodulated discs from young leaves yielded 0.4 to 6 pg of cytokinin (zeatin equivalents) per g fresh weight of leaf tissue, whereas nonnodulated discs from the same leaves yielded 0 to 0.003 pg per g fresh weight. These estimates probably include free-base cytokinins and, if present, any nucleoside cytokinins precipitable by acidic silver nitrate. Cytokinin concentrations in Psychotria leaf nodules appear to be higher than normally found in green leaves of other plants. In 1-butanol-acetic acid-water (12:3:5, v/v), the one peak of activity chromatographed with an RF similar to zeatin's, but both number and identity of the active substance(s) remain unknown. These findings suggest that a cytokinin is produced by bacteria in leaf nodules of P. punctata and that it is involved in the symbiosis.Miehe (10) Torrey (14,15) have found them in a medium in which pure cultures of Rhizobium had grown.The purposes of this investigation were to determine whether leaf nodules of Psychotria punctata contain cytokinin(s), to measure their concentrations, and to compare these concentrations with those in other portions of the same leaves.