When [8-14C]-N-(A'-isopentenyl) adenosine is incubated with the endosperm of corn (2 weeks after pollination), it is converted to ['4C]-N6-(4-hydroxy-3-methylbut-2-trans-enyl) adenosine, trans-ribosylzeatin. This biosynthetic step, N6-(A2-isopentenyl)adenosine to ribosylzeatin, also occurs in the mycorrhizal fungus, Rhizopogon roseolus.Cytokinin activity has been detected in extracts of various plants and plant organs. An active component in the extracts has been identified as trans-ribosylzeatin, trans-RZ2 (Fig. 1), in the case of immature fruit (16,17,20) and chicory root (2), and as its dihydro derivative in the case of Lupin (14) (see review by Kende (12) for a more detailed account). Recently Dyson and Hall (5) reported the presence of i6Ado in an extract of an autonomous strain of tobacco tissue. Of these naturally occurring cytokinins, trans-RZ exhibits the greatest activity in the tobacco callus bioassay system (9).As part of a study of the metabolism of cytokinins, we have investigated the biosynthesis of trans-RZ, and in this paper we describe its synthesis from the precursor, i6Ado. We chose systems known to contain free trans-RZ, a mycorrhizal fungus, Rhizopogon roseolus (22) and corn kernels (16,20