CYTOKININ CONTENT AND TISSUE DISTRIBUTION IN PLANTS TRANSFORMED BY A RECONSTRUCTED ISOPENTENYL TRANSFERASE GENE

The cytokinin gene, isopentenyl transferase (ipt), was placed under the control of a heat-inducible promoter from the Drosophila melanogaster hsp70 gene and introduced into Nicotiana plumbaginifolia by cocultivation with Agrobacterium tumefaciens. Transformants were analyzed for organ-specific expression, cytokinin levels and effects on plant development before and after heat induction. The ipt gene transcripts were detected in leaves and stems but not roots of transgenic plants following a 2 hour, 45-degrees-C treatment. Maximum mRNA levels observed occurred 2 hours after heat treatment and 46 hours later were detected only in leaves. Zeatin and zeatinriboside concentrations 2 hours after heat shock ranged from over 900 to 2000 pmol/g, representing a greater than 140- to 200-fold increase over uninduced levels. After 46 hours, approximately 50% of the cytokinins are still present in the leaves as opposed to much reduced levels in the stems. Transgenic plants were greener, shorter, had an underdeveloped root system, reduced leaf width, and increased growth of axillary buds. After a single heat treatment, plants exhibited a darker green pigment and continued growth of lateral buds. Transient accumulations of endogenous cytokinins following thermal induction did not appear to alter the plant's preprogrammed pattern of differentiation.