ZEA3: A Negative Modulator of Cytokinin Responses in Plant Seedlings

In Nicotiana plumbaginifolia cytokinins affect seedling development by inhibiting root growth and hypocotyl elongation and by stimulating cotyledon expansion. The zea3.1 mutant was selected for its inability to grow in conditions of low nitrogen and for its ability to grow independently on inhibitory concentrations of zeatin (J.D. Faure, M. Jullien, M. Caboche [1994] Plant J 5: 481-491). The zea3.1 growth response to cytokinins is reflected by an increase in cotyledon expansion due to cell division and by a swelling of the hypocotyl due to cell enlargement. An analysis of the seedling's root length and fresh weight over a wide range of benzyladenine concentrations showed that zea3.1 plants exhibit a higher sensitivity and an amplified response to cytokinins. A similar response of zea3.1 to benzyladenine was also seen in the expression of msr1, a cytokinin-regulated gene. Regulation of msr1 expression by protein phosphorylation was unaffected by the zea3.1 mutation. No significant differences in cytokinin and auxin levels were found between zea3.1 and wild-type seedlings, suggesting that the mutant phenotype is not caused by an alteration of these hormone levels. The data presented suggest that ZEA3 negatively modulates cytokinin responses and may function as a broad regulator of seedling development.