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Inhibition of growth of cultured tobacco cells at low concentrations of lovastatin is reversed by cytokinin.

De novo synthesis of mevalonic acid, which is catalyzed by 3-hydroxy-3-methylglutaryl coenzyme A reductase, is the first committed step in the formation of isoprenoid compounds. Various studies have shown that mevalonic acid-derived compounds are required for growth of plant and animal cells, a conclusion supported by the observation that cells treated with lovastatin (a potent inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase) cease growth. We show that Nicotiana tabacum BY-2 cells, which require exogenous auxin for growth in culture but do not require exogenous cytokinin, are growth inhibited by 1 mum lovastatin. However, these cells are capable of growing in the presence of 1 mum lovastatin if 8 mum zeatin is supplied in the medium. Furthermore, benzyladenine, kinetin, and thidiazuron effectively reverse the inhibition of growth of these cells at 1 mum lovastatin, whereas adenine and 6-methyladenine have no effect. These results demonstrate that restoration of growth to lovastatin-treated cells is cytokinin specific and is not caused by metabolism of cytokinin into other isoprenoid compounds. Cytokinin does not effectively reverse the effects of higher concentrations of lovastatin, but mevalonic acid does, consistent with the hypothesis that cytokinin biosynthesis is more sensitive to lovastatin than the biosynthesis of other essential isoprenoid compounds in tobacco cells. This observation suggests that lovastatin can be used to induce cytokinin dependence in cytokinin-autonomous tobacco cell cultures.