ABSTRACT
Previous work has shown that an increase in cytosolic pH plays an important role in the induction of phosphoenolpyruvate carboxylase (PEPc) kinase by light in C(4) plants. The potential involvement of a similar effect in the induction of PEPc kinase in the Crassulacean acid metabolism (CAM) plant Kalanchoë fedtschenkoi was assessed using leaf disks. Treatment of disks with the weak base NH(4)Cl did not affect induction of the kinase. Prolonged treatment of disks with weak acids prevented both decarboxylation of malate during the day and induction of the kinase in the following night, but short treatments had no effect. The data are consistent with the view that a high cytosolic malate content can prevent induction of phosphoenolpyruvate carboxylase kinase. Changes in cytosolic pH may affect induction of the kinase in CAM plants by a secondary effect rather than as part of a primary signalling pathway.
ABSTRACT
Phosphoenolpyruvate carboxylase (PEPc) catalyzes the primary fixation of CO(2) in Crassulacean acid metabolism plants. Flux through the enzyme is regulated by reversible phosphorylation. PEPc kinase is controlled by changes in the level of its translatable mRNA in response to a circadian rhythm. The physiological significance of changes in the levels of PEPc-kinase-translatable mRNA and the involvement of metabolites in control of the kinase was investigated by subjecting Kalanchoë daigremontiana leaves to anaerobic conditions at night to modulate the magnitude of malate accumulation, or to a rise in temperature at night to increase the efflux of malate from vacuole to cytosol. Changes in CO(2) fixation and PEPc kinase activity reflected those in kinase mRNA. The highest rates of CO(2) fixation and levels of kinase mRNA were observed in leaves subjected to anaerobic treatment for the first half of the night and then transferred to ambient air. In leaves subjected to anaerobic treatment overnight and transferred to ambient air at the start of the day, PEPc-kinase-translatable mRNA and activity, the phosphorylation state of PEPc, and fixation of atmospheric CO(2) were significantly higher than those for control leaves for the first 3 h of the light period. A nighttime temperature increase from 19 degrees C to 27 degrees C led to a rapid reduction in kinase mRNA and activity; however, this was not observed in leaves in which malate accumulation had been prevented by anaerobic treatment. These data are consistent with the hypothesis that a high concentration of malate reduces both kinase mRNA and the accumulation of the kinase itself.