The Effect of Light on the Tricarboxylic Acid Cycle in Green Leaves: I. Relative Rates of the Cycle in the Dark and the Light

Excised green leaves of mung bean (Phaseolus aureus L. var. Mungo) were used to determine the effect of light on the rate of endogenous respiration via the tricarboxylic acid cycle. Illumination with white light at an intensity of 0.043 gram calories cm(-2)min(-1) (approximately 8600 lux) of visible radiation (400-700 nm) gave a rate of apparent photosynthesis, measured as net CO(2) uptake, of 21 mg CO(2) dm(-2)hr(-1) which was about 11-fold greater than the rate of dark respiration. The feeding of (14)CO(2) or (14)C-labeled acids of the tricarboxylic acid cycle in the dark for 2 hours was established as a suitable method for labeling mitochondrial pools of cycle intermediates.At a concentration of 0.1 mm 3-(3,4-dichlorophenyl)-1,1-dimethylurea, apparent photosynthesis was inhibited 82%, and the refixation of (14)CO(2) derived internally from endogenous respiration was largely prevented. In the presence of this inhibitor endogenous respiration, measured as (14)CO(2) evolution, continued in the light at a rate comparable to that in the dark. Consequently, under these conditions light-induced nonphotosynthetic processes have no significant effect on endogenous dark respiration. Inhibitors of the tricarboxylic acid cycle, malonate and fluoroacetate, were used to determine the relative rates of carbon flux through the cycle in the dark and in the light by measuring the rate of accumulation of (14)C in either succinate or citrate. Results were interpreted to indicate that the tricarboxylic acid cycle functions in the light at a rate similar to that in the dark except for a brief initial inhibition on transition from dark to light. Evidence was obtained that succinate dehydrogenase as well as aconitase, was inhibited in the presence of fluoroacetate.