Effect of ethephon and daminozide on the respiration of isolated leaf cells

A combined foliar application of ethephon (2-chloroethylphosphonic acid) at 0.8 kg/ha and daminozide (butanedioic acid mono (2,2 dimethylhydrazide) at 3.2 kg/ha inhibited the vegetative growth of Black Valentine bean (Phaseolus vulgaris L.) without the leaf chlorosis and necrosis caused by ethephon alone. This antagonistic interaction was further evaluated by examining the effect of ethephon and daminozide on respiration and lipid synthesis of isolated leaf cells. Ethephon (1.0 mM) promoted¹⁴CO₂ evolution from cells incubated with¹⁴C-glucose for 14 h by approximately 75%. Characterization of this response with Black Valentine bean mitochondria indicated that the observed stimulation could not be attributed to the existence of a major cyanide insensitive pathway or the possibility of ethephon acting as an uncoupler, which supports the view that ethephon (or ethylene) acts in the cytosol rather than in mitochondria. Daminozide at 30.0 and 60.0 mM inhibited¹⁴CO₂ evolution of isolated cells by 30 and 70%, respectively. Ethephon in combination with daminozide (1.0+60 mM) resulted in a 32% inhibition of respiration. Daminozide (60.0 mM) inhibited the incorporation of¹⁴C-glucose into chloroform-methanol soluble products by 47%, but did not affect the incorporation of¹⁴C-acetate. The results suggest that daminozide may reduce or overcome any stimulatory effect of ethephon on respiration and support an active inhibitory site for daminozide in mitochondria.