The rate of dark 02 uptake of Elodea canadensis leaves was titrated with either cyanide or sulfide in the presence and in the absence of 5 millimolar salicylhydroxamic acid (SHAM), an inhibitor of the altemative oxidase. The inhibition of 02 uptake by SHAM alone was very small (3-6%), suggesting that actual respiration mainly occurred through the cytochrome pathway. 02 uptake was slightly stimulated by cyanide at concentrations of 50 micromolar or higher, but in the presence of SHAM respiration was strongly suppressed. The effects of sulfide on 02 uptake were similar to those of cyanide, except that the percent stimulation of 02 uptake by sulfide alone was somewhat higher than that of cyanide. However, the estimates of the capacity of the altemative pathway were similar with both inhibitors. Another difference is that maximal inhibition of respiration in the presence of SHAM was observed with lower concentrations of sulfide (50 micromolar) than cyanide (250 micromolar). The results suggest that sulfide can be used as a suitable inhibitor of cytochrome c oxidase in studies with intact plant tissues, and that sulfide does not apparently inhibit the altemative oxidase.Cyanide and sulfide are noncompetitive inhibitors of Cyt c oxidase and their comparative effects have been widely studied at the enzyme level (5, 10, 12). However, cyanide has been the inhibitor of Cyt c oxidase most frequently used to estimate the so-called "cyanide-resistant respiration" (8, 9). Sulfide has been little used in works on plant respiration, mainly in some ecophysiological studies on the effects of sulfide as an environmental pollutant (1, 4, 6, 7). The results suggest that usually there is little inhibition of respiration by sulfide, that is, a considerable "sulfide-resistant respiration" exists in plants. However, as far as we know, there have been no attempts to relate the use of sulfide with the expression of the different mitochondrial electron transport pathways in intact plant tissue, and this will be the main objective of this communication.The results obtained in the present work suggested that the effects of sulfide and cyanide on respiration of Elodea canadensis leaves are comparable, that is, both compounds inhibited the Cyt pathway, and elicited the expression of the 1249 nonphosphorylating alternative pathway. The higher activity of the alternative pathway in the presence of sulfide should be taken into account in the interpretation ofecophysiological studies on sulfide pollution in relation to respiration.
MATERIAL AND METHODS Plant MaterialPlants of Elodea canadensis Michx were bought from an aquarium shop and were further grown for a few days in a cabinet under constant conditions: 20C (day and night), and a quantum flux density (400-700 nm) of 100 to 200 1AE/m2. s, supplied by Sylvania cool-white VHO (215 W) fluorescent tubes for 13 h/d. Plants were placed in an aerated nutrient solution containing KNO3, 1 mM; Ca(NO3)2, 1 mM; MgSO4, 0.375 mM; NaH2PO4, 0.33 mM; EDTA FeNa, 13.5 ,uM.
Measurement of Respiration02 u...