A plasmamembrane redox system and proton transport in isolated mesophyll cells

Potassium ferricyanide (K(3)Fe[CN](6)) was added to aerated and stirred nonbuffered suspensions of mechanically isolated photosynthetically competent Asparagus sprengeri Regel mesophyll cells. Rates of Fe(CN)(6) (3-) reduction and H(+) efflux were measured with or without illumination. On the addition of 1 millimolar Fe(CN)(6) (3-) to nonilluminated cell suspensions acidification of the medium indicated an H(+) efflux of 1.54 nanomoles H(+)/10(6) cells per minute. Simultaneous Fe(CN)(6) (3-) reduction occurred at a rate of 1.55 nanomoles Fe(CN)(6) (3-)/10(6) cells per minute. Illumination stimulated these rates 14 to 17 times and corresponding values were 26.1 nanomoles H(+)/10(6) cells per minute and 22.9 nanomoles Fe(CN)(6) (3-)/10(6) cells per minute. These two processes appeared to be tightly coupled and were rapidly inhibited when illuminated suspensions were transferred to darkness or treated with 1 micromolar 3-(3,4-dichlorophenyl)-1,1 dimethylurea. Addition of 0.1 millimolar diethylstilbestrol eliminated ATP dependent H(+) efflux in illuminated or nonilluminated cells but had no influence on Fe(CN)(6) (3-) dependent H(+) efflux. Recent reports indicate that a transmembrane redox system spans the plasma membrane of root cells and is coupled to the efflux of H(+). The present report extends these observations to photosynthetically competent mesophyll cells. The results indicate a transport process independent of ATP driven H(+) efflux which operates with a H(+)/e(-) stoichiometry of one.