The effect of an applied electric field has been measured for the quantum yield of formation of the P+H-, P+Q(A)-, and 3P states of nonoriented Rb. sphaeroides reaction centers at 80 K in poly(vinyl alcohol) (PVA) films and in glycerol-buffer glasses. Both the shape and magnitude of the field dependence are very similar for all three observables in both media where the data overlap. This result suggests that the reduction of the quantum yield occurs at a very early step in the charge separation process. More than 50% reduction in the quantum yield is observed at fields of 2.5 x 10(6) V/cm in PVA films. Possible mechanisms include field-dependent nonradiative decay of 1P, a field-dependent rate of formation or decay of P+B- if P+B- is important, and field-dependent formation or decay of an unrelaxed form of P+H-. Small electric field effects are observed on the recombination rate of the radical pair P+H- and the magnetic field dependence of the 3P quantum yield. This is likely because those orientational subpopulations of reaction centers which survive to become P+H- and 3p in an electric field are those which did not experience quantum yield failure at an early step. The field-induced effects provide a basis for a possible mechanism to regulate photosynthetic charge separation by transmembrane potentials.