Fusicoccin (FC), like indoleacetic acid (IAA), causes Golgi-localized B-1,4-glucan synthase (GS) activity to increase when applied to pea third internode segments whose GS activity has declined after isolation from the plant. This suggests that GS activity is modulated by H' extrusion; in agreement, vanadate and nigericin inhibit the GS response. The GS response is not due to acidification of the cell wall. Treatment of tissue with heavy water, which in effect raises intracellular pH, mimics the MATERIALS AND METHODS Pea (Pisum sativum cv Alaska) seedlings were grown and third internode segments 8 mm long were cut as described (27), except that for all experiments other than that ofTable I, each internode was abraded (14) by stroking with an aqueous paste of No. 305 emery powder (Edmund Scientific, Barrington, NJ) which was then rinsed off by vigorous agitation in water, before cutting the segment. The segments were kept 2 to 3 h at 35C in moist air to deplete their GS activity (27).Lots of 30 segments, each in a small Stender dish, were pretreated usually for 30 to 60 min at 35°C in a medium containing 30 mm sucrose, 0.8 mm citric acid/Pipes (Tris salt) buffer pH 6.5 (unless specified otherwise in figure or table legends), containing usually 1 mM KCI, plus indicated additions. This pretreatment medium was removed and 2.0 ml of a similar incubation medium, with or without IAA or FC as indicated, the initial pH of which had been carefully measured to 0.01 pH unit with a semimicro glass electrode, were added, and the dishes were kept normally at 35C for 60 or 90 min (the incubation period). The incubation medium was removed and its pH was remeasured, while the segments were rinsed with ice water and held on ice until they were homogenized to prepare membranes for enzyme assay. H+ extrusion was estimated from the measured drop in pH (ApH) of the medium during incubation, using the postincubation titration curve