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Addition of 100 millimolar KCl, NaCl, or Na(2)SO(4) strongly promoted acidification of the medium by cells of Nicotiana tabacum/gossii in suspension culture. Acidification was greater in the case of NaCl-adapted than in that of wild type cells, and strikingly so in KCl medium when fusicoccin (FC) was present. Back-titration indicated that net proton secretion in KCl medium was increased 4-fold by FC treatment in the case of adapted cells; but was not even doubled in wild type cells. Membrane potential was higher in NaCl-adapted cells. FC treatment hyperpolarized wild, but not NaCl-adapted cells, suggesting a higher degree of coupling between H(+) efflux and K(+) influx in adapted cells; FC enhanced net K(+) uptake in adapted but not in wild cells. Acidification by cells suspended in 10 millimolar KCl was highly sensitive to vanadate, but that after addition of 100 millimolar KCl or NaCl was much less sensitive. Addition of 100 millimolar NaCl to wild type cells already provided with 10 millimolar KCl briefly accelerated, then slowed down the rate of acidification. If the addition was made after acidification had already ceased, alkalization was observed, particularly in the presence of FC. The results are consistent with the operation of a Na(+)-H(+) antiporter.

Proton Fluxes as a Response to External Salinity in Wild Type and NaCl-Adapted Nicotiana Cell Lines