Salt-induced Inhibition of Phosphate Transport and Release of Membrane Proteins from Barley Roots

Osmotic shock with sequential 30-minute treatments in ice-cold saline solutions and H₂O released proteins from excised barley roots and inhibited the subsequent uptake of orthophosphate (Pi). The amount of protein released increased sharply at NaCl concentrations above 0.05 molar, approximately the threshold concentration above which Pi uptake was increasingly suppressed. About 60% of the nearly 100 micrograms of protein per gram fresh weight of roots that was eluted in 0.16 molar NaCl treatments apparently had no function in Pi transport, since it was eluted at NaCl concentrations (≤0.05 molar) that did not affect Pi uptake. Although 0.16 molar NaCl completely inhibited Pi uptake, active uptake resumed at about 60% of control rates within 1 to 2 hours. The presence of either puromycin or cycloheximide greatly reduced the recovery of Pi uptake activity after the NaCl treatment. Mannitol and various monovalent and divalent salts at concentrations isosmotic with NaCl also inhibited Pi uptake, but CaCl₂ was consistently the least inhibitory. The correlation between the concentration of the osmotic treatments and the simultaneous loss of protein and Pi uptake activity, together with the evidence that uptake recovery requires protein synthesis, support the hypothesis that the proteins eluted are required for active Pi transport.