NaCl increases the activity of the plasma membrane H+-ATPase in C3 halophyte Suaeda salsa callus

Abstract

Suaeda salsa calli treated with different concentrations of NaCl were used to examine the response of the plasma membrane (PM) H+-ATPase to NaCl and its role in salt tolerance. The optimum concentration of NaCl for growth of the calli was 50 mM, while growth was significantly inhibited at 250 mM NaCl. The ion and organic solute contents of calli increased with increasing NaCl. Activity of the PM H+-ATPase increased when the calli were treated with NaCl over a certain concentration range (0–150 mM NaCl). However, the activity reached its maximum with 150 mM NaCl. Immunoblotting analysis of the PM H+-ATPase protein from calli cultures with anti-Zea mays H+-ATPase serum (monoclonal 46E5B11D5) identified a single polypeptide of ~90 kDa. The peptide levels increased in the calli treated with NaCl at 150 mM NaCl compared to control, but the increase at 50 mM NaCl was less pronounced. Northern blot analysis showed that the expression of the PM H+-ATPase also increased after the calli were treated with NaCl. These results suggest that the increase in PM H+-ATPase activity is due to both an increase in the amount of PM H+-ATPase protein and an up-regulation of the PM H+-ATPase gene, which is involved in the salt tolerance of S. salsa calli.