Salt‐induced changes in protein composition in light‐grown callus of Mesembryanthemum crystallinum

Abstract

The halophyte Mesembryanthemum crystallinum (ice plant) has been suggested as a model for salt‐tolerance in higher plants. To investigate salt‐induced changes in polypeptide patterns at the cellular level, a light‐grown callus of M. crystallinum with substantial chlorophyll content, was established and the effect of NaCl on the composition of phenol‐extracted protein was examined by SDS‐ and 2D‐polyacrylamide gel electrophoresis (PAGE). SDS‐PAGE showed the accumulation of five polypeptides with estimated molecular masses of 40, 34, 32, 29 and 14 kDa was enhanced by the addition of 200 mM NaCl to the culture media. The addition of ABA (10 μM) or mannitol (400 mM) did not elicit the same degree of accumulation of these salt‐specific proteins. These polypeptides were classified into two groups according to their course of induction: early responsive (40, 34, 29 kDa) and late‐responsive (32, 14 kDa) proteins. In addition, two polypeptides (20, 18 kDa) were transiently accumulated during salt treatment. Further separation of soluble proteins by 2‐D gel electrophoresis, either isoelectric focusing (IEF) or non‐equilibrium pH‐gradient electrophoresis (NEPHGE) followed by SDS‐PAGE, showed more alterations in accumulation of polypeptides by NaCl than 1‐D gel electrophoresis. Overall, levels of more than 30% of basic polypeptides, detected by NEPHGE/SDS‐PAGE, were altered by 200 mM NaCl treatment, while only 10% of neutral and acidic polypeptides, detected by IEF/SDS‐PAGE, were changed. The enhanced expression of these proteins by salt in cultured cells is most likely related to the cellular responses to salinity, and not to the mechanism of CAM induction in this facultative halophyte.