Osmoadaptation in Rhizobia: Ectoine-induced salt tolerance

After having shown that ectoine (a tetrahydropyrimidine) displays osmoprotective properties towards Escherichia coli (M. Jebbar, R. Talibart, K. Gloux, T. Bernard, and C. Blanco, J. Bacteriol. 174:5027-5035, 1992), we have investigated the involvement of this molecule in the osmotic adaptation of Rhizobium meliloti. Ectoine appeared almost as effective as glycine betaine in improving the growth of R. meliloti under adverse osmotic conditions (0.5 M NaCl). Moreover, improvement of growth of rhizobial strains insensitive to glycine betaine was also observed. Ectoine transport proved inducible, periplasmic protein dependent, and, as shown by competition experiments, distinct from the transport of glycine betaine. Medium osmolarity had little effect on the uptake characteristics, since the rate of influx increased from 12 to only 20 nmol min(-1) mg of protein(-1) when NaCl concentrations were raised from 0 to 0.3 or 0.5 M, with a constant of transport of 80 micromolar. Natural-abundance (13)C-nuclear magnetic resonance and radiolabelling assays showed that ectoine, unlike glycine betaine, is not intracellularly accumulated and, as a consequence, does not repress the synthesis of endogenous compatible solutes (glutamate, N-acetylglutaminylglutamine amide, and trehalose). Furthermore, the strong rise in glutamate content in cells osmotically stressed in the presence of ectoine suggests that, instead of being involved in osmotic balance restoration, ectoine should play a key role in triggering the synthesis of endogenous osmolytes. Hence, we believe that there are at least two distinct classes of osmoprotectants: those such as glycine betaine or glutamate, which act as genuine osmolytes, and those such as ectoine, which act as chemical mediators