Growth characteristics at low Na+ concentration and the stability of the Na+ requirement of a marine bacterium

Abstract

Studies of the marine bacterium Alteromonas haloplanktis 214 (formerly referred to as marine pseudomonad B-16) showed that as the Na+ concentration in the growth medium decreased from 230 to 34 mM, the lowest concentration permitting growth, the length of the lag period preceding exponential growth increased. Once growth had begun, except for a slight reduction in rate of growth at 34 mM Na+, the generation time and extent of growth remained essentially constant over the range of Na+ concentrations tested. Plate counts showed that during the lag period the numbers of viable cells introduced as inoculum into a complex medium containing 33 mM Na+ decreased exponentially before increasing. Repeated subculture of the cells at 33 mM Na+ failed to eliminate the lag period or reduce the loss of viability of the cells. The viability loss and the lag period could be eliminated either by raising the NaCl concentration to 130 mM or by adding sufficient sucrose to make the osmotic pressure of the medium equal to that obtained by adding 130 mM NaCl. In a chemically defined medium, sucrose added to maintain tonicity reduced but did not eliminate the lag periods obtained at suboptimal Na+ concentrations. Increasing the number of cells plated on trypticase agar medium reduced the Na+ concentration required to permit growth. Evidence was obtained of a requirement of A. haloplanktis for Ca2+ for growth. Ca2+ spared to a small extent the requirement for Na+ for growth. Some 1010 cells of a histidine-requiring, streptomycin-resistant mutant of A. haloplanktis 214, still viable after treatment with N-methyl-N′-nitro-N-nitrosoguanidine, were screened for capacity to grow in the absence of Na+. Since no non-Na+-requiring mutants were isolated, the requirement of this organism for Na+ would appear to be extremely stable.