Differential effects of growth temperature on ice nuclei active at different temperatures that are produced by cells...

The temperature at which ice-nucleating bacteria are grown causes differences of 100- to 10,000-fold in the fraction of cells that nucleate ice at a given temperature (ice nucleation frequency). Ice nucleation frequencies of cells of Pseudomonas syringae grown at temperatures that ranged from 9 to 33 degrees C were examined in order to more accurately characterize physiological effects on ice nuclei active at temperatures of from about -2 to -10 degrees C, the temperature range for this phenotype. Large differences in ice nucleation frequency occurred at all but the lowest assay temperatures in cells of P. syringae grown in the temperature range of 15 to 33 degrees C. These differences in ice nucleation frequency may be attributed, at least in part, to post-translational factors. Because other studies have indicated that ice nuclei active at the lowest assay temperatures may reflect the amount of ice nucleation protein produced, while higher nucleation temperatures reflect aggregates of this ice nucleation protein, data was normalized to the frequency of ice nuclei active at the lowest ice nucleation temperatures (which also correspond to the most abundant nuclei). This was done in order to develop a baseline of comparison for cells grown at different temperatures that more clearly shows possible post-translational effects such as aggregation of the nucleation protein. After this normalization was performed, and in contrast to the results noted above, the number of ice nuclei in cells grown at 9, 15, and 20 degrees C that were active at different assay temperatures was very similar. Differences in ice nucleation frequency that occurred over all assay temperatures in cells grown between 9 and 20 degrees C may be attributed to differences in the total number of nuclei present in the population of cells. The large effects of growth temperature on nucleation frequency have important implications for estimating numbers of ice nucleating bacteria in environmental samples by determining the number of bacterial ice nuclei in such samples.