Distinct survival, growth lag, and ribosomal RNA degradation kinetics during long-term starvation for carbon or phosphate

Abstract

AbstractStationary phase is the general term for the state a bacterial culture reaches when no further increase in cell number occurs due to the exhaustion of nutrients in the growth medium. Depending on the type of nutrient that is first depleted, the metabolic state of the stationary phase cells may vary greatly, and the subsistence strategies that best support cell survival may differ. As ribosomes play a central role in bacterial growth and energy expenditure, ribosome preservation is a key element of such strategies. To investigate the degree of ribosome preservation during long-term starvation, we compared the dynamics of ribosomal RNA (rRNA) levels of carbon-starved and phosphorus-starvedEscherichia colicultures for up to 28 days. The starved cultures’ contents of full-length 16S and 23S rRNA decreased exponentially and phosphorus starvation resulted in much more rapid rRNA degradation than carbon starvation. Bacterial survival kinetics were also quantified over the starvation period. Upon replenishment of the nutrient in question, carbon-starved cells resumed growth faster than cells starved for phosphate for the equivalent amount of time, and for both conditions, the lag time increased with the starvation time. While these results are in accordance with the hypothesis that cells with a larger ribosome pool recover more readily upon replenishment of nutrients, we also observed that the lag time kept increasing with increasing starvation time, also when the amount of rRNA per viable cell remained constant.ImportanceBacteria grow exponentially consuming nutrients, and then starve until the next nutrient is added. To elucidate the survival kinetics of the cells under starvation, we performed month-long, carbon and phosphorus starvation experiments ofEscherichia colimonitoring ribosomal RNA levels and survival of the cells. The starved cultures’ concentration of ribosomal RNA dropped exponentially with time, and the speed of degradation was much quicker under the phosphorus starvation than the carbon starvation. We have also quantified the lag time, i.e., the time needed to resume growth when the starved cells are transferred into fresh media. The observation revealed that the lag time increases with starvation time and the phosphorus starvation has a greater impact on the increase of the lag time.