Substrate and electron donor limitation induce phenotypic heterogeneity in different metabolic activities in a green...

Populations of genetically identical cells can display marked variation in phenotypic traits; such variation is termed phenotypic heterogeneity. Here, we investigate the effect of substrate and electron donor limitation on phenotypic heterogeneity in N₂ and CO₂ fixation in the green sulphur bacterium Chlorobium phaeobacteroides. We grew populations in chemostats and batch cultures and used stable isotope labelling combined with nanometer-scale secondary ion mass spectrometry (NanoSIMS) to quantify phenotypic heterogeneity. Experiments in H₂ S (i.e. electron donor) limited chemostats show that varying levels of NH4+ limitation induce heterogeneity in N₂ fixation. Comparison of phenotypic heterogeneity between chemostats and batch (unlimited for H₂ S) populations indicates that electron donor limitation drives heterogeneity in N₂ and CO₂ fixation. Our results demonstrate that phenotypic heterogeneity in a certain metabolic activity can be driven by different modes of limitation and that heterogeneity can emerge in different metabolic processes upon the same mode of limitation. In conclusion, our data suggest that limitation is a general driver of phenotypic heterogeneity in microbial populations.