Global phylogeography of pelagic Polynucleobacter bacteria: Restricted geographic distribution of subgroups, isolation by distance and influence of climate†

The free-living planktonic freshwater bacterium Polynucleobacter necessarius subspecies asymbioticus (> 99% 16S rRNA similarity) represents a taxon with a cosmopolitan distribution and apparently ubiquitous occurrence in lentic freshwater habitats. We tested for intra-taxon biogeographic patterns by combining cultivation-independent and cultivation methods. A culture collection of 204 strains isolated from globally distributed freshwater habitats (Arctic to Antarctica) was investigated for phylogeographic patterns based on sequences of two markers, the 16S-23S internal transcribed spacers and the glutamine synthetase gene (glnA). Genetic distance between isolates showed significant geographic distance-decay patterns for both markers, suggesting that an isolation-by-distance mechanism influences the global phylogeography. Furthermore, a couple of subgroups showed restricted geographic distributions. Strains of one subgroup were exclusively obtained from tropical sites on four continents (pantropical subgroup). Cultivation-independent methods were used to confirm the restricted geographic distributions of two subgroups. The pantropical taxon could be detected in 63% of investigated tropical habitats but not in any of 121 European freshwater samples. Physiological tests indicated that almost all strains of the pantropical subgroup failed to grow at temperatures of 4°C, while strains affiliated with other subgroups showed good growth at this temperature. This suggests that thermal adaptation is involved in phylogeographic structuring of the global Polynucleobacter population.