Discovery and evidence of nitrogen fixation by thermophilic heterotrophs in hot springs

A previously unknown phenomenon, nitrogenase activity at high temperatures by nonphotosynthetic prokaryotes in thermal springs, is reported. This activity was revealed by dark-bottle incubations during cyanobacterial nitrogenase studies in hot springs of Yellowstone National Park. Benthic mat cores from alkaline thermal streams reduced acetylene in the dark from 30° to 60°C. the bacterial activity was distinguished from that of sympatric cyanobacteria by its sensitivity to oxygen (light bottles), heterotrophic response to organic amendments, and distinctive spatial distribution. Field tests for other biotic or abiotic explanations of the activity were negative. Anaerobic enrichment cultures supported the field evidence. Fourteen morphological types of presumptive diazotrophs, including sulfate-reducing and spore-forming rods. have been observed in these cultures.