In situ hydrogen dynamics in a hot spring microbial mat during a diel cycle

Microbes can produce molecular hydrogen (H2) via fermentation, dinitrogen fixation or direct photolysis, yet the H2 dynamics in cyanobacterial communities has only been explored in a few natural systems and mostly in the laboratory. In this study, we investigated the diel in situ H2 dynamics in a hot spring microbial mat, where various ecotypes of unicellular cyanobacteria (Synechococcus sp.) are the only oxygenic phototrophs. In the evening, H2 accumulated rapidly after the onset of darkness reaching peak values of up to 30 μmol H2 L(-1) at about 1 mm depth below the mat surface, slowly decreasing to about 11 μmol H2 L(-1) just before sunrise. Another pulse of H2 production reaching a peak concentration of 46 μmol H2 L(-1) was found in the early morning under dim light conditions too low to induce accumulation of O2 in the mat. The light stimulation of H2 accumulation indicated nitrogenase activity to be an important source of H2 during the morning. This is in accordance with earlier findings of a distinct early morning peak in N2 fixation and expression of Synechococcus nitrogenase genes in mat samples from the same location. Fermentation might have contributed to the formation of H2 during the night, where accumulation of other fermentation products lowered the pH in the mat to IMPORTANCE: Hydrogen is a key intermediate in anaerobic metabolism, and with the development of a sulfide-insensitive microsensor for H2 it is now possible to study the microdistribution of H2 in stratified microbial communities such as the photosynthetic microbial mat investigated here. The capability to measure H2 profiles within the mat as compared to previous measurements of H2 emission gives much more detailed information about sources and sinks of H2 in such communities, and it was demonstrated that high rates of H2 formation in the early morning when the mat was exposed to low light intensities could be explained by nitrogen fixation where H2 is formed as a by-product.