Inferences on soil biogeochemical processes based on metagenomic profiles is a challenging task due to enormous diversity of soil microbes and the fragile linkage between gene abundance and functioning. Here we used the biological sink of H2 as a case study to test the hypothesis that [NiFe]-hydrogenase gene distribution and expression profiles explain variations in H2 oxidation rate measured in soil collected in poplar monoculture, larch plantation and farmland. Shotgun metagenomic and metatranscriptomic analyses of soil samples exposed to elevated or low H2 concentration led to the identification of 45 genes encoding the large subunit of [NiFe]-hydrogenases belonging to 8 distinct phyla. Our results indicate that despite significant sequencing effort, retrieved hydrogenase sequences are not in themselves adequate surrogates of H2 oxidation activity in these soils. In fact, land-use exerted a greater influence than H2 exposure on both hydrogenase gene distribution and expression though expression of certain genes responded to H2. We argue that approaches relying on PCR/RT-PCR amplicon sequencing or quantification combined with physicochemical parameters are currently the best option to infer the activity of H2-oxidizing bacteria and probably other specialist functional guilds with similar population size in soil.

The enrichment of H₂-oxidizing bacteria (HOB) by H₂ generated by nitrogen-fixing nodules has been shown to have a fertilization effect on several different crops. The benefit of HOB is attributed to their production of plant growth-promoting factors, yet their interactions with other members of soil microbial communities have received little attention. Here we report that the energy potential of H₂, when supplied to soil, alters ecological niche partitioning of bacteria and fungi, with multifaceted consequences for both generalist and specialist microbial functions. We used dynamic microcosms to expose soil to the typical atmospheric H₂ mixing ratio (0.5 ppmv) permeating soils, as well as mixing ratios comparable to those found at the soil-nodule interface (10,000 ppmv). Elevated H₂ exposure exerted direct effects on two HOB subpopulations distinguished by their affinity for H₂ while enhancing community level carbon substrate utilization potential and lowering CH₄ uptake activity in soil. We found that H₂ triggered changes in the abundance of microorganisms that were reproducible yet inconsistent across soils at the taxonomic level and even among HOB. Overall, H₂ exposure altered microbial process rates at an intensity that depends upon soil abiotic and biotic features. We argue that further examination of direct and indirect effects of H₂ on soil microbial communities will lead to a better understanding of the H₂ fertilization effect and soil biogeochemical processes.IMPORTANCE An innovative dynamic microcosm chamber system was used to demonstrate that H₂ diffusing in soil triggers changes in the distribution of HOB and non-HOB. Although the response was uneven at the taxonomic level, an unexpected coordinated response of microbial functions was observed, including abatement of CH₄ oxidation activity and stimulation of carbon turnover. Our work suggests that elevated H₂ rewires soil biogeochemical structure through a combination of direct effects on the growth and persistence of HOB and indirect effects on a variety of microbial processes involving HOB and non-HOB.