Despite progress in understanding microbial biogeography of surface soils, few studies have investigated depth-dependent distributions of terrestrial microorganisms in subsoils. We leveraged high-throughput sequencing of 16S rRNA genes obtained from soils collected from the RARE: Charitable Research Reserve (Cambridge, ON, Canada) to assess the influence of depth on bacterial communities across various land-use types. Although bacterial communities were strongly influenced by depth across all sites, the magnitude of this influence was variable and demonstrated that land-use attributes also played a significant role in shaping soil bacterial communities. Soil pH exhibited a large gradient across samples and strongly influenced shifts in bacterial communities with depth and across different land-use systems, especially considering that physicochemical conditions showed generally consistent trends with depth. We observed significant (p ≤ 0.001) and strongly correlated taxa with depth and pH, with a strong predominance of positively depth-correlated OTUs without cultured representatives. These findings highlight the importance of depth in soil biogeographical surveys and that subsurface soils harbour understudied bacterial members with potentially unique and important functions in deeper soil horizons that remain to be characterized.

Although Thaumarchaeota are important contributors to ammonia oxidation in terrestrial habitats, distributions of ammonia oxidizers along soil depth profiles are poorly understood, especially in relation to distinct land usages. Leveraging the close proximity of forest, field and agricultural plots at the RARE: Charitable Research Reserve, we examined soil thaumarchaeotal biogeography at three different depths (0-15, 15-30 and 30-45 cm) from plots within areas of contrasting land usage. Data from high-throughput sequencing of thaumarchaeotal 16S rRNA gene sequences demonstrated that OTU richness was affected significantly by depth and land-use type. Specifically, thaumarchaeotal diversity was higher in soils from forest sites than from field sites, and lower within 0-15 cm soils than either 15-30 cm or 30-45 cm soils. Soil land-use type influenced the relative abundance of the Soil Crenarchaeota Group (SCG), with a lower relative abundance of SCG in forest sites compared to field sites. At the OTU level, thaumarchaeotal communities changed with increasing soil depth for agricultural soils, in contrast to homogeneous depth profiles generated from forest site samples. Soil pH was the strongest factor impacting thaumarchaeotal community composition and, importantly, the evenness of archaeal taxa. Nitrogen, carbon and soil texture shaped thaumarchaeotal community composition among field site samples.

The soil microbial community (SMC) and soil organic matter (SOM) are inherently related and are sensitive to land‐use changes. Microorganisms regulate essential soil functions that are key to SOM dynamics, whereas SOM dynamics define the SMC. To expand our understanding of soil health, we evaluated biological and SOM indicators in long‐term (18‐yr) continuous silage corn (Zea mays L.), continuous soybean [Glycine max (L.) Merr.], and perennial grass ecosystems in Ontario, Canada. The SMC was evaluated via ester‐linked fatty acid methyl ester (EL‐FAME) and amplicon sequencing. Soil organic matter was evaluated via a new combined enzyme assay that provides a single biogeochemical cycling value for C, N, P, and S cycling activity (CNPS), as well as loss‐on‐ignition, permanganate oxidizable C (POXC), and total C and N. Overall, soil health indicators followed the trend of grasses > corn > soybean. Grass systems had up to 8.1 times more arbuscular mycorrhizal fungi, increased fungal/bacteria ratios (via EL‐FAME), and higher microbial diversity (via sequencing). The POXC was highly variable within treatments and did not significantly differ between systems. The novel CNPS activity assay, however, was highly sensitive to management (up to 2.2 and 3.2 times higher under grasses than corn and soybean, respectively) and was positively correlated (ρ > .92) to SOM, total C, and total N. Following the “more is better” model, where higher values of the measured parameters indicate a healthier soil, our study showed decreased soil health under monocultures, especially soybean, and highlights the need to implement sustainable agriculture practices that maintain soil health.

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