Knowledge of how interactions of clay minerals and extracellular enzymes (EEs) influence organic matter turnover in soils are still under discussion. We studied the effect of different montmorillonite contents on EE activities, using two experiments—(1) an adsorption experiment with a commercially available enzyme (α-glucosidase) and (2) an incubation experiment (10 days) where microorganisms were stimulated to produce enzymes through organic carbon (OC) addition (starch and cellulose). Soil mixtures with different montmorillonite contents were created in four levels to a sandy soil: +0% (control), +0.1%, +1%, and +10%. The potential enzyme activity (pEA) of four enzymes, α-glucosidase, β-glucosidase, cellobiohydrolase, and aminopeptidase, involved in the soil carbon and nitrogen cycle were analysed. The adsorption experiment revealed a reduction in the catalytic activity of α-glucosidase by up to 76% with increasing montmorillonite contents. However, the incubation experiment showed an inhibitory effect on pEA only directly after the stimulation of in-situ EE production by OC addition. At later incubation stages, higher pEA was found in soils with higher montmorillonite contents. This mismatch between both experiments, with a transient reduction in catalytic activity for the incubation experiments, points to the continuous production of enzymes by soil microorganisms. It is conceivable that microbial adaptation is characterized by higher investment in EEs production induced by increasing clay contents and a stabilisation of the EEs by clay minerals. Our results point to the need to better understand EE-clay mineral-OC interactions regarding potential microbial adaptations and EE stabilisation with potentially prolonged activities.

We investigated the effects of substrate (cellulose or starch) and different clay contents on the production of microbial extracellular polymeric substances (EPS) and concomitant development of stable soil aggregates. Soils were incubated with different amounts of montmorillonite (+ 0.1%, + 1%, + 10%) both with and without two substrates of contrasting quality (starch and cellulose). Microbial respiration (CO2), biomass carbon (C), EPS-protein, and EPS-polysaccharide were determined over the experimental period. The diversity and compositional shifts of microbial communities (bacteria/archaea) were analysed by sequencing 16S rRNA gene fragments amplified from soil DNA. Soil aggregate size distribution was determined and geometric mean diameter calculated for aggregate formation. Aggregate stabilities were compared among 1–2-mm size fraction. Starch amendment supported a faster increase than cellulose in both respiration and microbial biomass. Microbial community structure and composition differed depending on the C substrate added. However, clay addition had a more pronounced effect on alpha diversity compared to the addition of starch or cellulose. Substrate addition resulted in an increased EPS concentration only if combined with clay addition. At high clay addition, starch resulted in higher EPS concentrations than cellulose. Where additional substrate was not provided, EPS-protein was only weakly correlated with aggregate formation and stability. The relationship became stronger with addition of substrate. Labile organic C thus clearly plays a role in aggregate formation, but increasing clay content was found to enhance aggregate stability and additionally resulted in the development of distinct microbial communities and increased EPS production.

Adsorption of extracellular enzymes to soil minerals is assumed to protect them against degradation, while modifying their activities at the same time. However, the persistence of the activity of adsorbed enzymes remains poorly understood. Therefore, we studied the persistence of cellulase and α-amylase activities after adsorption to soil amended with various amounts (+1, +5, and +10 wt.%) of three typical soil minerals, montmorillonite, kaolinite, and goethite. Soil without mineral addition (pure soil), pure minerals, and pure dissolved enzymes were used as references. Soil mineral–enzyme complexes were prepared and then incubated for 100 days; temporal changes in enzyme activities were analyzed after 0, 0.1, 1, 10, and 100 days. The specific enzyme activities (activities normalized to protein content) and their persistence (activities relative to activities at day 0) were compared to enzyme activities in solution and after sorption to the control soil. Amylase adsorption to pure minerals increased in the following order: montmorillonite > kaolinite > goethite. That of cellulase increased in the following order: goethite > montmorillonite > kaolinite. Adsorption of enzymes to soils did not increase in the same order of magnitude as the addition of reactive binding sites. Based on inverse relationships between the amount of enzyme adsorbed and the specific enzyme activity and their persistency, we showed that a limited availability of sorption sites is important for high specific activity and persistence of the enzymes. This is probably the consequence of less and weaker bonds, as compared to a high availability of sorption sites, resulting in a smaller impact on the active sites of the enzyme. Hence, we suppose that the soil mineral phase supports microorganisms in less-sorptive environments by saving energy on enzyme production, since small enzyme release could already result in sufficient activities to degrade respective target carbon substrates.

Abstract. Microbial processes are one of the key factors driving carbon (C) and nutrient cycling in terrestrial ecosystems, and are strongly driven by the equilibrium between resource availability and demand. In deeply weathered tropical rainforest soils of Africa, it remains unclear whether patterns of microbial processes differ between soils developed from geochemically contrasting parent materials. Here we show that resource availability across soil depths and regions from mafic to felsic geochemistry shape patterns of soil microbial processes. During a 120-day incubation experiment, we found that microbial biomass C and extracellular enzyme activity were highest in the mafic region. Microbial C limitation was highest in the mixed sedimentary region and lowest in the felsic region, which we propose is related to the strength of contrasting C stabilization mechanisms and varying C quality. None of the investigated regions and soil depths showed signs of nitrogen (N) limitation for microbial processes. Microbial phosphorus (P) limitation increased with soil depth but was similar across geochemical regions, indicating that subsoils in the investigated soils were depleted in rock-derived nutrients and are therefore dependent on efficient biological recycling of nutrients. Microbial C limitation was lowest in subsoils, indicating that subsoil microbes can significantly participate in C cycling and limit C storage if increased oxygen availability is prevalent. Using multivariable regressions, we demonstrate that microbial biomass C normalized to soil organic C content (MBCSOC) is controlled by soil geochemistry and substrate quality, while microbial biomass C normalized to soil weight (MBCSoil) is predominantly driven by resource distribution. We conclude that due to differences in resource availability, microbial processes in deeply weathered tropical rainforest soils greatly vary across geochemical regions which must be considered when assessing soil microbial processes in organic matter turnover models.

Background: Soil microbes are key drivers of carbon (C) and nutrient cycling in terrestrial ecosystems, and their properties are influenced by the relationship between resource demand and availability. Aims:Our objective was to investigate patterns of microbial properties and their controls to understand whether they differ between soils derived from geochemically contrasting parent material in tropical montane forests. Methods:We measured microbial biomass C (MBC Soil ), potential extracellular enzyme activity (pEEA), and assessed microbial investments in C and nutrient acquisition at the beginning and end of a 120-day laboratory incubation experiment using soils developed from three geochemically contrasting parent material (i.e., mafic, mixed sediment, and felsic) and three soil depths (0-70 cm). Results:We found that MBC Soil and pEEA were highest in soils developed from the mafic parent material. Microbial investment in C acquisition was highest in soils developed from mixed sedimentary rocks and lowest in soils developed from the felsic parent material.We propose that our findings are related to the strength of contrasting mineral-related C stabilization mechanisms and varying C quality. No predominant microbial investment in nitrogen (N) acquisition was observed, whereas investment in phosphorus (P) acquisition was highest in subsoils. We found lower microbial investment in C acquisition in subsoils indicating relatively high C availability, and that microbes in subsoils can substantially participate in C cycling and limit C storage if moisture and oxygen conditions are suitable.Geochemical soil properties and substrate quality were important controls on MBC Soil per unit soil organic C (MBC SOC ), particularly after the exhaustion of labile and fast cycling C, that is, at the end of the incubation. Conclusion:Although a laboratory incubation experiment cannot reflect real-world conditions, it allowed us to understand how soil properties affect microbial properties. We conclude that parent material is an important driver of microbial properties in tropical montane forests despite the advanced weathering degree of soils.