Soils hold a globally important carbon pool that is generally more persistent than the carbon stored in plant biomass. However, this carbon is becoming increasingly vulnerable to disturbances such as soil warming, fire, and erosion. Managing land to increase soil carbon sequestration and persistence may therefore improve long-term soil carbon storage and contribute to climate change mitigation. It has been hypothesized that grazing by large herbivores may enhance the persistence of soil carbon by increasing the amount of soil organic matter forming more stable associations with mineral particles (mineral-associated organic matter). We compared sheep-grazed and ungrazed plots within the Gibson Grazing and Successional Experiment located in the Upper Seeds calcareous grassland in Wytham Woods, Oxfordshire, using organic matter fractionation to estimate the surface (0-5 cm) carbon stocks in the mineral-associated and particulate organic matter fractions. Counter to predictions, after 35 years sheep grazing had not increased mineral-associated organic matter carbon stocks relative to ungrazed plots. We hypothesize that this indicates the saturation of mineral surfaces in both grazed and ungrazed treatments and the inability of grazing to increase soil nitrogen stocks and decrease pH to levels conducive for mineral-associated carbon sequestration. Only one of twelve soil properties examined showed statistically detectable responses to grazing: spring-grazing increased the C:N ratio in the mineral-associated organic matter. While the number of tests performed (24) means this may be a false-positive result, if genuine it would be consistent with a more direct pathway from plant exudates to mineral-associated organic matter formation due to compensatory growth in response to spring-grazing. Overall, the results of this long-term experiment do not support the hypothesis that grazing can improve the persistence of the soil carbon pool. ### Competing Interest Statement The authors have declared no competing interest.

Abstract Climate warming and summer droughts alter soil microbial activity, affecting greenhouse gas (GHG) emissions in Arctic and alpine regions. However, the long-term effects of warming, and implications for future microbial resilience, are poorly understood. Using one alpine and three Arctic soils subjected to in situ long-term experimental warming, we simulated drought in laboratory incubations to test how microbial functional-gene abundance affects fluxes in three GHGs: carbon dioxide, methane, and nitrous oxide. We found that responses of functional gene abundances to drought and warming are strongly associated with vegetation type and soil carbon. Our sites ranged from a wet, forb dominated, soil carbon-rich systems to a drier, soil carbon-poor alpine site. Resilience of functional gene abundances, and in turn methane and carbon dioxide fluxes, was lower in the wetter, carbon-rich systems. However, we did not detect an effect of drought or warming on nitrous oxide fluxes. All gene–GHG relationships were modified by vegetation type, with stronger effects being observed in wetter, forb-rich soils. These results suggest that impacts of warming and drought on GHG emissions are linked to a complex set of microbial gene abundances and may be habitat-specific.

There is widespread concern that cessation of grazing in historically grazed ecosystems is causing biotic homogenization and biodiversity loss. We used 12 montane grassland sites along an 800 km north–south gradient across the UK, to test whether cessation of grazing affects local α- and β-diversity of below-ground food webs. We show cessation of grazing leads to strongly decreased α-diversity of most groups of soil microbes and fauna, particularly of relatively rare taxa. By contrast, the β-diversity varied between groups of soil organisms. While most soil microbial communities exhibited increased homogenization after cessation of grazing, we observed decreased homogenization for soil fauna after cessation of grazing. Overall, our results indicate that exclusion of domesticated herbivores from historically grazed montane grasslands has far-ranging negative consequences for diversity of below-ground food webs. This underscores the importance of grazers for maintaining the diversity of below-ground communities, which play a central role in ecosystem functioning.

It is widely accepted that the measurement of organic and inorganic forms of carbon (C) and nitrogen (N) in soils should be performed on fresh extracts taken from fresh soil samples. However, this is often not possible, and it is common practice to store samples (soils and/or extracts), despite a lack of guidance on best practice. We utilised a case study on a temperate grassland soil taken from different depths to demonstrate how differences in soil and/or soil extract storage temperature (4 or −20 ∘C) and duration can influence sample integrity for the quantification of soil-dissolved organic C and N (DOC and DON), extractable inorganic nitrogen (NH4+ and NO3-) and microbial biomass C and N (MBC and MBN). The appropriateness of different storage treatments varied between topsoils and subsoils, highlighting the need to consider appropriate storage methods based on soil depth and soil properties. In general, we found that storing soils and extracts by freezing at −20 ∘C was least effective at maintaining measured values of fresh material, whilst refrigerating (4 ∘C) soils for less than a week for DOC and DON and up to a year for MBC and MBN and refrigerating soil extracts for less than a week for NH4+ and NO3- did not jeopardise sample integrity. We discuss and provide the appropriate tools to ensure researchers consider best storage practice methods when designing and organising ecological research involving assessments of soil properties related to C and N cycling. We encourage researchers to use standardised methods where possible and to report their storage treatment (i.e. temperature, duration) when publishing findings on aspects of soil and ecosystem functioning. In the absence of published storage recommendations for a given soil type, we encourage researchers to conduct a pilot study and publish their findings.

Mangrove ecosystems are important for carbon storage due to their high productivity and low decomposition rates. Waterways have experienced increased nutrient loads as a result of anthropogenic activities and it is unclear how this may affect carbon and nutrient cycles in downstream mangroves that receive these nutrient-rich waters. Using a laboratory-based incubation experiment, this study aimed to assess the effects of nutrient addition on the diversity and structure of mangrove soil bacterial communities, as well as biomass and activity of the soil microbial community, under different oxygen conditions. Bacterial community diversity and composition was characterised using 16S rRNA gene sequencing and microbial activity was examined through the measurement of microbial respiration and the activities of enzymes associated with organic matter decomposition. Nitrogen addition caused clear shifts in bacterial community composition, with decreases in bacterial diversity and the abundance of sulfate-reducing bacteria. Microbial biomass also decreased with nitrogen addition under reduced oxygen incubations. Changes in bacterial community structure were accompanied by changes in the activity of some enzymes involved in carbon, nitrogen, and phosphorus cycling. Under reduced oxygen conditions, nitrogen addition resulted in a significant increase in the microbial metabolic quotient but no accompanying change in microbial respiration, which was explained by a decrease in microbial biomass. The findings of this study indicate that nitrogen loading has potential implications for microbial communities and carbon and nutrient cycling in mangrove environments that warrant further investigation under field conditions.