Current controversies on mechanisms controlling soil carbon storage: implications for interactions with practitioners and policy-makers. A review.

There is currently an intense debate about the potential for additional organic carbon storage in soil, the strategies by which it may be accomplished and what the actual benefits might be for agriculture and the climate. Controversy forms an essential part of the scientific process, but on the topic of soil carbon storage, it may confuse the agricultural community and the general public and may delay actions to fight climate change. In an attempt to shed light on this topic, the originality of this article lies in its intention to provide a balanced description of contradictory scientific opinions on soil carbon storage and to examine how the scientific community can support decision-making despite the controversy. In the first part, we review and attempt to reconcile conflicting views on the mechanisms controlling organic carbon dynamics in soil. We discuss the divergent opinions about chemical recalcitrance, the microbial or plant origin of persistent soil organic matter, the contribution of particulate organic matter to additional organic carbon storage in soil, and the spatial and energetic inaccessibility of soil organic matter to decomposers. In the second part, we examine the advantages and limitations of big data management and modeling, which are essential tools to link the latest scientific theories with the actions taken by stakeholders. Finally, we show how the analysis and discussion of controversies can guide scientists in supporting stakeholders for the design of (i) appropriate trade-offs for biomass use in agriculture and forestry and (ii) climate-smart management practices, keeping in mind their still unresolved effects on soil carbon storage.

High Microbial Diversity Promotes Soil Ecosystem Functioning.

In soil, the link between microbial diversity and carbon transformations is challenged by the concept of functional redundancy. Here, we hypothesized that functional redundancy may decrease with increasing carbon source recalcitrance and that coupling of diversity with C cycling may change accordingly. We manipulated microbial diversity to examine how diversity decrease affects the decomposition of easily degradable (i.e., allochthonous plant residues) versus recalcitrant (i.e., autochthonous organic matter) C sources. We found that a decrease in microbial diversity (i) affected the decomposition of both autochthonous and allochthonous carbon sources, thereby reducing global CO2 emission by up to 40%, and (ii) shaped the source of CO2 emission toward preferential decomposition of most degradable C sources. Our results also revealed that the significance of the diversity effect increases with nutrient availability. Altogether, these findings show that C cycling in soil may be more vulnerable to microbial diversity changes than expected from previous studies, particularly in ecosystems exposed to nutrient inputs. Thus, concern about the preservation of microbial diversity may be highly relevant in the current global-change context assumed to impact soil biodiversity and the pulse inputs of plant residues and rhizodeposits into the soil.IMPORTANCE With hundreds of thousands of taxa per gram of soil, microbial diversity dominates soil biodiversity. While numerous studies have established that microbial communities respond rapidly to environmental changes, the relationship between microbial diversity and soil functioning remains controversial. Using a well-controlled laboratory approach, we provide empirical evidence that microbial diversity may be of high significance for organic matter decomposition, a major process on which rely many of the ecosystem services provided by the soil ecosystem. These new findings should be taken into account in future studies aimed at understanding and predicting the functional consequences of changes in microbial diversity on soil ecosystem services and carbon storage in soil.

Grassland-Cropping Rotations: An Avenue for Agricultural Diversification to Reconcile High Production with Environmental Quality.

A need to increase agricultural production across the world to ensure continued food security appears to be at odds with the urgency to reduce the negative environmental impacts of intensive agriculture. Around the world, intensification has been associated with massive simplification and uniformity at all levels of organization, i.e., field, farm, landscape, and region. Therefore, we postulate that negative environmental impacts of modern agriculture are due more to production simplification than to inherent characteristics of agricultural productivity. Thus by enhancing diversity within agricultural systems, it should be possible to reconcile high quantity and quality of food production with environmental quality. Intensification of livestock and cropping systems separately within different specialized regions inevitably leads to unacceptable environmental impacts because of the overly uniform land use system in intensive cereal areas and excessive N-P loads in intensive animal areas. The capacity of grassland ecosystems to couple C and N cycles through microbial-soil-plant interactions as a way for mitigating the environmental impacts of intensive arable cropping system was analyzed in different management options: grazing, cutting, and ley duration, in order to minimize trade-offs between production and the environment. We suggest that integrated crop-livestock systems are an appropriate strategy to enhance diversity. Sod-based rotations can temporally and spatially capture the benefits of leys for minimizing environmental impacts, while still maintaining periods and areas of intensive cropping. Long-term experimental results illustrate the potential of such systems to sequester C in soil and to reduce and control N emissions to the atmosphere and hydrosphere.

Potential impacts of climate change on the productivity and soil carbon stocks of managed grasslands.

Rain-fed pastoral systems are tightly connected to meteorological conditions. It is, therefore, likely that climate change, including changing atmospheric CO2 concentration, temperature, precipitation and patterns of climate extremes, will greatly affect pastoral systems. However, exact impacts on the productivity and carbon dynamics of these systems are still poorly understood, particularly over longtime scales. The present study assesses the potential effects of future climatic conditions on productivity and soil organic carbon (SOC) stocks of mowed and rotationally grazed grasslands in France. We used the CenW ecosystem model to simulate carbon, water, and nitrogen cycles in response to changes in environmental drivers and management practices. We first evaluated model responses to individual changes in each key meteorological variable to get better insights into the role and importance of each individual variable. Then, we used 3 sets of meteorological variables corresponding to 3 Representative Concentration Pathways (RCP 2.6, RCP 4.5 and RCP 8.5) for long-term model runs from 1975 to 2100. Finally, we used the same three RCPs to analyze the responses of modelled grasslands to extreme climate events. We found that increasing temperature slightly increased grasslands productivities but strongly reduced SOC stocks. A reduction in precipitation led to reductions of biomass and milk production but increased SOC. Conversely, doubling CO2 concentration strongly increased biomass and milk production and marginally reduced SOC. These SOC trends were unexpected. They arose because both increasing precipitation and CO2 increased photosynthetic carbon gain, but they had an even greater effect on the proportion of biomass that could be grazed. The amount of carbon remaining on site and able to contribute to SOC formation was actually reduced under both higher precipitation and CO2. The simulations under the three RCPs indicated that grassland productivity was increased, but that required higher N fertilizer application rates and also led to substantial SOC losses. We thus conclude that, while milk productivity may continue at current rates under climate change, or even increase slightly, there could be some soil C losses over the 21st century. In addition, under the highest-emission scenario, the increasing importance of extreme climate conditions (heat waves and droughts) might render conditions at our site in some years as unsuitable for milk production. It highlights the importance of tailoring farming practices to achieve the dual goals of maintaining agricultural production while safeguarding soil C stocks.

Is plant biomass input driving soil organic matter formation processes in grassland soil under contrasting management?

Grassland management practices vary in stocking rates and plant removal strategies (grazing versus mowing). They influence organic matter (OM) inputs, which were postulated as main controls of soil organic carbon (SOC) sequestration and might therefore control SOC stabilization. The aim of this study was to test this hypothesis by investigating the impacts of grassland harvesting regimes on parameters related to soil microbial functioning and soil organic matter (SOM) formation processes. We used a thirteen-year experiment in Central France under contrasting management (unmanaged, grazing with two intensities, mowing, bare fallow) to establish a carbon input gradient based on biomass leftovers after harvest. We investigated microbial biomass, basal respiration and enzyme activities as indicators of microbial functioning, and amino sugar content and composition as indicator of persistent SOM formation and origin through necromass accumulation. Responses of these parameters to carbon input along the gradient were contrasting and in most cases unrelated. Only the microbial C/N ratio and amino sugar contents showed a linear response indicating that they are influenced by plant-derived OM input. Other parameters were most probably more influenced by root activity, presence of herbivores, and/or physicochemical changes following management activities impacting soil microbial functioning. Grassland harvesting strategies influence SOC sequestration not only by changing carbon input quantity, but also through their effects on belowground processes possibly related to changing carbon input types and physiochemical soil properties.

Land-use intensification differentially affects bacterial, fungal and protist communities and decreases microbiome network complexity.

BACKGROUND: Soil microbial communities are major drivers of cycling of soil nutrients that sustain plant growth and productivity. Yet, a holistic understanding of the impact of land-use intensification on the soil microbiome is still poorly understood. Here, we used a field experiment to investigate the long-term consequences of changes in land-use intensity based on cropping frequency (continuous cropping, alternating cropping with a temporary grassland, perennial grassland) on bacterial, protist and fungal communities as well as on their co-occurrence networks. RESULTS: We showed that land use has a major impact on the structure and composition of bacterial, protist and fungal communities. Grassland and arable cropping differed markedly with many taxa differentiating between both land use types. The smallest differences in the microbiome were observed between temporary grassland and continuous cropping, which suggests lasting effects of the cropping system preceding the temporary grasslands. Land-use intensity also affected the bacterial co-occurrence networks with increased complexity in the perennial grassland comparing to the other land-use systems. Similarly, co-occurrence networks within microbial groups showed a higher connectivity in the perennial grasslands. Protists, particularly Rhizaria, dominated in soil microbial associations, as they showed a higher number of connections than bacteria and fungi in all land uses. CONCLUSIONS: Our findings provide evidence of legacy effects of prior land use on the composition of the soil microbiome. Whatever the land use, network analyses highlighted the importance of protists as a key element of the soil microbiome that should be considered in future work. Altogether, this work provides a holistic perspective of the differential responses of various microbial groups and of their associations to agricultural intensification.

Does the higher root carbon contribution to soil under cropping cycles following grassland conversion also increase shoot biomass?

This study tested the possible root biomass improvements in crop rotations after the conversion of grasslands, and crop samples from maize, winter wheat, and winter barley were collected during 2011-2013 from a long-term experimental site in Lusignan, France (http://www.soere-acbb). Root biomass C quantification was performed using δ13C isotopic signatures to determine the presence of both C3 and C4 plants. We also calculated the recovery rate of maize root biomass C. The results showed that after crop rotations, 0-60 cm root biomass C values were 44.1, 34.2, and 18.7 g C m-2 for maize, winter wheat, and winter barley respectively. The Root biomass C of crops after conversion to grassland was approximately 2-3 times those observed after crop rotations. However, incorporating ley grassland duration into crop rotations showed limited improvements in shoot biomass C and grain yield of the crops, regardless of the decreased rate of N fertilizer for maize. Moreover, root biomass C had a significant relationship with N supply from residues (P < .05). Nevertheless, shoot biomass C of only maize showed significance in its relationships with N supply and root biomass C. In addition, in each 30 cm soil layer (0-30 cm, 30-60 cm, and 60-90 cm), the recovery rate of maize roots decreased to approximately 15% when root biomass C increased to 10 g C m-2. However, further increases in root biomass C had little impact on the recovery rate. In conclusion, compared with continuous cropland, incorporating ley grassland duration into crop rotations increases root biomass C of crops, but this change may not be a significant increase of the shoot biomass C or grain yield. This finding simply indicates the improved C input from crops and the potential to increase soil organic C, as well as providing a model for the sustainability of crop rotations.

Legacy effects of temporary grassland in annual crop rotation on soil ecosystem services.

The introduction of temporary grassland into an annual crop rotation is recognized to improve soil ecosystem services, and resulting legacies can be beneficial for the following crops. In this context, the aim of the present study was to evaluate legacy effects of introducing temporary grassland into an annual crop rotation on five ecosystem services (i) soil structure maintenance (aggregate stability), (ii) water regulation (saturated hydraulic conductivity), (iii) biodiversity conservation (microbial biomass and microbial metabolic activity, as well as microorganism, enchytraeid, springtail and earthworm communities), (iv) pathogen regulation (soil suppressiveness to Verticillium dahliae), and (v) forage production and quality. Three crop rotation schemes, maintained for twelve years, were compared in four random blocks, one being an annual crop rotation without grassland (0%), another with a medium percentage of grassland (50%, corresponding to 3 years of continuous grassland in the crop rotation), and a third one with a high percentage of grassland in the crop rotation (75%, corresponding to 6 years of continuous grassland in the crop rotation). The results showed that the grassland introduction into an annual crop rotation improved, whatever the duration of the grassland, soil structure maintenance and biodiversity conservation, while it decreased pathogen regulation and did not modify water regulation. Comparing the two crop rotations that included grassland, indicated a stronger beneficial grassland legacy effect for the higher proportion of grassland concerning soil structure maintenance and biodiversity conservation. By contrast, water regulation, pathogen regulation and forage production were not affected by the legacy of the 75% grassland during the rotation. Overall, our findings demonstrated the extent to which grassland legacies are affecting the current state of soil properties and possible ecosystem services provided. To improve ecosystem services, soil management should take legacy effects into account and consider longer timeframes to apply beneficial practices.

Combining eddy covariance measurements with process-based modelling to enhance understanding of carbon exchange rates of dairy pastures.

Many temperate grasslands are used for dairying, and ongoing research aims to better understand these systems in order to increase animal production and soil organic carbon (SOC) stocks. However, it is difficult to fully understand management effects on SOC because most changes are slow and difficult to distinguish from natural variability, even if changes are important over years to decades. Eddy covariance (EC) measurements can overcome this problem by continuously measuring net carbon exchange from pastures, but net balances are very sensitive to even small systematic measurement errors. Combining EC measurements with detailed process-based modelling can reduce the risks inherent in total reliance on EC measurements. Modelling can also reveal information about the underlying processes that drive observed fluxes. Here, we describe carbon exchange patterns of five paddocks situated at four different locations in New Zealand and France where EC data and detailed physiological modelling were available. The work showed that respiration by grazing animals was often only incompletely captured in EC measurements. This was most problematic when fluxes were based on gap-filling, which could have estimated incorrect fluxes during grazing periods based on observations from periods without grazing. We then aimed to extract plant physiological insights from these studies. We found appreciable carbon uptake rates even at temperatures below 0 °C. After grazing, carbon uptake was reduced for up to 2 weeks. This reduction was larger than expected from reduced leaf area after grazing, but the factors contributing to that difference have not yet been identified. Detailed physiological models can also extrapolate findings to new management regimes, environmental conditions or plant attributes. This overcomes the limitation of experimental studies, which are necessarily restricted to actual site and weather conditions allowing models to make further progress on predicting management effects on SOC.

Temperature sensitivity of decomposition decreases with increasing soil organic matter stability.

Evaluation of the temperature sensitivity of soil organic matter (SOM) decomposition is critical for forecasting whether soils in a warming world will lose or gain carbon and, therefore, accelerate or mitigate climate warming. It is usually described, using Arrhenius kinetics, as increasing with the stability of the substrate in laboratory conditions, where substrate availability is non-limiting and where chemical recalcitrance, therefore, predominantly regulates stability. However, conditions of non-limiting subtrate availability are rare in the undisturbed soil, where physicochemical protection of substrates may control their stability. The aim of this study was to assess the temperature sensitivity of decomposition of SOM with contrasting stability in the field. Our conceptual approach was based on in situ measurements of soil CO2 efflux at a range of temperatures from root exclusion plots of increasing age (1 month and three decades) and, therefore, with SOM of increasing stability. From a set of short-term measurements in spring, using diurnal temperature variation, the relative temperature sensitivity of SOM decomposition decreased significantly (p < 0.0001) with increasing SOM stability, and was weak (Q10 < 1.3) in long-term root exclusion plots. This result was confirmed in a similar set of short-term measurements repeated later in the year, in summer, as well as from an analysis perfomed at the seasonal timscale. We provide direct field evidence that the temperature sensitivity of SOM decomposition decreases with increasing stability, in direct contrast with Arrhenius kinetics prediction, and therefore show that stability of SOM in the field cannot be the sole result of chemical recalcitrance. We conclude that the physicochemical protection of SOM, which controls SOM stability in the field, constrains the temperature sensitivity of SOM decomposition under field conditions.

Ozone flux in plant ecosystems: new opportunities for long-term monitoring networks to deliver ozone-risk assessments.

Ozone (O3) is a photochemically formed reactive gas responsible for a decreasing carbon assimilation in plant ecosystems. Present in the atmosphere in trace concentrations (less than 100 ppbv), this molecule is capable of inhibiting carbon assimilation in agricultural and forest ecosystems. Ozone-risk assessments are typically based on manipulative experiments. Present regulations regarding critical ozone levels are mostly based on an estimated accumulated exposure over a given threshold concentration. There is however a scientific consensus over flux estimates being more accurate, because they include plant physiology analyses and different environmental parameters that control the uptake-that is, not just the exposure-of O3. While O3 is a lot more difficult to measure than other non-reactive greenhouse gases, UV-based and chemiluminescence sensors enable precise and fast measurements and are therefore highly desirable for eddy covariance studies. Using micrometeorological techniques in association with latent heat flux measurements in the field allows for the partition of ozone fluxes into the stomatal and non-stomatal sinks along the soil-plant continuum. Long-term eddy covariance measurements represent a key opportunity in estimating carbon assimilation at high-temporal resolutions, in an effort to study the effect of climate change on photosynthetic mechanisms. Our aim in this work is to describe potential of O3 flux measurement at the canopy level for ozone-risk assessment in established long-term monitoring networks.

Modelling nitrous oxide emissions from mown-grass and grain-cropping systems: Testing and sensitivity analysis of DailyDayCent using high frequency measurements.

The DailyDayCent biogeochemical model was used to simulate nitrous oxide (N2O) emissions from two contrasting agro-ecosystems viz. a mown-grassland and a grain-cropping system in France. Model performance was tested using high frequency measurements over three years; additionally a local sensitivity analysis was performed. Annual N2O emissions of 1.97 and 1.24kgNha-1year-1 were simulated from mown-grassland and grain-cropland, respectively. Measured and simulated water filled pore space (r=0.86, ME=-2.5%) and soil temperature (r=0.96, ME=-0.63°C) at 10cm soil depth matched well in mown-grassland. The model predicted cumulative hay and crop production effectively. The model simulated soil mineral nitrogen (N) concentrations, particularly ammonium (NH4+), reasonably, but the model significantly underestimated soil nitrate (NO3-) concentration under both systems. In general, the model effectively simulated the dynamics and the magnitude of daily N2O flux over the whole experimental period in grain-cropland (r=0.16, ME=-0.81gNha-1day-1), with reasonable agreement between measured and modelled N2O fluxes for the mown-grassland (r=0.63, ME=-0.65gNha-1day-1). Our results indicate that DailyDayCent has potential for use as a tool for predicting overall N2O emissions in the study region. However, in-depth analysis shows some systematic discrepancies between measured and simulated N2O fluxes on a daily basis. The current exercise suggests that the DailyDayCent may need improvement, particularly the sub-module responsible for N transformations, for better simulating soil mineral N, especially soil NO3- concentration, and N2O flux on a daily basis. The sensitivity analysis shows that many factors such as climate change, N-fertilizer use, input uncertainty and parameter value could influence the simulation of N2O emissions. Sensitivity estimation also helped to identify critical parameters, which need careful estimation or site-specific calibration for successful modelling of N2O emissions in the study region.

Meta-barcoded evaluation of the ISO standard 11063 DNA extraction procedure to characterize soil bacterial and fungal community diversity and composition.

This study was designed to assess the influence of three soil DNA extraction procedures, namely the International Organization for Standardization (ISO-11063, GnS-GII and modified ISO procedure (ISOm), on the taxonomic diversity and composition of soil bacterial and fungal communities. The efficacy of each soil DNA extraction method was assessed on five soils, differing in their physico-chemical characteristics and land use. A meta-barcoded pyrosequencing approach targeting 16S and 18S rRNA genes was applied to characterize soil microbial communities. We first observed that the GnS-GII introduced some heterogeneity in bacterial composition between replicates. Then, although no major difference was observed between extraction procedures for soil bacterial diversity, we saw that the number of fungal genera could be underestimated by the ISO-11063. In particular, this procedure underestimated the detection in several soils of the genera Cryptococcus, Pseudallescheria, Hypocrea and Plectosphaerella, which are of ecological interest. Based on these results, we recommend using the ISOm method for studies focusing on both the bacterial and fungal communities. Indeed, the ISOm procedure provides a better evaluation of bacterial and fungal communities and is limited to the modification of the mechanical lysis step of the existing ISO-11063 standard.

Land Use History Shifts In Situ Fungal and Bacterial Successions following Wheat Straw Input into the Soil.

Soil microbial communities undergo rapid shifts following modifications in environmental conditions. Although microbial diversity changes may alter soil functioning, the in situ temporal dynamics of microbial diversity is poorly documented. Here, we investigated the response of fungal and bacterial diversity to wheat straw input in a 12-months field experiment and explored whether this response depended on the soil management history (grassland vs. cropland). Seasonal climatic fluctuations had no effect on the diversity of soil communities. Contrastingly fungi and bacteria responded strongly to wheat regardless of the soil history. After straw incorporation, diversity decreased due to the temporary dominance of a subset of copiotrophic populations. While fungi responded as quickly as bacteria, the resilience of fungal diversity lasted much longer, indicating that the relative involvement of each community might change as decomposition progressed. Soil history did not affect the response patterns, but determined the identity of some of the populations stimulated. Most strikingly, the bacteria Burkholderia, Lysobacter and fungi Rhizopus, Fusarium were selectively stimulated. Given the ecological importance of these microbial groups as decomposers and/or plant pathogens, such regulation of the composition of microbial successions by soil history may have important consequences in terms of soil carbon turnover and crop health.

Stability of soil microbial structure and activity depends on microbial diversity.

Despite the central role of microbes in soil processes, empirical evidence concerning the effect of their diversity on soil stability remains controversial. Here, we addressed the ecological insurance hypothesis by examining the stability of microbial communities along a gradient of soil microbial diversity in response to mercury pollution and heat stress. Diversity was manipulated by dilution extinction approach. Structural and functional stabilities of microbial communities were assessed from patterns of genetic structure and soil respiration after the stress. Dilution led to the establishment of a consistent diversity gradient, as revealed by 454 sequencing of ribosomal genes. Diversity stability was enhanced in species-rich communities whatever the stress whereas functional stability was improved with increasing diversity after heat stress, but not after mercury pollution. This discrepancy implies that the relevance of ecological insurance for soil microbial communities might depend on the type of stress. Our results also suggest that the significance of microbial diversity for soil functional stability might increase with available soil resources. This could have strong repercussions in the current 'global changes' context because it suggests that the combined increased frequencies of extreme climatic events, nutrient loading and biotic exploitation may amplify the functional consequences of diversity decrease.

Current status, uncertainty and future needs in soil organic carbon monitoring.

Increasing human demands on soil-derived ecosystem services requires reliable data on global soil resources for sustainable development. The soil organic carbon (SOC) pool is a key indicator of soil quality as it affects essential biological, chemical and physical soil functions such as nutrient cycling, pesticide and water retention, and soil structure maintenance. However, information on the SOC pool, and its temporal and spatial dynamics is unbalanced. Even in well-studied regions with a pronounced interest in environmental issues information on soil carbon (C) is inconsistent. Several activities for the compilation of global soil C data are under way. However, different approaches for soil sampling and chemical analyses make even regional comparisons highly uncertain. Often, the procedures used so far have not allowed the reliable estimation of the total SOC pool, partly because the available knowledge is focused on not clearly defined upper soil horizons and the contribution of subsoil to SOC stocks has been less considered. Even more difficult is quantifying SOC pool changes over time. SOC consists of variable amounts of labile and recalcitrant molecules of plant, and microbial and animal origin that are often operationally defined. A comprehensively active soil expert community needs to agree on protocols of soil surveying and lab procedures towards reliable SOC pool estimates. Already established long-term ecological research sites, where SOC changes are quantified and the underlying mechanisms are investigated, are potentially the backbones for regional, national, and international SOC monitoring programs.

Evaluation of the ISO standard 11063 DNA extraction procedure for assessing soil microbial abundance and community structure.

Soil DNA extraction has become a critical step in describing microbial biodiversity. Historically, ascertaining overarching microbial ecological theories has been hindered as independent studies have used numerous custom and commercial DNA extraction procedures. For that reason, a standardized soil DNA extraction method (ISO-11063) was previously published. However, although this ISO method is suited for molecular tools such as quantitative PCR and community fingerprinting techniques, it has only been optimized for examining soil bacteria. Therefore, the aim of this study was to assess an appropriate soil DNA extraction procedure for examining bacterial, archaeal and fungal diversity in soils of contrasting land-use and physico-chemical properties. Three different procedures were tested: the ISO-11063 standard; a custom procedure (GnS-GII); and a modified ISO procedure (ISOm) which includes a different mechanical lysis step (a FastPrep ®-24 lysis step instead of the recommended bead-beating). The efficacy of each method was first assessed by estimating microbial biomass through total DNA quantification. Then, the abundances and community structure of bacteria, archaea and fungi were determined using real-time PCR and terminal restriction fragment length polymorphism approaches. Results showed that DNA yield was improved with the GnS-GII and ISOm procedures, and fungal community patterns were found to be strongly dependent on the extraction method. The main methodological factor responsible for differences between extraction procedure efficiencies was found to be the soil homogenization step. For integrative studies which aim to examine bacteria, archaea and fungi simultaneously, the ISOm procedure results in higher DNA recovery and better represents microbial communities.