The importance of accounting method and sampling depth to estimate changes in soil carbon stocks.

BACKGROUND: As interest in the voluntary soil carbon market surges, carbon registries have been developing new soil carbon measurement, reporting, and verification (MRV) protocols. These protocols are inconsistent in their approaches to measuring soil organic carbon (SOC). Two areas of concern include the type of SOC stock accounting method (fixed-depth (FD) vs. equivalent soil mass (ESM)) and sampling depth requirement. Despite evidence that fixed-depth measurements can result in error because of changes in soil bulk density and that sampling to 30 cm neglects a significant portion of the soil profile's SOC stock, most MRV protocols do not specify which sampling method to use and only require sampling to 30 cm. Using data from UC Davis's Century Experiment ("Century") and UW Madison's Wisconsin Integrated Cropping Systems Trial (WICST), we quantify differences in SOC stock changes estimated by FD and ESM over 20 years, investigate how sampling at-depth (> 30 cm) affects SOC stock change estimates, and estimate how crediting outcomes taking an empirical sampling-only crediting approach differ when stocks are calculated using ESM or FD at different depths. RESULTS: We find that FD and ESM estimates of stock change can differ by over 100 percent and that, as expected, much of this difference is associated with changes in bulk density in surface soils (e.g., r = 0.90 for Century maize treatments). This led to substantial differences in crediting outcomes between ESM and FD-based stocks, although many treatments did not receive credits due to declines in SOC stocks over time. While increased variability of soils at depth makes it challenging to accurately quantify stocks across the profile, sampling to 60 cm can capture changes in bulk density, potential SOC redistribution, and a larger proportion of the overall SOC stock. CONCLUSIONS: ESM accounting and sampling to 60 cm (using multiple depth increments) should be considered best practice when quantifying change in SOC stocks in annual, row crop agroecosystems. For carbon markets, the cost of achieving an accurate estimate of SOC stocks that reflect management impacts on soils at-depth should be reflected in the price of carbon credits.

Persistent soil carbon enhanced in Mollisols by well-managed grasslands but not annual grain or dairy forage cropping systems.

Intensive crop production on grassland-derived Mollisols has liberated massive amounts of carbon (C) to the atmosphere. Whether minimizing soil disturbance, diversifying crop rotations, or re-establishing perennial grasslands and integrating livestock can slow or reverse this trend remains highly uncertain. We investigated how these management practices affected soil organic carbon (SOC) accrual and distribution between particulate (POM) and mineral-associated (MAOM) organic matter in a 29-y-old field experiment in the North Central United States and assessed how soil microbial traits were related to these changes. Compared to conventional continuous maize monocropping with annual tillage, systems with reduced tillage, diversified crop rotations with cover crops and legumes, or manure addition did not increase total SOC storage or MAOM-C, whereas perennial pastures managed with rotational grazing accumulated more SOC and MAOM-C (18 to 29% higher) than all annual cropping systems after 29 y of management. These results align with a meta-analysis of data from published studies comparing the efficacy of soil health management practices in annual cropping systems on Mollisols worldwide. Incorporating legumes and manure into annual cropping systems enhanced POM-C, microbial biomass, and microbial C-use efficiency but did not significantly increase microbial necromass accumulation, MAOM-C, or total SOC storage. Diverse, rotationally grazed pasture management has the potential to increase persistent soil C on Mollisols, highlighting the key role of well-managed grasslands in climate-smart agriculture.

The soil microbial carbon pump: From conceptual insights to empirical assessments.

The global soil carbon (C) pool is massive, so relatively small changes in soil organic carbon (SOC) stocks can significantly alter atmospheric C and global climate. The recently proposed concept of the soil microbial carbon pump (MCP) emphasizes the active role of soil microbes in SOC storage by integrating the continual microbial transformation of organic C from labile to persistent anabolic forms. However, the concept has not been evaluated with data. Here, we combine datasets, including microbial necromass biomarker amino sugars and SOC, from two long-term agricultural field studies conducted by large United States bioenergy research programs. We interrogate the soil MCP concept by investigating the asynchronous responses of microbial necromass and SOC to land-use change. Microbial necromass appeared to preferentially accumulate in soil and be the dominant contributor to SOC accrual in diversified perennial bioenergy crops. Specifically, ~92% of the additional SOC enhanced by plant diversity was estimated to be microbial necromass C, and >76% of the additional SOC enhanced by land-use transition from annual to perennial crops was estimated to be microbial necromass. This suggests that the soil MCP was stimulated in diversified perennial agroecosystems. We further delineate and suggest two parameters-soil MCP capacity and efficacy-reflecting the conversion of plant C into microbial necromass and the contribution of microbial necromass to SOC, respectively, that should serve as valuable metrics for future studies evaluating SOC storage under alternative management in changing climates.

Community Structure of Arbuscular Mycorrhizal Fungi in Soils of Switchgrass Harvested for Bioenergy.

Learning more about the biodiversity and composition of arbuscular mycorrhizal fungi (AMF) under alternative agricultural management scenarios may be important to the sustainable intensification of switchgrass grown as a bioenergy crop. Using PacBio single-molecule sequencing and taxonomic resolution to the level of amplicon sequence variant (ASV), we assessed the effects of nitrogen amendment on AMF associating with switchgrass and explored relationships between AMF and switchgrass yield across three sites of various productivities in Wisconsin. Nitrogen amendment had little effect on AMF diversity metrics or community composition. While AMF ASV diversity was not correlated with switchgrass yield, AMF family richness and switchgrass yield had a strong, positive relationship at one of our three sites. Each of our sites was dominated by unique ASVs of the species Paraglomus brasilianum, indicating regional segregation of AMF at the intraspecific level. Our molecular biodiversity survey identified putative core members of the switchgrass microbiome, as well as novel clades of AMF, especially in the order Paraglomerales and the genus Nanoglomus Furthermore, our phylogenies unite the cosmopolitan, soil-inhabiting clade deemed GS24 with Pervetustaceae, an enigmatic family prevalent in stressful environments. Future studies should isolate and characterize the novel genetic diversity found in switchgrass agroecosystems and explore the potential yield benefits of AMF richness.IMPORTANCE We assessed the different species of beneficial fungi living in agricultural fields of switchgrass, a large grass grown for biofuels, using high-resolution DNA sequencing. Contrary to our expectations, the fungi were not greatly affected by fertilization. However, we found a positive relationship between plant productivity and the number of families of beneficial fungi at one site. Furthermore, we sequenced many species that could not be identified with existing reference databases. One group of fungi was highlighted in an earlier study for being widely distributed but of unknown taxonomy. We discovered that this group belonged to a family called Pervetustaceae, which may benefit switchgrass in stressful environments. To produce higher-yielding switchgrass in a more sustainable manner, it could help to study these undescribed fungi and the ways in which they may contribute to greater switchgrass yield in the absence of fertilization.

Soil nitrate leaching under grazed cool-season grass pastures of the North Central US.

Nitrate leaching from agricultural cropping systems contributes to widespread and devastating eutrophication of water bodies globally. In the North Central USA, this problem is acute, with millions of dollars spent annually in efforts to clean up recreational and drinking water. The frequent soil disturbance and exogenous nitrogen (N) amendments applied in annual cropping systems make them major sources of ground- and surface-water nitrate pollution. Perennial grasslands under managed livestock grazing have been touted for their ability to retain soils and nutrients while simultaneously providing milk and meat to society. The present study provides an evaluation of the peer-reviewed literature addressing nitrate leaching loads beneath corn, pasture and prairie in temperate humid and sub-humid regions of the US, with a focus on cool-season grass pastures. Inputs of exogenous N to these agroecosystems comes from wet and dry deposition, livestock manure from imported feed, biological fixation and inorganic N fertilizer. Nitrate loads were highest beneath corn and lowest beneath restored prairie and switchgrass managed for bioenergy. Cool-season grass pastures had relatively low levels of nitrate leaching loads where little or no N was applied. However, where grazed perennial grasslands had inorganic N applied, nitrate leaching loads rivaled those of corn in some cases. When producing milk and meat from livestock, grazed perennial cool-season grass pastures should reduce nitrate leaching loads compared to growing corn that is used to feed livestock in confinement. However, cool-season grass pastures can lose significant nitrate to leaching with moderate- to high-levels of exogenous N inputs. © 2020 Society of Chemical Industry.

Empirical Evidence for the Potential Climate Benefits of Decarbonizing Light Vehicle Transport in the U.S. with Bioenergy from Purpose-Grown Biomass with and without BECCS.

Climate mitigation scenarios limiting global temperature increases to 1.5 °C rely on decarbonizing vehicle transport with bioenergy production plus carbon capture and storage (BECCS), but climate impacts for producing different bioenergy feedstocks have not been directly compared experimentally or for ethanol vs electric light-duty vehicles. A field experiment at two Midwest U.S. sites on contrasting soils revealed that feedstock yields of seven potential bioenergy cropping systems varied substantially within sites but little between. Bioenergy produced per hectare reflected yields: miscanthus > poplar > switchgrass > native grasses ≈ maize stover (residue) > restored prairie ≈ early successional. Greenhouse gas emission intensities for ethanol vehicles ranged from 20 to -179 g CO2e MJ-1: maize stover ≫ miscanthus ≈ switchgrass ≈ native grasses ≈ poplar > early successional ≥ restored prairie; direct climate benefits ranged from ∼80% (stover) to 290% (restored prairie) reductions in CO2e compared to petroleum and were similar for electric vehicles. With carbon capture and storage (CCS), reductions in emission intensities ranged from 204% (stover) to 416% (restored prairie) for ethanol vehicles and from 329 to 558% for electric vehicles, declining 27 and 15%, respectively, once soil carbon equilibrates within several decades of establishment. Extrapolation based on expected U.S. transportation energy use suggests that, once CCS potential is maximized with CO2 pipeline infrastructure, negative emissions from bioenergy with CCS for light-duty electric vehicles could capture >900 Tg CO2e year-1 in the U.S. In the future, as other renewable electricity sources become more important, electricity production from biomass would offset less fossil fuel electricity, and the advantage of electric over ethanol vehicles would decrease proportionately.

Inorganic N addition replaces N supplied to switchgrass (Panicum virgatum) by arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal fungi (AMF) provide many benefits in agroecosystems including improved soil tilth, carbon sequestration, and water and nutrient transfer to plants. AMF are known to affect plant nitrogen (N) dynamics and transfer N to plants, but there have been few studies addressing whether the amount of N transferred to plants by AMF is agronomically relevant. We used δ15 N natural abundance methods and δ15 N mass balance equations to estimate the amount of plant N derived from AMF transfer in perennial grasses managed for bioenergy production under different N addition treatments (0, 56, and 196 kg N/ha). Differentiation of δ15 N among plant, soil N, and AMF pools was higher than anticipated leading to calculations of 34-55% of plant N transferred by AMF in the treatments receiving no N addition to 6-22% of plant N transferred to plants in high-N addition treatments. AMF extra-radical hyphae biomass was significantly reduced in the high-N (196 kg N/ha) addition treatments, which was negatively correlated to enriched plant δ15 N. Our results suggest that N addition decreases AMF N transfer to plants. When N was limiting to plant growth, AMF supplied agronomically significant amounts of plant N, and a higher proportion of overall plant N. Because differentiation between N pools was greater than expected, stable isotope measurements can be used to estimate N transfer to AMF plant hosts.

Microbial Community Structure and Functional Potential in Cultivated and Native Tallgrass Prairie Soils of the Midwestern United States.

The North American prairie covered about 3.6 million-km2 of the continent prior to European contact. Only 1-2% of the original prairie remains, but the soils that developed under these prairies are some of the most productive and fertile in the world, containing over 35% of the soil carbon in the continental United States. Cultivation may alter microbial diversity and composition, influencing the metabolism of carbon, nitrogen, and other elements. Here, we explored the structure and functional potential of the soil microbiome in paired cultivated-corn (at the time of sampling) and never-cultivated native prairie soils across a three-states transect (Wisconsin, Iowa, and Kansas) using metagenomic and 16S rRNA gene sequencing and lipid analysis. At the Wisconsin site, we also sampled adjacent restored prairie and switchgrass plots. We found that agricultural practices drove differences in community composition and diversity across the transect. Microbial biomass in prairie samples was twice that of cultivated soils, but alpha diversity was higher with cultivation. Metagenome analyses revealed denitrification and starch degradation genes were abundant across all soils, as were core genes involved in response to osmotic stress, resource transport, and environmental sensing. Together, these data indicate that cultivation shifted the microbiome in consistent ways across different regions of the prairie, but also suggest that many functions are resilient to changes caused by land management practices - perhaps reflecting adaptations to conditions common to tallgrass prairie soils in the region (e.g., soil type, parent material, development under grasses, temperature and rainfall patterns, and annual freeze-thaw cycles). These findings are important for understanding the long-term consequences of land management practices to prairie soil microbial communities and their genetic potential to carry out key functions.

Associative nitrogen fixation (ANF) in switchgrass (Panicum virgatum) across a nitrogen input gradient.

Associative N fixation (ANF), the process by which dinitrogen gas is converted to ammonia by bacteria in casual association with plants, has not been well-studied in temperate ecosystems. We examined the ANF potential of switchgrass (Panicum virgatum L.), a North American prairie grass whose productivity is often unresponsive to N fertilizer addition, via separate short-term 15N2 incubations of rhizosphere soils and excised roots four times during the growing season. Measurements occurred along N fertilization gradients at two sites with contrasting soil fertility (Wisconsin, USA Mollisols and Michigan, USA Alfisols). In general, we found that ANF potentials declined with long-term N addition, corresponding with increased soil N availability. Although we hypothesized that ANF potential would track plant N demand through the growing season, the highest root fixation rates occurred after plants senesced, suggesting that root diazotrophs exploit carbon (C) released during senescence, as C is translocated from aboveground tissues to roots for wintertime storage. Measured ANF potentials, coupled with mass balance calculations, suggest that ANF appears to be an important source of N to unfertilized switchgrass, and, by extension, to temperate grasslands in general.

Carbon storage potential increases with increasing ratio of C4 to C3 grass cover and soil productivity in restored tallgrass prairies.

Long-term soil carbon (C) storage is essential for reducing CO2 in the atmosphere. Converting unproductive and environmentally sensitive agricultural lands to grasslands for bioenergy production may enhance C storage. However, a better understanding of the interacting effects of grass functional composition (i.e., relative abundance of C4 and C3 grass cover) and soil productivity on C storage will help guide sustainable grassland management. Our objective was to examine the relationship between grass functional composition and potential C storage and how it varies with potential soil productivity. We estimated C inputs from above- and belowground net primary productivity (ANPP and BNPP), and heterotrophic respiration (R H) to calculate net ecosystem production (NEP), a measure of potential soil C storage, in grassland plots of relatively high- and low-productivity soils spanning a gradient in the ratio of C4 to C3 grass cover (C4:C3). NEP increased with increasing C4:C3, but only in potentially productive soils. The positive relationship likely stemmed from increased ANPP, rather than BNPP, which was possibly related to efficient resource-use and physiological/anatomical advantages of C4 plants. R H was negatively correlated with C4:C3, possibly because of changes in microclimate or plant-microbe interactions. It is possible that in potentially productive soils, C storage can be enhanced by favoring C4 over C3 grasses through increased ANPP and BNPP and reduced R H. Results also suggest that potential C storage gains from C4 productivity would not be undermined by a corresponding increase in R H.

Increased duration of aquatic resource pulse alters community and ecosystem responses in a subarctic plant community.

Allochthonous resource movement across ecosystem boundaries creates episodic linkages between ecosystems. The sensitivity of the community to external resources of varying duration can alter the baseline upon which future pulses of allochthony can act. We explored the terrestrial ecosystem response to pulsed inputs of lake-derived resources with a manipulative experiment in a subarctic heathland where we assessed plant community and nutrient availability responses to additions of midge carcasses (Diptera: Chironomidae). Insect carcasses were added as either a one-time pulse or a 4-yr press to simulate differing durations of allochthony, which is common in the area. We found that midge pulses significantly elevated soil inorganic [N] in the first year (7× over background levels) but were significantly diminished (1.5×) by the second year after the initial pulse. The press treatment continued to elevate total soil inorganic [N] to 13× over background levels by the fourth year of midge additions, but then declined to 3.6× background in year 5 when experimental midge additions had ceased. In contrast to the soil inorganic N response, plant biomass was similar in pulse-addition and control plots over the course of the experiment. However, by the second year of the study plant biomass in press-addition plots were significantly higher than controls (>50%), and continued to increase over the 4 yr of the press treatment. Midge addition stimulated dominance of graminoids and thatch litter in plots that had previously been primarily heathland vegetation, a response that persisted 4 yr post-midge addition. Our findings suggest that soil and plant community responses to persistent insect carcass deposition (e.g., press) into heathland vegetation has the potential to carry forward in a way that modifies the baseline ecosystem conditions upon which additional allochthony may act.

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples.

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial communities, a relatively precise but effort-intensive technique to assay microbial community composition is phospholipid fatty acid (PLFA) analysis. PLFA was developed to analyze phospholipid biomarkers, which can be used as indicators of microbial biomass and the composition of broad functional groups of fungi and bacteria. It has commonly been used to compare soils under alternative plant communities, ecology, and management regimes. The PLFA method has been shown to be sensitive to detecting shifts in microbial community composition. An alternative method, fatty acid methyl ester extraction and analysis (MIDI-FA) was developed for rapid extraction of total lipids, without separation of the phospholipid fraction, from pure cultures as a microbial identification technique. This method is rapid but is less suited for soil samples because it lacks an initial step separating soil particles and begins instead with a saponification reaction that likely produces artifacts from the background organic matter in the soil. This article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples. The method combines the accuracy achieved through PLFA profiling by extracting and concentrating soil lipids as a first step, and a reduction in effort by saponifying the organic material extracted and processing with the MIDI-FA method as a second step.

Cellulosic biofuel contributions to a sustainable energy future: Choices and outcomes.

Cellulosic crops are projected to provide a large fraction of transportation energy needs by mid-century. However, the anticipated land requirements are substantial, which creates a potential for environmental harm if trade-offs are not sufficiently well understood to create appropriately prescriptive policy. Recent empirical findings show that cellulosic bioenergy concerns related to climate mitigation, biodiversity, reactive nitrogen loss, and crop water use can be addressed with appropriate crop, placement, and management choices. In particular, growing native perennial species on marginal lands not currently farmed provides substantial potential for climate mitigation and other benefits.

Comparing herbaceous plant communities in active and passive riparian restoration.

Understanding the efficacy of passive (reduction or cessation of environmental stress) and active (typically involving planting or seeding) restoration strategies is important for the design of successful revegetation of degraded riparian habitat, but studies explicitly comparing restoration outcomes are uncommon. We sampled the understory herbaceous plant community of 103 riparian sites varying in age since restoration (0 to 39 years) and revegetation technique (active, passive, or none) to compare the utility of different approaches on restoration success across sites. We found that landform type, percent shade, and summer flow helped explain differences in the understory functional community across all sites. In passively restored sites, grass and forb cover and richness were inversely related to site age, but in actively restored sites forb cover and richness were inversely related to site age. Native cover and richness were lower with passive restoration compared to active restoration. Invasive species cover and richness were not significantly different across sites. Although some of our results suggest that active restoration would best enhance native species in degraded riparian areas, this work also highlights some of the context-dependency that has been found to mediate restoration outcomes. For example, since the effects of passive restoration can be quite rapid, this approach might be more useful than active restoration in situations where rapid dominance of pioneer species is required to arrest major soil loss through erosion. As a result, we caution against labeling one restoration technique as better than another. Managers should identify ideal restoration outcomes in the context of historic and current site characteristics (as well as a range of acceptable alternative states) and choose restoration approaches that best facilitate the achievement of revegetation goals.

Does Plant Biomass Manipulation in Static Chambers Affect Nitrous Oxide Emissions from Soils?

One of the most widespread approaches for measurement of greenhouse gas emissions from soils involves the use of static chambers. This method is relatively inexpensive, is easily replicated, and is ideally suited to plot-based experimental systems. Among its limitations is the loss of detection sensitivity with increasing chamber height, which creates challenges for deployment in systems including tall vegetation. It is not always possible to avoid inclusion of plants within chambers or to extend chamber height to fully accommodate plant growth. Thus, in many systems, such as perennial forages and biomass crops, plants growing within static chambers must either be trimmed or folded during lid closure. Currently, data on how different types of biomass manipulation affect measured results is limited. Here, we compare the effects of cutting vs. folding of biomass on nitrous oxide measurements in switchgrass ( L.) and alfalfa ( L.) systems. We report only limited evidence of treatment effects during discrete sampling events and little basis for concern that effects may intensify over time as biomass manipulation is repeatedly imposed. However, nonsignificant treatment effects that were consistently present amounted to significant overall trends in three out of the four systems studied. Such minor disparities in flux could amount to considerable quantities over time, suggesting that caution should be exercised when comparing cumulative emission values from studies using different biomass manipulation strategies.

Nitrogen Fertilization Effects on Productivity and Nitrogen Loss in Three Grass-Based Perennial Bioenergy Cropping Systems.

Nitrogen (N) fertilization can greatly improve plant productivity but needs to be carefully managed to avoid harmful environmental impacts. Nutrient management guidelines aimed at reducing harmful forms of N loss such as nitrous oxide (N2O) emissions and nitrate (NO3(-)) leaching have been tailored for many cropping systems. The developing bioenergy industry is likely to make use of novel cropping systems, such as polycultures of perennial species, for which we have limited nutrient management experience. We studied how a switchgrass (Panicum virgatum) monoculture, a 5-species native grass mixture and an 18-species restored prairie responded to annual fertilizer applications of 56 kg N ha(-1) in a field-scale agronomic trial in south-central Wisconsin over a 2-year period. We observed greater fertilizer-induced N2O emissions and sub-rooting zone NO3(-) concentrations in the switchgrass monoculture than in either polyculture. Fertilization increased aboveground net primary productivity in the polycultures, but not in the switchgrass monoculture. Switchgrass was generally more productive, while the two polycultures did not differ from each other in productivity or N loss. Our results highlight differences between polycultures and a switchgrass monoculture in responding to N fertilization.