Enhanced weathering in the US Corn Belt delivers carbon removal with agronomic benefits.

Terrestrial enhanced weathering (EW) of silicate rocks, such as crushed basalt, on farmlands is a promising scalable atmospheric carbon dioxide removal (CDR) strategy that urgently requires performance assessment with commercial farming practices. We report findings from a large-scale replicated EW field trial across a typical maize-soybean rotation on an experimental farm in the heart of the United Sates Corn Belt over 4 y (2016 to 2020). We show an average combined loss of major cations (Ca2+ and Mg2+) from crushed basalt applied each fall over 4 y (50 t ha-1 y-1) gave a conservative time-integrated cumulative CDR potential of 10.5 ± 3.8 t CO2 ha-1. Maize and soybean yields increased significantly (P < 0.05) by 12 to 16% with EW following improved soil fertility, decreased soil acidification, and upregulation of root nutrient transport genes. Yield enhancements with EW were achieved with significantly (P < 0.05) increased key micro- and macronutrient concentrations (including potassium, magnesium, manganese, phosphorus, and zinc), thus improving or maintaining crop nutritional status. We observed no significant increase in the content of trace metals in grains of maize or soybean or soil exchangeable pools relative to controls. Our findings suggest that widespread adoption of EW across farming sectors has the potential to contribute significantly to net-zero greenhouse gas emissions goals while simultaneously improving food and soil security.

Iron Chelation in Soil: Scalable Biotechnology for Accelerating Carbon Dioxide Removal by Enhanced Rock Weathering.

Enhanced rock weathering (EW) is an emerging atmospheric carbon dioxide removal (CDR) strategy being scaled up by the commercial sector. Here, we combine multiomics analyses of belowground microbiomes, laboratory-based dissolution studies, and incubation investigations of soils from field EW trials to build the case for manipulating iron chelators in soil to increase EW efficiency and lower costs. Microbial siderophores are high-affinity, highly selective iron (Fe) chelators that enhance the uptake of Fe from soil minerals into cells. Applying RNA-seq metatranscriptomics and shotgun metagenomics to soils and basalt grains from EW field trials revealed that microbial communities on basalt grains significantly upregulate siderophore biosynthesis gene expression relative to microbiomes of the surrounding soil. Separate in vitro laboratory incubation studies showed that micromolar solutions of siderophores and high-affinity synthetic chelator (ethylenediamine-N,N'-bis-2-hydroxyphenylacetic acid, EDDHA) accelerate EW to increase CDR rates. Building on these findings, we develop a potential biotechnology pathway for accelerating EW using the synthetic Fe-chelator EDDHA that is commonly used in agronomy to alleviate the Fe deficiency in high pH soils. Incubation of EW field trial soils with potassium-EDDHA solutions increased potential CDR rates by up to 2.5-fold by promoting the abiotic dissolution of basalt and upregulating microbial siderophore production to further accelerate weathering reactions. Moreover, EDDHA may alleviate potential Fe limitation of crops due to rising soil pH with EW over time. Initial cost-benefit analysis suggests potassium-EDDHA could lower EW-CDR costs by up to U.S. $77 t CO2 ha-1 to improve EW's competitiveness relative to other CDR strategies.

Improved net carbon budgets in the US Midwest through direct measured impacts of enhanced weathering.

Terrestrial enhanced weathering (EW) through the application of Mg- or Ca-rich rock dust to soil is a negative emission technology with the potential to address impacts of climate change. The effectiveness of EW was tested over 4 years by spreading ground basalt (50 t ha-1 year-1 ) on maize/soybean and miscanthus cropping systems in the Midwest US. The major elements of the carbon budget were quantified through measurements of eddy covariance, soil carbon flux, and biomass. The movement of Mg and Ca to deep soil, released by weathering, balanced by a corresponding alkalinity flux, was used to measure the drawdown of CO2 , where the release of cations from basalt was measured as the ratio of rare earth elements to base cations in the applied rock dust and in the surface soil. Basalt application stimulated peak biomass and net primary production in both cropping systems and caused a small but significant stimulation of soil respiration. Net ecosystem carbon balance (NECB) was strongly negative for maize/soybean (-199 to -453 g C m-2 year-1 ) indicating this system was losing carbon to the atmosphere. Average EW (102 g C m-2 year-1 ) offset carbon loss in the maize/soybean by 23%-42%. NECB of miscanthus was positive (63-129 g C m-2 year-1 ), indicating carbon gain in the system, and EW greatly increased inorganic carbon storage by an additional 234 g C m-2 year-1 . Our analysis indicates a co-deployment of a perennial biofuel crop (miscanthus) with EW leads to major wins-increased harvested yields of 29%-42% with additional carbon dioxide removal (CDR) of 8.6 t CO2 ha-1 year-1 . EW applied to maize/soybean drives a CDR of 3.7 t CO2 ha-1 year-1 , which partially offsets well-established carbon losses from soil from this crop rotation. EW applied in the US Midwest creates measurable improvements to the carbon budgets perennial bioenergy crops and conventional row crops.

Climate vs Energy Security: Quantifying the Trade-offs of BECCS Deployment and Overcoming Opportunity Costs on Set-Aside Land.

Bioenergy with carbon capture and storage (BECCS) sits at the nexus of the climate and energy security. We evaluated trade-offs between scenarios that support climate stabilization (negative emissions and net climate benefit) or energy security (ethanol production). Our spatially explicit model indicates that the foregone climate benefit from abandoned cropland (opportunity cost) increased carbon emissions per unit of energy produced by 14-36%, making geologic carbon capture and storage necessary to achieve negative emissions from any given energy crop. The toll of opportunity costs on the climate benefit of BECCS from set-aside land was offset through the spatial allocation of crops based on their individual biophysical constraints. Dedicated energy crops consistently outperformed mixed grasslands. We estimate that BECCS allocation to land enrolled in the Conservation Reserve Program (CRP) could capture up to 9 Tg C year-1 from the atmosphere, deliver up to 16 Tg CE year-1 in emissions savings, and meet up to 10% of the US energy statutory targets, but contributions varied substantially as the priority shifted from climate stabilization to energy provision. Our results indicate a significant potential to integrate energy security targets into sustainable pathways to climate stabilization but underpin the trade-offs of divergent policy-driven agendas.

Assessing the Returns to Land and Greenhouse Gas Savings from Producing Energy Crops on Conservation Reserve Program Land.

Using land already enrolled in the Conservation Reserve Program (CRP) in the eastern region of the U.S. for producing energy crops for bioenergy while reducing land rental payments offers the potential for lowering the program costs, increasing returns to CRP landowners, and displacing greenhouse gas (GHG) emissions from fossil fuels. We develop an integrated modeling approach to analyze the combination of biomass prices and CRP land rental payment reductions that can incentivize energy crop production on CRP land and its potential to increase soil carbon stocks and displace fossil fuel emissions. We find that conversion of 3.4 million ha in the CRP can be economically viable at a minimum biomass price of $75 Mg-1 with full CRP land rental payment or at $100 Mg-1 with 75% of this land rental payment; this conversion can result in savings of 0.52 and 1.25 billion Mg CO2-eq in life-cycle emissions through the displacement of energy-equivalent fossil fuels and coal-based electricity, respectively, and an additional 0.11 billion Mg CO2-eq soil carbon sequestration relative to the status quo, with CRP left unharvested over the 2016-2030 period. The soil carbon debt due to the transition from unharvested CRP land to energy crops is short-lived and more than offset by the reduction in fossil fuel emissions. The net discounted benefits from producing energy crops on CRP land through a reduced need for government payments to maintain existing enrollment, higher returns to CRP landowners, and the value of the reduction in GHG emissions could be as high as $16-$30 billion by using them for cellulosic biofuels to displace gasoline and $35-$68 billion by displacing coal-based electricity over the 2016-2030 period if biomass prices are $75-$125 Mg-1 and land rental payments are reduced by 25%.

Soybean photosynthetic and biomass responses to carbon dioxide concentrations ranging from pre-industrial to the distant future.

Increasing atmospheric carbon dioxide concentration ([CO2]) directly impacts C3 plant photosynthesis and productivity, and the rate at which [CO2] is increasing is greater than initially predicted by worst-case scenario climate models. Thus, it is increasingly important to assess the physiological responses of C3 plants, especially those that serve as important crops, to [CO2] beyond the mid-range levels used in traditional experiments. Here, we grew the C3 crop soybean (Glycine max) at eight different [CO2] levels spanning subambient (340 ppm) to the highest level thought plausible (~2000 ppm) in chambers for 5 weeks. Physiological development was delayed and plant height and total leaf area increased at [CO2] levels higher than ambient conditions, with very little difference in these parameters among the elevated [CO2] treatments >900 ppm. Daily photosynthesis initially increased with rising [CO2] but began to level off at ~1000 ppm CO2. Similar results occurred in biomass accumulation. Thus, as [CO2] continues to match or exceed the worst-case emission scenarios, these results indicate that carbon gain, growth, and potentially yield increases will diminish, thereby ultimately constraining the positive impact that continuing increases in atmospheric [CO2] could have on crop productivity and global terrestrial carbon sinks.

Soils' dirty little secret: Depth-based comparisons can be inadequate for quantifying changes in soil organic carbon and other mineral soil properties.

Quantifying changes in soil organic carbon (SOC) stocks and other soil properties is essential for understanding how soils will respond to land management practices and global change. Although they are widely used, comparisons of SOC stocks at fixed depth (FD) intervals are subject to errors when changes in bulk density or soil organic matter occur. The equivalent soil mass (ESM) method has been recommended in lieu of FD for assessing changes in SOC stocks in mineral soils, but ESM remains underutilized for SOC stocks and has rarely been used for other soil properties. In this paper, we draw attention to the limitations of the FD method and demonstrate the advantages of the ESM approach. We provide illustrations to show that the FD approach is susceptible to errors not only for quantifying SOC stocks but also for soil mass-based properties such as SOC mass percent, C:N mass ratio, and δ13 C. We describe the ESM approach and show how it mitigates the FD method limitations. Using bulk density change simulations applied to an empirical dataset from bioenergy cropping systems, we show that the ESM method provides consistently lower errors than FD when quantifying changes in SOC stocks and other soil properties. To simplify the use of ESM, we detail how the method can be integrated into sampling schemes, and we provide an example R computer script that can perform ESM calculations on large datasets. We encourage future studies, whether temporal or comparative, to utilize sampling methods that are amenable to the ESM approach. Overall, we agree with previous recommendations that ESM should be the standard method for evaluating SOC stock changes in mineral soils, but we further suggest that ESM may also be preferred for comparisons of other soil properties including mass percentages, elemental mass ratios, and stable isotope composition.

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.

Black walnut alley cropping is economically competitive with row crops in the Midwest USA.

The maize-soybean rotation (MSR) dominates the Midwest United States and degrades many ecological functions. Black walnut (Juglans nigra L.) plantation forestry (PF) and alley cropping (AC) are two alternative land-uses that can enhance productivity and restore ecosystem services. Given the lack of robust market mechanisms to monetize ecosystems services, we tested whether the profitability of PF and AC could drive adoption in the Midwest. Publically available data on black walnut soil suitability, timber prices, crop productivity, and cash rents were combined in a high-resolution spatial analysis to identify regions where these alternatives can outcompete MSR. To avoid selecting an arbitrary discount rate at which to make comparisons, we determined the threshold discount rate necessary to make PF or AC economically competitive with MSR. We show that, with a 5% discount rate, PF and AC could be more profitable on 17.0% and 23.4% of MSR land, respectively. Contrary to the common assumption that woody agricultural alternatives should first be adopted in marginal row crop areas, the economic competitiveness of PF and AC was not correlated with MSR productivity. Instead, black walnut growth rate was the central driver of PF and AC competitiveness, underscoring a necessary shift away from the current MSR-centric perspective in defining target regions for land-use alternatives. Results reveal major opportunities for landowners and investors to increase profitability by investing in PF and AC on both "marginal" and productive MSR land.

Mediation of Impacts of Elevated CO2 and Light Environment on Arabidopsis thaliana (L.) Chemical Defense against Insect Herbivory Via Photosynthesis.

Elevated CO2 alters C3 plant tolerance to insect herbivory, as well as the induction kinetics of defense hormones salicylic acid (SA) and jasmonic acid (JA), but the underlying physiological mechanism causing this response is not well understood. In principle, SA could be induced under elevated CO2 by reactive oxygen signals generated in photosynthesis, ultimately influencing chemical defense. To test whether the effects of elevated CO2 on C3 plant chemical defense against herbivorous insects are modulated by photosynthesis, Arabidopsis thaliana var. Col-0 plants were grown in two 2 × 2 × 2 nested factorial combinations of ambient (400 ppm) and elevated (800 ppm) CO2, and two dimensions of light regimes comprising intensity ('mild' 150 µmol E m-2 s-1 vs. 'low' light, 75 µmol E m-2 s-1) and periodicity ('continuous', 150 µmol E m-2 s-1 vs. 'dynamic', in which lights were turned off, then on, for 15 min every 2 h). Plants were challenged with herbivore damage from third instar Trichoplusia ni (cabbage looper). Consistent with experimental predictions, elevated CO2 interacted with light as well as herbivory to induce foliar concentration of SA, while JA was suppressed. Under dynamic light, foliar content of total glucosinolates was reduced. Under combination of elevated CO2 and dynamic light, T. ni removed significantly more leaf tissue relative to control plants. The observations that CO2 and light interactively modulate defense against T. ni in A. thaliana provide an empirical argument for a role of photosynthesis in C3 plant chemical defense.

Frontiers in alley cropping: Transformative solutions for temperate agriculture.

Annual row crops dominate agriculture around the world and have considerable negative environmental impacts, including significant greenhouse gas emissions. Transformative land-use solutions are necessary to mitigate climate change and restore critical ecosystem services. Alley cropping (AC)-the integration of trees with crops-is an agroforestry practice that has been studied as a transformative, multifunctional land-use solution. In the temperate zone, AC has strong potential for climate change mitigation through direct emissions reductions and increases in land-use efficiency via overyielding compared to trees and crops grown separately. In addition, AC provides climate change adaptation potential and ecological benefits by buffering alley crops to weather extremes, diversifying income to hedge financial risk, increasing biodiversity, reducing soil erosion, and improving nutrient- and water-use efficiency. The scope of temperate AC research and application has been largely limited to simple systems that combine one timber tree species with an annual grain. We propose two frontiers in temperate AC that expand this scope and could transform its climate-related benefits: (i) diversification via woody polyculture and (ii) expanded use of tree crops for food and fodder. While AC is ready now for implementation on marginal lands, we discuss key considerations that could enhance the scalability of the two proposed frontiers and catalyze widespread adoption.

Grazing alters net ecosystem C fluxes and the global warming potential of a subtropical pasture.

The impact of grazing on C fluxes from pastures in subtropical and tropical regions and on the environment is uncertain, although these systems account for a substantial portion of global C storage. We investigated how cattle grazing influences net ecosystem CO2 and CH4 exchange in subtropical pastures using the eddy covariance technique. Measurements were made over several wet-dry seasonal cycles in a grazed pasture, and in an adjacent pasture during the first three years of grazer exclusion. Grazing increased soil wetness but did not affect soil temperature. By removing aboveground biomass, grazing decreased ecosystem respiration (Reco ) and gross primary productivity (GPP). As the decrease in Reco was larger than the reduction in GPP, grazing consistently increased the net CO2 sink strength of subtropical pastures (55, 219 and 187 more C/m2 in 2013, 2014, and 2015). Enteric ruminant fermentation and increased soil wetness due to grazers, increased total net ecosystem CH4 emissions in grazed relative to ungrazed pasture (27-80%). Unlike temperate, arid, and semiarid pastures, where differences in CH4 emissions between grazed and ungrazed pastures are mainly driven by enteric ruminant fermentation, our results showed that the effect of grazing on soil CH4 emissions can be greater than CH4 produced by cattle. Thus, our results suggest that the interactions between grazers and soil hydrology affecting soil CH4 emissions play an important role in determining the environmental impacts of this management practice in a subtropical pasture. Although grazing increased total net ecosystem CH4 emissions and removed aboveground biomass, it increased the net storage of C and decreased the global warming potential associated with C fluxes of pasture by increasing its net CO2 sink strength.

Elevated CO2 and temperature increase soil C losses from a soybean-maize ecosystem.

Warming temperatures and increasing CO2 are likely to have large effects on the amount of carbon stored in soil, but predictions of these effects are poorly constrained. We elevated temperature (canopy: +2.8 °C; soil growing season: +1.8 °C; soil fallow: +2.3 °C) for 3 years within the 9th-11th years of an elevated CO2 (+200 ppm) experiment on a maize-soybean agroecosystem, measured respiration by roots and soil microbes, and then used a process-based ecosystem model (DayCent) to simulate the decadal effects of warming and CO2 enrichment on soil C. Both heating and elevated CO2 increased respiration from soil microbes by ~20%, but heating reduced respiration from roots and rhizosphere by ~25%. The effects were additive, with no heat × CO2 interactions. Particulate organic matter and total soil C declined over time in all treatments and were lower in elevated CO2 plots than in ambient plots, but did not differ between heat treatments. We speculate that these declines indicate a priming effect, with increased C inputs under elevated CO2 fueling a loss of old soil carbon. Model simulations of heated plots agreed with our observations and predicted loss of ~15% of soil organic C after 100 years of heating, but simulations of elevated CO2 failed to predict the observed C losses and instead predicted a ~4% gain in soil organic C under any heating conditions. Despite model uncertainty, our empirical results suggest that combined, elevated CO2 and temperature will lead to long-term declines in the amount of carbon stored in agricultural soils.

The impact of water management practices on subtropical pasture methane emissions and ecosystem service payments.

Pastures are an extensive land cover type; however, patterns in pasture greenhouse gas (GHG) exchange vary widely depending on climate and land management. Understanding this variation is important, as pastures may be a net GHG source or sink depending on these factors. We quantified carbon dioxide (CO2 ) and methane (CH4 ) fluxes from subtropical pastures in south Florida for three wet-dry seasonal cycles using eddy covariance, and estimated two annual budgets of CO2 , CH4 , and GHG equivalent emissions. We also estimated the impact of water retention practices on pasture GHG emissions and assessed the impact of these emissions on stakeholder payments for water retention services in a carbon market framework. The pastures were net CO2 sinks sequestering up to 163 ± 54 g CO2 -C·m-2 ·yr-1 (mean ± 95% CI), but were also strong CH4 sources emitting up to 23.5 ± 2.1 g CH4 -C·m-2 ·yr-1 . Accounting for the increased global warming potential of CH4 , the pastures were strong net GHG sources emitting up to 584 ± 78 g CO2 -C eq.·m-2 ·yr-1 , and all CO2 uptake was offset by wet season CH4 emissions from the flooded landscape. Our analysis suggests that CH4 emissions due to increased flooding from water management practices is a small component of the pasture GHG budget, and water retention likely contributes 2-11% of net pasture GHG emissions. These emissions could reduce water retention payments by up to ~12% if stakeholders were required to pay for current GHG emissions in a carbon market. It would require at least 93.7 kg CH4 -C emissions per acre-foot water storage (1 acre-foot = 1233.48 m3 ) for carbon market costs to exceed water retention payments, and this scenario is highly unlikely as we estimate current practices are responsible for 11.3 ± 7.2 kg CH4 -C emissions per acre-foot of water storage. Our results demonstrate that water retention practices aimed at reducing nutrient loading to the Everglades are likely only responsible for a minor increase in pasture GHG emissions and would have a small economic consequence in a carbon market.

Potential of global croplands and bioenergy crops for climate change mitigation through deployment for enhanced weathering.

Conventional row crop agriculture for both food and fuel is a source of carbon dioxide (CO2) and nitrous oxide (N2O) to the atmosphere, and intensifying production on agricultural land increases the potential for soil C loss and soil acidification due to fertilizer use. Enhanced weathering (EW) in agricultural soils-applying crushed silicate rock as a soil amendment-is a method for combating global climate change while increasing nutrient availability to plants. EW uses land that is already producing food and fuel to sequester carbon (C), and reduces N2O loss through pH buffering. As biofuel use increases, EW in bioenergy crops offers the opportunity to sequester CO2 while reducing fossil fuel combustion. Uncertainties remain in the long-term effects and global implications of large-scale efforts to directly manipulate Earth's atmospheric CO2 composition, but EW in agricultural lands is an opportunity to employ these soils to sequester atmospheric C while benefitting crop production and the global climate.

The influence of drought and heat stress on long-term carbon fluxes of bioenergy crops grown in the Midwestern USA.

Perennial grasses are promising feedstocks for bioenergy production in the Midwestern USA. Few experiments have addressed how drought influences their carbon fluxes and storage. This study provides a direct comparison of ecosystem-scale measurements of carbon fluxes associated with miscanthus (Miscanthus × giganteus), switchgrass (Panicum virgatum), restored native prairie and maize (Zea mays)/soybean (Glycine max) ecosystems. The main objective of this study was to assess the influence of a naturally occurring drought during 2012 on key components of the carbon cycle and plant development relative to non-extreme years. The perennials reached full maturity 3-5 years after establishment. Miscanthus had the highest gross primary production (GPP) and lowest net ecosystem exchange (NEE) in 2012 followed by similar values for switchgrass and prairie, and the row crops had the lowest GPP and highest NEE. A post-drought effect was observed for miscanthus. Over the duration of the experiment, perennial ecosystems were carbon sinks, as indicated by negative net ecosystem carbon balance (NECB), while maize/soybean was a net carbon source. Our observations suggest that perennial ecosystems, and in particular miscanthus, can provide a high yield and a large potential for CO2 fixation even during drought, although drought may negatively influence carbon uptake in the following year, questioning the long-term consequence of its maintained productivity.

Alteration of forest succession and carbon cycling under elevated CO2.

Regenerating forests influence the global carbon (C) cycle, and understanding how climate change will affect patterns of regeneration and C storage is necessary to predict the rate of atmospheric carbon dioxide (CO2 ) increase in future decades. While experimental elevation of CO2 has revealed that young forests respond with increased productivity, there remains considerable uncertainty as to how the long-term dynamics of forest regrowth are shaped by elevated CO2 (eCO2 ). Here, we use the mechanistic size- and age- structured Ecosystem Demography model to investigate the effects of CO2 enrichment on forest regeneration, using data from the Duke Forest Free-Air Carbon dioxide Enrichment (FACE) experiment, a forest chronosequence, and an eddy-covariance tower for model parameterization and evaluation. We find that the dynamics of forest regeneration are accelerated, and stands consistently hit a variety of developmental benchmarks earlier under eCO2 . Because responses to eCO2 varied by plant functional type, successional pathways, and mature forest composition differed under eCO2 , with mid- and late-successional hardwood functional types experiencing greater increases in biomass compared to early-successional functional types and the pine canopy. Over the simulation period, eCO2 led to an increase in total ecosystem C storage of 9.7 Mg C ha(-1) . Model predictions of mature forest biomass and ecosystem-atmosphere exchange of CO2 and H2 O were sensitive to assumptions about nitrogen limitation; both the magnitude and persistence of the ecosystem response to eCO2 were reduced under N limitation. In summary, our simulations demonstrate that eCO2 can result in a general acceleration of forest regeneration while altering the course of successional change and having a lasting impact on forest ecosystems.

Nitrogen deposition and greenhouse gas emissions from grasslands: uncertainties and future directions.

Increases in atmospheric nitrogen deposition (Ndep) can strongly affect the greenhouse gas (GHG; CO2, CH4, and N2O) sink capacity of grasslands as well as other terrestrial ecosystems. Robust predictions of the net GHG sink strength of grasslands depend on how experimental N loads compare to projected Ndep rates, and how accurately the relationship between GHG fluxes and Ndep is characterized. A literature review revealed that the vast majority of experimental N loads were higher than levels these ecosystems are predicted to experience in the future. Using a process-based biogeochemical model, we predicted that low levels of Ndep either enhanced or reduced the net GHG sink strength of most grasslands, but as experimental N loads continued to increase, grasslands transitioned to a N saturation-decline stage, where the sensitivity of GHG exchange to further increases in Ndep declined. Most published studies represented treatments well into the N saturation-decline stage. Our model results predict that the responses of GHG fluxes to N are highly nonlinear and that the N saturation thresholds for GHGs varied greatly among grasslands and with fire management. We predict that during the 21st century some grasslands will be in the N limitation stage where others will transition into the N saturation-decline stage. The linear relationship between GHG sink strength and N load assumed by most studies can overestimate or underestimate predictions of the net GHG sink strength of grasslands depending on their N baseline status. The next generation of global change experiments should be designed at multiple N loads consistent with future Ndep rates to improve our empirical understanding and predictive ability.

Leaf-galling phylloxera on grapes reprograms host metabolism and morphology.

Endoparasitism by gall-forming insects dramatically alters the plant phenotype by altering growth patterns and modifying plant organs in ways that appear to directly benefit the gall former. Because these morphological and physiological changes are linked to the presence of the insect, the induced phenotype is said to function as an extension of the parasite, albeit by unknown mechanisms. Here we report the gall-forming aphid-like parasite phylloxera, Daktulosphaira vitifoliae, induces stomata on the adaxial surface of grape leaves where stomata typically do not occur. We characterized the function of the phylloxera-induced stomata by tracing transport of assimilated carbon. Because induction of stomata suggests a significant manipulation of primary metabolism, we also characterized the gall transcriptome to infer the level of global reconfiguration of primary metabolism and the subsequent changes in downstream secondary metabolism. Phylloxera feeding induced stomata formation in proximity to the insect and promoted the assimilation and importation of carbon into the gall. Gene expression related to water, nutrient, and mineral transport; glycolysis; and fermentation increased in leaf-gall tissues. This shift from an autotrophic to a heterotrophic profile occurred concurrently with decreased gene expression for nonmevalonate and terpenoid synthesis and increased gene expression in shikimate and phenylpropanoid biosynthesis, secondary metabolite systems that alter defense status in grapes. These functional insect-induced stomata thus comprise part of an extended phenotype, whereby D. vitifoliae globally reprograms grape leaf development to alter patterns of primary metabolism, nutrient mobilization, and defense investment in favor of the galling habit.