Benefitting productivity and the environment: Current and future maize cropping systems improve yield while reducing nitrate load.

Increases in cereal crop yield per area have increased global food security. "Era" studies compare historical and modern crop varieties in controlled experimental settings and are routinely used to understand how advances in crop genetics and management affect crop yield. However, to date, no era study has explored how advances in maize (Zea mays L.) genetics and management (i.e., cropping systems) have affected environmental outcomes. Here, we developed a cropping systems era study in Iowa, USA, to examine how yield and nitrate losses have changed from "Old" systems common in the 1990s to "Current" systems common in the 2010s, and to "Future" systems projected to be common in the 2030s. We tested the following hypothesis: If maize yield and nitrogen use efficiency have improved over previous decades, Current and Future maize systems will have benefits to water quality compared to Old systems. We show that not only have maize yield and nitrogen use efficiency (kg grain kg-1 N), on average, improved over time but also yield-scaled nitrate load + soil nitrate was reduced by 74% and 91% from Old to Current and Future systems, respectively. Continuing these trajectories of improvement will be critical to meet the needs of a growing and more affluent population while reducing deleterious effects of agricultural systems on ecosystem services.

Soil depth and geographic distance modulate bacterial ß-diversity in deep soil profiles throughout the U.S. Corn Belt.

Understanding how microbial communities are shaped across spatial dimensions is of fundamental importance in microbial ecology. However, most studies on soil biogeography have focused on the topsoil microbiome, while the factors driving the subsoil microbiome distribution are largely unknown. Here we used 16S rRNA amplicon sequencing to analyse the factors underlying the bacterial ß-diversity along vertical (0-240 cm of soil depth) and horizontal spatial dimensions (~500,000 km2 ) in the U.S. Corn Belt. With these data we tested whether the horizontal or vertical spatial variation had stronger impacts on the taxonomic (Bray-Curtis) and phylogenetic (weighted Unifrac) ß-diversity. Additionally, we assessed whether the distance-decay (horizontal dimension) was greater in the topsoil (0-30 cm) or subsoil (in each 30 cm layer from 30-240 cm) using Mantel tests. The influence of geographic distance versus edaphic variables on the bacterial communities from the different soil layers was also compared. Results indicated that the phylogenetic ß-diversity was impacted more by soil depth, while the taxonomic ß-diversity changed more between geographic locations. The distance-decay was lower in the topsoil than in all subsoil layers analysed. Moreover, some subsoil layers were influenced more by geographic distance than any edaphic variable, including pH. Although different factors affected the topsoil and subsoil biogeography, niche-based models explained the community assembly of all soil layers. This comprehensive study contributed to elucidating important aspects of soil bacterial biogeography including the major impact of soil depth on the phylogenetic ß-diversity, and the greater influence of geographic distance on subsoil than on topsoil bacterial communities in agroecosystems.

Silver lining to a climate crisis in multiple prospects for alleviating crop waterlogging under future climates.

Extreme weather events threaten food security, yet global assessments of impacts caused by crop waterlogging are rare. Here we first develop a paradigm that distils common stress patterns across environments, genotypes and climate horizons. Second, we embed improved process-based understanding into a farming systems model to discern changes in global crop waterlogging under future climates. Third, we develop avenues for adapting cropping systems to waterlogging contextualised by environment. We find that yield penalties caused by waterlogging increase from 3-11% historically to 10-20% by 2080, with penalties reflecting a trade-off between the duration of waterlogging and the timing of waterlogging relative to crop stage. We document greater potential for waterlogging-tolerant genotypes in environments with longer temperate growing seasons (e.g., UK, France, Russia, China), compared with environments with higher annualised ratios of evapotranspiration to precipitation (e.g., Australia). Under future climates, altering sowing time and adoption of waterlogging-tolerant genotypes reduces yield penalties by 18%, while earlier sowing of winter genotypes alleviates waterlogging by 8%. We highlight the serendipitous outcome wherein waterlogging stress patterns under present conditions are likely to be similar to those in the future, suggesting that adaptations for future climates could be designed using stress patterns realised today.

County-scale crop yield prediction by integrating crop simulation with machine learning models.

Crop yield prediction is of great importance for decision making, yet it remains an ongoing scientific challenge. Interactions among different genetic, environmental, and management factors and uncertainty in input values are making crop yield prediction complex. Building upon a previous work in which we coupled crop modeling with machine learning (ML) models to predict maize yields for three US Corn Belt states, here, we expand the concept to the entire US Corn Belt (12 states). More specifically, we built five new ML models and their ensemble models, considering the scenarios with and without crop modeling variables. Additional input values in our models are soil, weather, management, and historical yield data. A unique aspect of our work is the spatial analysis to investigate causes for low or high model prediction errors. Our results indicated that the prediction accuracy increases by coupling crop modeling with machine learning. The ensemble model overperformed the individual ML models, having a relative root mean square error (RRMSE) of about 9% for the test years (2018, 2019, and 2020), which is comparable to previous studies. In addition, analysis of the sources of error revealed that counties and crop reporting districts with low cropland ratios have high RRMSE. Furthermore, we found that soil input data and extreme weather events were responsible for high errors in some regions. The proposed models can be deployed for large-scale prediction at the county level and, contingent upon data availability, can be utilized for field level prediction.

Photosynthetic decline in aging perennial grass is not fully explained by leaf nitrogen.

Aging in perennial plants is traditionally observed in terms of changes in end-of-season biomass; however, the driving phenological and physiological changes are poorly understood. We found that 3-year-old (mature) stands of the perennial grass Miscanthus×giganteus had 19-30% lower Anet than 1-year-old M.×giganteus (juvenile) stands; 10-34% lower maximum carboxylation rates of Rubisco and 34% lower light-saturated Anet (Asat). These changes could be related to nitrogen (N) limitations, as mature plants were larger and had 14-34% lower leaf N on an area basis (Na) than juveniles. However, N fertilization restored Na to juvenile levels but compensated only 50% of the observed decline in leaf photosynthesis with age. Comparison of leaf photosynthesis per unit of leaf N (PNUE) showed that mature stands had at least 26% lower PNUE than juvenile stands across all N fertilization rates, suggesting that other factors, besides N, may be limiting photosynthesis in mature stands. We hypothesize that sink limitations in mature stands could be causing feedback inhibition of photosynthesis which is associated with the age-related decline in photosynthesis.

Exploring Trade-Offs Between Profit, Yield, and the Environmental Footprint of Potential Nitrogen Fertilizer Regulations in the US Midwest.

Multiple strategies are available that could reduce nitrogen (N) fertilizer use in agricultural systems, ranging from voluntary adoption of new N management practices by farmers to government regulations. However, these strategies have different economic and political costs, and their relative effectiveness in decreasing N leaching has not been evaluated at scale, particularly concerning potential trade-offs in crop yield and profitability. To inform policy efforts in the US Midwest, we quantified the effects of four policy scenarios designed to reduce fertilizer N inputs without sacrificing maize yields below 95%. A simulated dataset for economically optimum N rates and corresponding leaching losses was developed using a process-based crop model across 4,030 fields over 30 years. Policy scenarios were (1) higher N prices, (2) N leaching fee, (3) N balance fee, and (4) voluntary reduction of N use by farmers, each implemented under a range of sub-levels (low to high severity). Aggregated results show that all policies decreased N rates and N leaching, but this was associated with an exponential increase in economic costs. Achieving an N leaching reduction target of 20% has an estimated pollution control cost of 30-37 US$/ha, representing 147 million US$/year when scaled up to the state level, which is in the range of current government payments for existing conservation programs. Notably, such control of N losses would reduce the environmental impact of agriculture on water quality (externalities) by an estimated 524 million US$/year, representing an increase in society welfare of 377 million US$/year. Among the four policies, directly charging a fee on N leaching helped mitigate economic losses while improving the point source reduction effect (i.e., targeting fields that were leaching hotspots) and better internalization effect (i.e., targeting fields with higher environmental impact costs). This study provides actionable data to inform the development of cost-effective N fertilizer regulations by integrating changes in crop productivity and N losses in economic terms at the field level.

Climate Change and Management Impacts on Soybean N Fixation, Soil N Mineralization, N2O Emissions, and Seed Yield.

Limited knowledge about how nitrogen (N) dynamics are affected by climate change, weather variability, and crop management is a major barrier to improving the productivity and environmental performance of soybean-based cropping systems. To fill this knowledge gap, we created a systems understanding of agroecosystem N dynamics and quantified the impact of controllable (management) and uncontrollable (weather, climate) factors on N fluxes and soybean yields. We performed a simulation experiment across 10 soybean production environments in the United States using the Agricultural Production Systems sIMulator (APSIM) model and future climate projections from five global circulation models. Climate change (2020-2080) increased N mineralization (24%) and N2O emissions (19%) but decreased N fixation (32%), seed N (20%), and yields (19%). Soil and crop management practices altered N fluxes at a similar magnitude as climate change but in many different directions, revealing opportunities to improve soybean systems' performance. Among many practices explored, we identified two solutions with great potential: improved residue management (short-term) and water management (long-term). Inter-annual weather variability and management practices affected soybean yield less than N fluxes, which creates opportunities to manage N fluxes without compromising yields, especially in regions with adequate to excess soil moisture. This work provides actionable results (tradeoffs, synergies, directions) to inform decision-making for adapting crop management in a changing climate to improve soybean production systems.

Maize Leaf Appearance Rates: A Synthesis From the United States Corn Belt.

The relationship between collared leaf number and growing degree days (GDD) is crucial for predicting maize phenology. Biophysical crop models convert GDD accumulation to leaf numbers by using a constant parameter termed phyllochron (°C-day leaf-1) or leaf appearance rate (LAR; leaf oC-day-1). However, such important parameter values are rarely estimated for modern maize hybrids. To fill this gap, we sourced and analyzed experimental datasets from the United States Corn Belt with the objective to (i) determine phyllochron values for two types of models: linear (1-parameter) and bilinear (3-parameters; phase I and II phyllochron, and transition point) and (ii) explore whether environmental factors such as photoperiod and radiation, and physiological variables such as plant growth rate can explain variability in phyllochron and improve predictability of maize phenology. The datasets included different locations (latitudes between 48° N and 41° N), years (2009-2019), hybrids, and management settings. Results indicated that the bilinear model represented the leaf number vs. GDD relationship more accurately than the linear model (R 2 = 0.99 vs. 0.95, n = 4,694). Across datasets, first phase phyllochron, transition leaf number, and second phase phyllochron averaged 57.9 ± 7.5°C-day, 9.8 ± 1.2 leaves, and 30.9 ± 5.7°C-day, respectively. Correlation analysis revealed that radiation from the V3 to the V9 developmental stages had a positive relationship with phyllochron (r = 0.69), while photoperiod was positively related to days to flowering or total leaf number (r = 0.89). Additionally, a positive nonlinear relationship between maize LAR and plant growth rate was found. Present findings provide important parameter values for calibration and optimization of maize crop models in the United States Corn Belt, as well as new insights to enhance mechanisms in crop models.

Nitrate losses across 29 Iowa watersheds: Measuring long-term trends in the context of interannual variability.

In the U.S. Corn Belt, annual croplands are the primary source of nitrate loading to waterways. Long periods of fallow cause most nitrate loss, but there is extreme interannual variability in the magnitude of nitrate loss due to weather. Using mean annual (2001-2018) flow-weighted nitrate-N concentration (FWNC; mg NO3 - -N L-1 ), load (kg NO3 - -N), and yield (kg NO3 - -N ha-1 cropland) for 29 watersheds, our objectives were (a) to quantify the magnitude and interannual variability of 5-yr moving average FWNC, load, and yield; (2) to estimate the probability of measuring 41% reductions in nitrate loss after isolating the effect of weather on nitrate loss by quantifying the interannual variability of nitrate loss in watersheds where there was no trend in 5-yr moving average nitrate loss (Iowa targets a 41% nitrate loss reduction from croplands); and (c) to identify factors that, in the absence of long-term trends in nitrate loss, best explain the interannual variability in nitrate loss. Averaged across all watersheds, the mean probability of measuring a statistically significant 41% reduction in FWNC across 15 yr, should it occur, was 96%. However, the probabilities of measuring 41% reductions in nitrate load and yield were only 44 and 32%. Across watersheds, soil organic matter, tile drainage, interannual variability of precipitation, and watershed area accounted for interannual variability in these nitrate loss indices. Our results have important implications for setting realistic timelines to measure nitrate loss reductions against the background of interannual weather variation and can help to target monitoring intensity across diverse watersheds.

Simulated dataset of corn response to nitrogen over thousands of fields and multiple years in Illinois.

Nitrogen (N) fertilizer recommendations for corn (Zea mays L.) in the US Midwest have been a puzzle for several decades, without agreement among stakeholders for which methodology is the best to balance environmental and economic outcomes. Part of the reason is the lack of long-term data of crop responses to N over multiple fields since trial data is often limited in the number of soils and years it can explore. To overcome this limitation, we designed an analytical platform based on crop simulations run over millions of farming scenarios over extensive geographies. The database was calibrated and validated using data from more than four hundred trials in the region. This dataset can have an important role for research and education in N management, machine leaching, and environmental policy analysis. The calibration and validation procedure provides a framework for future gridded crop model studies. We describe dataset characteristics and provide thorough descriptions of the model setup.

Crop traits enabling yield gains under more frequent extreme climatic events.

Climate change (CC) in central China will change seasonal patterns of agricultural production through increasingly frequent extreme climatic events (ECEs). Breeding climate-resilient wheat (Triticum aestivum L.) genotypes may mitigate adverse effects of ECEs on crop productivity. To reveal crop traits conducive to long-term yield improvement in the target population of environments, we created 8,192 virtual genotypes with contrasting but realistic ranges of phenology, productivity and waterlogging tolerance. Using these virtual genotypes, we conducted a genotype (G) by environment (E) by management (M) factorial analysis (G×E×M) using locations distributed across the entire cereal cropping zone in mid-China. The G×E×M invoked locally-specific sowing dates under future climates that were premised on shared socioeconomic pathways SSP5-8.5, with a time horizon centred on 2080. Across the simulated adaptation landscape, productivity was primarily driven by yield components and phenology (average grain yield increase of 6-69% across sites with optimal combinations of these traits). When incident solar radiation was not limiting carbon assimilation, ideotypes with higher grain yields were characterised by earlier flowering, higher radiation-use efficiency and larger maximum kernel size. At sites with limited solar radiation, crops required longer growing periods to realise genetic yield potential, although higher radiation-use efficiency and larger maximum kernel size were again prospective traits enabling higher rates of yield gains. By 2080, extreme waterlogging stress in some regions of mid-China will impact substantially on productivity, with yield penalties of up to 1,010 kg ha-1. Ideotypes with optimal G×M could mitigate yield penalty caused by waterlogging by up to 15% under future climates. These results help distil promising crop trait by best management practice combinations that enable higher yields and robust adaptation to future climates and more frequent extreme climatic events, including flash flooding and soil waterlogging.

Corrigendum: Exploring trade-offs between profit, yield, and the environmental footprint of potential nitrogen fertilizer regulations in the US Midwest.

[This corrects the article DOI: 10.3389/fpls.2022.852116.].

Revisiting Biological Nitrogen Fixation Dynamics in Soybeans.

Biological nitrogen (N) fixation is the most relevant process in soybeans (Glycine max L.) to satisfy plant N demand and sustain seed protein formation. Past studies describing N fixation for field-grown soybeans mainly focused on a single point time measurement (mainly toward the end of the season) and on the partial N budget (fixed-N minus seed N removal), overlooking the seasonal pattern of this process. Therefore, this study synthesized field datasets involving multiple temporal measurements during the crop growing season to characterize N fixation dynamics using both fixed-N (kg ha-1) and N derived from the atmosphere [Ndfa (%)] to define: (i) time to the maximum rate of N fixation (ß2), (ii) time to the maximum Ndfa (α2), and (iii) the cumulative fixed-N. The main outcomes of this study are that (1) the maximum rate of N fixation was around the beginning of pod formation (R3 stage), (2) time to the maximum Ndfa (%) was after full pod formation (R4), and (3) cumulative fixation was positively associated with the seasonal vapor-pressure deficit (VPD) and growth cycle length but negatively associated with soil clay content, and (4) time to the maximum N fixation rate (ß2) was positively impacted by season length and negatively impacted by high temperatures during vegetative growth (but positively for VPD, during the same period). Overall, variation in the timing of the maximum rate of N fixation occurred within a much narrower range of growth stages (R3) than the timing of the maximum Ndfa (%), which varied broadly from flowering (R1) to seed filing (R5-R6) depending on the evaluated studies. From a phenotyping standpoint, N fixation determinations after the R4 growth stage would most likely permit capturing both maximum fixed-N rate and maximum Ndfa (%). Further investigations that more closely screen the interplay between N fixation with soil-plant-environment factors should be pursued.

An interaction regression model for crop yield prediction.

Crop yield prediction is crucial for global food security yet notoriously challenging due to multitudinous factors that jointly determine the yield, including genotype, environment, management, and their complex interactions. Integrating the power of optimization, machine learning, and agronomic insight, we present a new predictive model (referred to as the interaction regression model) for crop yield prediction, which has three salient properties. First, it achieved a relative root mean square error of 8% or less in three Midwest states (Illinois, Indiana, and Iowa) in the US for both corn and soybean yield prediction, outperforming state-of-the-art machine learning algorithms. Second, it identified about a dozen environment by management interactions for corn and soybean yield, some of which are consistent with conventional agronomic knowledge whereas some others interactions require additional analysis or experiment to prove or disprove. Third, it quantitatively dissected crop yield into contributions from weather, soil, management, and their interactions, allowing agronomists to pinpoint the factors that favorably or unfavorably affect the yield of a given location under a given weather and management scenario. The most significant contribution of the new prediction model is its capability to produce accurate prediction and explainable insights simultaneously. This was achieved by training the algorithm to select features and interactions that are spatially and temporally robust to balance prediction accuracy for the training data and generalizability to the test data.

Corn Yield Prediction With Ensemble CNN-DNN.

We investigate the predictive performance of two novel CNN-DNN machine learning ensemble models in predicting county-level corn yields across the US Corn Belt (12 states). The developed data set is a combination of management, environment, and historical corn yields from 1980 to 2019. Two scenarios for ensemble creation are considered: homogenous and heterogenous ensembles. In homogenous ensembles, the base CNN-DNN models are all the same, but they are generated with a bagging procedure to ensure they exhibit a certain level of diversity. Heterogenous ensembles are created from different base CNN-DNN models which share the same architecture but have different hyperparameters. Three types of ensemble creation methods were used to create several ensembles for either of the scenarios: Basic Ensemble Method (BEM), Generalized Ensemble Method (GEM), and stacked generalized ensembles. Results indicated that both designed ensemble types (heterogenous and homogenous) outperform the ensembles created from five individual ML models (linear regression, LASSO, random forest, XGBoost, and LightGBM). Furthermore, by introducing improvements over the heterogenous ensembles, the homogenous ensembles provide the most accurate yield predictions across US Corn Belt states. This model could make 2019 yield predictions with a root mean square error of 866 kg/ha, equivalent to 8.5% relative root mean square and could successfully explain about 77% of the spatio-temporal variation in the corn grain yields. The significant predictive power of this model can be leveraged for designing a reliable tool for corn yield prediction which will in turn assist agronomic decision makers.

Environmental Factors Associated With Nitrogen Fixation Prediction in Soybean.

Biological nitrogen (N)-fixation is the most important source of N for soybean [Glycine max (L.) Merr.], with considerable implications for sustainable intensification. Therefore, this study aimed to investigate the relevance of environmental factors driving N-fixation and to develop predictive models defining the role of N-fixation for improved productivity and increased seed protein concentration. Using the elastic net regularization of multiple linear regression, we analyzed 40 environmental factors related to weather, soil, and crop management. We selected the most important factors associated with the relative abundance of ureides (RAU) as an indicator of the fraction of N derived from N-fixation. The most relevant RAU predictors were N fertilization, atmospheric vapor pressure deficit (VPD) and precipitation during early reproductive growth (R1-R4 stages), sowing date, drought stress during seed filling (R5-R6), soil cation exchange capacity (CEC), and soil sulfate concentration before sowing. Soybean N-fixation ranged from 60 to 98% across locations and years (n = 95). The predictive model for RAU showed relative mean square error (RRMSE) of 4.5% and an R2 value of 0.69, estimated via cross-validation. In addition, we built similar predictive models of yield and seed protein to assess the association of RAU and these plant traits. The variable RAU was selected as a covariable for the models predicting yield and seed protein, but with a small magnitude relative to the sowing date for yield or soil sulfate for protein. The early-reproductive period VPD affected all independent variables, namely RAU, yield, and seed protein. The elastic net algorithm successfully depicted some otherwise challenging empirical relationships to assess with bivariate associations in observational data. This approach provides inference about environmental variables while predicting N-fixation. The outcomes of this study will provide a foundation for improving the understanding of N-fixation within the context of sustainable intensification of soybean production.

A time-dependent parameter estimation framework for crop modeling.

The performance of crop models in simulating various aspects of the cropping system is sensitive to parameter calibration. Parameter estimation is challenging, especially for time-dependent parameters such as cultivar parameters with 2-3 years of lifespan. Manual calibration of the parameters is time-consuming, requires expertise, and is prone to error. This research develops a new automated framework to estimate time-dependent parameters for crop models using a parallel Bayesian optimization algorithm. This approach integrates the power of optimization and machine learning with prior agronomic knowledge. To test the proposed time-dependent parameter estimation method, we simulated historical yield increase (from 1985 to 2018) in 25 environments in the US Corn Belt with APSIM. Then we compared yield simulation results and nine parameter estimates from our proposed parallel Bayesian framework, with Bayesian optimization and manual calibration. Results indicated that parameters calibrated using the proposed framework achieved an 11.6% reduction in the prediction error over Bayesian optimization and a 52.1% reduction over manual calibration. We also trained nine machine learning models for yield prediction and found that none of them was able to outperform the proposed method in terms of root mean square error and R2. The most significant contribution of the new automated framework for time-dependent parameter estimation is its capability to find close-to-optimal parameters for the crop model. The proposed approach also produced explainable insight into cultivar traits' trends over 34 years (1985-2018).

An integrated framework reinstating the environmental dimension for GWAS and genomic selection in crops.

Identifying mechanisms and pathways involved in gene-environment interplay and phenotypic plasticity is a long-standing challenge. It is highly desirable to establish an integrated framework with an environmental dimension for complex trait dissection and prediction. A critical step is to identify an environmental index that is both biologically relevant and estimable for new environments. With extensive field-observed complex traits, environmental profiles, and genome-wide single nucleotide polymorphisms for three major crops (maize, wheat, and oat), we demonstrated that identifying such an environmental index (i.e., a combination of environmental parameter and growth window) enables genome-wide association studies and genomic selection of complex traits to be conducted with an explicit environmental dimension. Interestingly, genes identified for two reaction-norm parameters (i.e., intercept and slope) derived from flowering time values along the environmental index were less colocalized for a diverse maize panel than for wheat and oat breeding panels, agreeing with the different diversity levels and genetic constitutions of the panels. In addition, we showcased the usefulness of this framework for systematically forecasting the performance of diverse germplasm panels in new environments. This general framework and the companion CERIS-JGRA analytical package should facilitate biologically informed dissection of complex traits, enhanced performance prediction in breeding for future climates, and coordinated efforts to enrich our understanding of mechanisms underlying phenotypic variation.

Coupling machine learning and crop modeling improves crop yield prediction in the US Corn Belt.

This study investigates whether coupling crop modeling and machine learning (ML) improves corn yield predictions in the US Corn Belt. The main objectives are to explore whether a hybrid approach (crop modeling + ML) would result in better predictions, investigate which combinations of hybrid models provide the most accurate predictions, and determine the features from the crop modeling that are most effective to be integrated with ML for corn yield prediction. Five ML models (linear regression, LASSO, LightGBM, random forest, and XGBoost) and six ensemble models have been designed to address the research question. The results suggest that adding simulation crop model variables (APSIM) as input features to ML models can decrease yield prediction root mean squared error (RMSE) from 7 to 20%. Furthermore, we investigated partial inclusion of APSIM features in the ML prediction models and we found soil moisture related APSIM variables are most influential on the ML predictions followed by crop-related and phenology-related variables. Finally, based on feature importance measure, it has been observed that simulated APSIM average drought stress and average water table depth during the growing season are the most important APSIM inputs to ML. This result indicates that weather information alone is not sufficient and ML models need more hydrological inputs to make improved yield predictions.

The Effects of Soil Depth on the Structure of Microbial Communities in Agricultural Soils in Iowa, USA.

This study investigated the differences in microbial community abundance, composition and diversity throughout the depth profiles in soils collected from corn and soybean fields in lowa, USA using 16S rRNA amplicon sequencing. The results revealed decreased richness and diversity in microbial communities at increasing soil depth. Soil microbial community composition differed due to crop type only in the top 60 cm and due to location only in the top 90 cm. While the relative abundance of most phyla decreased in deep soils, the relative abundance of the phylum Proteobacteria increased and dominated agricultural soils below the depth of 90 cm. Although soil depth was the most important factor shaping microbial communities, edaphic factors including soil organic matter, soil bulk density and the length of time that deep soils were saturated with water were all significant factors explaining the variation in soil microbial community composition. Soil organic matter showed the highest correlation with the exponential decrease in bacterial abundance with depth. A greater understanding of how soil depth influences the diversity and composition of soil microbial communities is vital for guiding sampling approaches in agricultural soils where plant roots extend beyond the upper soil profile. In the long term a greater knowledge of the influence of depth on microbial communities should contribute to new strategies that enhance the sustainability of soil which is a precious resource for food security.IMPORTANCE Determining how microbial properties change across different soils and within the soil depth profile, will be potentially beneficial to understanding the long-term processes that are involved in the health of agricultural ecosystems. Most literature on soil microbes has been restricted to the easily accessible surface soils. However, deep soils are important in soil formation, carbon sequestration, and in providing nutrients and water for plants. In the most productive agricultural systems in the USA where soybean and corn are grown, crop plant roots extend into the deeper regions of soils (> 100 cm), but little is known about the taxonomic diversity or the factors that shape deep soil microbial communities. The findings reported here highlight the importance of soil depth in shaping microbial communities, provide new information about edaphic factors that influence the deep soil communities and reveal more detailed information on taxa that exist in deep agricultural soils.