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Abstract Agricultural researchers are increasingly encouraged to engage with stakeholders to improve the usefulness of their projects, but iterative research on the design and assessment of stakeholder engagement is scarce. The USDA Long‐Term Agroecosystem Research (LTAR) Network recognizes the importance of effective engagement in increasing the utility of information and technologies for future agriculture. Diverse stakeholders and researchers at the Kellogg Biological Station (KBS) LTAR site co‐designed the KBS LTAR Aspirational Cropping System Experiment, a process that provides a testing ground and interdisciplinary collaborations to develop theory‐driven assessment protocols for continuous stakeholder engagement. Informed by prior work, we designed an assessment protocol that aims to measure participant preferences, experiences, and perceived benefits at various stages of this long‐term project. Two online surveys were conducted in 2021 and 2022 among participants of LTAR engagement events at KBS, using a pre‐post design, resulting in 125 total responses. Survey respondents had positive perceptions of the collaboratively designed research experiment. They had a strong expectation that the research would generate conservation and environmental advances while also informing policy and programs. Respondents also indicated a desire to network with other stakeholders. The research team noted the significant role of a long‐term stakeholder engagement specialist in inviting participants from diverse backgrounds and creating an open and engaging experience. Overall, results highlight an interdisciplinary path of intentional and iterative engagement and evaluation to build a program that is adaptive and responsive to stakeholder needs. 

Abstract Mitigating the effects of climate stress on crops is important for global food security. The microbiome associated with plant roots, the rhizobiome, can harbor beneficial microbes that alleviate stress, but the factors influencing their recruitment are unclear. We conducted a greenhouse experiment using field soil with a legacy of growing switchgrass and common bean to investigate the impact of short-term drought severity on the recruitment of active bacterial rhizobiome members. We applied 16S rRNA and 16S rRNA gene sequencing for both crops and metabolite profiling for switchgrass. We included planted and unplanted conditions to distinguish environment- versus plant-mediated rhizobiome drivers. Differences in community structure were observed between crops and between drought and watered and planted and unplanted treatments within crops. Despite crop-specific communities, drought rhizobiome dynamics were similar across the two crops. The presence of a plant more strongly explained the rhizobiome variation in bean (17%) than in switchgrass (3%), with a small effect of plant mediation during drought observed only for the bean rhizobiome. The switchgrass rhizobiome was stable despite changes in rhizosphere metabolite profiles between planted and unplanted treatments. We conclude that rhizobiome responses to short-term drought are crop-specific, with possible decoupling of plant exudation from rhizobiome responses. 

Abstract BackgroundRoot and soil microbial communities constitute the below-ground plant microbiome, are drivers of nutrient cycling, and affect plant productivity. However, our understanding of their spatiotemporal patterns is confounded by exogenous factors that covary spatially, such as changes in host plant species, climate, and edaphic factors. These spatiotemporal patterns likely differ across microbiome domains (bacteria and fungi) and niches (root vs. soil). ResultsTo capture spatial patterns at a regional scale, we sampled the below-ground microbiome of switchgrass monocultures of five sites spanning > 3 degrees of latitude within the Great Lakes region. To capture temporal patterns, we sampled the below-ground microbiome across the growing season within a single site. We compared the strength of spatiotemporal factors to nitrogen addition determining the major drivers in our perennial cropping system. All microbial communities were most strongly structured by sampling site, though collection date also had strong effects; in contrast, nitrogen addition had little to no effect on communities. Though all microbial communities were found to have significant spatiotemporal patterns, sampling site and collection date better explained bacterial than fungal community structure, which appeared more defined by stochastic processes. Root communities, especially bacterial, were more temporally structured than soil communities which were more spatially structured, both across and within sampling sites. Finally, we characterized a core set of taxa in the switchgrass microbiome that persists across space and time. These core taxa represented < 6% of total species richness but > 27% of relative abundance, with potential nitrogen fixing bacteria and fungal mutualists dominating the root community and saprotrophs dominating the soil community. ConclusionsOur results highlight the dynamic variability of plant microbiome composition and assembly across space and time, even within a single variety of a plant species. Root and soil fungal community compositions appeared spatiotemporally paired, while root and soil bacterial communities showed a temporal lag in compositional similarity suggesting active recruitment of soil bacteria into the root niche throughout the growing season. A better understanding of the drivers of these differential responses to space and time may improve our ability to predict microbial community structure and function under novel conditions. 

Abstract Long‐term experiments are critical for understanding ecological processes, but their management comes with unique challenges. As time passes, projects may encounter unavoidable changes due to external factors, like availability of materials, affecting aspects of their research methodology. At the Kellogg Biological Station Long‐Term Ecological Research Site, one of the many National Science Foundation‐funded long‐term research stations, a three‐decade project recently experienced a supply‐chain‐induced change in insect sampling methodology in their lady beetle observation study. Since 1989, lady beetles (Coleoptera: Coccinellidae) have been sampled weekly over the growing season using yellow sticky cards. In 2021, the original sticky traps were discontinued by the manufacturer and replaced with a similar, but not identical trap. We conducted a 3‐year study while the new traps were phased in to examine how the trap change would impact the observed biodiversity patterns at the site. We examined community metrics and individual taxa captures to examine within‐year and between‐year differences in performance between the card types. Overall, we noted several small but statistically detectable differences in capture patterns between the two trap types. After accounting for other sources of variation, we observed a difference in Shannon diversity of insects captured on the two card types, but not richness or abundance, for the overall insect community. Yet, these differences were dwarfed by the magnitude of difference observed between years within card types. For individual taxa, similar patterns held: between trap differences could be detected statistically, but the number of differences in capture rate between trap types was less than the number of differences observed for the same trap, between years. Thus, we conclude that while subtle changes in methodology could impact data produced in long‐term experiments; in this case, the magnitude of this change is smaller than other factors such as time and plant treatment. However, if sustained changes in the capture rates of focal taxa are observed, future data users may use our observations to specifically quantify and correct for these shifting patterns related to the protocol change. 

Abstract ‘Marginal lands’ are low productivity sites abandoned from agriculture for reasons such as low or high soil water content, challenging topography, or nutrient deficiency. To avoid competition with crop production, cellulosic bioenergy crops have been proposed for cultivation on marginal lands, however on these sites they may be more strongly affected by environmental stresses such as low soil water content. In this study we used rainout shelters to induce low soil moisture on marginal lands and determine the effect of soil water stress on switchgrass growth and the subsequent production of bioethanol. Five marginal land sites that span a latitudinal gradient in Michigan and Wisconsin were planted to switchgrass in 2013 and during the 2018–2021 growing seasons were exposed to reduced precipitation under rainout shelters in comparison to ambient precipitation. The effect of reduced precipitation was related to the environmental conditions at each site and biofuel production metrics (switchgrass biomass yields and composition and ethanol production). During the first year (2018), the rainout shelters were designed with 60% rain exclusion, which did not affect biomass yields compared to ambient conditions at any of the field sites, but decreased switchgrass fermentability at the Wisconsin Central–Hancock site. In subsequent years, the shelters were redesigned to fully exclude rainfall, which led to reduced biomass yields and inhibited fermentation for three sites. When switchgrass was grown in soils with large reductions in moisture and increases in temperature, the potential for biofuel production was significantly reduced, exposing some of the challenges associated with producing biofuels from lignocellulosic biomass grown under drought conditions. 

Abstract Microorganisms can help plants and animals contend with abiotic stressors, but why they provide such benefits remains unclear. Here we investigated byproduct benefits, which occur when traits that increase the fitness of one species provide incidental benefits to another species with no direct cost to the provider. In a greenhouse experiment, microbial traits predicted plant responses to soil moisture such that bacteria with self‐beneficial traits in drought increased plant early growth, size at reproduction, and chlorophyll concentration under drought, while bacteria with self‐beneficial traits in well‐watered environments increased these same plant traits in well‐watered soils. Thus, microbial traits that promote microbial success in different moisture environments also promote plant success in these same environments. Our results demonstrate that byproduct benefits, a concept developed to explain the evolution of cooperation in pairwise mutualisms, can also extend to interactions between plants and nonsymbiotic soil microbes. 

Abstract To advance understanding of biodiversity and ecosystem function, ecologists seek widely applicable relationships among species diversity and other ecosystem characteristics such as species productivity, biomass, and abundance. These metrics vary widely across ecosystems and no relationship among any combination of them that is valid across habitats, taxa, and spatial scales, has heretofore been found. Here we derive such a relationship, an equation of state, among species richness, energy flow, biomass, and abundance by combining results from the Maximum Entropy Theory of Ecology and the Metabolic Theory of Ecology. It accurately captures the relationship among these state variables in 42 data sets, including vegetation and arthropod communities, that span a wide variety of spatial scales and habitats. The success of our ecological equation of state opens opportunities for estimating difficult-to-measure state variables from measurements of others, adds support for two current theories in ecology, and is a step toward unification in ecology. 

Abstract Cellulosic bioenergy is a primary land‐based climate mitigation strategy, with soil carbon (C) storage and nitrogen (N) conservation as important mitigation elements. Here, we present 13 years of soil C and N change under three cellulosic cropping systems: monoculture switchgrass (Panicum virgatumL.), a five native grasses polyculture, and no‐till corn (Zea maysL.). Soil C and N fractions were measured four times over 12 years. Bulk soil C in the 0–25 cm depth at the end of the study period ranged from 28.4 (± 1.4 se) Mg C ha⁻¹in no‐till corn, to 30.8 (± 1.4) Mg C ha⁻¹in switchgrass, and to 34.8 (± 1.4) Mg C ha⁻¹in native grasses. Mineral‐associated organic matter (MAOM) ranged from 60% to 90% and particulate organic matter (POM) from 10% to 40% of total soil C. Over 12 years, total C as well as both C fractions persisted under no‐till corn and switchgrass and increased under native grasses. In contrast, POM N stocks decreased 33% to 45% across systems, whereas MAOM N decreased only in no‐till corn and by less than 13%. Declining POM N stocks likely reflect pre‐establishment land use, which included alfalfa and manure in earlier rotations. Root production and large soil aggregate formation explained 69% (p < 0.001) and 36% (p = 0.024) of total soil C change, respectively, and 60% (p = 0.020) and 41% (p = 0.023) of soil N change, demonstrating the importance of belowground productivity and soil aggregates for producing and protecting soil C and conserving soil N. Differences between switchgrass and native grasses also indicate a dependence on plant diversity. Soil C and N benefits of bioenergy crops depend strongly on root productivity and pre‐establishment land use. 

abstract Long-term observations and experiments in diverse drylands reveal how ecosystems and services are responding to climate change. To develop generalities about climate change impacts at dryland sites, we compared broadscale patterns in climate and synthesized primary production responses among the eight terrestrial, nonforested sites of the United States Long-Term Ecological Research (US LTER) Network located in temperate (Southwest and Midwest) and polar (Arctic and Antarctic) regions. All sites experienced warming in recent decades, whereas drought varied regionally with multidecadal phases. Multiple years of wet or dry conditions had larger effects than single years on primary production. Droughts, floods, and wildfires altered resource availability and restructured plant communities, with greater impacts on primary production than warming alone. During severe regional droughts, air pollution from wildfire and dust events peaked. Studies at US LTER drylands over more than 40 years demonstrate reciprocal links and feedbacks among dryland ecosystems, climate-driven disturbance events, and climate change. 

Abstract Various soil health indicators that measure a chemically defined fraction of nitrogen (N) or a process related to N cycling have been proposed to quantify the potential to supply N to crops, a key soil function. We evaluated five N indicators (total soil N, autoclavable citrate extractable N, water‐extractable organic N, potentially mineralizable N, andN‐acetyl‐β‐D‐glucosaminidase activity) at 124 sites with long‐term experiments across North America evaluating a variety of managements. We found that 59%–81% of the variation in N indicators was among sites, with indicator values decreasing with temperature and increasing with precipitation and clay content. The N indicators increased from 6%–39% in response to decreasing tillage, cover cropping, retaining residue, and applying organic sources of nutrients. Overall, increasing the quantity of organic inputs, whether from increased residue retention, cover cropping, or rotations with higher biomass, resulted in higher values of the N indicators. Although N indicators responded to management in similar ways, the analysis cost and availability of testing laboratories is highly variable. Further, given the strong relationships of the N indicators with carbon (C) indicators, measuring soil organic C along with 24‐h potential C mineralization could be used as a proxy for N supply instead of measuring potentially mineralizable N or any other N indicator directly.