Colin W. Bell

Linking Microbial Community Structure and Function to Seasonal Differences in Soil Moisture and Temperature in a Chihuahuan Desert Grassland

Global and regional climate models predict higher air temperature and less frequent, but larger precipitation events in arid regions within the next century. While many studies have addressed the impact of variable climate in arid ecosystems on plant growth and physiological responses, fewer studies have addressed soil microbial community responses to seasonal shifts in precipitation and temperature in arid ecosystems. This study examined the impact of a wet (2004), average (2005), and dry (2006) year on subsequent responses of soil microbial community structure, function, and linkages, as well as soil edaphic and nutrient characteristics in a mid-elevation desert grassland in the Chihuahuan Desert. Microbial community structure was classified as bacterial (Gram-negative, Gram-positive, and actinomycetes) and fungal (saprophytic fungi and arbuscular mycorrhiza) categories using (fatty acid methyl ester) techniques. Carbon substrate use and enzymic activity was used to characterize microbial community function annually and seasonally (summer and winter). The relationship between saprophytic fungal community structure and function remained consistent across season independent of the magnitude or frequency of precipitation within any given year. Carbon utilization by fungi in the cooler winter exceeded use in the warmer summer each year suggesting that soil temperature, rather than soil moisture, strongly influenced fungal carbon use and structure and function dynamics. The structure/function relationship for AM fungi and soil bacteria notably changed across season. Moreover, the abundance of Gram-positive bacteria was lower in the winter compared to Gram-negative bacteria. Bacterial carbon use, however, was highest in the summer and lower during the winter. Enzyme activities did not respond to either annual or seasonal differences in the magnitude or timing of precipitation. Specific structural components of the soil microbiota community became uncoupled from total microbial function during different seasons. This change in the microbial structure/function relationship suggests that different components of the soil microbial community may provide similar ecosystem function, but differ in response to seasonal temperature and precipitation. As soil microbes encounter increased soil temperatures and altered precipitation amounts and timing that are predicted for this region, the ability of the soil microbial community to maintain functional resilience across the year may be reduced in this Chihuahuan Desert ecosystem.

Precipitation timing and magnitude differentially affect aboveground annual net primary productivity in three perennial species in a Chihuahuan Desert grassland

Plant productivity in deserts may be more directly responsive to soil water availability than to precipitation. However, measurement of soil moisture alone may not be enough to elucidate plant responses to precipitation pulses, as edaphic factors may influence productivity when soil moisture is adequate. The first objective of the study was to determine the responses of the aboveground annual net primary productivity (ANPP) of three perennial species (from different functional groups) in a Chihuahuan Desert grassland to variation in natural precipitation (annual and seasonal) and a 25% increase in seasonal precipitation (supplemental watering in summer and winter). Secondly, ANPP responses to other key environmental and soil parameters were explored during dry, average, and wet years over a 5-yr period. ANPP predictors for each species were dynamic. High ANPP in Dasylirion leiophyllum was positively associated with higher soil NH(4)-N and frequent larger precipitation events, while that in Bouteloua curtipendula was positively correlated with frequent small summer precipitation events with short inter-pulse periods and supplemental winter water. Opuntia phaeacantha was responsive to small precipitation events with short inter-pulse periods. Although several studies have shown ANPP increases with increases in precipitation and soil moisture in desert systems, this was not observed here as a universal predictor of ANPP, particularly in dry years.

High-throughput fluorometric measurement of potential soil extracellular enzyme activities.

Microbes in soils and other environments produce extracellular enzymes to depolymerize and hydrolyze organic macromolecules so that they can be assimilated for energy and nutrients. Measuring soil microbial enzyme activity is crucial in understanding soil ecosystem functional dynamics. The general concept of the fluorescence enzyme assay is that synthetic C-, N-, or P-rich substrates bound with a fluorescent dye are added to soil samples. When intact, the labeled substrates do not fluoresce. Enzyme activity is measured as the increase in fluorescence as the fluorescent dyes are cleaved from their substrates, which allows them to fluoresce. Enzyme measurements can be expressed in units of molarity or activity. To perform this assay, soil slurries are prepared by combining soil with a pH buffer. The pH buffer (typically a 50 mM sodium acetate or 50 mM Tris buffer), is chosen for the buffers particular acid dissociation constant (pKa) to best match the soil sample pH. The soil slurries are inoculated with a nonlimiting amount of fluorescently labeled (i.e. C-, N-, or P-rich) substrate. Using soil slurries in the assay serves to minimize limitations on enzyme and substrate diffusion. Therefore, this assay controls for differences in substrate limitation, diffusion rates, and soil pH conditions; thus detecting potential enzyme activity rates as a function of the difference in enzyme concentrations (per sample). Fluorescence enzyme assays are typically more sensitive than spectrophotometric (i.e. colorimetric) assays, but can suffer from interference caused by impurities and the instability of many fluorescent compounds when exposed to light; so caution is required when handling fluorescent substrates. Likewise, this method only assesses potential enzyme activities under laboratory conditions when substrates are not limiting. Caution should be used when interpreting the data representing cross-site comparisons with differing temperatures or soil types, as in situ soil type and temperature can influence enzyme kinetics.

Soil microbial and nutrient responses to 7 years of seasonally altered precipitation in a Chihuahuan Desert grassland

Soil microbial communities in Chihuahuan Desert grasslands generally experience highly variable spatiotemporal rainfall patterns. Changes in precipitation regimes can affect belowground ecosystem processes such as decomposition and nutrient cycling by altering soil microbial community structure and function. The objective of this study was to determine if increased seasonal precipitation frequency and magnitude over a 7-year period would generate a persistent shift in microbial community characteristics and soil nutrient availability. We supplemented natural rainfall with large events (one/winter and three/summer) to simulate increased precipitation based on climate model predictions for this region. We observed a 2-year delay in microbial responses to supplemental precipitation treatments. In years 3-5, higher microbial biomass, arbuscular mycorrhizae abundance, and soil enzyme C and P acquisition activities were observed in the supplemental water plots even during extended drought periods. In years 5-7, available soil P was consistently lower in the watered plots compared to control plots. Shifts in soil P corresponded to higher fungal abundances, microbial C utilization activity, and soil pH. This study demonstrated that 25% shifts in seasonal rainfall can significantly influence soil microbial and nutrient properties, which in turn may have long-term effects on nutrient cycling and plant P uptake in this desert grassland.

Extracellular enzymes in terrestrial, freshwater, and marine environments: perspectives on system variability and common research needs

Extracellular enzymes produced by heterotrophic microbial communities are major drivers of carbon and nutrient cycling in terrestrial, freshwater, and marine environments. Although carbon and nutrient cycles are coupled on global scales, studies of extracellular enzymes associated with terrestrial, freshwater, and marine microbial communities are not often compared across ecosystems. In part, this disconnect arises because the environmental parameters that control enzyme activities in terrestrial and freshwater systems, such as temperature, pH, and moisture content, have little explanatory power for patterns of enzyme activities in marine systems. Instead, factors such as the functional diversity of microbial communities may explain varying patterns of enzyme activities observed in the ocean to date. In any case, many studies across systems focus on similar issues that highlight the commonalities of microbial community organization. Examples include the effective lifetime of enzymes released into the environment; the extent to which microbial communities coordinate enzyme expression to decompose complex organic substrates; and the influence of microbial community composition on enzyme activities and kinetics. Here we review the often-disparate research foci in terrestrial, freshwater, and marine environments. We consider the extent to which environmental factors may regulate extracellular enzyme activities within each ecosystem, and highlight commonalities and current methodological challenges to identify research questions that may aid in integrating cross-system perspectives in the future.

Increased plant productivity and decreased microbial respiratory C loss by plant growth-promoting rhizobacteria under elevated CO2

Increased plant productivity and decreased microbial respiratory C loss can potentially mitigate increasing atmospheric CO2, but we currently lack effective means to achieve these goals. Soil microbes may play critical roles in mediating plant productivity and soil C/N dynamics under future climate scenarios of elevated CO2 (eCO2) through optimizing functioning of the root-soil interface. By using a labeling technique with 13C and 15N, we examined the effects of plant growth-promoting Pseudomonas fluorescens on C and N cycling in the rhizosphere of a common grass species under eCO2. These microbial inoculants were shown to increase plant productivity. Although strong competition for N between the plant and soil microbes was observed, the plant can increase its capacity to store more biomass C per unit of N under P. fluorescens addition. Unlike eCO2 effects, P. fluorescens inoculants did not change mass-specific microbial respiration and accelerate soil decomposition related to N cycling, suggesting these microbial inoculants mitigated positive feedbacks of soil microbial decomposition to eCO2. The potential to mitigate climate change by optimizing soil microbial functioning by plant growth-promoting Pseudomonas fluorescens is a prospect for ecosystem management.

New insights into enzymes in the environment

This article introduces the thematic special issue “Enzymes in the Environment: Incorporating Enzymes and Microbial Physiology into Biogeochemical Models”. This collection of review articles, original studies, and models stemmed from a workshop organized by the Enzymes in the Environment Research Coordination Network, which was held at Colorado State University in Fort Collins, CO, USA in May 2012. The meeting, themed “Incorporating Enzymes and Microbial Physiology into Biogeochemical Models” featured talks from both modelers and experimentalists with experience ranging across terrestrial, aquatic, and marine ecosystems. This special issue represents a series of integrative discussions that occurred among experimentalists and modelers at the meeting. This issue highlights that a better understanding of carbon cycling pathways and subsequent carbon availability dynamics among ecosystems may be achieved by simultaneously integrating aspects of terrestrial and aquatic carbon cycling and microbial physiology into biogeochemical cycling models.

Opposing effects of different soil organic matter fractions on crop yields

Soil organic matter is critical to sustainable agriculture because it provides nutrients to crops as it decomposes and increases nutrient- and water-holding capacity when built up. Fast- and slow-cycling fractions of soil organic matter can have different impacts on crop production because fast-cycling fractions rapidly release nutrients for short-term plant growth and slow-cycling fractions bind nutrients that mineralize slowly and build up water-holding capacity. We explored the controls on these fractions in a tropical agroecosystem and their relationship to crop yields. We performed physical fractionation of soil organic matter from 48 farms and plots in western Kenya. We found that fast-cycling, particulate organic matter was positively related to crop yields, but did not have a strong effect, while slower-cycling, mineral-associated organic matter was negatively related to yields. Our finding that slower-cycling organic matter was negatively related to yield points to a need to revise the view that stabilization of organic matter positively impacts food security. Our results support a new paradigm that different soil organic matter fractions are controlled by different mechanisms, potentially leading to different relationships with management outcomes, like crop yield. Effectively managing soils for sustainable agriculture requires quantifying the effects of specific organic matter fractions on these outcomes.

Phosphorus mobilizing consortium Mammoth P™ enhances plant growth

Phosphorus (P) is a critical nutrient used to maximize plant growth and yield. Current agriculture management practices commonly experience low plant P use efficiency due to natural chemical sorption and transformations when P fertilizer is applied to soils. A perplexing challenge facing agriculture production is finding sustainable solutions to deliver P more efficiently to plants. Using prescribed applications of specific soil microbial assemblages to mobilize soil bound—P to improve crop nutrient uptake and productivity has rarely been employed. We investigated whether inoculation of soils with a bacterial consortium developed to mobilize soil P, named Mammoth PTM, could increase plant productivity. In turf, herbs, and fruits, the combination of conventional inorganic fertilizer combined with Mammoth PTM increased productivity up to twofold compared to the fertilizer treatments without the Mammoth PTM inoculant. Jalapeño plants were found to bloom more rapidly when treated with either Mammoth P. In wheat trials, we found that Mammoth PTM by itself was able to deliver yields equivalent to those achieved with conventional inorganic fertilizer applications and improved productivity more than another biostimulant product. Results from this study indicate the substantial potential of Mammoth PTM to enhance plant growth and crop productivity.

Bridging the gap between modelers and experimentalists

Second International Enzymes in the Environment Research Coordination Network Workshop: Incorporating Enzymes and Microbial Physiology Into Biogeochemical Models; Colorado State University, Fort Collins, Colorado, 15–18 May 2012 Recent studies have provided new insights into the factors controlling the production and activities of extracellular enzymes, the proximate agents of organic matter decomposition. Enzymes and other aspects of microbial physiology have been incorporated into simple models but have not yet been explicitly incorporated into biogeochemical models capable of predicting fluxes of carbon and nutrients. One reason for this is that empirical data collected at one or few locations and time points do not meet the needs of modelers attempting to integrate across spatial and temporal scales. Furthermore, there is a disconnect between the types of data measured in observational and experimental studies and the data needed to parameterize biogeochemical models. For example, the most common approach is to measure maximum potential enzyme activities in laboratory assays, whereas models require measurements of enzyme kinetics when substrate is limiting.