Biochar amendment suppresses N2 O emissions but has no impact on 15 N site preference in an anaerobic soil.

RATIONALE: Biochar amendments often decrease N2 O gas production from soil, but the mechanisms and magnitudes are still not well characterized since N2 O can be produced via several different microbial pathways. We evaluated the influence of biochar amendment on N2 O emissions and N2 O isotopic composition, including 15 N site preference (SP) under anaerobic conditions. METHODS: An agricultural soil was incubated with differing levels of biochar. Incubations were conducted under anaerobic conditions for 10 days with and without acetylene, which inhibits N2 O reduction to N2 . The N2 O concentrations were measured every 2 days, the SPs were determined after 5 days of incubation, and the inorganic nitrogen concentrations were measured after the incubation. RESULTS: The SP values with acetylene were consistent with N2 O production by bacterial denitrification and those without acetylene were consistent with bacterial denitrification that included N2 O reduction to N2 . There was no effect of biochar on N2 O production in the presence of acetylene between day 3 and day 10. However, in the absence of acetylene, soils incubated with 4% biochar produced less N2 O than soils with no biochar addition. Different amounts of biochar amendment did not change the SP values. CONCLUSIONS: Our study used N2 O emission rates and SP values to understand biochar amendment mechanisms and demonstrated that biochar amendment reduces N2 O emissions by stimulating the last step of denitrification. It also suggested a possible shift in N2 O-reducing microbial taxa in 4% biochar samples.

Applying the principles of isotope analysis in plant and animal ecology to forensic science in the Americas.

The heart of forensic science is application of the scientific method and analytical approaches to answer questions central to solving a crime: Who, What, When, Where, and How. Forensic practitioners use fundamentals of chemistry and physics to examine evidence and infer its origin. In this regard, ecological researchers have had a significant impact on forensic science through the development and application of a specialized measurement technique-isotope analysis-for examining evidence. Here, we review the utility of isotope analysis in forensic settings from an ecological perspective, concentrating on work from the Americas completed within the last three decades. Our primary focus is on combining plant and animal physiological models with isotope analyses for source inference. Examples of the forensic application of isotopes-including stable isotopes, radiogenic isotopes, and radioisotopes-span from cotton used in counterfeit bills to anthrax shipped through the U.S. Postal Service and from beer adulterated with cheap adjuncts to human remains discovered in shallow graves. Recent methodological developments and the generation of isotope landscapes, or isoscapes, for data interpretation promise that isotope analysis will be a useful tool in ecological and forensic studies for decades to come.

Bryophyte gas-exchange dynamics along varying hydration status reveal a significant carbonyl sulphide (COS) sink in the dark and COS source in the light.

Carbonyl sulphide (COS) is a potential tracer of gross primary productivity (GPP), assuming a unidirectional COS flux into the vegetation that scales with GPP. However, carbonic anhydrase (CA), the enzyme that hydrolyses COS, is expected to be light independent, and thus plants without stomata should continue to take up COS in the dark. We measured net CO2 (AC ) and COS (AS ) uptake rates from two astomatous bryophytes at different relative water contents (RWCs), COS concentrations, temperatures and light intensities. We found large AS in the dark, indicating that CA activity continues without photosynthesis. More surprisingly, we found a nonzero COS compensation point in light and dark conditions, indicating a temperature-driven COS source with a Q10 (fractional change for a 10°C temperature increase) of 3.7. This resulted in greater AS in the dark than in the light at similar RWC. The processes underlying such COS emissions remain unknown. Our results suggest that ecosystems dominated by bryophytes might be strong atmospheric sinks of COS at night and weaker sinks or even sources of COS during daytime. Biotic COS production in bryophytes could result from symbiotic fungal and bacterial partners that could also be found on vascular plants.

Stable isotopes in leaf water of terrestrial plants.

Leaf water contains naturally occurring stable isotopes of oxygen and hydrogen in abundances that vary spatially and temporally. When sufficiently understood, these can be harnessed for a wide range of applications. Here, we review the current state of knowledge of stable isotope enrichment of leaf water, and its relevance for isotopic signals incorporated into plant organic matter and atmospheric gases. Models describing evaporative enrichment of leaf water have become increasingly complex over time, reflecting enhanced spatial and temporal resolution. We recommend that practitioners choose a model with a level of complexity suited to their application, and provide guidance. At the same time, there exists some lingering uncertainty about the biophysical processes relevant to patterns of isotopic enrichment in leaf water. An important goal for future research is to link observed variations in isotopic composition to specific anatomical and physiological features of leaves that reflect differences in hydraulic design. New measurement techniques are developing rapidly, enabling determinations of both transpired and leaf water δ(18) O and δ(2) H to be made more easily and at higher temporal resolution than previously possible. We expect these technological advances to spur new developments in our understanding of patterns of stable isotope fractionation in leaf water.

Shrubland resilience varies across soil types: implications for operationalizing resilience in ecological restoration.

In ecosystems with alternative stable states, restoration success can be thought of as overcoming the resilience of an undesirable state to promote an alternative state that yields greater ecosystem services. Since greater resilience of undesirable states translates into reduced restoration potential, quantifying differences in resilience can enhance restoration planning. In the context of shrub-encroached rangeland restoration, shrubland resilience is the capacity of a woody vegetated state to absorb management interventions designed to produce a more desirable grass-dominated state, and remain within its current regime. Therefore, differences in the resilience of a state can be quantified in a relative sense by measuring whether a state switches to an alternate state following perturbation or remains in its current stability domain. Here we designed an experimental manipulation to assess the contribution of soils to differences in the relative resilience of a shrub-invaded state. In this large-scale experiment, we repeated perturbations across a gradient of soil textures to inform restoration practitioners of differences in the relative resilience of shrubland occurring on different soil types to common rangeland restoration practices. On each soil type, we compared the relative ability of the shrubland state to withstand chemical and mechanical brush control treatments, commonly employed in this study region, to untreated controls. While the shrubland community composition did not differ prior to the study, its capacity to absorb and recover from brush removal treatments depended on soil type. Shrubland resilience to chemical and mechanical brush removal was highest on coarse soils. On these soils, brush removal temporarily restored grassland dominance, but woody plants quickly regained pretreatment levels of dominance. However, shrublands on fine soils did not recover following treatments, continuing to be grass-dominated for the duration of the study. This study highlights a simple approach for prioritizing restoration actions by mapping the locations of different soil attributes that support shrub-dominated states with differing levels of resilience to brush control. This experimental approach provides a basis for operationalizing resilience in restoration and prioritizing management actions across a range of environmental conditions, which is critical given the economic constraints associated with broad-scale mechanical and chemical interventions for rangeland restoration.

Contrasting water-use strategies revealed by species-specific transpiration dynamics in the Caatinga dry forest.

In forest ecosystems, transpiration (T) patterns are important for quantifying water and carbon fluxes and are major factors in predicting ecosystem change. Seasonal changes in rainfall and soil water content can alter the sensitivity of sap flux density to daily variations in vapor pressure deficit (VPD). This sensitivity is species-specific and is thought to be related to hydraulic strategies. The aim of this work is to better understand how the sap flux density of species with low versus high wood density differ in their sensitivity to VPD and soil water content and how potentially opposing water-use strategies influence T dynamics, and ultimately, correlations to evapotranspiration (ET). We use hysteresis area analysis to quantify the sensitivity of species-specific sap flux density to changes in the VPD, breakpoint-based models to determine the soil water content threshold instigating a T response and multiscalar wavelet coherency to correlate T to ET. We found that low wood density Commiphora leptophloeos (Mart.) Gillett had a more dynamic T pattern, a greater sensitivity to VPD at high soil water content, required a higher soil water content threshold for this sensitivity to be apparent, and had a significant coherency correlation with ET at daily to monthly timescales. This behavior is consistent with a drought avoidance strategy. High wood density Cenostigma pyramidale  (Tul.)  E. Gagnon & G. P. Lewis, conversely, had a more stable T pattern, responded to VPD across a range of soil water content, tolerated a lower soil water content threshold to T, and had a significant coherency correlation with ET at weekly timescales. This behavior is consistent with a drought-tolerant strategy. We build on previous research to show that these species have contrasting water-use strategies that should be considered in large-scale modeling efforts.

Nitrogen limitation constrains sustainability of ecosystem response to CO2.

Enhanced plant biomass accumulation in response to elevated atmospheric CO2 concentration could dampen the future rate of increase in CO2 levels and associated climate warming. However, it is unknown whether CO2-induced stimulation of plant growth and biomass accumulation will be sustained or whether limited nitrogen (N) availability constrains greater plant growth in a CO2-enriched world. Here we show, after a six-year field study of perennial grassland species grown under ambient and elevated levels of CO2 and N, that low availability of N progressively suppresses the positive response of plant biomass to elevated CO2. Initially, the stimulation of total plant biomass by elevated CO2 was no greater at enriched than at ambient N supply. After four to six years, however, elevated CO2 stimulated plant biomass much less under ambient than enriched N supply. This response was consistent with the temporally divergent effects of elevated CO2 on soil and plant N dynamics at differing levels of N supply. Our results indicate that variability in availability of soil N and deposition of atmospheric N are both likely to influence the response of plant biomass accumulation to elevated atmospheric CO2. Given that limitations to productivity resulting from the insufficient availability of N are widespread in both unmanaged and managed vegetation, soil N supply is probably an important constraint on global terrestrial responses to elevated CO2.

Extensive Variation in Drought-Induced Gene Expression Changes Between Loblolly Pine Genotypes.

Drought response is coordinated through expression changes in a large suite of genes. Interspecific variation in this response is common and associated with drought-tolerant and -sensitive genotypes. The extent to which different genetic networks orchestrate the adjustments to water deficit in tolerant and sensitive genotypes has not been fully elucidated, particularly in non-model or woody plants. Differential expression analysis via RNA-seq was evaluated in root tissue exposed to simulated drought conditions in two loblolly pine (Pinus taeda L.) clones with contrasting tolerance to drought. Loblolly pine is the prevalent conifer in southeastern U.S. and a major commercial forestry species worldwide. Significant changes in gene expression levels were found in more than 4,000 transcripts [drought-related transcripts (DRTs)]. Genotype by environment (GxE) interactions were prevalent, suggesting that different cohorts of genes are influenced by drought conditions in the tolerant vs. sensitive genotypes. Functional annotation categories and metabolic pathways associated with DRTs showed higher levels of overlap between clones, with the notable exception of GO categories in upregulated DRTs. Conversely, both differentially expressed transcription factors (TFs) and TF families were largely different between clones. Our results indicate that the response of a drought-tolerant loblolly pine genotype vs. a sensitive genotype to water limitation is remarkably different on a gene-by-gene level, although it involves similar genetic networks. Upregulated transcripts under drought conditions represent the most diverging component between genotypes, which might depend on the activation and repression of substantially different groups of TFs.

Plant functional types broadly describe water use strategies in the Caatinga, a seasonally dry tropical forest in northeast Brazil.

In seasonally dry tropical forests, plant functional type can be classified as deciduous low wood density, deciduous high wood density, or evergreen high wood density species. While deciduousness is often associated with drought-avoidance and low wood density is often associated with tissue water storage, the degree to which these functional types may correspond to diverging and unique water use strategies has not been extensively tested.We examined (a) tolerance to water stress, measured by predawn and mid-day leaf water potential; (b) water use efficiency, measured via foliar δ13C; and (c) access to soil water, measured via stem water δ18O.We found that deciduous low wood density species maintain high leaf water potential and low water use efficiency. Deciduous high wood density species have lower leaf water potential and variable water use efficiency. Both groups rely on shallow soil water. Evergreen high wood density species have low leaf water potential, higher water use efficiency, and access alternative water sources. These findings indicate that deciduous low wood density species are drought avoiders, with a specialized strategy for storing root and stem water. Deciduous high wood density species are moderately drought tolerant, and evergreen high wood density species are the most drought tolerant group.Synthesis. Our results broadly support the plant functional type framework as a way to understand water use strategies, but also highlight species-level differences.

Stable isotope models to predict geographic origin and cultivation conditions of marijuana.

Here we describe stable isotope based models using hydrogen and carbon isotope ratios to predict geographic region-of-origin and growth environment for marijuana, with the intent of applying these models to analyses of marijuana trafficking in the USA. The models were developed on the basis of eradication specimens and border specimens seized throughout the USA. We tested reliability of the geographic region-of-origin and growth environment models with a "blind" set of 60 marijuana eradication specimens obtained from counties throughout the USA. The two geographic region-of-origin model predictions were 60-67% reliable and cultivation environment model predictions were 86% accurate for the blind specimens. We demonstrate here that stable isotope ratio analysis of marijuana seizures can significantly improve our understanding of marijuana distribution networks and it is for that purpose that these models were developed.

Tree species fine-root demography parallels habitat specialization across a sandhill soil resource gradient.

Single species can substantially alter belowground processes in ecosystems via differential root production and death. However, information on species differences in fine-root demography is virtually absent for natural communities. In this field study, we recorded species-specific fine-root (<2 mm in diameter) demography in adults of four tree species (Pinus palustris, Quercus laevis, Q. incana, and Q. margaretta) that are distributed differentially along soil resource gradients in the fall-line sandhills of the southeastern United States. At a subxeric habitat where all four species co-occur, roots of individual trees of each species were isolated in rhizotrons and tracked individually for three years. Quercus species had similar fine-root morphology but differed substantially for fine-root demography and architecture. Quercus laevis and Q. incana, the species from xeric habitats, showed lower fine-root production, death, percentage mortality, turnover rates, and risk of death, and greater life span and mean root segment length (MRSL) than Q. margaretta, the species from subxeric habitats. Fine roots of P. palustris (a generalist) showed high production and intermediate mortality, turnover rate, longevity, and MRSL. Fine-root survival increased with root order (first to fourth in centripetal order), but the degree of change was species specific. Q. margaretta showed greater increases in survival with order, but all species had similar demography of third- and fourth-order roots. Mycorrhizal roots had greater longevity than non-mycorrhizal roots only in Q. laevis. Species differences were also seasonal. Although these Quercus species are leaf deciduous, some growth of fine roots occurred in Q. margaretta during the "leaf-dormant" season. In our narrow-scale species comparison, species differences in ecological distribution were consistent with the observed variation in fine-root demography and architecture with greater resolution than leaf characters or other root traits such as morphology. Our results also show that narrow-scale variation in fine-root demography (including intra-generic differences) can be as large as broad-scale variation across biomes and vegetation types. Hence, small shifts in community composition have the potential to produce substantial changes below ground.

Using δ13C and δ18O to analyze loblolly pine (Pinus taeda L.) response to experimental drought and fertilization.

Drought frequency and intensity are projected to increase throughout the southeastern USA, the natural range of loblolly pine (Pinus taeda L.), and are expected to have major ecological and economic implications. We analyzed the carbon and oxygen isotopic compositions in tree ring cellulose of loblolly pine in a factorial drought (~30% throughfall reduction) and fertilization experiment, supplemented with trunk sap flow, allometry and microclimate data. We then simulated leaf temperature and applied a multi-dimensional sensitivity analysis to interpret the changes in the oxygen isotope data. This analysis found that the observed changes in tree ring cellulose could only be accounted for by inferring a change in the isotopic composition of the source water, indicating that the drought treatment increased the uptake of stored moisture from earlier precipitation events. The drought treatment also increased intrinsic water-use efficiency, but had no effect on growth, indicating that photosynthesis remained relatively unaffected despite 19% decrease in canopy conductance. In contrast, fertilization increased growth, but had no effect on the isotopic composition of tree ring cellulose, indicating that the fertilizer gains in biomass were attributable to greater leaf area and not to changes in leaf-level gas exchange. The multi-dimensional sensitivity analysis explored model behavior under different scenarios, highlighting the importance of explicit consideration of leaf temperature in the oxygen isotope discrimination (Δ18Oc) simulation and is expected to expand the inference space of the Δ18Oc models for plant ecophysiological studies.

Plant diversity, CO2, and N influence inorganic and organic N leaching in grasslands.

In nitrogen (N)-limited systems, the potential to sequester carbon depends on the balance between N inputs and losses as well as on how efficiently N is used, yet little is known about responses of these processes to changes in plant species richness, atmospheric CO2 concentration ([CO2]), and N deposition. We examined how plant species richness (1 or 16 species), elevated [CO2] (ambient or 560 ppm), and inorganic N addition (0 or 4 g x m(-2) x yr(-1)) affected ecosystem N losses, specifically leaching of dissolved inorganic N (DIN) and organic N (DON) in a grassland field experiment in Minnesota, USA. We observed greater DIN leaching below 60 cm soil depth in the monoculture plots (on average 1.8 and 3.1 g N x m(-2) x yr(-1) for ambient N and N-fertilized plots respectively) than in the 16-species plots (0.2 g N x m(-2) x yr(-1) for both ambient N and N-fertilized plots), particularly when inorganic N was added. Most likely, loss of complementary resource use and reduced biological N demand in the monoculture plots caused the increase in DIN leaching relative to the high-diversity plots. Elevated [CO2] reduced DIN concentrations under conditions when DIN concentrations were high (i.e., in N-fertilized and monoculture plots). Contrary to the results for DIN, DON leaching was greater in the 16-species plots than in the monoculture plots (on average 0.4 g N x m(-2) x yr(-1) in 16-species plots and 0.2 g N x m(-2) x yr(-1) in monoculture plots). In fact, DON dominated N leaching in the 16-species plots (64% of total N leaching as DON), suggesting that, even with high biological demand for N, substantial amounts of N can be lost as DON. We found no significant main effects of elevated [CO2] on DIN or DON leaching; however, elevated [CO2] reduced the positive effect of inorganic N addition on DON leaching, especially during the second year of observation. Our results suggest that plant species richness, elevated [CO2], and N deposition alter DIN loss primarily through changes in biological N demand. DON losses can be as large as DIN loss but are more sensitive to organic matter production and turnover.

Geography and vintage predicted by a novel GIS model of wine delta18O.

Wine hydrogen and oxygen stable isotopes record the climatic conditions experienced by the grape vine and the isotopic composition of the vine's source water during berry development. As such, stable isotopes have been explored extensively for use in detecting wine adulteration or for independently verifying claims of origin. We present the results of a study designed to evaluate the relationships between wine water delta18O and spatial climate and precipitation delta18O patterns across the winegrape-growing regions of Washington, Oregon, and California. Retail wines produced from typically small vineyards across these regions were obtained from the 2002 vintage, and the delta18O of wine water was analyzed using a CO2 equilibration method. Significant correlations were observed between the measured wine water delta18O from 2002 and the long-term average precipitation delta18O and late season 2002 climate, based on a spatial join with continuous geographic information system (GIS) maps of these drivers. We then developed a regression model that was implemented spatially in a GIS. The GIS model is the first of its kind and allows spatially explicit predictions of wine delta18O across the region. Because high spatial resolution monthly climate layers are now available for many years, wine delta18O could be modeled for previous years. We therefore tested the model by executing it for specific years and comparing the model predictions with previously published results for wine delta18O from seven vintages from Napa and Livermore Valleys, California. With the exception of one year, an anomaly potentially related to the effects of El Niño on precipitation isotopic composition, the model predicted well the wine delta18O for both locations for all vintages and generally reflected the consistent enrichment of wine from Napa relative to Livermore. Our results suggest that wine water delta18O records both source water delta18O and climate during the late stages of winegrape ripening and that GIS models of wine water delta18O are useful tools for independently verifying claims of regional origin and vintage.

Forensic applications of light-element stable isotope ratios of Ricinus communis seeds and ricin preparations.

Seeds of the castor plant Ricinus communis are of forensic interest because they are the source of the poison ricin. We tested whether stable isotope ratios of castor seeds and ricin preparations can be used as a forensic signature. We collected over 300 castor seed samples worldwide and measured the C, N, O, and H isotope ratios of the whole seeds and oil. We prepared ricin by three different procedures, acetone extraction, salt precipitation, and affinity chromatography, and compared their isotope ratios to those of the source seeds. The N isotope ratios of the ricin samples and source seeds were virtually identical. Therefore, N isotope ratios can be used to correlate ricin prepared by any of these methods to source seeds. Further, stable isotope ratios distinguished >99% of crude and purified ricin protein samples in pairwise comparison tests. Stable isotope ratios therefore constitute a valuable forensic signature for ricin preparations.

Tracing retail cannabis in the United States: geographic origin and cultivation patterns.

BACKGROUND: Although cannabis is the most readily available and widely used illicit drug in the United States, there remains significant uncertainty about the importance of different production regions and trafficking patterns. METHODS: We analysed 628 "retail" cannabis seizures from over 50 municipalities across the United States for hydrogen and carbon isotope ratios to predict their growth locations and environments. RESULTS: Results are presented for 22 consolidated retail locations across the United States. Evaluation of specimens from within these retail areas suggested that cannabis seizures had region-dependent origins, often from both domestic and foreign sources, and although indoor growth was common in many areas, there was also regional dependence in the proportions cultivated under indoor versus outdoor conditions. CONCLUSION: Street-available cannabis exhibits region-specific trafficking patterns, both Mexican- and Canadian-grown cannabis are apparently widely available, and indoor-grown cannabis appears to be cultivated and trafficked in both warm and cool weather localities throughout the United States.

The stable isotope ratios of marijuana. II. Strontium isotopes relate to geographic origin.

Effectively addressing marijuana trade is aided by understanding marijuana geographic sources. We analyzed the (87)Sr/(86)Sr of marijuana samples grown in 79 counties across the United States to determine if a primary geologic signal is retained in marijuana, which could therefore be useful for geographic sourcing. The marijuana results were compared with modeled bedrock (87)Sr/(86)Sr values based on (87)Rb decay rates and a generalized geologic map of the U.S.A. A significant correlation was observed between marijuana (87)Sr/(86)Sr and modeled bedrock (87)Sr/(86)Sr. Although values clustered near the 1:1 relationship, there was a predominance of positive anomalies, perhaps attributable to carbonate bedrock. A small number of negative anomalies were also observed, which were generally associated with granitic bedrocks. These results suggest that strontium isotopes in marijuana record the geographic origins of marijuana, and that refinement of the base strontium map (or strontium isoscape) and improved understanding of other strontium sources would be productive.

Stable isotope ratios of marijuana. I. Carbon and nitrogen stable isotopes describe growth conditions.

There remains significant uncertainty in illicit marijuana cultivation. We analyzed the delta(13)C and delta(15)N of 508 domestic samples from known U.S.A. counties, 31 seized from a single location, 5 samples grown in Mexico and Colombia, and 10 northwest border seizures. For a subset, inflorescences and leaves were analyzed separately. These data revealed a strong correspondence, with inflorescences having slightly higher delta(13)C and delta(15)N values than leaves. A framework for interpreting these results is introduced and evaluated. Samples identified as outdoor-grown by delta(13)C were generally recorded as such by the Drug Enforcement Administration (DEA). DEA-classified indoor-grown samples had the most negative delta(13)C values, consistent with indoor cultivation, although many were also in the outdoor-grown domain. Delta(15)N indicated a wide range of fertilizers across the dataset. Samples seized at the single location suggested multiple sources. Northwest border delta(13)C values suggested indoor growth, whereas for the Mexican and Colombian samples they indicated outdoor growth.

A simplified GIS approach to modeling global leaf water isoscapes.

The stable hydrogen (delta(2)H) and oxygen (delta(18)O) isotope ratios of organic and inorganic materials record biological and physical processes through the effects of substrate isotopic composition and fractionations that occur as reactions proceed. At large scales, these processes can exhibit spatial predictability because of the effects of coherent climatic patterns over the Earth's surface. Attempts to model spatial variation in the stable isotope ratios of water have been made for decades. Leaf water has a particular importance for some applications, including plant organic materials that record spatial and temporal climate variability and that may be a source of food for migrating animals. It is also an important source of the variability in the isotopic composition of atmospheric gases. Although efforts to model global-scale leaf water isotope ratio spatial variation have been made (especially of delta(18)O), significant uncertainty remains in models and their execution across spatial domains. We introduce here a Geographic Information System (GIS) approach to the generation of global, spatially-explicit isotope landscapes (= isoscapes) of "climate normal" leaf water isotope ratios. We evaluate the approach and the resulting products by comparison with simulation model outputs and point measurements, where obtainable, over the Earth's surface. The isoscapes were generated using biophysical models of isotope fractionation and spatially continuous precipitation isotope and climate layers as input model drivers. Leaf water delta(18)O isoscapes produced here generally agreed with latitudinal averages from GCM/biophysical model products, as well as mean values from point measurements. These results show global-scale spatial coherence in leaf water isotope ratios, similar to that observed for precipitation and validate the GIS approach to modeling leaf water isotopes. These results demonstrate that relatively simple models of leaf water enrichment combined with spatially continuous precipitation isotope ratio and climate data layers yield accurate global leaf water estimates applicable to important questions in ecology and atmospheric science.