CeO2 nanoparticle dose and exposure modulate soybean development and plant-mediated responses in root-associated bacterial communities.

Agricultural soils are increasingly undergoing inadvertent and purposeful exposures to engineered CeO2 nanoparticles (NPs), which can impact crops and root-associated microbial communities. However, interactions between NP concentration and exposure duration on plant-mediated responses of root-associated bacterial communities are not well understood. Soybeans seedlings were grown in soil with uncoated NPs added at concentrations of 0, 1 or 100 mg kg-1. Total soil exposure durations were either 190 days, starting 106 days before planting or 84 days with NP amendments coinciding with planting. We assessed plant development, bacterial diversity, differential abundance and inferred functional changes across rhizosphere, rhizoplane, and root tissue compartments. Plant non-monotonic dose responses were mirrored in bacterial communities. Most notably, effects were magnified in the rhizoplane under low-dose, short-exposures. Enriched metabolic pathways were primarily related to biosynthesis and degradation/utilization/assimilation, rather than responses to metals or oxidative stress. Our results indicate that plant-mediated bacterial responses were greater than direct NP impacts. Also, we identify needs for modeling non-monotonic legume stress responses that account for coinfection with mutualistic and parasitic bacteroids. Our findings provide new insights regarding effects of applications of soil amendments such as biosolids containing NPs or nano-enabled formulations used in cultivation of legumes and other crops.

Ozone exposure-response relationships parametrized for sixteen tree species with varying sensitivity in the United States.

It is well known that exposure to ambient O3 can decrease growth in many tree species in the United States (US). Our study reports experimental data from outdoor open-top chamber (OTC) studies that quantify total biomass response changes for seedlings of 16 species native to western and eastern North America, which were exposed to several levels of elevated O3 for one or more years. The primary objective of this study is to establish a reference set of parameters for these seedling exposure-response relationships using a 3-month (92 day) 12-hr W126 O3 metric used by US Environmental Protection Agency and other agencies to assess risk to trees from O3 exposure. We classified the 16 species according to their sensitivity, based on the biomass loss response functions to protect from a 5% biomass loss. The three-month 12-h W126 estimated to result in a 5% biomass loss was 2.5-9.2 ppm-h for sensitive species, 20.8-25.2 ppm-h for intermediate species, and > 28.7 ppm-h for insensitive species. The most sensitive tree species include black cherry, ponderosa pine, quaking aspen, red alder, American sycamore, tulip poplar and winged sumac. These species are ecologically important and widespread across US. The effects of O3 on whole-plant biomass depended on exposure duration and dynamics and on the number of successive years of exposure. These species-specific exposure-response relationships will allow US agencies and other groups to better estimate biomass losses based on ozone exposures in North America and can be used in risk assessment and scenario analyses.

Focused Microbiome Shifts in Reconstructed Wetlands Correlated with Elevated Copper Concentrations Originating from Micronized Copper Azole-Treated Wood.

Micronized copper (Cu) azole (MCA) wood preservative formulations include Cu in nano form, and relatively little is known about longer term effects of Cu leached from MCA into wetland ecosystems. We tested the hypothesis that changes in soil microbiomes within reconstructed freshwater wetlands will be associated with exposure to elevated Cu concentrations originating from immersed MCA-treated wood stakes. Eight replicate communities were assembled with Willamette Valley (OR, USA) flood plain soil and clonally propagated wetland plants within mesocosms. Inundated communities were equilibrated for 5 months before installation of MCA or control southern yellow pine stakes (n = 4 communities/experimental group). Soil samples were collected for 16S and internal transcribed spacer amplicon sequencing to quantify responses in prokaryotes and eukaryotes, respectively, at 15 time points, spanning two simulated seasonal dry downs, for up to 678 days. Physiochemical properties of water and soil were monitored at 20 and 12 time points respectively, over the same period. For both taxonomic groups of organisms, phylogenetic diversity increased and was positively correlated with elapsed days. Furthermore, there was significant divergence among eukaryotes during the second year based on experimental group. Although the composition of taxa underwent succession over time, there was significantly reduced relative abundance of sequence variants from Gomphonema diatoms and Scutellinia fungi in communities where MCA wood stakes were present compared with the controls. These focused microbiome shifts were positively correlated with surface water Cu and soil Cu concentrations, which were significantly elevated in treated communities. The reconstructed communities were effective systems for assessing potential impacts to wetland microbiomes after exposure to released copper. The results further inform postcommercialization risk assessments on MCA-treated wood. Environ Toxicol Chem 2021;40:3351-3368. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.

Wheat exposure to cerium oxide nanoparticles over three generations reveals transmissible changes in nutrition, biochemical pools, and response to soil N.

This study investigated the effects of third generation exposure to cerium oxide nanoparticles (CeO2-NPs) on biomass, elemental and 15N uptake, and fatty acid contents of wheat (Triticum aestivum). At low or high nitrogen treatment (48 or 112 mg N), seeds exposed for two generations to 0 or 500 mg CeO2-NPs per kg soil treatment were cultivated for third year in soil amended with 0 or 500 mg CeO2-NPs per kg soil. The results showed that parental and current exposures to CeO2-NPs increased the root biomass in daughter plants with greater magnitude of increase at low N than high N. When wheat received CeO2-NPs in year 3, root elemental contents increased primarily at low N, suggesting an important role of soil N availability in altering root nutrient acquisition. The δ15N ratios, previously shown to be altered by CeO2-NPs, were only affected by current and not parental exposure, indicating effects on N uptake and/or metabolism are not transferred from one generation to the next. Seed fatty acid composition was also influenced both by prior and current exposure to CeO2-NPs. The results suggest that risk assessments of NP exposure may need to include longer-term, transgenerational effects on growth and grain quality of agronomic crops.

Shifts in N and δ15N in wheat and barley exposed to cerium oxide nanoparticles.

The effects of cerium oxide nanoparticles (CeO2-NPs) on 15N/14N ratio (δ15N) in wheat and barley were investigated. Seedlings were exposed to 0 and 500 mg CeO2-NPs/L (Ce-0 and Ce-500, respectively) in hydroponic suspension supplied with NH4NO3, NH4 +, or NO3 -. N uptake and δ15N discrimination (i.e. differences in δ15N of plant and δ15N of N source) were measured. Results showed that N content and 15N abundance decreased in wheat but increased in barley. Ce-500 only induced whole-plant δ15N discrimination (-1.48‰, P ≤ 0.10) with a simultaneous decrease (P ≤ 0.05) in whole-plant δ15N (-3.24‰) compared to Ce-0 (-2.74‰) in wheat in NH4 +. Ce-500 decreased (P ≤ 0.01) root δ15N of wheat in NH4NO3 and NH4 + (3.23 and -2.25‰, respectively) compared to Ce-0 (4.96 and -1.27‰, respectively), but increased (P ≤ 0.05) root δ15N of wheat in NO3 - (3.27‰) compared to Ce-0 (2.60‰). Synchrotron micro-XRF revealed the presence of CeO2-NPs in shoots of wheat and barley regardless of N source. Although the longer-term consequences of CeO2-NP exposure on N uptake and metabolism are unknown, the results clearly show the potential for ENMs to interfere with plant metabolism of critical plant nutrients such as N even when toxicity is not observed.

Douglas-Fir ( Pseudotsuga menziesii (Mirb.) Franco) Transcriptome Profile Changes Induced by Diesel Emissions Generated with CeO2 Nanoparticle Fuel Borne Catalyst.

It is important to understand molecular effects on plants exposed to compounds released from use of products containing engineered nanomaterials. Here, we present mRNA sequencing data on transcriptome impacts to Douglas-fir following 2 weeks of sublethal exposure to 30:1 diluted airborne emissions released from combustion of diesel fuel containing engineered CeO2 nanoparticle catalysts (DECe). Our hypothesis was that chamber exposure to DECe would induce distinct transcriptome changes in seedling needles compared with responses to conventional diesel exhaust (DE) or filtered DECe Gas Phase. Significantly increased uptake/binding of Ce in needles of DECe treated seedlings was 2.7X above background levels and was associated with altered gene expression patterns. All 225 Blast2GO gene ontologies (GOs) enriched by up-regulated DECe transcripts were nested within GOs for DE, however, 29 of 31 enriched GOs for down-regulated DECe transcripts were unique. MapMan analysis also identified three pathways enriched with DECe down-regulated transcripts. There was prominent representation of genes with attenuated expression in transferase, transporter, RNA regulation and protein degradation GOs and pathways. CeO2 nanoparticle additive decreased and shifted molecular impact of diesel emissions. Wide-spread use of such products and chronic environmental exposure to DECe may adversely affect plant physiology and development.

13C isotopic signature and C concentration of soil density fractions illustrate reduced C allocation to subalpine grassland soil under high atmospheric N deposition.

We followed soil C fluxes in a subalpine grassland system exposed to experimentally increased atmospheric N deposition for 7 years. Earlier we found that, different from the plant productivity response, the bulk soil C stock increase was highest at the medium, not the high N input as hypothesized. This implies that a smaller N-deposition rate has a greater potential to favor the biological greenhouse gas-sink. To help elucidate the mechanisms controlling those changes in SOC in response to N deposition, we produced four soil density fractions and analyzed soil organic C concentration [SOC], as well as δ13C signatures (δ13CSOC) of SOC components. Soil respired CO2 (δ13CCO2) was analyzed to better distinguish seasonal short term dynamics from N-deposition effects and to identify the predominant substrate of soil respiration. Both at the start of the experiment and after 7 years we found a strong, negative correlation between [SOC] and δ13CSOC of the soil density fractions in the control treatment, consistent with an advanced stage of microbial processing of SOC in fractions of higher density. During the experiment the [SOC] increased in the two lighter density fractions, but decreased in the two heavier fractions, suggesting a possible priming effect that accelerated decomposition of formerly recalcitrant (heavy) organic matter pools. The seasonal pattern of soil δ13CCO2 was affected by weather and canopy development, and δ13CCO2 values for the different N treatment levels indicated that soil respiration originated primarily from the lightest density fractions. Surprisingly, [SOC] increases were significantly higher under medium N deposition in the <1.8 fraction and in bulk soil, compared to the high N treatment. Analogously, the depletion of δ13CSOC was significantly higher in the medium compared to the high N treatment in the three lighter fractions. Thus, medium N deposition favored the highest C sequestration potential, compared to the low N control and the high N treatment. Clearly, our results show that it is inappropriate to use plant productivity N response as an indicator for shifts in SOC content in grassland ecosystems. Here, isotopic techniques illustrated why atmospheric N deposition of 14 kg N ha-1 yr-1 is below, and 54 kg N ha-1 yr-1 is above a threshold that tips the balance between new, assimilative gains and respiratory losses towards a net loss of [SOC] for certain soil fractions in the subalpine grassland.

A comprehensive framework for evaluating the environmental health and safety implications of engineered nanomaterials.

Engineered nanomaterials (ENM) are a growing aspect of the global economy, and their safe and sustainable development, use, and eventual disposal requires the capability to forecast and avoid potential problems. This review provides a framework to evaluate the health and safety implications of ENM releases into the environment, including purposeful releases such as for antimicrobial sprays or nano-enabled pesticides, and inadvertent releases as a consequence of other intended applications. Considerations encompass product life cycles, environmental media, exposed populations, and possible adverse outcomes. This framework is presented as a series of compartmental flow diagrams that serve as a basis to help derive future quantitative predictive models, guide research, and support development of tools for making risk-based decisions. After use, ENM are not expected to remain in their original form due to reactivity and/or propensity for hetero-agglomeration in environmental media. Therefore, emphasis is placed on characterizing ENM as they occur in environmental or biological matrices. In addition, predicting the activity of ENM in the environment is difficult due to the multiple dynamic interactions between the physical/chemical aspects of ENM and similarly complex environmental conditions. Others have proposed the use of simple predictive functional assays as an intermediate step to address the challenge of using physical/chemical properties to predict environmental fate and behavior of ENM. The nodes and interactions of the framework presented here reflect phase transitions that could be targets for development of such assays to estimate kinetic reaction rates and simplify model predictions. Application, refinement, and demonstration of this framework, along with an associated knowledgebase that includes targeted functional assay data, will allow better de novo predictions of potential exposures and adverse outcomes.

Intergenerational responses of wheat (Triticum aestivum L.) to cerium oxide nanoparticles exposure.

The intergenerational impact of engineered nanomaterials in plants is a key knowledge gap in the literature. A soil microcosm study was performed to assess the effects of multi-generational exposure of wheat (Triticum aestivum L.) to cerium oxide nanoparticles (CeO2-NPs). Seeds from plants that were exposed to 0, 125, and 500 mg CeO2-NPs/kg soil (Ce-0, Ce-125 or Ce-500, respectively) in first generation (S1) were cultivated in factorial combinations of Ce-0, Ce-125 or Ce-500 to produce second generation (S2) plants. The factorial combinations for first/second generation treatments in Ce-125 were S1-Ce-0/S2-Ce-0, S1-Ce-0/S2-Ce-125, S1-Ce-125/S2-Ce-0 and S1-Ce-125/S2-Ce-125, and in Ce-500 were S1-Ce-0/S2-Ce-0, S1-Ce-0/S2-Ce-500, S1-Ce-500/S2-Ce-0 and S1-Ce-500/S2-Ce-500. Agronomic, elemental, isotopic, and synchrotron X-ray fluorescence (XRF) and X-ray absorption near-edge spectroscopy (XANES) data were collected on second generation plants. Results showed that plants treated during the first generation only with either Ce-125 or Ce-500 (e.g. S1-Ce-125/S2-Ce-0 or S1-Ce-500/S2-Ce-0) had reduced accumulation of Ce (61 or 50%), Fe (49 or 58%) and Mn (34 or 41%) in roots, and δ15N (11 or 8%) in grains compared to the plants not treated in both generations (i.e. S1-Ce-0/S2-Ce-0). Plants treated in both generations with Ce-125 (i.e. S1-Ce-125/S2-Ce-125) produced grains that had lower Mn, Ca, K, Mg and P relative to plants treated in the second generation only (i.e. S1-Ce-0/S2-Ce-125). In addition, synchrotron XRF elemental chemistry maps of soil/plant thin-sections revealed limited transformation of CeO2-NPs with no evidence of plant uptake or accumulation. The findings demonstrated that first generation exposure of wheat to CeO2-NPs affects the physiology and nutrient profile of the second generation plants. However, the lack of concentration-dependent responses indicate that complex physiological processes are involved which alter uptake and metabolism of CeO2-NPs in wheat.

Molecular and physiological responses to titanium dioxide and cerium oxide nanoparticles in Arabidopsis.

Changes in tissue transcriptomes and productivity of Arabidopsis thaliana were investigated during exposure of plants to 2 widely used engineered metal oxide nanoparticles, titanium dioxide (nano-titania) and cerium dioxide (nano-ceria). Microarray analyses confirmed that exposure to either nanoparticle altered the transcriptomes of rosette leaves and roots, with comparatively larger numbers of differentially expressed genes found under nano-titania exposure. Nano-titania induced more differentially expressed genes in rosette leaves, whereas roots had more differentially expressed genes under nano-ceria exposure. MapMan analyses indicated that although nano-titania up-regulated overall metabolism in both tissues, metabolic processes under nano-ceria remained mostly unchanged. Gene enrichment analysis indicated that both nanoparticles mainly enriched ontology groups such as responses to stress (abiotic and biotic), and defense responses (pathogens), and responses to endogenous stimuli (hormones). Nano-titania specifically induced genes associated with photosynthesis, whereas nano-ceria induced expression of genes related to activating transcription factors, most notably those belonging to the ethylene responsive element binding protein family. Interestingly, there were also increased numbers of rosette leaves and plant biomass under nano-ceria exposure, but not under nano-titania. Other transcriptomic responses did not clearly relate to responses observed at the organism level, possibly because of functional and genomic redundancy in Arabidopsis, which may mask expression of morphological changes, despite discernable responses at the transcriptome level. In addition, transcriptomic changes often relate to transgenerational phenotypic development, and hence it may be productive to direct further experimental work to integrate high-throughput genomic results with longer term changes in subsequent generations. Environ Toxicol Chem 2017;36:71-82. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Germination and early plant development of ten plant species exposed to titanium dioxide and cerium oxide nanoparticles.

Ten agronomic plant species were exposed to different concentrations of nano-titanium dioxide (nTiO2 ) or nano-cerium oxide (nCeO2 ) (0 µg/mL, 250 µg/mL, 500 µg/mL, and 1000 µg/mL) to examine potential effects on germination and early seedling development. The authors modified a standard test protocol developed for soluble chemicals (OPPTS 850.4200) to determine if such an approach might be useful for screening engineered nanomaterials (ENMs) and whether there were differences in response across a range of commercially important plant species to 2 common metal oxide ENMs. Eight of 10 species responded to nTiO2 , and 5 species responded to nCeO2 . Overall, it appeared that early root growth may be a more sensitive indicator of potential effects from ENM exposure than germination. The observed effects did not always relate to the exposure concentration, indicating that mass-based concentration may not fully explain the developmental effects of these 2 ENMs. The results suggest that nTiO2 and nCeO2 have different effects on early plant growth of agronomic species, with unknown effects at later stages of the life cycle. In addition, standard germination tests, which are commonly used for toxicity screening of new materials, may not detect the subtle but potentially more important changes associated with early growth and development in terrestrial plants. Environ Toxicol Chem 2016;35:2223-2229. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

Phenotypic and genomic responses to titanium dioxide and cerium oxide nanoparticles in Arabidopsis germinants.

The effects of exposure to nanoparticles of titanium dioxide (nano-titanium) and cerium oxide (nano-cerium) on gene expression and growth in Arabidopsis thaliana germinants were studied by using microarrays and quantitative real-time polymerase chain reaction (qPCR), and by evaluating germinant phenotypic plasticity. Exposure to 12 d of either nano-titania or nano-ceria altered the regulation of 204 and 142 genes, respectively. Genes induced by the nanoparticles mainly include ontology groups annotated as stimuli responsive, including both abiotic (oxidative stress, salt stress, water transport) and biotic (respiratory burst as a defense against pathogens) stimuli. Further analysis of the differentially expressed genes indicates that both nanoparticles affected a range of metabolic processes (deoxyribonucleic acid [DNA] metabolism, hormone metabolism, tetrapyrrole synthesis, and photosynthesis). Individual exposures to the nanoparticles increased percentages of seeds with emergent radicles, early development of hypocotyls and cotyledons, and those with fully grown leaves. Although there were distinct differences between the nanoparticles in their affect on molecular mechanisms attributable to enhancing germinant growth, both particles altered similar suites of genes related to various pathways and processes related to enhanced growth.

Preferential interaction of Na+ over K+ with carboxylate-functionalized silver nanoparticles.

Elucidating mechanistic interactions between monovalent cations (Na(+)/K(+)) and engineered nanoparticle surfaces to alter particle stability in polar media have received little attention. We investigated relative preferential interaction of Na(+) and K(+) with carboxylate-functionalized silver nanoparticles (carboxylate-AgNPs) to determine if interaction preference followed the Hofmeister series (Na(+)>K(+)). We hypothesized that Na(+) will show greater affinity than K(+) to pair with carboxylates on AgNP surfaces, thereby destabilizing the colloidal system. Destabilization upon Na(+) or K(+) interacting with carboxylate-AgNPs was evaluated probing changes in multiple physicochemical characteristics: surface plasmon resonance/optical absorbance, electrical conductivity, pH, hydrodynamic diameter, electrophoretic mobility, surface charge, amount of Na(+)/K(+) directly associated with AgNPs, and Ag(+) dissociation kinetics. We show that Na(+) and K(+) react differently, indicating local Na(+) pairing with carboxylates on AgNP surfaces is kinetically faster and remarkably favored over K(+), thus supporting Hofmeister ordering. Our results suggest that AgNPs may transform into micron-size aggregates upon release into aqueous environments and that the fate of such aggregates may need consideration when assessing environmental risk.

Potential for metal contamination by direct sonication of nanoparticle suspensions.

While conducting toxicity tests with nano titanium dioxide, the authors found that test suspensions were being contaminated with aluminum and titanium from tip erosion during direct sonication. The contaminating alloy particles had a measurable size distribution and zeta potential using dynamic light scattering, which changed the measured characteristics of the suspensions. Caution should be used when employing direct sonication for preparing test suspensions due to potential interferences of these particles in toxicological assessments.

Below-ground carbon allocation in mature beech and spruce trees following long-term, experimentally enhanced O3 exposure in Southern Germany.

Canopies of adult European beech (Fagus sylvatica) and Norway spruce (Picea abies) were labeled with CO(2) depleted in (13)C to evaluate carbon allocation belowground. One-half the trees were exposed to elevated O(3) for 6 yrs prior to and during the experiment. Soil-gas sampling wells were placed at 8 and 15 cm and soil CO(2) was sampled during labeling in mid-late August, 2006. In beech, delta(13)CO(2) at both depths decreased approximately 50 h after labeling, reflecting rapid translocation of fixed C to roots and release through respiration. In spruce, label was detected in fine-root tissue, but there was no evidence of label in delta(13)CO(2). The results show that C fixed in the canopy rapidly reaches respiratory pools in beech roots, and suggest that spruce may allocate very little of recently-fixed carbon into root respiration during late summer. A change in carbon allocation belowground due to long-term O(3) exposure was not observed.

Belowground effects of enhanced tropospheric ozone and drought in a beech/spruce forest (Fagus sylvatica L./Picea abies [L.] Karst).

The effects of experimentally elevated O(3) on soil respiration rates, standing fine-root biomass, fine-root production and delta(13)C signature of newly produced fine roots were investigated in an adult European beech/Norway spruce forest in Germany during two subsequent years with contrasting rainfall patterns. During humid 2002, soil respiration rate was enhanced under elevated O(3) under beech and spruce, and was related to O(3)-stimulated fine-root production only in beech. During dry 2003, the stimulating effect of O(3) on soil respiration rate vanished under spruce, which was correlated with decreased fine-root production in spruce under drought, irrespective of the O(3) regime. delta(13)C signature of newly formed fine-roots was consistent with the differing g(s) of beech and spruce, and indicated stomatal limitation by O(3) in beech and by drought in spruce. Our study showed that drought can override the stimulating O(3) effects on fine-root dynamics and soil respiration in mature beech and spruce forests.

An approach for evaluating the effectiveness of various ozone air quality standards for protecting trees.

We demonstrate an approach for evaluating the level of protection attained using a variety of forms and levels of past, current, and proposed Air Quality Standards (AQSs). The U.S. Clean Air Act requires the establishment of ambient air quality standards to protect health and public welfare. However, determination of attainment of these standards is based on ambient pollutant concentrations rather than prevention of adverse effects. To determine if a given AQS protected against adverse effects on vegetation, hourly ozone concentrations were adjusted to create exposure levels that "just attain" a given standard. These exposures were used in combination with a physiologically-based tree growth model to account for the interactions of climate and ozone. In the evaluation, we used ozone concentrations from two 6-year time periods from the San Bernardino Mountains in California. There were clear differences in the level of vegetation protection achieved with the various AQSs. Based on modeled plant growth, the most effective standards were the California 8-hr average maximum of 70 ppb and a seasonal, cumulative, concentration-weighted index (SUM06), which if attained, resulted in annual growth reductions of 1% or less. Least effective was the 1-hr maximum of 120 ppb which resulted in a 7% annual reduction. We conclude that combining climate, exposure scenarios, and a process-based plant growth simulator was a useful approach for evaluating effectiveness of current or proposed air quality standards, or evaluating the form and/or level of a standard based on preventing adverse growth effects.

Ozone and natural systems: understanding exposure, response, and risk.

Research aimed at understanding the response of plants to ozone has been conducted for over four decades but little of it has addressed intact natural systems. Even so, there is sufficient scientific information at this time to support air quality standards that will protect natural terrestrial ecosystems from ozone. What is unknown is the risk associated with continued exposure of natural systems, including both above- and below-ground components, in combination with other stresses including changing temperature and precipitation, elevated carbon dioxide, pests and pathogens, invasive species, and other activities that may fragment the landscape. Research to support an assessment of the ecological risk associated with ozone as it exists, in a milieu of stresses, must include endpoints beyond those addressed in the past, primarily productivity and species composition. To estimate the risk to society of ozone impacts on natural systems, endpoints such as the integrity of soil food webs, the quantity and quality of water supplied from terrestrial ecosystems, wildlife and recreational values, and the transfer and fate of carbon, nutrients, and water within the systems must be quantified. Not only will this research provide the basis for a sound estimate of risk, but also it will improve our understanding of fundamental ecosystem processes.

Source-sink balance and carbon allocation below ground in plants exposed to ozone.

The role of tropospheric ozone in altering plant growth and development has been the subject of thousands of publications over the last several decades. Still, there is limited understanding regarding the possible effects of ozone on soil processes. In this review, the effects of ozone are discussed using the flow of carbon from the atmosphere, through the plant to soils, and back to the atmosphere as a framework. A conceptual model based on carbohydrate signaling is used to illustrate physiological changes in response to ozone, and to discuss possible feedbacks that may occur. Despite past emphasis on above-ground effects, ozone has the potential to alter below-ground processes and hence ecosystem characteristics in ways that are not currently being considered. Contents Summary 213 I. Introduction 213 II. Source-sink model: carbohydrate signaling 214 III. Effect of ozone on above-ground sources and sinks 216 IV. Decreased allocation below ground 218 V. Carbon flux to soils 220 VI. Soil food web 223 VII. Summary, conclusions and future research 223 Acknowledgements 223 References 223.

Role of carbohydrate supply in white and brown root respiration of ponderosa pine.

• Respiration of intact ponderosa pine (Pinus ponderosa) fine roots (< 2.5 mm) was measured to determine the role of recently fixed carbohydrate in maintaining root metabolism of growing white (WR) and recently suberized brown roots (BR). • The CO2 efflux and O2 uptake of individual roots were followed continuously over 24 h after carbohydrate supply was altered by exposing shoots to light/dark treatments and by root excision. • In situ respiration of individual WR and BR averaged 86.0 ± 2.6 and 21.1 ± 1.5 mol CO2  g-1  h-1 , respectively. Growth respiration was estimated to be approximately two-thirds the rate of WR respiration. Attached WR and BR respiration did not decline significantly over 24 h under continuous light. The WR respiration significantly decreased during a dark period. All roots maintained relatively constant respiration rates for at least 6 h after excision. Respiratory quotient (RQ; CO2  : O2 ) was not different between attached (0.84 ± 0.014) and detached (0.85 ± 0.017) roots. CO2 environment of the cuvette did not influence WR or BR respiration. • The WR appear to be more sensitive to supply of current photosynthate than BR. Shoot light environment needs to be considered when measuring root and soil CO2 efflux.