Evaluating Bacterial Nanocellulose Interfaces for Recording Surface Biopotentials from Plants.

The study of plant electrophysiology offers promising techniques to track plant health and stress in vivo for both agricultural and environmental monitoring applications. Use of superficial electrodes on the plant body to record surface potentials may provide new phenotyping insights. Bacterial nanocellulose (BNC) is a flexible, optically translucent, and water-vapor-permeable material with low manufacturing costs, making it an ideal substrate for non-invasive and non-destructive plant electrodes. This work presents BNC electrodes with screen-printed carbon (graphite) ink-based conductive traces and pads. It investigates the potential of these electrodes for plant surface electrophysiology measurements in comparison to commercially available standard wet gel and needle electrodes. The electrochemically active surface area and impedance of the BNC electrodes varied based on the annealing temperature and time over the ranges of 50 °C to 90 °C and 5 to 60 min, respectively. The water vapor transfer rate and optical transmittance of the BNC substrate were measured to estimate the level of occlusion caused by these surface electrodes on the plant tissue. The total reduction in chlorophyll content under the electrodes was measured after the electrodes were placed on maize leaves for up to 300 h, showing that the BNC caused only a 16% reduction. Maize leaf transpiration was reduced by only 20% under the BNC electrodes after 72 h compared to a 60% reduction under wet gel electrodes in 48 h. On three different model plants, BNC-carbon ink surface electrodes and standard invasive needle electrodes were shown to have a comparable signal quality, with a correlation coefficient of >0.9, when measuring surface biopotentials induced by acute environmental stressors. These are strong indications of the superior performance of the BNC substrate with screen-printed graphite ink as an electrode material for plant surface biopotential recordings.

Comment on "Soybean photosynthesis and crop yield are improved by accelerating recovery from photoprotection".

De Souza et al. (Research Articles, 19 Aug 2022, adc9831) recently claimed major soybean yield increases resulting from transformation of the nonphotochemical quenching mechanism of photosynthesis. However, there is little basis for the premise that such a transformation would result in yield increase. The field experiment was flawed and does not provide evidence for increases in crop yield.

Predominant Microbial Colonizers in the Root Endosphere and Rhizosphere of Turfgrass Systems: Pseudomonas veronii, Janthinobacterium lividum, and Pseudogymnoascus spp.

Microbes can colonize plant roots to modulate plant health and environmental fitness. Thus, using microbes to improve plant adaptation to biotic and abiotic stresses will be promising to abate the heavy reliance of management systems on synthetic chemicals and limited resource. This is particularly important for turfgrass systems because intensive management for plant available nutrients (e.g., nitrogen), water, and pest control is necessary to maintain a healthy and aesthetic landscape. However, little is known on microbial species and host compatibility in turfgrass root endosphere and rhizosphere. Here, by using marker gene high throughput sequencing approaches we demonstrated that a few bacterial and fungal species prevailed the root endosphere and rhizosphere and were of a broad host spectrum. Irrespective of turfgrass species (bermudagrass, ultradwarf bermudagrass, creeping bentgrass, and tall fescue), defoliation intensities (i.e., mowing height and frequency), turfgrass sites, and sampling time, Pseudomonas veronii was predominant in the root endosphere, constituting ∼38% of the total bacterial community, which was much higher than its presence in the bulk soil (∼0.5%) and rhizosphere (∼4.6%). By contrast, Janthinobacterium lividum and fungal species of the genus Pseudogymnoascus were more abundant in the rhizosphere, constituting ∼15 and ∼ 39% of the total bacterial and fungal community, respectively, compared to their respective presence in the bulk soil (∼ 0.1 and 5%) and root endosphere (∼ 0.8 and 0.3%). Such stark contrasts in the microbiome composition between the root endosphere, rhizosphere, and bulk soil were little influenced by turfgrass species, suggesting the broad turfgrass host compatibility of these bacterial and fungal species. Further, their dominance in respective niches were mutually unaffected, implying the possibility of developing a multiple species formula for coping turfgrass with environmental stresses. These species were likely involved in controlling pests, such as infectious nematodes and fungi, decomposing root debris, and helping turfgrass water and nutrient uptake; yet these possibilities need to be further examined.

Increasing Photosynthesis: Unlikely Solution For World Food Problem.

Increasing the photosynthesis rate of plants has been recently revitalized as an approach for increasing grain crop yields and solving world food crises. The idea that photosynthesis is the key to increasing grain crop yields is not new. Considerable research in the 1970s and 1980s showed that carbon input was not limiting for crop growth and yield. Instead, the availability and uptake of water and nutrients were found to be critical for increasing grain yield, and that conclusion still applies today. In this Opinion article, nitrogen limitation is given particular attention because of its quantitative linkage with vegetative and reproductive growth and its essential role as a quantitative component of seeds.

Leaf Traits That Contribute to Differential Ozone Response in Ozone-Tolerant and Sensitive Soybean Genotypes.

Ozone (O3) is a phytotoxic air pollutant that limits crop productivity. Breeding efforts to improve yield under elevated O3 conditions will benefit from understanding the mechanisms that contribute to O3 tolerance. In this study, leaf gas exchange and antioxidant metabolites were compared in soybean genotypes (Glycine max (L.) Merr) differing in ozone sensitivity. Mandarin (Ottawa) (O3-sensitive) and Fiskeby III (O3-tolerant) plants grown under charcoal-filtered (CF) air conditions for three weeks were exposed for five days to either CF conditions or 70 ppb O3 in continuously stirred tank reactors (CSTRs) in a greenhouse. In the CF controls, stomatal conductance was approximately 36% lower for Fiskeby III relative to Mandarin (Ottawa) while the two genotypes exhibited similar levels of photosynthesis. Ozone exposure induced significant foliar injury on leaves of Mandarin (Ottawa) associated with declines in both stomatal conductance (by 77%) and photosynthesis (by 38%). In contrast, O3 exposure resulted in minimal foliar injury on leaves of Fiskeby III with only a small decline in photosynthesis (by 5%), and a further decline in stomatal conductance (by 30%). There was a general trend towards higher ascorbic acid content in leaves of Fiskeby III than in Mandarin (Ottawa) regardless of treatment. The results confirm Fiskeby III to be an O3-tolerant genotype and suggest that reduced stomatal conductance contributes to the observed O3 tolerance through limiting O3 uptake by the plant. Reduced stomatal conductance was associated with enhanced water-use efficiency, providing a potential link between O3 tolerance and drought tolerance.

Limited-transpiration response to high vapor pressure deficit in crop species.

Water deficit under nearly all field conditions is the major constraint on plant yields. Other than empirical observations, very little progress has been made in developing crop plants in which specific physiological traits for drought are expressed. As a consequence, there was little known about under what conditions and to what extent drought impacts crop yield. However, there has been rapid progress in recent years in understanding and developing a limited-transpiration trait under elevated atmospheric vapor pressure deficit to increase plant growth and yield under water-deficit conditions. This review paper examines the physiological basis for the limited-transpiration trait as result of low plant hydraulic conductivity, which appears to be related to aquaporin activity. Methodology was developed based on aquaporin involvement to identify candidate genotypes for drought tolerance of several major crop species. Cultivars of maize and soybean are now being marketed specifically for arid conditions. Understanding the mechanism of the limited-transpiration trait has allowed a geospatial analyses to define the environments in which increased yield responses can be expected. This review highlights the challenges and approaches to finally develop physiological traits contributing directly to plant improvement for water-limited environments.

Leaf emergence (phyllochron index) and leaf expansion response to soil drying in cowpea genotypes.

Drought can result in severely decreased leaf area development, which impacts plant growth and yield. However, rarely is leaf emergence or leaf expansion separated to resolve the relative sensitivity to water-deficit of these two processes. Experiments were undertaken to impose drought over approximately 2 weeks for eight cowpea (Vigna unguiculata) genotypes grown in pots under controlled environmental conditions. Daily measures of phyllochron index (PI, leaf emergence) and leaf area increase (leaf expansion) were obtained. Each of these measures was referenced against volumetric soil water content, i.e. fraction transpirable soil water. Although there was no clear difference between leaf emergence and leaf expansion in sensitivity to drying soil, both processes were more sensitive to soil drying than plant transpiration rate. Genotypic differences in the soil water content at the initiation of the decline in PI were identified. However, no consistent difference in sensitivity to water-deficit in leaf expansion was found. The difference in leaf emergence among genotypes in sensitivity to soil drying can now be exploited to provide guidance for plant improvement and crop yield increase.

Pot binding as a variable confounding plant phenotype: theoretical derivation and experimental observations.

MAIN CONCLUSION: Theoretical derivation predicted growth retardation due to pot water limitations, i.e., pot binding. Experimental observations were consistent with these limitations. Combined, these results indicate a need for caution in high-throughput screening and phenotyping. Pot experiments are a mainstay in many plant studies, including the current emphasis on developing high-throughput, phenotyping systems. Pot studies can be vulnerable to decreased physiological activity of the plants particularly when pot volume is small, i.e., "pot binding". It is necessary to understand the conditions under which pot binding may exist to avoid the confounding influence of pot binding in interpreting experimental results. In this paper, a derivation is offered that gives well-defined conditions for the occurrence of pot binding based on restricted water availability. These results showed that not only are pot volume and plant size important variables, but the potting media is critical. Artificial potting mixtures used in many studies, including many high-throughput phenotyping systems, are particularly susceptible to the confounding influences of pot binding. Experimental studies for several crop species are presented that clearly show the existence of thresholds of plant leaf area at which various pot sizes and potting media result in the induction of pot binding even though there may be no immediate, visual plant symptoms. The derivation and experimental results showed that pot binding can readily occur in plant experiments if care is not given to have sufficiently large pots, suitable potting media, and maintenance of pot water status. Clear guidelines are provided for avoiding the confounding effects of water-limited pot binding in studying plant phenotype.

Limited transpiration under high vapor pressure deficits of creeping bentgrass by application of Daconil-Action.

MAIN CONCLUSION: First observation that chemical spray can induce limited-transpiration rate under high vapor pressure deficit. It appears that acibenzolar may be key in inducing this water conservation trait. Irrigation and water use have become major issues in management of turfgrasses. Plant health products that have been introduced into the turfgrass market have been observed to improve plant performance in water stress conditions. In this study, we evaluated whether a selection of common plant health products alter the ability of creeping bentgrass (Agrostis stolonifera L.) to control transpiration under high vapor pressure deficit (VPD). The plant health treatments--Daconil Action, Insignia, and Signature--were applied to plots on golf course putting greens located in Raleigh NC and in Scottsdale, AZ. Using intact cores removed from the putting greens, transpiration rates were measured over a range of VPDs in controlled conditions. In all cases stretching over a 3-year period, bentgrass cores from field plots treated with Daconil-Action limited transpiration under high VPD conditions, while check treatments with water, and others treated with Insignia or Signature did not. Transpiration control became engaged when VPDs reached values ranging from 1.39 to 2.50 kPa, and was not strongly influenced by the field temperature at which the bentgrass was growing. Because all plots in NC had been treated with chlorothalonil-the key ingredient in Daconil Action to control diseases-it was concluded that the likely chemical ingredient in Daconil Action triggering the transpiration control response was acibenzolar. This is the first evidence that the limited-transpiration trait can be induced by a chemical application, and it implies significant potential for ameliorating drought vulnerability in cool-season turfgrasses, and likely other plant species.

Reactive nitrogen in turfgrass systems: relations to soil physical, chemical, and biological properties.

Turfgrass systems contribute to the loading of reactive N to water and air via runoff, leaching, and gas emission. Yet, a comprehensive approach has never been developed to assess N loss potential from turfgrass systems. We used pools and production of reactive N (inorganic N, extractable organic N, and NO) to estimate N loss potential and hypothesized that this potential could be predicated by basic soil properties. A total of 68 soil samples were taken from 17 bermudagrass sites in North Carolina. Basic soil properties were analyzed, including soil C and N, C:N ratio, microbial biomass, moisture, pH, and percent silt/clay/sand. Soil samples varied most widely in texture, followed by soil C and N, microbial biomass, moisture, pH, and C:N ratio. The pools of extractable organic N and inorganic N were comparable, indicating that soluble organic N should be considered as a pathway of N loss from turfgrass. Turfgrass with large pools and production of reactive N was characterized by high soil C and N, microbial biomass, and moisture. Because soil C and N accumulate over time after turfgrass establishment, turfgrass age could be a suitable practical indicator of N loss potential and thus could be used to implement changes in management. Pools and production of reactive N in liquid and gas phases were well correlated, suggesting that if a turfgrass system has a high potential of N loss via leaching and runoff, it may also be of a high potential for NO emissions.

Temperature interactions with transpiration response to vapor pressure deficit among cultivated and wild soybean genotypes.

A key strategy in soybean drought research is increased stomatal sensitivity to high vapor pressure deficit (VPD), which contributes to the 'slow wilting' trait observed in the field. These experiments examined whether temperature of the growth environment affected the ability of plants to respond to VPD, and thus control transpiration rate (TR). Two soybean [Glycine max (L.) Merr.] and four wild soybean [Glycine soja (Sieb. and Zucc.)] genotypes were studied. The TR was measured over a range of VPD when plants were growing at 25 or 30°C, and again after an abrupt increase of 5°C. In G. max, a restriction of TR became evident as VPD increased above 2.0 kPa when temperature was near its growth optimum of 30°C. 'Slow wilting' genotype plant introduction (PI) 416937 exhibited greater TR control at high VPD compared with Hutcheson, and only PI 416937 restrained TR after the shift to 35°C. Three of the four G. soja genotypes exhibited control over TR with increasing VPD when grown at 25°C, which is near their estimated growth optimum. The TR control became engaged at lower VPD than in G. max and was retained to differing degrees after a shift to 30°C. The TR control systems in G. max and G. soja clearly were temperature-sensitive and kinetically definable, and more restrictive in the 'slow wilting' soybean genotype. For the favorable TR control traits observed in G. soja to be useful for soybean breeding in warmer climates, the regulatory linkage with lower temperatures must be uncoupled.

Arbuscular mycorrhizal fungi increase organic carbon decomposition under elevated CO2.

The extent to which terrestrial ecosystems can sequester carbon to mitigate climate change is a matter of debate. The stimulation of arbuscular mycorrhizal fungi (AMF) by elevated atmospheric carbon dioxide (CO(2)) has been assumed to be a major mechanism facilitating soil carbon sequestration by increasing carbon inputs to soil and by protecting organic carbon from decomposition via aggregation. We present evidence from four independent microcosm and field experiments demonstrating that CO(2) enhancement of AMF results in considerable soil carbon losses. Our findings challenge the assumption that AMF protect against degradation of organic carbon in soil and raise questions about the current prediction of terrestrial ecosystem carbon balance under future climate-change scenarios.

Temperature influences the ability of tall fescue to control transpiration in response to atmospheric vapour pressure deficit.

Water availability for turfgrass systems is often limited and is likely to become more so in the future. Here, we conducted experiments that examined the ability of tall fescue (Festuca arundinacea Schreb.) to control transpiration with increasing vapour pressure deficit (VPD) and determined whether control was influenced by temperature. The first study was under steady-state conditions at two temperatures (21 and 27°C) and two VPDs (1.2 and 1.8kPa). At the lower temperature, water use was similar at both VPDs, indicating a restriction of transpiration at high VPD. At 27°C, transpiration control at high VPD was weakened and root growth also declined; both responses increase susceptibility to water-deficit stress. Another series of experiments was used to examine the physiological stability of the transpiration control. Temperature and VPD were adjusted in a stepwise manner and transpiration measured across a range of VPD in the days following environmental shifts. Results indicated that VPD control acclimated to the growth environment, with adjustment to drier conditions becoming evident after ~1 week. Control was again more effective at cool than at hot temperatures. Collectively, the results indicate that transpiration control by this cool season grass is most effective in the temperature range where it is best adapted.

Soil microbial responses to elevated CO2 and O3 in a nitrogen-aggrading agroecosystem.

Climate change factors such as elevated atmospheric carbon dioxide (CO2) and ozone (O3) can exert significant impacts on soil microbes and the ecosystem level processes they mediate. However, the underlying mechanisms by which soil microbes respond to these environmental changes remain poorly understood. The prevailing hypothesis, which states that CO2- or O3-induced changes in carbon (C) availability dominate microbial responses, is primarily based on results from nitrogen (N)-limiting forests and grasslands. It remains largely unexplored how soil microbes respond to elevated CO2 and O3 in N-rich or N-aggrading systems, which severely hinders our ability to predict the long-term soil C dynamics in agroecosystems. Using a long-term field study conducted in a no-till wheat-soybean rotation system with open-top chambers, we showed that elevated CO2 but not O3 had a potent influence on soil microbes. Elevated CO2(1.5×ambient) significantly increased, while O3 (1.4×ambient) reduced, aboveground (and presumably belowground) plant residue C and N inputs to soil. However, only elevated CO2 significantly affected soil microbial biomass, activities (namely heterotrophic respiration) and community composition. The enhancement of microbial biomass and activities by elevated CO2 largely occurred in the third and fourth years of the experiment and coincided with increased soil N availability, likely due to CO2-stimulation of symbiotic N2 fixation in soybean. Fungal biomass and the fungi∶bacteria ratio decreased under both ambient and elevated CO2 by the third year and also coincided with increased soil N availability; but they were significantly higher under elevated than ambient CO2. These results suggest that more attention should be directed towards assessing the impact of N availability on microbial activities and decomposition in projections of soil organic C balance in N-rich systems under future CO2 scenarios.

The community composition of soil-denitrifying bacteria from a turfgrass environment.

Soil-denitrifying bacteria in highly-managed turfgrass systems were examined to assess their response to land-use change and time under management. Denitrifier community composition and diversity in a turfgrass chronosequence of 1 to 95-years-old were compared with those in an adjacent pine-dominant forest via molecular investigations of nirK and nosZ gene fragments. Both denaturing gradient gel electrophoresis and sequenced clone libraries revealed that the denitrifier community became more diverse after turf establishment, and the diversity was then preserved. Furthermore, the composition of the turfgrass denitrifier community was slightly affected by time under management. Meta-analysis of sequenced nirK and nosZ gene fragments from a variety of ecosystems showed that denitrifier communities in pine and turf were more similar to those in other environments than to each other, suggesting that land-use change substantially modified the composition and increased the diversity of denitrifiers. This study provides a useful baseline of nirK- and nosZ-type soil denitrifier communities to aid in the evaluation of ecological and environmental impacts of turfgrass systems.

Intensive management affects composition of betaproteobacterial ammonia oxidizers in turfgrass systems.

Turfgrass is a highly managed ecosystem subject to frequent fertilization, mowing, irrigation, and application of pesticides. Turf management practices may create a perturbed environment for ammonia oxidizers, a key microbial group responsible for nitrification. To elucidate the long-term effects of turf management on these bacteria, we assessed the composition of betaproteobacterial ammonia oxidizers in a chronosequence of turfgrass systems (i.e., 1, 6, 23, and 95 years old) and the adjacent native pines by using both 16S rRNA and amoA gene fragments specific to ammonia oxidizers. Based on the Shannon-Wiener diversity index of denaturing gradient gel electrophoresis patterns and the rarefaction curves of amoA clones, turf management did not change the relative diversity and richness of ammonia oxidizers in turf soils as compared to native pine soils. Ammonia oxidizers in turfgrass systems comprised a suite of phylogenetic clusters common to other terrestrial ecosystems. Nitrosospira clusters 0, 2, 3, and 4; Nitrosospira sp. Nsp65-like sequences; and Nitrosomonas clusters 6 and 7 were detected in the turfgrass chronosequence with Nitrosospira clusters 3 and 4 being dominant. However, both turf age and land change (pine to turf) effected minor changes in ammonia oxidizer composition. Nitrosospira cluster 0 was observed only in older turfgrass systems (i.e., 23 and 95 years old); fine-scale differences within Nitrosospira cluster 3 were seen between native pines and turf. Further investigations are needed to elucidate the ecological implications of the compositional differences.

Atmospheric vapor pressure deficit is critical in predicting growth response of "cool-season" grass Festuca arundinacea to temperature change.

There is a lack of information on plant response to multifactor environmental variability including the interactive response to temperature and atmospheric humidity. These two factors are almost always confounded because saturated vapor pressure increases exponentially with temperature, and vapor pressure deficit (VPD) could have a large impact on plant growth. In this study using climate controlled mini-greenhouses, we examined the interacting influence of temperature and VPD on long-term growth of tall fescue (Festuca arundinacea Schreb), a cool season grass. From past studies it was expected that growth of tall fescue would decline with warmer temperatures over the range of 18.5-27 degrees C, but growth actually increased markedly with increasing temperature when VPD was held constant. In contrast, growth declined in experiments where tall fescue was exposed to increasing VPD and temperature was held constant at 21 degrees C. The inhibited growth appears to be in response to a maximum transpiration rate that can be supported by the tall fescue plants. The sensitivity to VPD indicates that if VPD remains stable in future climates as it has in the past, growth of tall fescue could well be stimulated rather than decreased by global warming in temperate climate zones.

Gas exchange and carbon metabolism in two Prosopis species (Fabaceae) from semiarid habitats: effects of elevated CO2, N supply, and N source.

Predicting future plant and ecosystem responses to elevated CO(2) also requires an understanding of the role of other factors, especially soil nitrogen. This is particularly challenging for global aridlands where total N and the relative amounts of nitrate and ammonia vary both spatially and seasonally. We measured gas exchange and primary and secondary C metabolites in seedlings of two dominant aridland shrub species (Prosopis flexuosa [S America] and P. glandulosa [N America]) grown at ambient (350 ppm) or elevated (650 ppm) CO(2) and nitrogen at two levels (low [0.8 mM] and high [8.0 mM]) and at either 1 : 1 or 3 : 1 nitrate to ammonia. Whereas elevated CO(2) increased assimilation rate, water use efficiency, and primary carbon metabolites in both species, these increases were strongly contingent upon nitrogen availability. Elevated CO(2) did not increase secondary metabolites (i.e., phenolics). For these important aridland species, the effects of elevated CO(2) are strongly influenced by nitrogen availability and to a lesser extent by the relative amounts of nitrate and ammonia supplied, which underscores the importance of both the amount and chemical composition of soil nitrogen in mediating the potential responses of seedling growth and establishment of aridland plants under future CO(2)-enriched atmospheres.

Growth, productivity, and competitiveness of introgressed weedy Brassica rapa hybrids selected for the presence of Bt cry1Ac and gfp transgenes.

Concerns exist that transgenic crop x weed hybrid populations will be more vigorous and competitive with crops compared with the parental weed species. Hydroponic, glasshouse, and field experiments were performed to evaluate the effects of introgression of Bacillus thuringiensis (Bt) cry1Ac and green fluorescent protein (GFP) transgenes on hybrid productivity and competitiveness in four experimental Brassica rapa x transgenic Brassica napus hybrid generations (F1, BC1F1, BC2F1 and BC2F2). The average vegetative growth and nitrogen (N) use efficiency of transgenic hybrid generations grown under high N hydroponic conditions were lower than that of the weed parent (Brassica rapa, AA, 2n = 20), but similar to the transgenic crop parent, oilseed rape (Brassica napus, AACC, 2n = 38). No generational differences were detected under low N conditions. In two noncompetitive glasshouse experiments, both transgenic and nontransgenic BC2F2 hybrids had on average less vegetative growth and seed production than B. rapa. In two high intraspecific competition field experiments with varied herbivore pressure, BC2F2 hybrids produced less vegetative dry weight than B. rapa. The competitive ability of transgenic and nontransgenic BC2F2 hybrids against a neighbouring crop species were quantified in competition experiments that assayed wheat (Triticum aestivum) yield reductions under agronomic field conditions. The hybrids were the least competitive with wheat compared with parental Brassica competitors, although differences between transgenic and nontransgenic hybrids varied with location. Hybridization, with or without transgene introgression, resulted in less productive and competitive populations.

Growth, nitrogen uptake, and metabolism in two semiarid shrubs grown at ambient and elevated atmospheric CO2 concentrations: effects of nitrogen supply and source.

The effect of differences in nitrogen (N) availability and source on growth and nitrogen metabolism at different atmospheric CO(2) concentrations in Prosopis glandulosa and Prosopis flexuosa (native to semiarid regions of North and South America, respectively) was examined. Total biomass, allocation, N uptake, and metabolites (e.g., free NO(3)(-), soluble proteins, organic acids) were measured in seedlings grown in controlled environment chambers for 48 d at ambient (350 ppm) and elevated (650 ppm) CO(2) and fertilized with high (8.0 mmol/L) or low (0.8 mmol/L) N (N(level)), supplied at either 1 : 1 or 3 : 1 NO(3)(-) : NH(4)(+) ratios (N(source)). Responses to elevated CO(2) depended on both N(level) and N(source), with the largest effects evident at high N(level). A high NO(3)(-) : NH(4)(+) ratio stimulated growth responses to elevated CO(2) in both species when N was limiting and increased the responses of P. flexuosa at high N(level). Significant differences in N uptake and metabolites were found between species. Seedlings of both species are highly responsive to N availability and will benefit from increases in CO(2), provided that a high proportion of NO(3)- to NH(4)-N is present in the soil solution. This enhancement, in combination with responses that increase N acquisition and increases in water use efficiency typically found at elevated CO(2), may indicate that these semiarid species will be better able to cope with both nutrient and water deficits as CO(2) levels rise.