Comment on 'improving ecophysiological simulation models to predict the impact of elevated CO(2) concentration on crop productivity' by X. Yin.

SCOPE: The recent publication by Yin (2013; Annals of Botany 112: 465-475) referred to in the title above provides an excellent review of modelling approaches to predict the impact of elevated CO2 on crop productivity, as well as on the controversy regarding whether yield responses observed in free-air CO2 enrichment (FACE) experiments are indeed lower than those from chamber-based experiments. However, the wheat experiments in the example of fig. 1 in Yin's paper had a flaw as the control plots lacked blowers that were in the FACE plots, which warmed the FACE plots at night and hastened plant development. This Viewpoint seeks to highlight this fact, and to comment on the relative merits of FACE and enclosure experiments.

Do gastrointestinal taste receptors contribute to associative learning and foraging behavior?

Foraging behavior is an expression of learning, context, and experience arising from integration of sensory information obtained during feeding with postingestive consequences of food ingestion. Although it has been well established that gustatory and olfactory systems of the mouth and nose provide sensory information to the consumer (in the form of flavor), sweet and bitter taste receptors have recently been identified in the intestinal tract of humans and rodents. It remains possible that sensory information generated in the gut could contribute to the learning process. Thus, a series of experiments was conducted to determine if classical associative learning occurs when the conditional stimulus circumvents oronasal presentation via direct delivery to the gut or peritoneal cavity. Mice receiving an intragastric infusion of 5 mM sodium saccharin immediately followed by LiCl administration demonstrated a significant decrease in preference for 5 mM saccharin in 4 consecutive 23 h, 2-bottle preference tests versus water (P = 0.0053). Saccharin was highly preferred in mice receiving intragastric (IG) saccharin only or interperitoneal (i.p.) injection of LiCl only. This reduced preference indicated that mice "tasted" saccharin infused into the gut. However, efforts to replicate with a reduced infusion volume failed to result in decreased preference. To understand if there were alternative pathways for oral detection of infused saccharin, mice received intragastric infusions (5.4 mM) and i.p. injections (10.8 mM) of sodium fluorescein. Fluorescence was observed from the tongues and esophagi of mice infused with volumes of 0.5 mL or more or injected with volumes of 0.25 mL or greater. Interperitoneal injections of 5 mM saccharin in mice resulted in reduced preference for 5 mM saccharin presented orally in 2-bottle preference tests (P = 0.0287). Oral delivery of a 500-fold less concentration of saccharin (0.01 mM) during conditioning resulted in a similar preference expression as shown in the initial IG experiment. These results demonstrate that although compounds may be tasted in the mouth absent of oral contact, associative learning is attenuated. Therefore, intestinal taste receptors are unlikely to participate directly in learning and recognition of foods during foraging events.

Free-air CO2 enrichment of sorghum: soil carbon and nitrogen dynamics.

The positive impact of elevated atmospheric CO(2) concentration on crop biomass production suggests more carbon inputs to soil. Further study on the effect of elevated CO(2) on soil carbon and nitrogen dynamics is key to understanding the potential for long-term carbon storage in soil. Soil samples (0- to 5-, 5- to 10-, and 10- to 20-cm depths) were collected after 2 yr of grain sorghum [Sorghum bicolor (L.) Moench.] production under two atmospheric CO(2) levels: (370 [ambient] and 550 muL L(-1) [free-air CO(2) enrichment; FACE]) and two water treatments (ample water and limited water) on a Trix clay loam (fine, loamy, mixed [calcareous], hyperthermic Typic Torrifluvents) at Maricopa, AZ. In addition to assessing treatment effects on soil organic C and total N, potential C and N mineralization and C turnover were determined in a 60-d laboratory incubation study. After 2 yr of FACE, soil C and N were significantly increased at all soil depths. Water regime had no effect on these measures. Increased total N in the soil was associated with reduced N mineralization under FACE. Results indicated that potential C turnover was reduced under water deficit conditions at the top soil depth. Carbon turnover was not affected under FACE, implying that the observed increase in soil C with elevated CO(2) may be stable relative to ambient CO(2) conditions. Results suggest that, over the short-term, a small increase in soil C storage could occur under elevated atmospheric CO(2) conditions in sorghum production systems with differing water regimes.

Development of C4 photosynthesis in sorghum leaves grown under free-air CO2 enrichment (FACE).

The developmental pattern of C4 expression has been well characterized in maize and other C4 plants. However, few reports have explored the possibility that the development of this pathway may be sensitive to changes in atmospheric CO2 concentrations. Therefore, both the structural and biochemical development of leaf tissue in the fifth leaf of Sorghum bicolor plants grown at elevated CO2 have been characterized. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC) activities accumulate rapidly as the leaf tissue differentiates and emerges from the surrounding whorl. Rubisco was not expressed in a cell-specific manner in the youngest tissue at the base of the leaf, but did accumulate before PEPC was detected. This suggests that the youngest leaf tissue utilizes a C3-like pathway for carbon fixation. However, this tissue was in a region of the leaf receiving very low light and so significant rates of photosynthesis were not likely. Older leaf tissue that had emerged from the surrounding whorl into full sunlight showed the normal C4 syndrome. Elevated CO2 had no effect on the cell-specific localization of Rubisco or PEPC at any stage of leaf development, and the relative ratios of Rubisco to PEPC remained constant during leaf development. However, in the oldest tissue at the tip of the leaf, the total activities of Rubisco and PEPC were decreased under elevated CO2 implying that C4 photosynthetic tissue may acclimate to growth under elevated CO2.

The effect of long-term atmospheric CO2 enrichment on the intrinsic water-use efficiency of sour orange trees.

Every two months of 1992, as well as on three occasions in 1994-1995, we obtained leaf samples together with samples of surrounding air from eight well-watered and fertilized sour orange (Citrus aurantium L.) trees that were growing out-of-doors at Phoenix, Arizona, USA. These trees had been planted in the ground as small seedlings in July of 1987 and enclosed in pairs by four clear-plastic-wall open-top chambers of which two have been continuously maintained since November of that year at a CO2 concentration of 400 micromol mol(-1) and two have been maintained at 700 micromol mol(-1). In September 2000, we also extracted north-south and east-west oriented wood cores that passed through the center of each tree's trunk at a height of 45 cm above the ground. Stable-carbon isotope ratios (13C/12C) derived from these leaf, wood and air samples were used to evaluate each tree's intrinsic water-use efficiency (iWUE). The grand-average result was an 80% increase in this important plant parameter in response to the 300 micromol mol(-1) increase in atmospheric CO2 concentration employed in the study. This increase in sour orange tree iWUE is identical to the long-term CO2-induced increase in the trees' production of wood and fruit biomass, which suggests there could be little to no change in total water-use per unit land area for this species as the air's CO2 content continues to rise. It is also identical to the increase in the mean iWUE reported for 23 groups of naturally occurring trees scattered across western North America that was caused by the historical rise in the air's CO2 content that occurred between 1800 and 1985.

[Responses of agricultural crops of free-air CO2 enrichment].

Over the past decade, free-air CO2 enrichment (FACE) experiments have been conducted on several agricultural crops: wheat(Triticum aestivum L.), perennial ryegrass (Lolium perenne), and rice(Oryza sativa L.) which are C3 grasses; sorghum (Sorghum bicolor (L.) Möench), a C4 grass; white clover (Trifolium repens), a C3 legume; potato (Solanum tuberosum L.), a C3 forb with tuber storage; and cotton (Gossypium hirsutum L.) and grape (Vitis vinifera L.) which are C3 woody perennials. Using reports from these experiments, the relative responses of these crops was discussed with regard to photosynthesis, stomatal conductance, canopy temperature, water use, water potential, leaf area index, shoot and root biomass accumulation, agricultural yield, radiation use efficiency, specific leaf area, tissue nitrogen concentration, nitrogen yield, carbohydrate concentration, phenology, soil microbiology, soil respiration, trace gas emissions, and soil carbon sequestration. Generally, the magnitude of these responses varied with the functional type of plant and with the soil nitrogen and water status. As expected, the elevated CO2 increased photosynthesis and biomass production and yield substantially in C3 species, but little in C4, and it decreased stomatal conductance and transpiration in both C3 and C4 species and greatly improved water-use efficiency in all the crops. Growth stimulations were as large or larger under water-stress compared to well-watered conditions. Growth stimulations of non-legumes were reduced at low soil nitrogen, whereas elevated CO2 strongly stimulated the growth of the clover legume both at ample and under low N conditions. Roots were generally stimulated more than shoots. Woody perennials had larger growth responses to elevated CO2, while at the same time, their reductions in stomatal conductance were smaller. Tissue nitrogen concentrations went down while carbohydrate and some other carbon-based compounds went up due to elevated CO2, with leaves and foliage affected more than other organs. Phenology was accelerated slightly in most but not all species. Elevated CO2 affected some soil microbes greatly but not others, yet overall activity appears to be stimulated. Detection of statistically significant changes in soil organic carbon in any one study was impossible, yet combining results from several sites and years, it appears that elevated CO2 did increase sequestration of soil carbon. Whenever possible, comparisons were made between the FACE results and those from prior chamber-based experiments reviewed in the literature. Over all the data and parameters considered in this review, there are only two parameters for which the FACE- and chamber-based data appear to be inconsistent. One is that elevated CO2 from FACE appears to reduce stomatal conductance about one and a half times more than observed in prior chamber experiments. Similarly, elevated CO2 appears to have stimulated root growth relatively more than shoot growth under FACE conditions compared to chamber conditions. Nevertheless, for the most part, the FACE- and chamber-based results have been consistent, which gives confidence that conclusions drawn from both types of data are accurate. However, the more realistic FACE environment and the larger plot size have enabled more extensive robust multidisciplinary data sets to be obtained under conditions representative of open fields in the future high-CO2 world.

Determination of cabergoline by electrospray ionization tandem mass spectrometry: picogram detection via column focusing sample introduction.

An electrospray ionization tandem mass spectrometric method was developed for low-picogram detection of an ergot alkaloid, cabergoline, in coyote plasma extracts. Cabergoline is under investigation as an abortifacient in canid species. Central to the successful development of this method was the ability to introduce relatively large sample volumes into the mass spectrometer. This was achieved by focusing the analyte on a conventional high-performance liquid chromatography guard column prior to elution into the spectrometer. Volumes up to at least 900 microL could be injected onto the guard column using a 100% aqueous mobile phase. Cabergoline retained on the column was eluted as a discreet band into the mass spectrometer by the rapid addition of methanol (30%) to the mobile phase. As compared to flow injection sample introduction, the ability to inject larger sample volumes led to a greatly lowered detection limit. Using this technique and a modification of a previously reported extraction procedure, cabergoline could be determined in coyote plasma at concentrations as low as 9 pg of cabergoline/mL of plasma.

Bioavailability of metals in stream food webs and hazards to brook trout (Salvelinus fontinalis) in the upper Animas River watershed, Colorado.

The water quality, habitats, and biota of streams in the upper Animas River watershed of Colorado, USA, are affected by metal contamination associated with acid drainage. We determined metal concentrations in components of the food web of the Animas River and its tributaries-periphyton (aufwuchs), benthic invertebrates, and livers of brook trout (Salvelinus fontinalis)-and evaluated pathways of metal exposure and hazards of metal toxicity to stream biota. Concentrations of the toxic metals cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn) in periphyton, benthic invertebrates, and trout livers from one or more sites in the upper Animas River were significantly greater than those from reference sites. Periphyton from sites downstream from mixing zones of acid and neutral waters had elevated concentrations of aluminum (Al) and iron (Fe) reflecting deposition of colloidal Fe and Al oxides, and reduced algal biomass. Metal concentrations in benthic invertebrates reflected differences in feeding habits and body size among taxa, with greatest concentrations of Zn, Cu, and Cd in the small mayfly Rhithrogena, which feeds on periphyton, and greatest concentrations of Pb in the small stonefly Zapada, a detritivore. Concentrations of Zn and Pb decreased across each trophic linkage, whereas concentrations of Cu and Cd were similar across several trophic levels, suggesting that Cu and Cd were more efficiently transferred via dietary exposure. Concentrations of Cu in invertebrates and trout livers were more closely associated with impacts on trout populations and invertebrate communities than were concentrations of Zn, Cd, or Pb. Copper concentrations in livers of brook trout from the upper Animas River were substantially greater than background concentrations and approached levels associated with reduced brook trout populations in field studies and with toxic effects on other salmonids in laboratory studies. These results indicate that bioaccumulation and transfer of metals in stream food webs are significant components of metal exposure for stream biota of the upper Animas River watershed and suggest that chronic toxicity of Cu is an important factor limiting the distribution and abundance of brook trout populations in the watershed.

Determination of whole-body rotenone residues in the brown tree snake (Boiga irregularis).

The brown tree snake (Boiga irregularis) is an introduced pest in Guam, responsible for extensive agricultural damage, the extinction of several bird species, and severe and frequent electrical power outages. Rotenone, a naturally occurring pesticide, has been investigated as a possible chemical control agent. An analytical method was developed to assess whole body rotenone residues ranging in concentration from 0.035 to 250 microg g(-)(1) in snakes. The method employed ethyl acetate extraction of 2 g samples of cryogenically frozen, pulverized snakes, followed by silica and Florisil solid-phase extraction cleanup. Extract analysis was performed using a high-performance liquid chromatography system employing a cyanopropyl analytical column. Tissues fortified to concentrations of 0.035, 4.82, and 250 microg g(-)(1) yielded analyte recoveries of 85.1, 85.6, and 83.5%, respectively. The linear response of rotenone standard solutions was assessed from 0. 025 to 0.25 microg mL(-)(1) (r(2) = 0.9968) and from 0.250 to 125 microg mL(-)(1) (r(2) = 0.9999). The method was simple, rugged, and reliable.

Effects of atmospheric CO2 enrichment on the growth and development of Hymenocallis littoralis (Amaryllidaceae) and the concentrations of several antineoplastic and antiviral constituents of its bulbs.

Two 2-yr crops of tropical spider lily (Hymenocallis littoralis) plants were grown in field soil in clear-plastic-wall open-top enclosures in the Sonoran Desert environment of central Arizona. Half of the plants were exposed to ambient air of 400 ppm atmospheric CO(2) concentration and half of them were exposed to air of 700 ppm CO(2). This 75% increase in the air's CO(2) content resulted in a 48% increase in aboveground plant biomass and a 56% increase in belowground (bulb) biomass. It also increased the concentrations of five bulb constituents that have been demonstrated to possess anticancer and antiviral activities. Mean percentage increases in these concentrations were 6% for a two-constituent (1:1) mixture of 7-deoxynarciclasine and 7-deoxy-trans-dihydronarciclasine, 8% for pancratistatin, 8% for trans-dihydronarciclasine, and 28% for narciclasine, for a mean active ingredient percentage concentration increase of 12%. Combined with the 56% increase in bulb biomass, these percentage concentration increases resulted in a mean active ingredient increase of 75% for the 75% increase in the air's CO(2) concentration used in our experiments.

Development and testing of seven new synthetic coyote attractants.

Available evidence indicates that effective coyote attractants are blends of volatile substances. Typically, attractants are a combination of biological substances such as fermented glandular materials, urines, and rotted meats. Although effective, these attractants have several distinct disadvantages. Among these is the possibility that they are unnecessarily complex and variable and, thus, difficult to replicate from one batch to the next. Although attractants containing a few reagent grade materials are available, the chemicals selected and their concentrations are not derived from actual attractants. For this reason, commercially available coyote attractants were analyzed with the intention of developing relatively simple synthetic alternatives. Purge and trap headspace analysis with gas chromatography/mass selective detection was employed to identify the volatile components of known conventional and synthetic attractants. All identified compounds were grouped according to chemical functionality, and one compound from each functional group was chosen to represent the group. Using only these representative compounds, seven synthetic attractants were formulated. Bioassays with captive coyotes (Canis latrans) were conducted to compare behavioral responses elicited by the seven new attractants, a currently available synthetic attractant, and a control. The results indicated that the attractants elicited significantly different behavioral profiles.

Acclimation response of spring wheat in a free-air CO(2) enrichment (FACE) atmosphere with variable soil nitrogen regimes. 1. Leaf position and phenology determine acclimation response.

We have examined the photosynthetic acclimation of wheat leaves grown at an elevated CO(2) concentration, and ample and limiting N supplies, within a field experiment using free-air CO(2) enrichment (FACE). To understand how leaf age and developmental stage affected any acclimation response, measurements were made on a vertical profile of leaves every week from tillering until maturity. The response of assimilation (A) to internal CO(2) concentration (C(i)) was used to estimate the in vivo carboxylation capacity (Vc(max)) and maximum rate of ribulose-1,5-bisphosphate limited photosynthesis (A (sat)). The total activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), and leaf content of Rubisco and the Light Harvesting Chlorophyll a/b protein associated with Photosystem II (LHC II), were determined. Elevated CO(2) did not alter Vc(max) in the flag leaf at either low or high N. In the older shaded leaves lower in the canopy, acclimatory decline in Vc(max) and A (sat) was observed, and was found to correlate with reduced Rubisco activity and content. The dependency of acclimation on N supply was different at each developmental stage. With adequate N supply, acclimation to elevated CO(2) was also accompanied by an increased LHC II/Rubisco ratio. At low N supply, contents of Rubisco and LHC II were reduced in all leaves, although an increased LHC II/Rubisco ratio under elevated CO(2) was still observed. These results underscore the importance of leaf position, leaf age and crop developmental stage in understanding the acclimation of photosynthesis to elevated CO(2) and nutrient stress.

Acclimation response of spring wheat in a free-air CO(2) enrichment (FACE) atmosphere with variable soil nitrogen regimes. 2. Net assimilation and stomatal conductance of leaves.

Atmospheric CO(2) concentration continues to rise. It is important, therefore, to determine what acclimatory c hanges will occur within the photosynthetic apparatus of wheat (Triticum aestivum L. cv. Yecora Rojo) grown in a future high-CO(2) world at ample and limited soil N contents. Wheat was grown in an open field exposed to the CO(2) concentration of ambient air [370 mumol (CO(2)) mol(-1); Control] and air enriched to approximately 200 mumol (CO(2)) mol(-1) above ambient using a Free-Air CO(2) Enrichment (FACE) apparatus (main plot). A High (35 g m(-2)) or Low (7 and 1.5 g m(-2) for 1996 and 1997, respectfully) level of N was applied to each half of the main CO(2) treatment plots (split-plot). Under High-N, FACE reduced stomatal conductance (g (s)) by 30% at mid-morning (2 h prior to solar noon), 36% at midday (solar noon) and 27% at mid-afternoon (2.5 h after solar noon), whereas under Low-N, g (s) was reduced by as much as 31% at mid-morning, 44% at midday and 28% at mid-afternoon compared with Control. But, no significant CO(2) x N interaction effects occurred. Across seasons and growth stages, daily accumulation of carbon (A') was 27% greater in FACE than Control. High-N increased A' by 18% compared with Low-N. In contrast to results for g (s), however, significant CO(2) x N interaction effects occurred because FACE increased A' by 30% at High-N, but by only 23% at Low-N. FACE enhanced the seasonal accumulation of carbon (A'') by 29% during 1996 (moderate N-stress), but by only 21% during 1997 (severe N-stress). These results support the premise that in a future high-CO(2) world an acclimatory (down-regulation) response in the photosynthetic apparatus of field-grown wheat is anticipated. They also demonstrate, however, that the stimulatory effect of a rise in atmospheric CO(2) on carbon gain in wheat can be maintained if nutrients such as nitrogen are in ample supply.

Acclimation response of spring wheat in a free-air CO(2) enrichment (FACE) atmosphere with variable soil nitrogen regimes. 3. Canopy architecture and gas exchange.

The response of whole-canopy net CO(2) exchange rate (CER) and canopy architecture to CO(2) enrichment and N stress during 1996 and 1997 for open-field-grown wheat ecosystem (Triticum aestivum L. cv. Yecora Rojo) are described. Every Control (C) and FACE (F) CO(2) treatment (defined as ambient and ambient +200 mumol mol(-1), respectively) contained a Low- and High-N treatment. Low-N treatments constituted initial soil content amended with supplemental nitrogen applied at a rate of 70 kg N ha(-1) (1996) and 15 kg N ha(-1) (1997), whereas High-N treatments were supplemented with 350 kg N ha(-1) (1996 and 1997). Elevated CO(2) enhanced season-long carbon accumulation by 8% and 16% under Low-N and High-N, respectively. N-stress reduced season-long carbon accumulation 14% under ambient CO(2), but by as much as 22% under CO(2) enrichment. Averaging both years, green plant area index (GPAI) peaked approximately 76 days after planting at 7.13 for FH, 6.00 for CH, 3.89 for FL, and 3.89 for CL treatments. Leaf tip angle distribution (LTA) indicated that Low-N canopies were more erectophile than those of High-N canopies: 48 degrees for FH, 52 degrees for CH, and 58 degrees for both FL and CL treatments. Temporal trends in canopy greenness indicated a decrease in leaf chlorophyll content from the flag to flag-2 leaves of 25% for FH, 28% for CH, 17% for CL, and 33% for FL during 1997. These results indicate that significant modifications of canopy architecture occurs in response to both CO(2) and N-stress. Optimization of canopy architecture may serve as a mechanism to diminish CO(2) and N-stress effects on CER.

Two new oral chemical biomarkers for coyotes.

Pentachlorobenzene (PeCB) and 1,2,3,4-tetrachlorobenzene (TeCB) were evaluated as oral chemical biomarkers when administered to coyotes (Canis latrans) during the period of 31 January to 10 August 1994. Three coyotes each received 100 mg of PeCB and three received 100 mg of TeCB, each in a mineral oil formulation. Three additional coyotes received only the mineral oil carrier. Muscle and adipose tissues, blood serum, and fecal samples were evaluated by capillary gas chromatography with electron capture detection for 120 days following administration. Residues of PeCB were detected in serum, feces, and adipose and muscle tissues for 120 days post-treatment; TeCB residues were detected in feces and serum at 1 and 8 days post-treatment and in adipose tissue at 30 days post-treatment. Residues of TeCB were not detected in muscle tissue at any point in the study.

Increased Accumulation of Carbohydrates and Decreased Photosynthetic Gene Transcript Levels in Wheat Grown at an Elevated CO2 Concentration in the Field.

Repression of photosynthetic genes by increased soluble carbohydrate concentrations may explain acclimation of photosynthesis to elevated CO2 concentration. This hypothesis was examined in a field crop of spring wheat (Triticum aestivum L.) grown at both ambient (approximately 360 [mu]mol mol-1) and elevated (550 [mu]mol mol-1) atmospheric CO2 concentrations using free-air CO2 enrichment at Maricopa, Arizona. The correspondence of steady-state levels of mRNA transcripts (coding for the 83-kD photosystem I apoprotein, sedoheptulose-1,7-bisphosphatase, phosphoribulokinase, phosphoglycerokinase, and the large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase) with leaf carbohydrate concentrations (glucose-6-phosphate, glucose, fructose, sucrose, fructans, and starch) was examined at different stages of crop and leaf development and through the diurnal cycle. Overall only a weak correspondence between increased soluble carbohydrate concentrations and decreased levels for nuclear gene transcripts was found. The difference in soluble carbohydrate concentration between leaves grown at elevated and current ambient CO2 concentrations diminished with crop development, whereas the difference in transcript levels increased. In the flag leaf, soluble carbohydrate concentrations declined markedly with the onset of grain filling; yet transcript levels also declined. The results suggest that, whereas the hypothesis may hold well in model laboratory systems, many other factors modified its significance in this field wheat crop.

Herbivore avoidance of digitalis extracts is not mediated by cardiac glycosides.

This study was conducted to determine whether avoidance of digitalis (Digitalis purpurea) by mountain beaver (Aplodontia rufa) is induced by toxic cardiac glycosides. High-performance liquid chromatography and behavioral assays were used to relate animal responses with the presence of common cardiac glycosides in several digitalis extracts. Statistical analyses of multiple-choice tests showed no correlation between cardiac glycoside content and mountain beaver avoidance of apple cubes treated with digitalis extracts. Therefore, we concluded that known toxic cardiac glycosides were not responsible for chemosensory cues that inhibited intake of food treated with digitalis extracts. These results suggest that digitalis is a source of an effective nontoxic herbivore repellent.

Effects of Atmospheric CO(2) Enrichment on Photosynthesis, Respiration, and Growth of Sour Orange Trees.

Numerous net photosynthetic and dark respiratory measurements were made over a period of 4 years on leaves of 24 sour orange (Citrus aurantium) trees; 8 of them growing in ambient air at a mean CO(2) concentration of 400 microliters per liter, and 16 growing in air enriched with CO(2) to concentrations approaching 1000 microliters per liter. Over this CO(2) concentration range, net photosynthesis increased linearly with CO(2) by more than 200%, whereas dark respiration decreased linearly to only 20% of its initial value. These results, together with those of a comprehensive fine-root biomass determination and two independent aboveground trunk and branch volume inventories, suggest that a doubling of the air's current mean CO(2) concentration of 360 microliters per liter would enhance the growth of the trees by a factor of 3.8.