Acclimation of Two Tomato Species to High Atmospheric CO(2): II. Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase and Phosphoenolpyruvate Carboxylase

Lycopersicon esculentum Mill. cv Vedettos and Lycopersicon chmielewskii Rick, LA 1028, were exposed to two CO₂ concentrations (330 or 900 microliters per liter) for 10 weeks. The elevated CO₂ concentrations increased the initial ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity of both species for the first 5 weeks of treatment but the difference did not persist during the last 5 weeks. The activity of Mg²⁺-CO₂-activated Rubisco was higher in 900 microliters per liter for the first 2 weeks but declined sharply thereafter. After 10 weeks, leaves grown at 330 microliters per liter CO₂ had about twice the Rubisco activity compared with those grown at 900 microliters per liter CO₂. The two species showed the same trend to Rubisco declines under high CO₂ concentrations. The percent activation of Rubisco was always higher under high CO₂. The phosphoenolpyruvate carboxylase (PEPCase) activity measured in tomato leaves averaged 7.9% of the total Rubisco. PEPCase showed a similar trend with time as the initial Rubisco but with no significant difference between nonenriched and CO₂-enriched plants. Long-term exposure of tomato plants to high CO₂ was previously shown to induce a decline of photosynthetic efficiency. Based on the current study and on previous results, we propose that the decline of activated Rubisco is the main cause of the acclimation of tomato plants to high CO₂ concentrations.     

Acclimation to High CO(2) in Monoecious Cucumbers : II. Carbon Exchange Rates, Enzyme Activities, and Starch and Nutrient Concentrations

Carbon exchange capacity of cucumber (Cucumis sativus L.) germinated and grown in controlled environment chambers at 1000 microliters per liter CO₂ decreased from the vegetative growth stage to the fruiting stage, during which time capacity of plants grown at 350 microliters per liter increased. Carbon exchange rates (CERs) measured under growth conditions during the fruiting period were, in fact, lower in plants grown at 1000 microliters per liter CO₂ than those grown at 350. Progressive decreases in CERs in 1000 microliters per liter plants were associated with decreasing stomatal conductances and activities of ribulose bisphosphate carboxylase and carbonic anhydrase. Leaf starch concentrations were higher in 1000 microliters per liter CO₂ grown-plants than in 350 microliters per liter grown plants but calcium and nitrogen concentrations were lower, the greatest difference occurring at flowering. Sucrose synthase and sucrose-P-synthase activities were similar in 1000 microliters per liter compared to 350 microliters per liter plants during vegetative growth and flowering but higher in 350 microliters per liter plants at fruiting. The decreased carbon exchange rates observed in this cultivar at 1000 microliters per liter CO₂ could explain the lack of any yield increase (MM Peet 1986 Plant Physiol 80: 59-62) when compared with plants grown at 350 microliters per liter.     

Short-Term Effects of CO(2) on Gas Exchange of Leaves of Bigtooth Aspen (Populus grandidentata) in the Field

The short term effects of increased levels of CO₂ on gas exchange of leaves of bigtooth aspen (Populus grandidentata Michx.) were studied at the University of Michigan Biological Station, Pellston, MI. Leaf gas exchange was measured in situ in the upper half of the canopy, 12 to 14 meters above ground. In 1900 microliters per liter CO₂, maximum CO₂ exchange rate (CER) in saturating light was increased by 151% relative to CER in 320 microliters per liter CO₂. The temperature optimum for CER shifted from 25°C in 320 microliters per liter CO₂ to 37°C in 1900 microliters per liter CO₂. In saturating light, increasing CO₂ level over the range 60 to 1900 microliters per liter increased CER, decreased stomatal conductance, and increased leaf water use efficiency. The initial slope of the CO₂ response curve of CER was not significantly different at 20 and 30°C leaf temperatures, although the slope did decline significantly during leaf senescence. In 1900 microliters per liter CO₂, CER increased with increasing light. The light saturation point and maximum CER were higher in 30°C than in 20°C, although there was little effect of temperature in low light. The experimental results are consistent with patterns seen in laboratory studies of other C₃ species and define the parameters required by some models of aspen CER in the field.     

Effects of CO(2) Enrichment and Carbohydrate Content on the Dark Respiration of Soybeans

During the period of most active leaf expansion, the foliar dark respiration rate of soybeans (Glycine max cv Williams), grown for 2 weeks in 1000 microliters CO₂ per liter air, was 1.45 milligrams CO₂ evolved per hour leaf density thickness, and this was twice the rate displayed by leaves of control plants (350 microliters CO₂ per liter air). There was a higher foliar nonstructural carbohydrate level (e.g. sucrose and starch) in the CO₂ enriched compared with CO₂ normal plants. For example, leaves of enriched plants displayed levels of nonstructural carbohydrate equivalent to 174 milligrams glucose per gram dry weight compared to the 84 milligrams glucose per gram dry weight found in control plant leaves. As the leaves of CO₂ enriched plants approached full expansion, both the foliar respiration rate and carbohydrate content of the CO₂ enriched leaves decreased until they were equivalent with those same parameters in the leaves of control plants. A strong positive correlation between respiration rate and carbohydrate content was seen in high CO₂ adapted plants, but not in the control plants.

Mitochondria, isolated simultaneously from the leaves of CO₂ enriched and control plants, showed no difference in NADH or malate-glutamate dependent O₂ uptake, and there were no observed differences in the specific activities of NAD⁺ linked isocitrate dehydrogenase and cytochrome c oxidase. Since the mitochondrial O₂ uptake and total enzyme activities were not greater in young enriched leaves, the increase in leaf respiration rate was not caused by metabolic adaptations in the leaf mitochondria as a response to long term CO₂ enrichment. It was concluded, that the higher respiration rate in the enriched plant's foliage was attributable, in part, to a higher carbohydrate status.

     

Effect of changes in shoot carbon-exchange rate on soybean root nodule activity

The effect of short- and long-term changes in shoot carbon-exchange rate (CER) on soybean (Glycine max [L.] Merr.) root nodule activity was assessed to determine whether increases in photosynthate production produce a direct enhancement of symbiotic N₂ fixation. Shoot CER, root + nodule respiration, and apparent N₂ fixation (acetylene reduction) were measured on intact soybean plants grown at 700 microeinsteins per meter per second, with constant root temperature and a 14/10-hour light/dark cycle. There was no diurnal variation of root + nodule respiration or apparent N₂ fixation in plants assayed weekly from 14 to 43 days after planting. However, if plants remained in darkness following their normal dark period, a significant decline in apparent N₂ fixation was measured within 4 hours, and decreasing CO₂ concentration from 320 to 90 microliters CO₂ per liter produced diurnal changes in root nodule activity. Increasing shoot CER by 87, 84, and 76% in 2-, 3-, and 4-week-old plants, respectively, by raising the CO₂ concentration around the shoot from 320 to 1,000 microliters CO₂ per liter, had no effect on root + nodule respiration or acetylene-reduction rates during the first 10 hours of the increased CER treatment. When the CO₂-enrichment treatment was extended in 3-week-old plants, the only measured parameter that differed significantly after 3 days was shoot CER. After 5 days of continuous CO₂ enrichment, root + nodule respiration and acetylene reduction increased, but such changes reflected an increase in root nodule mass rather than greater specific root nodule activity. The results show that on a 24-hour basis the process of symbiotic N₂ fixation in soybean plants grown under controlled environmental conditions functioned at maximum capacity and was not limited by shoot CER. Whether N₂-fixation capacity was limited by photosynthate movement to root nodules or by saturation of metabolic processes in root nodules is not known.     

Short-Term and Long-Term Responses of Crassulacean Acid Metabolism Plants to Elevated CO(2)

For the leaf succulent Agave deserti and the stem succulent Ferocactus acanthodes, increasing the ambient CO₂ level from 350 microliters per liter to 650 microliters per liter immediately increased daytime net CO₂ uptake about 30% while leaving nighttime net CO₂ uptake of these Crassulacean acid metabolism (CAM) plants approximately unchanged. A similar enhancement of about 30% was found in dry weight gain over 1 year when the plants were grown at 650 microliters CO₂ per liter compared with 350 microliters per liter. Based on these results plus those at 500 microliters per liter, net CO₂ uptake over 24-hour periods and dry weight productivity of these two CAM succulents is predicted to increase an average of about 1% for each 10 microliters per liter rise in ambient CO₂ level up to 650 microliters per liter.     

Reversibility of Photosynthetic Inhibition in Cotton after Long-Term Exposure to Elevated CO(2) Concentrations

Cotton (Gossypium hirsutum L. cv Stoneville 213) was grown at 350 and 1000 microliters per liter CO₂. The plants grown at elevated CO₂ concentrations contained large starch pools and showed initial symptoms of visible physical damage. Photosynthetic rates were lower than expected based on instantaneous exposure to high CO₂.

A group of plants grown at 1000 microliters per liter CO₂ was switched to 350 microliters per liter CO₂. Starch pools and photosynthetic rates were monitored in the switched plants and in the two unswitched control groups. Photosynthetic rates per unit leaf area recovered to the level of the 350 microliters per liter CO₂ grown control group within four to five days. To assess only nonstomatal limitations to photosynthesis, a measure of photosynthetic efficiencies was calculated (moles CO₂ fixed per square meter per second per mole intercellular CO₂). Photosynthetic efficiency also recovered to the levels of the 350 microliters per liter CO₂ grown controls within three to four days.

Recovery was correlated to a rapid depletion of the starch pool, indicating that the inhibition of photosynthesis is primarily a result of feedback inhibition. However, complete recovery may involve the repair of damage to the chloroplasts caused by excessive starch accumulation. The rapid and complete reversal of photosynthetic inhibition suggests that the appearance of large, strong sinks at certain developmental stages could result in reduction of the large starch accumulations and that photosynthetic rates could recover to near the theoretical capacity during periods of high photosynthate demand.

     

Increases in Phosphorus Requirements for CO(2)-Enriched Pine Species

Pinus radiata D. Don (half-sib families 20010 and 20062) and Pinus caribaea var hondurensis (an open-pollinated family) were grown for 49 weeks at seven levels of phosphorus and at CO₂ concentrations of either 340 or 660 microliters per liter, to establish if the phosphorus requirements differed between the CO₂ concentrations and if mycorrhizal associations were affected. When soil phosphorus availability was low, phosphorus uptake was increased by elevated CO₂. This may have been related to changes in mycorrhizal competition. When the phosphorus concentration in the youngest fully expanded needles was above 600 milligrams per kilogram the shoot weight of all pine families was greater at high CO₂ due to increases in rates of photosynthesis. More dry weight was partitioned to the stems of P. radiata family 20010 and P. caribaea. At foliar phosphorus concentrations above 1000 milligrams per kilogram (P. radiata) and 700 milligrams per kilogram (P. caribaea), growth did not increase at 340 microliters of CO₂ per liter. Soluble sugar levels in the same needles mirrored the growth response, but the starch concentration declined with increasing phosphorus. At 660 microliters of CO₂ per liter, shoot weight and soluble sugar concentrations were still increasing up to a foliar P concentration of 1800 milligrams per kilogram for P. radiata and 1600 milligrams per kilogram for P. caribaea. The starch concentrations did not decline. These results indicate that higher foliar phosphorus concentrations are required to realize the maximum growth potential of pines at elevated CO₂.     

Effects of High Atmospheric CO(2) and Sink Size on Rates of Photosynthesis of a Soybean Cultivar

The effect of sink strength on photosynthetic rates under conditions of long-term exposure to high CO₂ has been investigated in soybean. Soybean plants (Merr. cv. Fiskeby V) were grown in growth chambers containing 350 microliters CO₂ per liter air until pod set. At that time, plants were trimmed to three trifoliolate leaves and either 21 pods (high sink treatment) or 6 pods (low sink treatment). Trimmed plants were either left in 350 microliters CO₂ per liter of air or placed in 1000 microliters CO₂ per liter of air (high CO₂ treatment) until pod maturity. Whole plant net photosynthetic rates of all plants were measured twice weekly, both at 350 microliters CO₂ per liter of air and 1000 microliters CO₂ per liter of air. Plants were also harvested at this time for dry weight measurements. Photosynthetic rates of high sink plants at both measurement CO₂ concentrations were consistently higher than those of low sink plants, and those of plants given the 350 microliter CO₂ per liter of air treatment were higher at both measurement CO₂ concentrations than those of plants given the 1000 microliters CO₂ per liter of air treatment. When plants were measured under treatment CO₂ levels, however, rates were higher in 1,000 microliter plants than 350 microliter CO₂ plants. Dry weights of all plant parts were higher in the 1,000 microliters CO₂ per liter air treatment than in the 350 microliters CO₂ per liter air treatment, and were higher in the low sink than in the high sink treatments.     

Use of Carbon Isotopes to Estimate Incorporation of Added CO(2) by Greenhouse-Grown Tomato Plants

A method is presented which uses the ¹³C and ¹⁴C isotope abundance in CO₂-enriched greenhouse crops to determine the percentage of plant organic carbon derived from artificially added CO₂. In a greenhouse experiment with CO₂ concentrations elevated to 1100 ± 100 microliters per liter during part of the daylight hours and maintained at normal atmospheric concentrations (340 microliters per liter) during the rest of the time, it was shown by ¹⁴C analysis that between 41% and 42% of the carbon in tomato plants (Lycopersicon esculentum var 4884) came from the artificially added CO₂. Similar results were obtained from ¹³C analyses when the CO₂ pressure-dependent isotope separation was taken into account.     

Acclimation to High CO(2) in Monoecious Cucumbers : I. Vegetative and Reproductive Growth

CO₂ concentrations of 1000 compared to 350 microliters per liter in controlled environment chambers did not increase total fruit weight or number in a monoecious cucumber (Cucumis sativus L. cv Chipper) nor did it increase biomass, leaf area, or relative growth rates beyond the first 16 days after seeding. Average fruit weight was slightly, but not significantly greater in the 1000 microliters per liter CO₂ treatment because fruit numbers were changed more than total weight. Plants grown at 1000 and 350 microliters per liter CO₂ were similar in distribution of dry matter and leaf area between mainstem, axillary, and subaxillary branches. Early flower production was greater in 1000 microliters per liter plants. Subsequent flower numbers were either lower in enriched plants or similar in the two treatments, except for the harvest at fruiting when enriched plants produced many more male flowers than the 350 microliters per liter treatments.     

Effects of CO(2) Concentration on Rubisco Activity, Amount, and Photosynthesis in Soybean Leaves

Growth at an elevated CO₂ concentration resulted in an enhanced capacity for soybean (Glycine max L. Merr. cv Bragg) leaflet photosynthesis. Plants were grown from seed in outdoor controlled-environment chambers under natural solar irradiance. Photosynthetic rates, measured during the seed filling stage, were up to 150% greater with leaflets grown at 660 compared to 330 microliters of CO₂ per liter when measured across a range of intercellular CO₂ concentrations and irradiance. Soybean plants grown at elevated CO₂ concentrations had heavier pod weights per plant, 44% heavier with 660 compared to 330 microliters of CO₂ per liter grown plants, and also greater specific leaf weights. Ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco) activity showed no response (mean activity of 96 micromoles of CO₂ per square meter per second expressed on a leaflet area basis) to short-term (~1 hour) exposures to a range of CO₂ concentrations (110-880 microliters per liter), nor was a response of activity (mean activity of 1.01 micromoles of CO₂ per minute per milligram of protein) to growth CO₂ concentration (160-990 microliters per liter) observed. The amount of rubisco protein was constant, as growth CO₂ concentration was varied, and averaged 55% of the total leaflet soluble protein. Although CO₂ is required for activation of rubisco, results indicated that within the range of CO₂ concentrations used (110-990 microliters per liter), rubisco activity in soybean leaflets, in the light, was not regulated by CO₂.     

Downward Regulation of Photosynthesis and Growth at High CO(2) Levels : No Evidence for Either Phenomenon in Three-Year Study of Sour Orange Trees

Numerous photosynthesis and growth measurements of sour orange (Citrus aurantium L.) trees maintained in ambient air and air enriched with an extra 300 microliters per liter of CO₂ have revealed the CO₂-enriched trees to have consistently sequestered approximately 2.8 times more carbon than the control trees over a period of three full years. Under field conditions in the natural environment, plants may not experience the downward regulation of photosynthetic capacity typically observed in long-term CO₂ enrichment experiments with plants growing in pots.     

Acclimation of Two Tomato Species to High Atmospheric CO(2): I. Sugar and Starch Concentrations

Lycopersicon esculentum Mill. cv Vedettos and Lycopersicon chmielewskii Rick, LA 1028, were exposed to two CO₂ concentrations (330 or 900 microliters per liter) for 10 weeks. Tomato plants grown at 900 microliters per liter contained more starch and more sugars than the control. However, we found no significant accumulation of starch and sugars in the young leaves of L. esculentum exposed to high CO₂. Carbon exchange rates were significantly higher in CO₂-enriched plants for the first few weeks of treatment but thereafter decreased as tomato plants acclimated to high atmospheric CO₂. This indicates that the long-term decline of photosynthetic efficiency of leaf 5 cannot be attributed to an accumulation of sugar and/or starch. The average concentration of starch in leaves 5 and 9 was always higher in L. esculentum than in L. chmielewskii (151.7% higher). A higher proportion of photosynthates was directed into starch for L. esculentum than for L. chmielewskii. However, these characteristics did not improve the long-term photosynthetic efficiency of L. chmielewskii grown at high CO₂ when compared with L. esculentum. The chloroplasts of tomato plants exposed to the higher CO₂ concentration exhibited a marked accumulation of starch. The results reported here suggest that starch and/or sugar accumulation under high CO₂ cannot entirely explain the loss of photosynthetic efficiency of high CO₂-grown plants.     

Effects of Atmospheric CO(2) Enrichment on the Growth and Mineral Nutrition of Quercus alba Seedlings in Nutrient-Poor Soil

One-year-old dormant white oak (Quercus alba L.) seedlings were planted in a nutrient-deficient forest soil and grown for 40 weeks in growth chambers at ambient (362 microliters per liter) or elevated (690 microliters per liter) levels of CO₂. Although all of the seedlings became severely N deficient, CO₂ enrichment enhanced growth by 85%, with the greatest enhancement in root systems. The growth enhancement did not increase the total water use per plant, so water-use efficiency was significantly greater in elevated CO₂. Total uptake of N, S, and B was not affected by CO₂, therefore, tissue concentrations of these nutrients were significantly lower in elevated CO₂. An increase in nutrient-use efficiency with respect to N was apparent in that a greater proportion of the limited N pool in the CO₂-enriched plants was in fine roots and leaves. The uptake of other nutrients increased with CO₂ concentration, and P and K uptake increased in proportion to growth. Increased uptake of P by plants in elevated CO₂ may have been a result of greater proliferation of fine roots and associated mycorrhizae and rhizosphere bacteria stimulating P mineralization. The results demonstrate that a growth response to CO₂ enrichment is possible in nutrient-limited systems, and that the mechanisms of response may include either increased nutrient supply or decreased physiological demand.     

Drought Stress and Elevated CO(2) Effects on Soybean Ribulose Bisphosphate Carboxylase Activity and Canopy Photosynthetic Rates

Soybean (Glycine max [L.] cv Bragg) was grown at 330 or 660 microliters CO₂ per liter in outdoor, controlled-environment chambers. When the plants were 50 days old, drought stress was imposed by gradually reducing irrigation each evening so that plants wilted earlier each succeeding day. On the ninth day, as the pots ran out of water CO₂ exchange rate (CER) decreased rapidly to near zero for the remainder of the day. Both CO₂-enrichment and drought stress reduced the total (HCO₃⁻/Mg²⁺-activated) extractable ribulose-1,5-bisphosphate carboxylase (RuBPCase) activity, as expressed on a chlorophyll basis. In addition, drought stress when canopy CER values and leaf water potentials were lowest, reduced the initial (nonactivated) RuBPCase activity by 50% compared to the corresponding unstressed treatments. This suggests that moderate to severe drought stress reduces the in vivo activation state of RuBPCase, as well as lowers the total activity. It is hypothesized that stromal acidification under drought stress causes the lowered initial RuBPCase activities. The K_m(CO₂) values of activated RuBPCase from stressed and unstressed plants were similar; 15.0 and 12.6 micromolar, respectively. RuBP levels were 10 to 30% lower in drought stressed as compared to unstressed treatments. However, RuBP levels increased from near zero at night to around 150 to 200 nanomoles per milligram chlorophyll during the day, even as water potentials and canopy CERs decreased. This suggests that the rapid decline in canopy CER cannot be attributed to drought stress induced limitations in the RuBP regeneration capability. Thus, in soybean leaves, a nonstomatal limitation of leaf photosynthesis under drought stress conditions appears due, in part, to a reduction of the in vivo activity of RuBPCase. Because initial RuBPCase activities were not reduced as much as canopy CER values, this enzymic effect does not explain entirely the response of soybean photosynthesis to drought stress.     

Interaction of water supply and N in wheat

The purpose of this study was to investigate effects of N nutrition and water stress on stomatal behavior and CO₂ exchange rate in wheat (Triticum aestivum L. cv Olaf). Wheat plants were grown hydroponically with high (100 milligrams per liter) and low (10 milligrams per liter) N. When plants were 38 days old, a 24-day water stress cycle was begun. A gradual increase in nutrient solution osmotic pressure from 0.03 to 1.95 mega Pascals was achieved by incremental additions of PEG-6,000. Plants in both N treatments adjusted osmotically, although leaf water potential was consistently lower and relative water content greater for low N plants in the first half of the stress cycle. Leaf conductance of high N plants appeared greater than that of low N plants at high water potentials, but showed greater sensitivity to reductions in water potential as indicated by earlier stomatal closure during the stress cycle. The apparent greater stomatal sensitivity of high N plants was associated with a curvilinear relationship between leaf conductance and leaf water potential; low N plants exhibited more of a threshold response. Trends in [CO₂]_INT throughout the stress cycle indicated nonstomatal effects of water stress on CO₂ exchange rate were greater in high N plants. Although estimates of [CO₂]_INT were generally lower in high N plants, they were relatively insensitive to leaf water potential-induced changes in leaf conductance. In contrast, [CO₂]_INT of low N plants dropped concomitantly with leaf conductance at low leaf water potentials. Oxygen response of CO₂ exchange rate for both treatments was affected less by reductions in water potential than was CO₂ exchange rate at 2.5% O₂, suggesting that CO₂ assimilation capacity of the leaves was affected more by reductions in leaf water potential than were processes related to photorespiration.     

Minor Physiological Response to Elevated CO(2) by the CAM Plant Agave vilmoriniana

One-year-old plants of the CAM leaf succulent Agave vilmoriniana Berger were grown outdoors at Riverside, California. Potted plants were acclimated to CO₂-enrichment (about 750 microliters per liter) by growth for 2 weeks in an open-top polyethylene chamber. Control plants were grown nearby where the ambient CO₂ concentration was about 370 microliters per liter. When the plants were well watered, CO₂-induced differences in stomatal conductances and CO₂ assimilation rates over the entire 24-hour period were not large. There was a large nocturnal acidification in both CO₂ treatments and insignificant differences in leaf chlorophyll content. Well watered plants maintained water potentials of −0.3 to −0.4 megapascals. When other plants were allowed to dry to water potentials of −1.2 to −1.7 megapascals, stomatal conductances and CO₂ uptake rates were reduced in magnitude, with the biggest difference in Phase IV photosynthesis. The minor nocturnal response to CO₂ by this species is interpreted to indicate saturated, or nearly saturated, phosphoenolpyruvate carboxylase activity at current atmospheric CO₂ concentrations. CO₂-enhanced diurnal activity of ribulose bisphosphate carboxylase activity remains a possibility.     

Effects of decreased net carbon exchange on carbohydrate metabolism in sugar beet source leaves

The relationship between CO₂ concentration and starch synthesis and degradation was studied by measuring leaf starch content and disappearance of ¹⁴C-starch. At a concentration of 340 microliters CO₂ per liter, starch accumulated without degradation of previously synthesized starch. Degradation of starch began when CO₂ concentration was lowered, but its synthesis continued. At 120 microliters CO₂ per liter rates of synthesis and degradation were equal. Even at the CO₂ compensation point, synthesis of starch continued. Concomitant starch synthesis and mobilization supported export from the leaf. Changes in starch metabolism that occur when photosynthesis is CO₂-limited provide a means to study regulation of starch metabolism and carbon allocation in translocating leaves.     

Growth Response of a Succulent Plant, Agave vilmoriniana, to Elevated CO(2)

Large (about 200 grams dry weight) and small (about 5 grams dry weight) specimens of the leaf succulent Agave vilmoriniana Berger were grown outdoors at Phoenix, Arizona. Potted plants were maintained in open-top chambers constructed with clear, plastic wall material. Four CO₂ concentrations of 350, 560, 675, and 885 microliters per liter were used during two growth periods and two water treatments. Small and large plants were grown for 6 months, while a few large plants were grown for 1 year. Wet-treatment plants received water twice weekly, whereas dry-treatment plants received slightly more water than they would under natural conditions. Plant growth rates in all treatments were significantly different between small and large specimens, but not between 6 month and 1 year large plants. Only the dry-treatment plants exhibited statistically different growth rates between the CO₂ treatments. This productivity response was equivalent to a 28% and 3-fold increase when mathematically interpolated between CO₂ concentrations of 300 and 600 microliters per liter for large and small plants, respectively.