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.     

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.     

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.

     

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.     

Prechilling of Xanthium strumarium L. Reduces Net Photosynthesis and, Independently, Stomatal Conductance, While Sensitizing the Stomata to CO(2)

Greenhouse-grown plants of Xanthium strumarium L. were exposed in a growth cabinet to 10 C during days and 5 C during nights for periods of up to 120 hours. Subsequently, CO₂ exchange, transpiration, and leaf temperature were measured on attached leaves and in leaf sections at 25 or 30 C, 19 C dew point of the air, 61 milliwatts per square centimeter irradiance, and CO₂ concentrations between 0 and 1000 microliters per liter ambient air. Net photosynthesis and stomatal conductance decreased and dark respiration increased with increasing duration of prechilling. The reduction in net photosynthesis was not a consequence of decreased stomatal conductance because the intercellular CO₂ concentration in prechilled leaves was equal to or greater than that in greenhouse-grown controls. The intercellular CO₂ concentration at which one-half maximum net photosynthesis occurred remained the same in prechilled leaves and controls (175 to 190 microliters per liter). Stomata of the control plants responded to changes in the CO₂ concentration of the air only slightly. Prechilling for 24 hours or more sensitized stomata to CO₂; they responded to changes in CO₂ concentration in the range from 100 to 1000 microliters per liter.     

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.

     

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.     

Effect of oxygen and temperature on the efficiency of photosynthetic carbon assimilation in two microscopic algae

The CO₂ compensation points of Coccochloris peniocystis, a blue-green alga and Chlamydomonas reinhardtii, a green alga, were determined at pH 8.0 in a closed system by a gas chromatographic technique. The compensation point of Chlamydomonas increased markedly with temperature, rising from 0.79 microliter per liter CO₂ at 15 C to 2.5 microliters per liter CO₂ at 35 C. In contrast, the compensation point of Coccochloris at 20 C was 0.71 microliter per liter CO₂ and rose to only 0.95 microliter per liter CO₂ at 40 C.     

Changes in Levels of Intermediates of the C(4) Cycle and Reductive Pentose Phosphate Pathway under Various Light Intensities in Maize Leaves

The rate of CO₂ assimilation and levels of metabolites of the C₄ cycle and reductive pentose phosphate pathway in attached leaves of maize (Zea mays L.) were measured over a range of light intensity from 0 to 1,900 microEinsteins per square meter per second under a saturated CO₂ concentration of 350 microliters per liter and a limiting CO₂ concentration of 133 microliters per liter. The level of ribulose 1,5-bisphosphate (RuBP) stayed almost constant (around 60 nanomoles per milligram chlorophyll [Chl]) from low to high light intensities under 350 microliters per liter. Levels of 3-phosphoglycerate (PGA) increased from 100 to 650 nanomoles per milligram Chl under 350 microliters per liter CO₂ with increasing light intensity. The calculated RuBP concentration of 6 millimolar (corresponded to 60 nanomoles per milligram Chl) was about two times above the estimated RuBP binding-site concentration on ribulose bisphosphate carboxylase-oxygenase (Rubisco) of ~2.6 millimolar in maize bundle sheath chloroplasts in the light. The ratio of RuBP/PGA increased with decreasing light intensity under 350 microliters per liter CO₂. These results suggest that RuBP carboxylation is under control of light intensity possibly due to a limited supply of CO₂ to Rubisco through the C₄ cycle whose activity is highly dependent on light intensity. Pyruvate level increased with increasing light intensity as long as photosynthesis rate increased. A positive relationship between levels of PGA and those of pyruvate during steady-state photosynthesis under various conditions suggests that an elevated concentration of PGA increases the carbon input into the C₄ cycle through the conversion of PGA to PEP and consequently the level of total intermediates of the C₄ cycle can be raised to mediate higher photosynthesis rate.     

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.     

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 CO₂ concentration of 400 microliters per liter, and 16 growing in air enriched with CO₂ to concentrations approaching 1000 microliters per liter. Over this CO₂ concentration range, net photosynthesis increased linearly with CO₂ 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 CO₂ concentration of 360 microliters per liter would enhance the growth of the trees by a factor of 3.8.     

Interactions between Ethylene, CO(2), and ABA on GA(3)-Induced Amylase Synthesis in Barley Aleurone Tissue

Gibberellic acid-induced synthesis and release of α-amylase in barley aleurone tissue was inhibited by abscisic acid. This inhibition was relieved by simultaneous application of ethylene ranging in concentration from 0.1 to 100 microliters per liter. When CO₂ was applied, it eliminated the effect of 0.1 microliter per liter ethylene and reimposed the abscisic acid inhibition. All concentrations of CO₂ tested from 400 to 10⁵ microliters per liter counteracted the effect of 0.1 microliter per liter ethylene, but had no observable effect on any higher concentration of ethylene. The results indicate that some processes necessary for embryo growth may be subject to regulation by ethylene and carbon dioxide at naturally occurring concentrations of the gases.     

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.     

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.     

C(4) Acid Metabolism and Dark CO(2) Fixation in a Submersed Aquatic Macrophyte (Hydrilla verticillata)

The CO₂ compensation point of the submersed aquatic macrophyte Hydrilla verticillata varied from high (above 50 microliters per liter) to low (10 to 25 microliters per liter) values, depending on the growth conditions. Plants from the lake in winter or after incubation in an 11 C/9-hour photoperiod had high values, whereas summer plants or those incubated in a 27 C/14-hour photoperiod had low values. The plants with low CO₂ compensation points exhibited dark ¹⁴CO₂ fixation rates that were up to 30% of the light fixation rates. This fixation reduced respiratory CO₂ loss, but did not result in a net uptake of CO₂ at night. The low compensation point plants also showed diurnal fluctuations in titratable acid, such as occur in Crassulacean acid metabolism plants. However, dark fixation and diurnal acid fluctuations were negligible in Hydrilla plants with high CO₂ compensation points.     

Effects of SO(2) and O(3) on Allocation of C-Labeled Photosynthate in Phaseolus vulgaris

A series of laboratory exposures of two varieties of bush bean (Phaseolus vulgaris L., var 274 and var 290) was conducted to determine the sensitivity of [¹⁴C]photosynthate allocation patterns to alteration by SO₂ and O₃. Experiments with the pollution-resistant 274 variety demonstrated short-term changes in both ¹⁴C and biomass allocation to roots of ¹⁴CO₂-labeled plants but no significant effect on yield by up to 40 hours of exposure to SO₂ at 0.50 microliters per liter or 4 hours of O₃ at 0.40 microliters per liter. Subsequent experiments with the more sensitive 290 variety demonstrated significant alteration of photosynthesis, translocation, and partitioning of photosynthate between plant parts including developing pods. Significant increases in foliar retention of photosynthate (+40%) occurred after 8 hours of exposure to SO₂ at 0.75 microliters per liter (6.0 microliters per liter-hour) and 11 hours of exposure to O₃ at 0.30 microliters per liter-hour (3.3 microliters-hours). Time series sampling of labeled tissues after ¹⁴CO₂ uptake showed that the disruption of translocation patterns was persistent for at least 1 week after exposures ceased. Subsequent longer-term exposures at lower concentrations of both O₃ (0.0, 0.10, 0.15, and 0.20 microliters per liter) and SO₂ (0.0, 0.20, and 0.40 microliters per liter) demonstrated that O₃ more effectively altered allocation than SO₂, that primary leaves were generally more sensitive than trifoliates, and that responses of trifoliate leaves varied with plant growth stage. Altered rates of allocation of photosynthate by leaves were generally associated with alterations of similar magnitude and opposite direction in developing pods. Collectively, these experiments suggest that allocation patterns can provide sensitive indices of incipient growth responses of pollution-stressed vegetation.     

Growth Kinetics, Carbohydrate, and Leaf Phosphate Content of Clover (Trifolium subterraneum L.) after Transfer to a High CO(2) Atmosphere or to High Light and Ambient Air

Intact air-grown (photosynthetic photon flux density, 400 microeinsteins per square meter per second) clover plants (Trifolium subterraneum L.) were transfered to high CO₂ (4000 microliters CO₂ per liter; photosynthetic photon flux density, 400 microeinsteins per square meter per second) or to high light (340 microliters CO₂ per liter; photosynthetic photon flux density, 800 microeinsteins per square meter per second) to similarly stimulate photosynthetic net CO₂ uptake. The daily increment of net CO₂ uptake declined transiently in high CO₂, but not in high light, below the values in air/standard light. After about 3 days in high CO₂, the daily increment of net CO₂ uptake increased but did not reach the high light values. Nightly CO₂ release increased immediately in high light, whereas there was a 3-day lag phase in high CO₂. During this time, starch accumulated to a high level, and leaf deterioration was observed only in high CO₂. After 12 days, starch was two- to threefold higher in high CO₂ than in high light, whereas sucrose was similar. Leaf carbohydrates were determined during the first and fourth day in high CO₂. Starch increased rapidly throughout the day. Early in the day, sucrose was low and similar in high CO₂ and ambient air (same light). Later, sucrose increased considerably in high CO₂. The findings that (a) much more photosynthetic carbon was partitioned into the leaf starch pool in high CO₂ than in high light, although net CO₂ uptake was similar, and that (b) rapid starch formation occurred in high CO₂ even when leaf sucrose was only slightly elevated suggest that low sink capacity was not the main constraint in high CO₂. It is proposed that carbon partitioning between starch (chloroplast) and sucrose (cytosol) was perturbed by high CO₂ because of the lack of photorespiration. Total phosphate pools were determined in leaves. Concentrations based on fresh weight of orthophosphate, soluble esterified phosphate, and total phosphate markedly declined during 13 days of exposure of the plants to high CO₂ but changed little in high light/ambient air. During this time, the ratio of orthophosphate to soluble esterified phosphate decreased considerably in high CO₂ and increased slightly in high light/ambient air. It appears that phosphate uptake and growth were similarly stimulated by high light, whereas the coordination was weak in high CO₂.     

Carbon Dioxide Exchange in Lichens : RELATIONSHIP BETWEEN NET PHOTOSYNTHETIC RATE AND CO(2) CONCENTRATION

The relationship between net photosynthesis and CO₂ concentration was investigated for four species of lichen using an infrared gas analyzer operating in a closed loop system. All species showed a linear relationship at low CO₂ levels (100 microliters per liter) with CO₂ saturation levels being in excess of 400 microliters per liter. Detailed studies of Sticta latifrons showed a strong influence of thallus water content which resulted in the net photosynthetic response at high water contents still being nearly linear at 1000 microliters per liter CO₂. Very low CO₂ compensation values (5 microliters per liter) were obtained under some conditions but the value varied between thalli and with thallus water content. The results differ from previous studies which reported low CO₂ saturation levels (200 microliters per liter) and no apparent effect of water content. It is suggested that some of these differences may result from the use of a discrete sampling injection infrared gas analyzer system in the earlier studies and an assessment is made of the influence of nonsaturating CO₂ levels, lack of cuvette ventilation, and data presentation for this technique.     

Effects of Water Stress on Photosynthesis and Carbon Partitioning in Soybean (Glycine max [L.] Merr.) Plants Grown in the Field at Different CO(2) Levels

The effects of water stress and CO₂ enrichment on photosynthesis, assimilate export, and sucrose-P synthase activity were examined in field grown soybean plants. In general, leaves of plants grown in CO₂-enriched atmospheres (300 microliters per liter above unenriched control, which was 349 ± 12 microliters per liter between 0500 and 1900 hours EST over the entire season) had higher carbon exchange rates (CER) compared to plants grown at ambient CO₂, but similar rates of export and similar activities of sucrose-P synthase. On most sample dates, essentially all of the extra carbon fixed as a result of CO₂ enrichment was partitioned into starch. CO₂-enriched plants had lower transpiration rates and therefore had a higher water use efficiency (milligrams CO₂ fixed per gram H₂O transpired) per unit leaf area compared to nonenriched plants. Water stress reduced CER in nonenriched plants to a greater extent than in CO₂-enriched plants. As CER declined, stomatal resistance increased, but this was not the primary cause of the decrease in assimilation because internal CO₂ concentration remained relatively constant. Export of assimilates was less affected by water stress than was CER. When CERs were low as a result of the imposed stress, export was supported by mobilization of reserves (mainly starch). Export rate and leaf sucrose concentration were related in a curvilinear manner. When sucrose concentration was above about 12 milligrams per square decimeter, obtained with nonstressed plants at high CO₂, there was no significant increase in export rate. Assimilate export rate was also correlated positively with SPS activity and the quantitative relationship varied with CER. Thus, export rate was a function of both CER and carbon partitioning.     

Abscission: the phytogerontological effects of ethylene

The role of ethylene in the aging of bean (Phaseolus vulgaris L. cv. Red Kidney) petiole abscission zone explants was examined. The data indicate that ethylene does accelerate aging in addition to inducing changes in break strength. Application of ethylene during the aging stage (stage 1) promoted abscission when followed by a second ethylene treatment during the cell separating stage (stage 2). The half-maximal effective concentration of ethylene to induce aging was around 0.3 microliter per liter; 10 microliters per liter was a saturating dose. CO₂ reversal of ethylene action during stage 1 was incomplete and gave ambiguous results. CO₂ (10%) reversed the effect of 10 microliters per liter ethylene but not 1 microliter per liter ethylene. The possibility that ethylene not only accelerated aging but was also a requirement for it was tested, and experimental evidence in favor of this idea was obtained. It was concluded that ethylene plays a dual role in the abscission of bean petiole explants: a phytogerontological effect and a cellulase-inducing effect.