PHYSIOLOGICAL RESPONSES TO RAINFALL IN OPUNTIA BASILARIS (CACTACEAE)

An immediate, marked response to small amounts of rainfall occurs in Opuntia basilaris, despite previous drought conditions. The effect of rainfall is upon plant water potential, which is the single most important parameter influencing stomatal opening, CO₂ assimilation, and organic acid synthesis. Nocturnal stomatal opening is initiated following rainfall, and stomata remain open during the daytime. Decreasing stomatal and mesophyll resistances correlate with increasing rates of nocturnal assimilation of ¹⁴CO₂. Photosynthetic rates of ¹⁴CO₂ assimilation are low, despite high plant water potentials and low stomatal diffusion resistances. The decreased mesophyll resistances and increased rates of nocturnal ¹⁴CO₂ assimilation correlate with the increases of nocturnal efficiency of water use and CO₂ assimilation. The diurnal efficiency of water use and CO₂ assimilation is lower than the nocturnal gas exchange efficiency values.     

Induction of Acid Metabolism in Portulacaria afra

Portulacaria afra, a succulent plant, shifts from a predominantly C₃ mode of gas exchange to a typical Crassulacean acid metabolism type CO₂ uptake in response to water or NaCl stress. Control plants in the absence of water stress assimilated CO₂ during the light (about 7-8 mg CO₂ dm⁻² hr⁻¹), transpiration (about 1.5 g dm⁻² hr⁻¹) was predominantly during the day, stomates were open during the day, and there was little diurnal organic acid fluctuation. Stressed plants showed only dark CO₂ uptake and dark water loss, nocturnal stomatal opening, and an increased diurnal fluctuation of titratable acidity. Within 2 weeks after rewatering, stressed plants returned to the control acid fluctuation levels indicating that the response to stress was reversible.     

Responses of succulents to plant water stress

Experiments were performed to test the hypothesis that succulents “shift” their method of photosynthetic metabolism in response to environmental change. Our data showed that there were at least three different responses of succulents to plant water status. When plant water status of Portulacaria afra (L.) Jacq. was lowered either by withholding water or by irrigating with 2% NaCl, a change from C₃-photosynthesis to Crassulacean acid metabolism (CAM) occurred. Fluctuation of titratable acidity and nocturnal CO₂ uptake was induced in the stressed plants. Stressed Peperomia obtusifolia A. Dietr. plants showed a change from C₃-photosynthesis to internal cycling of CO₂. Acid fluctuation commenced in response to stress but exogenous CO₂ uptake did not occur. Zygocactus truncatus Haworth plants showed a pattern of acid fluctuation and nocturnal CO₂ uptake typical of CAM even when well irrigated. The cacti converted from CAM to an internal CO₂ cycle similar to Peperomia when plants were water-stressed. Reverse phase gas exchange in succulents results in low water loss to carbon gain. Water is conserved and low levels of metabolic activity are maintained during drought periods by complete stomatal closure and continual fluctuation of organic acids.     

Occurrence of Crassulacean acid metabolism in Cissus trifoliata L. (Vitaceae)

Summary Evidence for the operation of CAM in the deciduous climber, Cissus trifoliata L., was obtained in field and laboratory studies. Under natural conditions, diurnal oscillations of titratable acidity and colorimetric measurements of night CO₂ fixation, determined for a period of two and a half years, suggested that acid accumulation was related to plant water status, assessed through the daily courses of stomatal resistance and xylem water potential during dry and rainy seasons. These findings were confirmed by gas exchange studies under controlled conditions which showed that the plant fixed all its CO₂ during the day when it was well irrigated; as water stress increased, dark CO₂ uptake gradually replaced fixation during the day until the plant only performed dark fixation. In severe water stress, even the rate of the latter process decreased until leaves fell.Abbreviations CAM Crassulacean acid metabolism - FW leaf fresh weight - SWC relative soil water content - PAR photosynthetically active radiation - TR total radiation; r, leaf diffusive resistance - WSD water saturation deficit (leaf-air vapour concentration difference) - RWC relative water content of leaves     

Heterogenous stomatal closure in response to leaf water deficits is not a universal phenomenon

The extent and occurrence of water stress-induced “patchy” CO₂ uptake across the surface of leaves was evaluated in a number of plant species. Leaves, while still attached to a plant, were illuminated and exposed to air containing [¹⁴C]CO₂ before autoradiographs were developed. Plant water deficits that caused leaf water potential depression to −1.1 megapascals during a 4-day period did result in heterogenous CO₂ assimilation patterns in bean (Phaseolus vulgaris). However, when the same level of stress was imposed more gradually (during 17 days), no patchy stomatal closure was evident. The patchy CO₂ assimilation pattern that occurs when bean plants are subjected to a rapidly imposed stress could induce artifacts in gas exchange studies such that an effect of stress on chloroplast metabolism is incorrectly deduced. This problem was characterized by examining the relationship between photosynthesis and internal [CO₂] in stressed bean leaves. When extent of heterogenous CO₂ uptake was estimated and accounted for, there appeared to be little difference in this relationship between control and stressed leaves. Subjecting spinach (Spinacea oleracea) plants to stress (leaf water potential depression to −1.5 megapascals) did not appear to cause patchy stomatal closure. Wheat (Triticum aestivum) plants also showed homogenous CO₂ assimilation patterns when stressed to a leaf water potential of −2.6 megapascals. It was concluded that water stress-induced patchy stomatal closure can occur to an extent that could influence the analysis of gas exchange studies. However, this phenomenon was not found to be a general response. Not all stress regimens will induce patchiness; nor will all plant species demonstrate this response to water deficits.     

Calculation of CO2 gas phase diffusion in leaves and its relation to stomatal resistance

A new theory and experimental method was developed to measure the diffusion resistance to CO₂ in the gas phase of mesophyll leaf tissue. Excised leaves were placed in a chamber and their net evaporation and CO₂ assimilation rates measured at two different ambient pressures. These data were used to calculate CO₂ gas phase diffusion resistances. A variety of field grown leaves were tested and the effects of various experimental errors considered. Increasing the gas phase diffusion resistance decreased transpiration more than it decreased CO₂ assimilation. It was concluded that gas phase diffusion resistance associated with CO₂ assimilation may sometimes be 100 or 200 s·m^-1 greater than the resistance implied by transpiration rates. This may be due to longer path lengths for the CO₂ diffusion, constricted in places by the shape and arrangement of mesophyll cells.     

Seasonal Shifts of Photosynthesis in Portulacaria afra (L.) Jacq

Portulacaria afra (L.) Jacq., a perennial facultative Crassulacean acid metabolism (CAM) species, was studied under natural photoperiods and temperatures in San Diego, California. The plants were irrigated every fourth day throughout the study period. Measurements of ¹⁴CO₂ uptake, stomatal resistance, and titratable acidity were made periodically from July 1981 through May 1982. P. afra maintained C₃ photosynthesis during the winter and the spring. Diurnal acid fluctuations were low and maximal ¹⁴CO₂ uptake occurred during the day. The day/night ratio of carbon uptake varied from 5 to 10 and indicated little nocturnal CO₂ uptake. CAM photosynthesis occurred during the summer and a mixture of both C₃ and CAM during the fall. Large acid fluctuations of 100 to 200 microequivalents per gram fresh weight were observed and maximal ¹⁴CO₂ uptake shifted to the late night and early morning hours. Daytime stomatal closure was evident. A reduction in the day/night ratio of carbon uptake to 2 indicated a significant contribution of nocturnal CO₂ uptake to the overall carbon gain of the plant. The seasonal shift from C₃ to CAM was facilitated by increasing daytime temperature and accompanied by reduced daytime CO₂ uptake despite irrigation.     

Carbon Flow and Metabolic Specialization in the Tissue Layers of the Crassulacean Acid Metabolism Plant, Peperomia camptotricha

Leaves of Peperomia camptotricha contain three distinct upper tissue layers and a one-cell thick lower epidermis. Light and dark CO₂ fixation rates and the activity of ribulose bisphosphate carboxylase/oxygenase and several C₄ enzymes were determined in the three distinct tissue layers. The majority of the C₄ enzyme activity and dark CO₂ fixation was associated with the spongy mesophyll, including the lower epidermis; and the least activity was found in the median palisade mesophyll. In contrast, the majority of the C₃ activity, that is ribulose bisphosphate carboxylase/oxygenase and light CO₂ fixation, was located in the palisade mesophyll. In addition, the diurnal flux in titratable acidity was greatest in the spongy mesophyll and lowest in the palisade mesophyll. The spatial separation of the C₃ and C₄ phases of carbon fixation in P. camptotricha suggests that this Crassulacean acid metabolism plant may have low photorespiratory rates when it exhibits daytime gas exchange (that is, when it is well watered). The results also indicate that this plant may be on an evolutionary path between a true Crassulacean acid metabolism plant and a true C₄ plant.     

Resistance Analysis of Nocturnal Carbon Dioxide Uptake by a Crassulacean Acid Metabolism Succulent, Agave deserti

Nocturnal CO₂ uptake by a Crassulacean acid metabolism succulent, Agave deserti Engelm. (Agavaceae), was measured so that the resistance properties of the mesophyll chlorenchyma cells and their CO₂ concentrations could be determined. Two equivalents of acidity were produced at night per mole of CO₂ taken up. The nocturnal CO₂ uptake became light-saturated at 3.5 mEinsteins cm⁻² of photosynthetically active radiation (400-700 nm) incident during the preceding day; at least 46 Einsteins were required per mole of CO₂ fixed. Variations in the daytime leaf temperature between 20 and 37 C had little effect on nocturnal CO₂ uptake. After the first few hours in the dark, the leaf liquid phase CO₂ resistance (r^liq_CO2) and the CO₂ concentration in the chlorenchyma cells (cⁱ_CO2) both increased, the latter usually reaching the ambient external CO₂ level at the end of the dark period. Increasing the leaf surface temperature above 15 C at night markedly increased the stomatal resistance, r^liq_CO2, and cⁱ_CO2.

The minimum r^liq_CO2 at night was about 1.6 seconds cm⁻¹. Based on the ratio of chlorenchyma surface area to total leaf surface area of 82, this r^liq_CO2 corresponded to a minimum cellular resistance of approximately 130 seconds cm⁻¹, comparable to values for mesophyll cells of C₃ plants. The contribution of the carboxylation reaction and/or other biochemical steps to r^liq_CO2 may increase appreciably as the nighttime temperature shifts a few degrees from the optimum or after a few hours in the dark, both of which caused large increases in r^liq_CO2. This necessitates a large internal leaf area for CO₂ diffusion into the chlorenchyma to support moderate nocturnal CO₂ uptake rates by these succulent leaves.

     

Elevated CO2 reduces the drought effect on nitrogen metabolism in barley plants during drought and subsequent recovery

The objective of this study was to determine the response of nitrogen metabolism to drought and recovery upon rewatering in barley (Hordeum vulgare L.) plants under ambient (350 μmol mol⁻¹) and elevated (700 μmol mol⁻¹) CO₂ conditions. Barley plants of the cv. Iranis were subjected to drought stress for 9, 13, or 16 days. The effects of drought under each CO₂ condition were analysed at the end of each drought period, and recovery was analysed 3 days after rewatering 13-day droughted plants. Soil and plant water status, protein content, maximum (NR_max) and actual (NR_act) nitrate reductase, glutamine synthetase (GS), and aminant (NADH-GDH) and deaminant (NAD-GDH) glutamate dehydrogenase activities were analysed. Elevated CO₂ concentration led to reduced water consumption, delayed onset of drought stress, and improved plant water status. Moreover, in irrigated plants, elevated CO₂ produced marked changes in plant nitrogen metabolism. Nitrate reduction and ammonia assimilation were higher at elevated than at ambient CO₂, which in turn yielded higher protein content. Droughted plants showed changes in water status and in foliar nitrogen metabolism. Leaf water potential (Ψ_w) and nitrogen assimilation rates decreased after the onset of water deprivation. NR_act and NR_max activity declined rapidly in response to drought. Similarly, drought decreased GS whereas NAD-GDH rose. Moreover, protein content fell dramatically in parallel with decreased leaf Ψ_w. In contrast, elevated CO₂ reduced the water stress effect on both nitrate reduction and ammonia assimilation coincident with a less-steep decrease in Ψ_w. On the other hand, Ψ_w practically reached control levels after 3 days of rewatering. In parallel with the recovery of plant water status, nitrogen metabolism was also restored. Thus, both NR_act and NR_max activities were restored to about 75-90% of control levels when water supply was restored; the GS activity reached 80-90% of control values; and GDH activities and protein content were similar to those of control plants. The recovery was always faster and slightly higher in plants grown under elevated CO₂ conditions compared to those grown in ambient CO₂, but midday Ψ_w dropped to similar values under both CO₂ conditions. The results suggest that elevated CO₂ improves nitrogen metabolism in droughted plants by maintaining better water status and enhanced photosynthesis performance, allowing superior nitrate reduction and ammonia assimilation. Ultimately, elevated CO₂ mitigates many of the effects of drought on nitrogen metabolism and allows more rapid recovery following water stress.     

C(3) Photosynthesis and Crassulacean Acid Metabolism in a Kansas Rock Outcrop Succulent, Talinum calycinum Engelm. (Portulacaceae)

The potential for Crassulacean acid metabolism (CAM) was investigated in the sandstone outcrop succulent Talinum calycinum in central Kansas. Field studies revealed CAM-like diurnal acid fluctuations in these plants. These fluctuations persisted under all moisture and temperature regimes in the laboratory. Despite this CAM-like acid metabolism, simultaneous gravimetric determinations of day- and nighttime transpiration rates indicated the presence of a C₃ gas exchange pattern. Subsequent analyses of diurnal CO₂ and H₂O exchange patterns under well-watered conditions and after 3, 5, and 7 days of drought confirmed these findings, though low rates of nocturnal CO₂ uptake were observed on the fifth night after continuous drought. Finally, the δ¹³C/¹²C value of this succulent, −27.8‰, emphasizes the insignificance of any nocturnal CO₂ uptake in the lifelong accumulation of carbon in this species. Thus, it is proposed that T. calycinum is a C₃ plant with some CAM characteristics, including the ability to re-fix respiratory CO₂ at night under all moisture regimes, potentially resulting in a conservation of carbon, and occasionally to fix atmospheric CO₂ at night. These findings may prove to be common among rock outcrop succulents.     

Stimulation of CAM Photosynthesis in Kalanchoë blossfeldiana by Transferring to Nitrogen-Deficient Conditions

Kalanchoë blossfeldiana Poelln. cv Hikan plants were grown hydroponically with nutrient solution containing 5 millimolar NO₃⁻ (or NH₄⁺) for 1 to 2 months and then transferred to nutrient solution containing no nitrogen. CO₂ uptake at night, nocturnal increase in titratable acidity, and activity of phosphoenolpyruvate carboxylase increased after the transfer. Thus, transfer to nitrogen-deficient conditions stimulates Crassulacean acid metabolism (CAM photosynthesis) in K. blossfeldiana. The importance of the plant nitrogen status (nitrogen-withdrawal status) for induction and stimulation of CAM photosynthesis is discussed.     

Water Relations and Photosynthesis of a Desert CAM Plant, Agave deserti

The water relations and photosynthesis of Agave deserti Engelm., a plant exhibiting Crassulacean acid metabolism, were measured in the Colorado desert. Although no natural stomatal opening of A. deserti occurred in the summer of 1975, it could be induced by watering. The resistance for water vapor diffusion from a leaf (R_WV) became less than 20 sec cm⁻¹ when the soil water potential at 10 cm became greater than −3 bars, as would occur after a 7-mm rainfall. As a consequence of its shallow root system (mean depth of 8 cm), A. deserti responded rapidly to the infrequent rains, and the succulent nature of its leaves allowed stomatal opening to continue for up to 8 days after the soil became drier than the plant. When the leaf temperature at night was increased from 5 to 20 C, R_WV increased 5-fold, emphasizing the importance of cool nighttime temperatures for gas exchange by this plant. Although most CO₂ uptake occurred at night, a secondary light-dependent rise in CO₂ influx generally occurred after dawn. The transpiration ratio (mass of water transpired/mass of CO₂ fixed) had extremely low values of 18 for a winter day, and approximately 25 for an entire year.     

Characterization and Partial Purification of Aldose-6-phosphate Reductase (Alditol-6-Phosphate:NADP 1-Oxidoreductase) from Apple Leaves

The Pereskia are morphologically primitive, leafed members of the Cactaceae. Gas exchange characteristics using a dual isotope porometer to monitor ¹⁴CO₂ and tritiated water uptake, diurnal malic acid fluctuations, phosphoenolpyruvate carboxylase, and malate dehydrogenase activities were examined in two species of the genus Pereskia, Pereskia grandifolia and Pereskia aculeata. Investigations were done on well watered (control) and water-stressed plants. Nonstressed plants showed a CO₂ uptake pattern indicating C₃ carbon metabolism. However, diurnal fluctuations in titratable acidity were observed similar to Crassulacean acid metabolism. Plants exposed to 10 days of water stress exhibited stomatal opening only during an early morning period. Titratable acidity, phosphoenolpyruvate carboxylase activity, and malate dehydrogenase activity fluctuations were magnified in the stressed plants, but showed the same diurnal pattern as controls. Water stress causes these cacti to shift to an internal CO₂ recycling (“idling”) that has all attributes of Crassulacean acid metabolism except nocturnal stomata opening and CO₂ uptake. The consequences of this shift, which has been observed in other succulents, are unknown, and some possibilities are suggested.     

Influence of Photoperiod and Leaf Age on Crassulacean Acid Metabolism in Portulacaria afra (L.) Jacq

The possibility that Crassulacean acid metabolism (CAM) is subject to long day photoperiodic control in Portulacaria afra (L.) Jacq., a facultative CAM plant, was studied. Periodic measurements of ¹⁴CO₂ uptake, stomatal resistance, and titratable acidity were made on plants exposed to long and short day photoperiods. Results indicates that waterstressed P. afra had primarily nocturnal CO₂ uptake, daytime stomatal closure, and a large diurnal acid fluctuation in either photoperiod. Mature leaf tissue from nonstressed plants under long days exhibited a moderate diurnal acid fluctuation and midday stomatal closure. Under short days, there was a reduced diurnal acid fluctuation in mature leaf tissue. Young leaf tissue taken from nonstressed plants did not utilize the CAM pathway under either photoperiod as indicated by daytime CO₂ uptake, lack of diurnal acid fluctuation, and incomplete daytime stomatal closure.

The induction of CAM in P. afra appears to be related to the water status of the plant and the age of the leaf tissue. The photosynthetic metabolism of mature leaves may be partly under the control of water stress and of photoperiod, where CAM is favored under long days.

     

Photosynthesis and Transpiration as a Function of Gaseous Diffusive Resistances in Orange Leaves

The CO₂ and H₂O vapour exchange of single attached orange, Citrus sinensis (L.), leaves was measured under laboratory conditions using infrared gas analysis. Gaseous diffusive resistances were derived from measurements at a saturating irradiance and at a leaf temperature optimum for photosynthesis. Variation in leaf resistance (within the range 1.6 to 60 s cm^-1) induced by moisture status, or by cyclic oscillations in stomatal aperture, was associated with changes in both photosynthesis and transpiration. At low leaf resistance (ri less than 10 s cm^-1) the ratio of transpiration to photosynthesis declined with reduced stomatal aperture, indicating a tighter stomatal control over H₂O vapour loss than over CO₂ assimilation. At higher leaf resistance (ri greater than 10 s cm^-1) changes in transpiration and photosynthesis were linearly related, but leaf resistance and mesophyll resistance were also positively correlated, so that strictly stomatal control of photosynthesis became more apparent than real. This evidence, combined with direct measurements of CO₂ diffusive resistances (in a -O₂ gas stream) emphasised the presence of a significant mesophyll resistance; i.e., an additional and rate limiting resistance to CO₂ assimilation over and above that encountered by H₂O vapour escaping from the leaf.     

Leaf anatomy,water relations and crassulacean acid metabolism in the chlorenchyma and colourless internal water-storage tissue of Carpobrotus edulis and Senecio ?mandraliscae

Both Carpobrotus edulis and Senecio ?mandraliscae possess leaves with a peripheral chlorenchyma and colourless internal water-storage tissue. Water stress in C. edulis growing under semi-natural conditions resulted in the induction of weak Crassulacean acid metabolism (CAM) whereas well-watered plants of S. ?mandraliscae exhibited a similar degree of CAM. Titratable acidity in the separated water-storage tissue was substantially lower than in the chlorenchyma in both species but, nevertheless, increased during the night and decreased during the day either when sampled from the intact plant or from incubated tissue slices. Indeed, the increase in nocturnal titratable acidity produced by the water-storage tissue in situ accounted for approx. 30% of total acidification on a per-leaf basis. It appears that during the night the water-storage tissue in these species is able to fix CO₂ which is subsequently released during the day to enter the photosynthetic carbon-reduction cycle of the chlorenchyma. Diurnal rhythms of water potential (Ψ) and osmotic potential (Ψs) were measured in separated chlorenchyma and water-storage tissue by thermocouple psychrometry. Both parameters increased during the latter part of the daytime and initial nocturnal period and decreased during the rest of the night and into the post-dawn period. The chlorenchyma of water-stressed plants of C. edulis appeared to possess a marked negative turgor pressure (as determined from Ψ-Ψs) but this was caused by a severe underestimation in the measurement of the chlorenchyma Ψ. It is suggested that this artefact arose from release of colloidal polysaccharide mucilage, or possibly tannins, from broken tannin cells producing a lowering of water activity when measured using thermocouple psychrometry.     

Effects of irradiance and leaf water deficit on net carbon dioxide assimilation and mesophyll and transport resistances

Rate of net CO₂ assimilation by soil-grown soybean plants were studied over a range of relative leaf water contents at each of four levels of irradiance. There was a large interaction between light level and leaf water deficit on the rate of CO₂ assimilation. The effect of leaf water deficit on assimilation became larger as irradiance increased. Both stomatal resistance to CO₂ transport and mesophyll resistance to CO₂ assimilation increased as leaf-water deficit increased. The increase in both resistance with changing leaf-water content was largest at high irradiance and became smaller as irradiance decreased. Relief of soil-moisture stress by watering induced large oscillations of CO₂ assimilation, stomatal resistance, and mesophyll resistance. The oscillation of the mesophyll resistance occurred in the absence of changes in relative water content and appeared to be related to oscillations in leaf temperature. The observed increase in mesophyll resistance with decreasing leaf-water content under nonoscillative conditions may be caused by changes in leaf temperature rather than leaf water content.     

Water relations and photosynthesis of a barrel cactus,Ferocactus acanthodes,in the Colorado desert

Summary The structural characteristics, water relations, and photosynthesis of Ferocactus acanthodes (Lemaire) Britton and Rose, a barrel cactus exhibiting Crassulacean acid metabolism (CAM), were examined in its native habitat in the western Colorado desert. Water storage in its succulent stem permitted nighttime stomatal opening ot continue for about 40 days after the soil water potential became less than that of the stem, a period whe the plant would be unable to extract water from the soil. After 7 months of drought and consequent unreplenished water loss from a plant, diurnal stomatal activity was not observed and the stem osmotic pressure was 6.4 bars, more than double the value measured during wet periods with nighttime stomatal opening. F. acanthodes had a shallow root system (mean depth of 8 cm) which responded within 24 h to rainfall.When the nocturnal stem surface temperature was raised from 8.0° C to 35.0° C, the stomatal resistance increased 4-fold, indicating that cool nighttime temperatures are advantageous for gas exchange by F. acanthodes. Moreover, the optimal temperature for CO₂ uptake in the dark was only 12.6° C. CO₂ uptake at night became maximal for 3.0 mEinsteins cm^-2 of photosynthetically active radiation incident during the preceding day, and the minimum number of incident quanta absorbed per CO₂ fixed was 68. The transpiration ratio (mass of water transpired/mass of CO₂ fixed) had the relatively low value of 70 for an entire year, consistent with values obtained for other CAM plants. The total amount of water annually diverted to the floral structures was about 6% of the stem wet weight. The annual growth increment estimated from the net CO₂ assimilation corresponded to about 10% of the stem mass for barrel cacti 34 cm tall, in agreement with measured dimension changes, and indicated that such plants were about 26 years old.     

Juvenile tank-bromeliads lacking tanks: do they engage in CAM photosynthesis?

In the epiphytic tillandsioids, Guzmania monostachia, Werauhia sanguinolenta, and Guzmania lingulata (Bromeliaceae), juvenile plants exhibit an atmospheric habit, whereas in adult plants the leaf bases overlap and form water-holding tanks. CO₂ gas-exchange measurements of the whole, intact plants and δ¹³C values of mature leaves demonstrated that C₃ photosynthesis was the principal pathway of CO₂ assimilation in juveniles and adults of all three species. Nonetheless, irrespective of plant size, all three species were able to display features of facultative CAM when exposed to drought stress. The capacity for CAM was the greatest in G. monostachia, allowing drought-stressed juvenile and adult plants to exhibit net CO₂ uptake at night. CAM expression was markedly lower in W. sanguinolenta, and minimal in G. lingulata. In both species, low-level CAM merely sufficed to reduce nocturnal respiratory net loss of CO₂. δ¹³C values were generally less negative in juveniles than in adult plants, probably indicating increased diffusional limitation of CO₂ uptake in juveniles.