Drought-induced shifts in daily CO2 uptake patterns for leafy cacti

For cacti with persistent, relatively large leaves, most shoot CO₂ uptake under well-watered conditions occurs by the leaves using the C₃ pathway. For three species in the primitive subfamily Pereskioideae, droughts of 7 or 14 days decreased leaf daytime net CO₂ uptake by an average of 49 and 88%, respectively; these species always had a net CO₂ release at night by the leaves and both at night and during the day by the stems. For three leafy species in subfamily Opuntioideae, 7 and 14 days of drought reduced leaf daytime net CO₂ uptake by 90 and 100%, respectively. Although drought reduced the total CO₂ uptake over 24 h, the average percentage occurring at night by the leaves of these species increased from 5% under wet conditions to 71% after 7 days of drought to 99% after 14 days of drought. For two of the three species of Opuntioideae, 7 days of drought caused the small net CO₂ uptake by the sterns to shift from the daytime to the nighttime, while for the third species drought caused a reduction of its stem nocturnal net CO₂ uptake. Thus, shifts from predominantly daytime to predominantly nighttime net CO₂ uptake can be induced by drought for the leaves and the stems of leafy cacti in subfamily Opuntioideae, indicating a high degree of biochemical versatility.     

Physiological responses of Opuntia ficus-indica to growth temperature

The influences of various day/night air temperatures on net CO₂ uptake and nocturnal acid accumulation were determined for Opuntia ficus-indica, complementing previous studies on the water relations and responses to photosynthetically active radiation (PAR) for this widely cultivated cactus. As for other Crassulacean acid metabolism (CAM) plants, net nocturnal CO₂ uptake had a relatively low optimal temperature, ranging from 11°C for plants grown at day/night air temperatures of 10°C/0°C to 23°C at 45°C/35°C. Stomatal opening, which occurred essentially only at night and was measured by changes in water vapor conductance, progressively decreased as the measurement temperature was raised. The CO₂ residual conductance, which describes chlorenchyma properties, had a temperature optimum a few degrees higher than the optimum for net CO₂ uptake at all growth temperatures. Nocturnal CO₂ uptake and acid accumulation summed over the whole night were maximal for growth temperatures near 25°C/15°C, CO₂ uptake decreasing more rapidly than acid accumulation as the growth temperature was raised. At day/night air temperatures that led to substantial nocturnal acid accumulation (25°C/15°C.). 90% saturation of acid accumulation required a higher total daily PAR than at non-optimal growth temperatures (10°C/0°C and 35°C/25°C). Also, the optimal temperature of net CO₂ uptake shifted downward when the plants were under drought conditions at all three growth temperatures tested, possibly reflecting an increased fractional importance of respiration at the higher temperatures during drought. Thus, water status, ambient PAR, and growth temperatures must all be considered when predicting the temperature response of gas exchange for O. ficus-indica and presumably for other CAM plants.     

Characteristics of Crassulacean Acid Metabolism in the Succulent C(4) Dicot, Portulaca oleracea L

Crassulacean acid metabolism (CAM) was investigated in leaves and stems of the succulent C₄ dicot Portulaca oleracea L. Diurnal acid fluctuations, CO₂ gas exchange, and leaf resistance were monitored under various photoperiod and watering regimes. No CAM activity was seen in well watered plants grown under 16-hour days. Under 8-hour days, however, well watered plants showed a CAM-like pattern of acid fluctuation with amplitudes of 102 and 90 microequivalents per gram fresh weight for leaves and stems, respectively. Similar patterns were also observed in detached leaves and defoliated stems. Leaf resistance values indicated that stomata were open during part of the dark period, but night acidification most likely resulted from refixation of respiratory CO₂. In water-stressed plants maximum acid accumulations were reduced under both long and short photoperiods. At night, these plants showed short periods of net CO₂ uptake and stomatal opening which continued all night long during preliminary studies under natural environmental conditions. Greatest acid fluctuations, in P. oleracea, with amplitudes of 128 microequivalents per gram fresh weight, were observed in water-stressed plants which had been rewatered, especially when grown under short days. No net CO₂ uptake took place, but stomata remained open throughout the night under these conditions. These results indicate that under certain conditions, such as water stress or short photoperiods, P. oleracea is capable of developing an acid metabolism with many similarities to CAM.     

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.     

Environmental influences on the productivity of three desert succulents in the south-western United States

Abstract Net CO₂ uptake over 24 h periods for shoots of Agave deserti, Ferocactus acanthodes, and Opuntia ficus-indica was measured under the ranges of water status, air temperature, and photo-synthetically active radiation (PAR) that occur in the south-western U.S.A. An environmental productivity index (EPI) was constructed indicating the overall influence of these three factors on net CO₂ uptake. Using growth chambers whose conditions were changed monthly to simulate the environmental conditions at a field site, the observed shoot dry weight increases per unit surface area changed in concert with monthly changes in EPI. The observed dry weight gain of the shoot was 17–19% lower than the predicted shoot net CO₂ uptake, which could be accounted for by carbon diversion to the roots. Mean monthly EPI was also predicted for 21 sites in the south-western U.S.A. All three species had low EPIs in the Colorado River basin, which has low annual rainfall and high summer temperatures, and in the north-central part of the region, which has low temperatures and low PAR during winter when water is available. The two native species, A. deserti and F. acanthodes, had high EPIs beyond their range in coastal southern California, where competition by other vegetation for PAR may limit net CO₂ uptake. Such regions as well as south-central California and south-central Arizona had high EPIs for all three species, indicating that these areas would be appropriate for the cultivation of O. ficus-indica.     

Relationships between Photosynthetically Active Radiation, Nocturnal Acid Accumulation, and CO(2) Uptake for a Crassulacean Acid Metabolism Plant, Opuntia ficus-indica

The influences of photosynthetically active radiation (PAR) and water status on nocturnal Crassulacean acid metabolism (CAM) were quantitatively examined for a widely cultivated cactus, Opuntia ficus-indica (L.) Miller. When the total daily PAR was maintained at 10 moles photons per square meter per day but the instantaneous PAR level varied, the rate of nocturnal H⁺ accumulation (tissue acidification) became 90% saturated near 700 micromoles per square meter per second, a PAR level typical for similar light saturation of C₃ photosynthesis. The total nocturnal H⁺ accumulation and CO₂ uptake reached 90% of maximum for a total daily PAR of about 22 moles per square meter per day. Light compensation occurred near 0 moles per square meter per day for nocturnal H⁺ accumulation and 4 moles per square meter per day for CO₂ uptake. Above a total daily PAR of 36 moles per square meter per day or for an instantaneous PAR of 1150 micromoles per square meter per second for more than 6 hours, the nocturnal H⁺ accumulation actually decreased. This inhibition, which occurred at PAR levels just above those occurring in the field, was accompanied by a substantial decrease in chlorophyll content over a 1-week period.

A minimum ratio of H⁺ accumulated to CO₂ taken up of 2.5 averaged over the night occurred for a total daily PAR of 31 moles per square meter per day under wet conditions. About 2 to 6 hours into the night under such conditions, a minimum H⁺-to-CO₂ ratio of 2.0 was observed. Under progressively drier conditions, both nocturnal H⁺ accumulation and CO₂ uptake decreased, but the H⁺-to-CO₂ ratio increased. A ratio of two H⁺ per CO₂ is consistent with the H⁺ production accompanying the conversion of starch to malic acid, and it apparently occurs for O. ficus-indica when CAM CO₂ uptake is strongly favored over respiratory activity.

     

Crassulacean Acid Metabolism in the Succulent C(4) Dicot, Portulaca oleracea L Under Natural Environmental Conditions

Crassulacean acid metabolism (CAM) was examined under natural environmental conditions in the succulent C₄ dicot Portulaca oleracea L. Two groups of plants were monitored; one was watered daily (well watered), while the other received water once every 3 to 4 weeks to produce a ψ of −8 bars (drought stressed). Gas exchange, transpiration rate, and titratable acidity were measured for 24-hour periods during the growing season. CAM activity was greatest in drought-stressed plants during late August which had 13 hour days and day/night temperatures of 35/15°C. Under these conditions net CO₂ uptake occurred slowly throughout the night. Diurnal fluctuations of titratable acidity took place in both leaves and stems with amplitudes of 17 and 47 microequivalents per gram fresh weight, respectively. Transpiration data indicated greater opening of stomata during the night than the day. CAM was less pronounced in drought-stressed P. oleracea plants in July and September; neither dark CO₂ uptake nor positive carbon balance occurred during the July measurements. In contrast, well-watered plants appeared to rely on C₄ photosynthesis throughout the season, although some acid fluctuations occurred in stems of these plants during September.     

Gas Exchange Characteristics of the Submerged Aquatic Crassulacean Acid Metabolism Plant, Isoetes howellii

The submerged aquatic plant Isoetes howellii Engelmann possesses Crassulacean acid metabolism (CAM) comparable to that known from terrestrial CAM plants. Infrared gas analysis of submerged leaves showed Isoetes was capable of net CO₂ uptake in both light and dark. CO₂ uptake rates were a function of CO₂ levels in the medium. At 2,500 microliters CO₂ per liter (gas phase, equivalent to 1.79 milligrams per liter aqueous phase), Isoetes leaves showed continuous uptake in both the light and dark. At this CO₂ level, photosynthetic rates were light saturated at about 10% full sunlight and were about 3-fold greater than dark CO₂ uptake rates. In the dark, CO₂ uptake rates were also a function of length of time in the night period. Measurements of dark CO₂ uptake showed that, at both 2,500 and 500 microliters CO₂ per liter, rates declined during the night period. At the higher CO₂ level, dark CO₂ uptake rates at 0600 h were 75% less than at 1800 h. At 500 microliters CO₂ per liter, net CO₂ uptake in the dark at 1800 h was replaced by net CO₂ evolution in the dark at 0600 h. At both CO₂ levels, the overnight decline in net CO₂ uptake was marked by periodic bursts of accelerated CO₂ uptake. CO₂ uptake in the light was similar at 1% and 21% O₂, and this held for leaves intact as well as leaves split longitudinally. Estimating the contribution of light versus dark CO₂ uptake to the total carbon gain is complicated by the diurnal flux in CO₂ availability under field conditions.     

Basal cactus phylogeny: implications of Pereskia (Cactaceae) paraphyly for the transition to the cactus life form

The cacti are well-known desert plants, widely recognized by their specialized growth form and essentially leafless condition. Pereskia, a group of 17 species with regular leaf development and function, is generally viewed as representing the "ancestral cactus," although its placement within Cactaceae has remained uncertain. Here we present a new hypothesis of phylogenetic relationships at the base of the Cactaceae, inferred from DNA sequence data from five gene regions representing all three plant genomes. Our data support a basal split in Cactaceae between a clade of eight Pereskia species, centered around the Caribbean basin, and all other cacti. Two other Pereskia clades, distributed mostly in the southern half of South America, are part of a major clade comprising Maihuenia plus Cactoideae, and Opuntioideae. This result highlights several events in the early evolution of the cacti. First, during the transition to stem-based photosynthesis, the evolution of stem stomata and delayed bark formation preceded the evolution of the stem cortex into a specialized photosynthetic tissue system. Second, the basal split in cacti separates a northern from an initially southern cactus clade, and the major cactus lineages probably originated in southern or west-central South America.     

Recovery of photosynthetic reactions after high-temperature treatments of a heat-tolerant cactus

Inhibition and recovery of net CO₂ uptake and three photosynthetic electron transport reactions as well as plant survival following high-temperature treatments were investigated for Opuntia ficus-indica. For plants maintained at 30°C/20°C day/night air temperatures, treatment at 60°C for 1 h irreversibly inhibited net CO₂ uptake and photosynthetic electron transport, resulting in plant death in about 60 days. When a plant maintained at 30°C/20°C was treated at 55°C for 1 h, net CO₂ uptake was completely inhibited 1 d after the treatment but fully recovered in 60 d. Differential inactivation of photosystem (PS) I, PSII, and whole chain electron transport activities occurred; PSI was the most tolerant of 55°C and took the least time (45 d) for total recovery. All 30°C/20°C plants survived a 1-h treatment at 55°C, although some pale green areas were observed on the cladode surfaces. In contrast to growing at 30°C/20°C, plants acclimated to 45°C/35°C survived 60°C for 1 h without showing any necrotic or pale green areas on the cladode surfaces. When such a plant was transferred to 30°C/20°C following the high-temperature treatment, recovery in net CO₂ uptake began in 1 d and progressed to complete recovery by 30 d. Growth temperatures thus influence the possibility for recovery of photosynthetic reactions and ultimately the survival of O. ficus-indica following a high-temperature exposure.Abbreviations DCPIP 2,6-dichlorophenol indophenol - MV methyl viologen - PAR photosynthetically active radiation - PSI or PSII photosystem I or II - WC whole chain     

Contribution of stem CO2 fixation to whole-plant carbon balance in nonsucculent species

In many plant species that remain leafless part of the year, CO₂ fixation occurring in green stems represents an important carbon gain. Traditionally, a distinction has been made between stem photosynthesis and corticular photosynthesis. All stem photosynthesis is, sensu stricto, cortical, since it is carried out largely by the stem cortex. We proposed the following nomenclature: stem net photosynthesis (SNP), which includes net CO₂ fixation by stems with stomata in the epidermis and net corticular CO₂ fixation in suberized stems, and stem recycling photosynthesis (SRP), which defines CO₂ ling in suberized stems. The proposed terms should reflect differences in anatomical and physiological traits. SNP takes place in the chlorenchyma below the epidermis with stomata, where the net CO₂ uptake occurs, and it resembles leaf photosynthesis in many characteristics. SRP is found in species where the chlorenchyma is beneath a well-developed stomata-free periderm and where reassimilation of internally respired CO₂ occurs. SNP is common in plants from desert ecosystems, rates reaching up to 60% of the leaf photosynthetic rate. SRP has been demonstrated in trees from temperate forests and it offsets partially a carbon loss by respiration of stem nonphotosynthetic tissues. Reassimilation can vary between 7 and 123% of respired CO₂, the latter figure implying net CO₂ uptake from the atmosphere. Both types of stem photosynthesis contribute positively to the carbon economy of the species, in which they occur; they are advantageous to the plant because they allow the maintenance of physiological activity during stress, an increase of integrated water use efficiency, and they provide the carbon source used in the production of new organs.     

Relationship between Respiration and CAM-Cycling in Peperomia camptotricha

Mature leaves of well-watered Peperomia camptotricha show Crassulacean acid metabolism (CAM). Young leaves show CAM-cycling in which CO₂ uptake occurs during the day concomitant with a marked diurnal fluctuation of organic acids as in CAM. Evidence is presented suggesting that respiration is the source of CO₂ for nocturnal acid synthesis in leaves exhibiting CAM-cycling. Respiratory quotients for these leaves were consistently much less than unity despite the fact that the leaves metabolize starch. The conservation of CO₂ by refixation into acids at night represents about 17% of the total photosynthetically fixed CO₂ and about 50% of the total respiratory CO₂.     

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.     

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.     

Frerea indica,a stem succulent CAM plant with deciduous C3 leaves

Summary During the winter in greenhouse culture, Frerea indica(Asclepiadaceae) is a leafless stem succulent resembling the other members of the Stapelieae subfamily. However, in spring it produces leaves which persist during the summer period. CO₂ exchange measurements were carried out with Frerea indica at its different seasonal states of development. The leafless stems in winter as well as the defoliated ones in summer show all characteristics of Crassulacean acid metabolism. The leaves, on the other hand, fix CO₂ with high rates according to the C₃ pathway; no diurnal malate fluctuations are detectable. This feature can be interpreted as an ecological adaptation where a seasonal plant dimorphism enables a functional change in the prevailing mode of CO₂ metabolism. In the leafless state, which probably represents the appearance of the plant during the dry period in nature, it is saving water by pursuing CAM. The season with enough water available can be used for high photosynthetic gain by C₃ leaves.     

Growth, CO2 uptake, and responses of the carboxylating enzymes to inorganic carbon in two highly productive CAM species at current and doubled CO2 concentrations

In Agave salmiana Otto ex Salm. var. salmiana grown for 4½ months in open-top chambers, 55% more leaves unfolded and 52% more fresh mass was produced at 730 than at 370μmol CO₂ mol^?1. A doubling of the CO₂ concentration also stimulated growth in another highly productive CAM species, Opuntia ficus-indica (L.) Miller, leading to earlier initial ion and 37% more daughter cladodes. Substantial net CO₂ uptake occurred earlier in the afternoon and lasted longer through the night for A. salmiana at 730 than at 370μmol CO₂ mol^?1, resulting in 59% more total daily net CO₂ uptake. The Michaelis constant (HCO₃^?) for PEPCasc was 15% lower for A. salmiana and 44% lower for O. ficus-indica when the CO₂ concentration was doubled; the percentage of Rubisco in the activated state in vivo was on average 64% higher at the doubled CO₂ concentration. Thus the substantial increases in net CO₂ uptake and biomass production that occurred in these two CAM species when the ambient CO₂ concentration was doubled resulted mainly from higher inorganic carbon levels for their carboxylating enzymes, a greater substrate affinity for PEPCase, and a greater percentage of Rubisco in the activated state.     

BRANCHING PATTERNS OF COLUMNAR CACTI: INFLUENCES ON PAR INTERCEPTION AND CO2 UPTAKE

Field measurements and a computer model were used to determine how stem shape and arrangement of stems in space affect interception of photosynthetically active radiation (PAR) and CO₂ uptake under otherwise optimal conditions for four species of columnar cacti (Carnegiea gigantea, Lophocereus schottii, Pachycereus pringlei, and Stenocereus thurberi). In simulations where the number of widely spaced stems was increased from 1 to 19 but plant volume remained constant, surface area and PAR interception increased, leading to 3-fold increases in whole-plant CO₂ uptake. Increasing the distance between stems from 0 cm to infinity decreased self-shading and increased predicted CO₂ uptake 4-fold. Stem length, diam, ribbing characteristics, and spine coverage also influenced PAR interception. The model indicated that the observed higher frequency of branches on the south side of the trunk of C. gigantea had only a slight, though positive, effect on CO₂ uptake for single-branched plants. Because of its greater surface area (A), a five-stemmed plant of C. gigantea typical for a field site near Tucson, Arizona was predicted to have 52% more CO₂ uptake than a single-stemmed plant of the same volume (V). Although large A/V decreases water storage per unit transpiring area, whole-plant CO₂ uptake can be increased when A/V is increased by branching for these constant-volume plants. However, the stems must be arranged to avoid excessive self-shading and thus keep the area below PAR compensation small.     

Light effects on leaf development and photosynthetic capacity of Hydrocotyle bonariensis Lam.

Rates of net CO₂ uptake were examined in developing leaves of Hydrocotyle bonariensis. Leaves that developed under high photosynthetically active radiation (48 mol m^-2 day^-1 PAR) were smaller, thicker, and reached maximum size sooner than did leaves that developed under low PAR (4.8 mol m^-2 day^-1). Maximum net CO₂ uptake rates were reached after 5 to 6 days expansion for both the low and the high PAR leaves. Leaves grown at high PAR had higher maximum photosynthetic rates and a higher PAR required for light saturation but showed a more rapid decline in rate with age than did low PAR leaves. To assess the basis for the difference observed in photosynthetic rates, CO₂ diffusion conductances and the mesophyll surface available for CO₂ absorption were examined for mature leaves. Stomatal conductance was the largest conductance in all treatments and did not vary appreciably with growth PAR. Mesophyll conductance progressively increased with growth PAR (up to 48 mol m^-2 day^-1) as did the mesophyll surface area per unit leaf area, but the cellular conductance exhibited most of its increase at low PAR (up to 4.8 mol m^-2 day^-1).     

Carbon balance during CAM: an assessment of respiratory CO2 recycling in the epiphytic bromeliads Aechmea nudicaulis and Aechmea fendleri

Abstract The regulation of crassulacean acid metabolism (CAM) under controlled environmental conditions has been investigated for two tropical epiphytes, relating plant water and carbon balance to growth form and habitat preference under natural conditions. Aechmea fendleri is restricted to wet, upper montane regions of Trinidad, while A. nudicaulis has a wider distribution extending into more arid regions of the island. Morphological characteristics of these plants are related to habitat preference in terms of leaf succulence (0.44 and 0.94 kg m^?2 for the two species respectively) and a distinct layer of water storage parenchyma in A. nudicaulis In contrast, the thinner leaves of A. fendleri contain little water-storage parenchyma and less chlorenchyma per unit area, but the plants have a more open leaf rosette. The two species differ in expression of CAM, since the proportion of respiratory CO₂ recycled as part of CAM had been found to be much lower in A. fendleri This study compared the efficiency of water use and role of respiratory CO₂ recycling under two PAR regimes (300 and 120 μnol m^?2 s^?1) and three night temperatures (12, 18 and 25 °C). Dark CO₂ uptake rates for both species were comparable to plants in the field (maximum of 2.3 ± 0.2 μmol m^?2s^?1± SD, n= 3). Total net CO₂ uptake at night increased on leaf area basis with temperature for both species under higher PAR, although under the low PAR regime CO₂ uptake was maximal at 18 °C. Water-use efficiency (WUE) increased at 18 °C and 25 °C during dark CO₂ uptake (Phase I) and also during late afternoon photosynthesis (Phase IV) in both species. For A. fendleri, dawn to dusk changes in titrable acidity (ΔH ⁺) were similar under high and low PAR, although ΔH⁺ was correlated to night temperature and PAR in A. nudicaulis. The proportion of ΔH⁺ derived from respiratory CO₂ also varied with experimental conditions. Thus percentage recycling was lower in A. fendleri under high PAR (0–10%), but was only reduced at 18 °C under low PAR. Recycling by A. nudicaulis ranged from 32–42% under high PAR, but was also reduced to 6% under low PAR at 18 °C; at 12 °C and 25 °C, recycling was 37% and 52% respectively. Previous studies have suggested a relationship between the proportion of recycling and degree of water stress. This study indicated that CAM as a CO₂ concentrating mechanism regulates both water-use efficiency and plant carbon balance in these epiphytes, in response to PAR and night temperature. However, the precise relationship between respiratory processes and the balance between external and internal sources of CO₂ is as yet unresolved.